Critical Reviews in Plant Sciences, 25:235–265, 2006

Copyright
c Taylor & Francis Group, LLC

ISSN: 0735-2689 print / 1549-7836 online
DOI: 10.1080/07352680600611543

Biofumigation and Enhanced Biodegradation: Opportunity

and Challenge in Soilborne Pest and Disease Management

John N. Matthiessen
CSIRO Entomology, Private Bag 5, P.O. Wembley, WA, Australia, 6913

John A. Kirkegaard
CSIRO Plant Industry, GPO Box 1600, Canberra, ACT, Australia, 2601

Table of Contents
I. PROLOGUE ................................................................................................................................................... 236

II. INTRODUCTION: BACKGROUND AND CONTEXT ................................................................................... 237

A. Recognition of Unique Challenges .............................................................................................................. 237
1. Integrated Pest Management Principles .............................................................................................. 237
2. Constraints on Choice and Implementation ........................................................................................ 238
B. Methyl Bromide Replacement .................................................................................................................... 238
C. Appropriate Targeting ................................................................................................................................ 239
D. The Biofumigation Concept ........................................................................................................................ 239
1. True Fumigants and Synthetic Isothiocyanate ..................................................................................... 239
2. Naturally Occurring Isothiocyanates .................................................................................................. 239
E. Enhanced Biodegradation ........................................................................................................................... 240
F. Scope and Purpose ..................................................................................................................................... 241

III. DEVELOPMENT OF THE BIOFUMIGATION CONCEPT ........................................................................... 241

A. Chemistry and Biological Effects of Glucosinolates and Derivatives ............................................................... 241
1. The Glucosinolate-Myrosinase System .............................................................................................. 241
B. Green Manuring and Biofumigation ............................................................................................................ 242
1. Green Manures ................................................................................................................................ 243
2. Trap Crops and Non-Hosting ............................................................................................................ 243
3. Non-Glucosinolate or Isothiocyanate-Related Effects .......................................................................... 243
4. Processed Brassica Amendments ...................................................................................................... 244
C. Biofumigation Potential—Beyond Empiricism ............................................................................................. 244
1. Glucosinolate Profiles ...................................................................................................................... 244
2. Effects of Plant Growth and Environment .......................................................................................... 248
3. Biological Activity of Various Isothiocyanates ................................................................................... 248
4. Implications for Adoption ................................................................................................................ 249
D. Realizing Biofumigation Potential ............................................................................................................... 249
1. Beyond In Vitro Studies—Activity of Isothiocyanates in Soil ............................................................... 249
2. Implications for Biofumigation ......................................................................................................... 250
3. Achieving High Release of Isothiocyanates ........................................................................................ 251
E. Field Implementation ................................................................................................................................. 252
1. Key Factors in Field Implementation ................................................................................................. 252
2. Farming Systems Fit ........................................................................................................................ 252
3. Case Study 1—Commercial Implementation in Potatoes ..................................................................... 253
4. Case Study 2—Bacterial Wilt ........................................................................................................... 253
Address correspondence to John N. Matthiessen, CSIRO Entomology, Private Bag 5, P.O. Wembley, WA, Australia, 6913. E-mail:
John.Matthiessen@csiro.au

235
236 J. N. MATTHIESSEN AND J. A. KIRKEGAARD

IV. FUTURE DIRECTIONS ................................................................................................................................. 254

A. Capitalizing on Results and Maintaining Momentum .................................................................................... 254
1. Revisit Previous Work and Adopt a “Best Bet” Approach .................................................................... 254
2. New Research to Increase Use of Biofumigation ................................................................................ 254
3. Environmental Issues—Fate of Isothiocyanates and Glucosinolates in Soil ........................................... 255

V. ENHANCED BIODEGRADATION OF SOIL-APPLIED PESTICIDES .......................................................... 255

A. Background and History ............................................................................................................................. 255
1. Environmental Fate .......................................................................................................................... 255
2. Microbially-Mediated Transformations .............................................................................................. 255
3. Determination of Enhanced Biodegradation ....................................................................................... 256
B. Mechanisms and Modifying Factors ............................................................................................................ 256
1. Soil pH ........................................................................................................................................... 256
2. Soil Calcium Level .......................................................................................................................... 257
3. Soil Type ........................................................................................................................................ 257
4. Overcoming Enhanced Biodegradation .............................................................................................. 258
C. Link to Biofumigation and Methyl Bromide ................................................................................................. 258
1. Enhanced Biodegradation of Metam Sodium ...................................................................................... 258
2. Cross-Degradation ........................................................................................................................... 258
3. Methyl Bromide Alternatives ............................................................................................................ 259

VI. CONCLUSIONS ............................................................................................................................................. 260

ACKNOWLEDGMENT ............................................................................................................................................. 261

REFERENCES .......................................................................................................................................................... 261

for self-sustaining further development and market penetration

Management of soilborne pests and diseases in cropping sys-
tems is often highly challenging—in implementation of accept-
able methodologies and in dealing with secondary problems. The
phase-out of methyl bromide brings this into particularly sharp
focus. There is a need for diversified options and alternatives to fill
different roles across the soilborne pest and disease management
spectrum, but flexibility is limited, as practicalities demand that
they fit into a prophylactic methodology. It is against such a back-
drop that expectations and promotion of alternatives must be set.
There is also a need to recognize potentially serious problems that
may have been masked under historical management regimes, but
for which the nature of the system offers little scope to avoid or
manage.
Biofumigation is the beneficial use of Brassica green manures
that release isothiocyanates chemically similar to methyl isoth-
iocyanate, the active agent from the synthetic fumigant metam
sodium, which is used as a substitute for methyl bromide in some
systems. A systematic approach to research into biofumigation,
specifically aimed at overcoming a long history of empiricism, has
seen significant recent advances in both basic and applied knowl-
edge. A key development has been achievement of maximal biofu-
migation potential through greatly enhanced release of appropriate
isothiocyanates into soil. These advances have led to commercial
adoption, demonstrating that biofumigation, when applied to ap-
propriate production systems, can have efficacy and offer cost
savings. Crucially, these systematically derived research and de-
velopment findings and their adoption now provide the impetus
of the concept. Despite this success, biofumigation is not seen as
being sufficiently powerful or practical in implementation to be an
alternative to methyl bromide on a broad scale and misdirection
in that regard could be counterproductive to more appropriately
targeted further development.
Enhanced microbial biodegradation is a cryptic phenomenon
that can diminish the efficacy of soil-applied pesticides, including
isothiocyanates and most other currently available methyl bromide
substitutes. Because methyl bromide is not susceptible, the phe-
nomenon has potentially serious implications in intensive produc-
tion systems switching from methyl bromide to reliance on other
compounds that are. It is an intractable problem once induced.
Avoidance of its onset is the only feasible management strategy.
This has been aided for some particularly vulnerable environments
by recent clarification of key risk factors associated with soil type,
soil pH, and calcium content.
Keywords Biofumigation, Brassica, isothiocyanate, glucosinolate,
metam sodium, soil, pest, disease, enhanced biodegrada-
tion, methyl bromide
I. PROLOGUE
In recent years the phasing-out of methyl bromide has had
the effect of bringing into sharp focus the need for alternative
strategies for the management of soilborne pests and diseases,
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
not just for users of methyl bromide but in a general sense.
Less generally realized is the prospect that problems related
to maintaining efficacy following the repeated application of
different pesticides to soil are likely to arise where previously
they did not occur. Although methyl bromide replacement is
not the subject of this review, it provides a topical and relevant
foundation for setting the context and theme.
Among the alternative control methods being touted to re-
place methyl bromide are the use of other fumigant-like pes-
ticides that have a long history of use, such as the methyl
isothiocyanate generator metam sodium, 1,3-dichloropropene
(1,3-D), chloropicrin, or mixtures of these, and various
biologically-based options. But can these agents and methods
achieve the efficacy and all-important consistency of perfor-
mance of the robust and forgiving methyl bromide in the highly
intensive production systems where it has been used for so long?
Additionally, are there issues with using such alternatives where
history can provide warnings of potential problems that need to
be recognized and managed?
Biofumigation, the use of isothiocyanate-generating brassi-
cas as biologically-active green manures to emulate the use of the
synthetic pesticide metam sodium, has a genesis deep in history,
but a level of adoption that belies a heavily documented potential.
Similarly, the broadly related issue of enhanced biodegradation
of soil-applied pesticides, to which metam sodium is susceptible,
has been well documented. However, the warnings from history
seem to have been rarely heeded. Why are these responses so?
The intent of this review is to be more than a repetitious
description of the history and mechanisms of the concept of
biofumigation as a potential control method, and of enhanced
biodegradation as a problem, in soilborne pest and disease man-
agement. Its theme is more about the conceptualization and con-
duct of research directed at forward-reaching development and
delivery of practical outcomes than it is about the reiteration of
core underlying concepts. In the case of biofumigation it aims to
provide an illustration of how research needs to be undertaken
within a systematic framework, be appropriately targeted, and
be based on realistic expectations and goals in order to have
reasonable prospects of harnessing the opportunity to deliver on
potential and achieve beneficial results. In the case of enhanced
biodegradation of soil-applied pesticides, it seeks to warn of a
long-known but poorly recognized and managed phenomenon
and to indicate how new information may help meet the chal-
lenge of its prevention and management in some situations.
II. INTRODUCTION: BACKGROUND AND CONTEXT
A. Recognition of Unique Challenges
Soilborne pests and diseases present serious challenges in
many plant production systems. These challenges have unique
characteristics that need to be recognized, particularly when en-
deavoring to devise, promote and implement “softer” manage-
ment options. Their recognition forms a contextual backdrop
for this review. There is generally little scope for radical change
237
in the underlying approach to management of soilborne pests
and diseases. The challenges arise directly through the capac-
ity of such organisms to reduce product yield or quality, and
in significant indirect ways not encountered when dealing with
above-ground pests and diseases. For example, there is often
considerable practical difficulty determining the threat and in
implementing specifically targeted control that may be more be-
nign than some of the conventional pesticide-based approaches.
Additional difficulties can arise in avoiding or overcoming some
serious problems that may occur through repeated pesticide ap-
plication to soil. Developing cost-effective solutions where none
have existed before can also present significant challenges.
1. Integrated Pest Management Principles
Generally, the contemporary search for solutions to any pest,
disease, and weed threats is at least idealistically founded on
the concept of integrated pest management (IPM). The under-
lying objective of IPM is the minimization of broad-spectrum
biocide use to avoid or reduce such problems as environmental
contamination, disruption of natural enemies, and induction of
resistance. That goal is achieved particularly through the avoid-
ance of prophylactic or scheduled application of such agents; any
intervention should be needs-based and specifically targeted. Im-
plementation is founded on monitoring for the occurrence and
abundance of noxious organisms and the concomitant risk of
economic injury.
The concepts and practices of IPM have been formulated
and developed into sometimes finely-tuned systems entirely in
above-ground cropping systems, including strategies to manage
resistance to biocides. In such systems monitoring (“scouting”),
if not always easy, is at least feasible and treatments, should the
system’s criteria deem intervention necessary, can be readily im-
plemented. Furthermore, the results of the intervention can be as
readily assessed as the threat. Controlling soilborne pest and dis-
eases is far more complicated and challenging than controlling
foliar ones. For producers confronting such threats there is an
array of impediments that render IPM concepts and strategies,
as classically conceived in above-ground cropping systems, ab-
stract idealizations that cannot be readily implemented. Apart
from the obvious major logistical and economic constraint of
sampling and monitoring in the dense and heterogeneous soil
medium, there are several other limitations that compound the
difficulties.
Commonly, many soilborne noxious organisms are micro-
scopic, such as fungi, bacteria, and nematodes, making moni-
toring intrinsically difficult and thus costly. Furthermore, par-
ticularly in the case of fungi, the organism may be a resting
life-stage for much of the time, the activation of which is highly
dependent on a set of complex environmental conditions that
render its noxiousness both qualitatively and quantitatively un-
predictable. The advent of commercial DNA probe diagnostic
systems shows the presence of organisms but cannot predict the
extent to which that potential will develop (Anon., 2005). In
the case of some soil insects, damaging population abundance
238 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
levels may be near limits of reliable detection (e.g., <1 insect
m−2 for two beetle species infesting potato crops in Australia
(Matthiessen and Learmonth, 1995)). This makes population
monitoring, the setting of economic injury levels and action
thresholds for control intervention impractical. In other cases,
soil-dwelling pest insects may be present as overwintering eggs
when crops are sown, with later life stages subsequently caus-
ing damage during crop growth, or they may invade crops by
flight during growth. All these factors make traditional IPM-
based monitoring and needs-based decision-making logistically
and economically unfeasible.
To further compound the difficulties presented by the at-
tributes of individual soilborne noxious organisms, a range of
them that individually may be susceptible to different specific
or more benign control treatments may occur simultaneously in
the often biologically rich and complex soil environment. Con-
versely, there is no doubt that there are many instances where
there has been an absence of pest or disease threats but control
measures have nevertheless been employed as insurance against
the risk of a perceived threat, although this is rarely if ever sci-
entifically tested. Despite this array of seemingly intractable
biological challenges, perhaps the most significant practical im-
pediment to implementation of integrated management princi-
ples for soilborne noxious organisms is simply that satisfactory
intervention during crop growth is generally impossible because
of the physical disruption necessary to place the control agent
into the soil at the site of the pest or disease. This aspect can be
exacerbated by the often phytotoxic nature of broad-spectrum
control agents.
2. Constraints on Choice and Implementation
As a consequence of these practical difficulties in implement-
ing management strategies, producers are either at the mercy of
the system or they are induced to seek and utilize more broadly
effective control measures than is likely to be the case in above-
ground crops. By the very nature of the challenges being faced,
the most feasible and effective management methodology for
soilborne organisms is application of an agent as a prophylac-
tic before sowing the crop. These agents are either extremely
broad-spectrum to eliminate the greatest possible range of ex-
isting threats or highly residual to control latent or subsequent
threats from external sources. Some residual forms of control
may need to last for several years, such as against soil insects
in multiple-cycle crops like sugar cane (Robertson et al., 1998).
All of this means that management methods coming under the
popularly-used IPM or “clean and green” mantra must still con-
form to a prophylactic approach, with some constraints that nec-
essarily entails.
Ultimately, the capacity to make a choice of whether to imple-
ment a pesticide-based management strategy against soilborne
organisms is driven by the economics of the production system.
Extensive cropping systems of relatively low productivity are
unlikely to face catastrophic failures over the whole crop or, if
there were a consistent threat of that nature, it is unlikely that
the cropping system could have developed or persisted in such
an environment in any event. Furthermore, extensive cropping
systems like cereals are seasonal and are also likely to be part
of a diversified rotation that brings intrinsic benefits through its
temporal and plant-type discontinuities. At the other extreme,
intensive cropping systems producing crops of high value, such
as strawberries, very often have almost continuous production
interrupted only by sufficient time to virtually sterilize the soil
against pathogens, pests, and weeds with the powerful fumigant
methyl bromide and allow for its dissipation before planting the
next crop.
B. Methyl Bromide Replacement
A highly significant factor making the implementation of al-
ternatives, especially “soft” options to methyl bromide particu-
larly problematic is that the whole production system (e.g., cul-
tivar development, nutrient management) has evolved over the
last fifty years around the universal use of methyl bromide soil
fumigation as a fundamental component of the system. It is the
factor causing the mostly mysterious, but universally observed
yield-boosting “increased growth response” in the absence of
any detectable pathogen (Munnecke, 1967; Duniway, 2002;
Ajwa et al., 2003; Martin, 2003). This evolved yield-focused
process, founded so strongly on the use of methyl bromide,
has worked against the development of alternative approaches
such as hardier disease tolerant cultivars. Methyl bromide is also
noted for being immensely robust and highly forgiving in use,
which ensures that methods derived in one system or region
are functional without modification in other systems or loca-
tions. The long accumulated experience with such a convenient
and robust agent has set at a very high level the benchmark
against which alternatives are measured. It is also likely that it
contributes to forming mind-sets where less conventional tech-
niques are potentially dismissed out of hand because they are
seen as sure to be less effective or too difficult to implement.
Because of these characteristics of methyl bromide-based
production systems, the implementation of less robust alterna-
tives is likely to need adjunct changes in the system or they may
need modifications in different locations or climatic conditions
for them to be efficacious and economic. The widespread use
and effectiveness of methyl bromide as part of an evolved system
also works against easy implementation of alternatives because
there is often a paucity of data on the noxious organisms (if any)
that are constraining production. An important aspect in this
regard, with particular relevance to the substitution of methyl
bromide with other compounds is the lack of knowledge of the
concentration × time dose required to control particular noxious
organisms. It has also meant that pesticide-independent options,
most of which take a long time to implement, such as selection
of resistant germplasm and biological and cultural control, have
generally been underexplored.
All this serves to demonstrate that replacement of methyl bro-
mide with more benign options is far from a trivial issue. Great
 BIOFUMIGATION AND ENHANCED BIODEGRADATION 239

care needs to be taken not to make unrealistic or sweeping claims ticide metam sodium (sodium N -methyldithiocarbamate) has
about the potential of various methodologies to be alternatives been widely used since the 1950s world-wide to control a range
without a good understanding of these intensive production sys- of noxious soilborne organisms in potato and similar moderately
tems. To do so risks ridicule and out-of-hand dismissal at the intensive production systems (van Berkum and Hoestra, 1979).
pragmatic producer level and ultimately irrelevance at the sci- Metam sodium generates the compound methyl isothio-
entific level. cyanate upon contact with moist soil (Turner and Corden, 1963;
Munnecke, 1967; Smelt and Leistra, 1974) and it is methyl
C. Appropriate Targeting isothiocyanate that possesses broad-spectrum biological activ-
ity against nematodes, fungal pathogens, insects, and weeds
Between the extensive and intensive agricultural system ex-
(Munnecke et al., 1962; Richardson and Thorn, 1969). Methyl
tremes lie many plant production systems facing varying levels
isothiocyanate is not gaseous at ambient temperatures and with a
of threat from noxious soilborne organisms. Choices made will
distribution ratio in the soil water and air phases of around 99:1,
depend on the cost of inputs, possible acceptance by the producer
it principally diffuses through soil dissolved in water (Smelt and
of a lower input/lower yield approach that remains economic, or
Leistra, 1974; Lembright, 1990). This relatively high diffusibil-
a producer’s personal or market-driven desire to move an exist-
ity, which becomes more uniform when metam sodium is applied
ing production system away from “artificial” control techniques
in irrigation (Gerstl et al., 1977) and promotes high efficacy deep
towards more “natural” methods. Some highly intensive methyl
into sandy soil profiles in particular (Santo and Qualls, 1984), is
bromide-dependent industries are now facing considerable diffi-
probably largely what has led to metam sodium being described
culties in finding alternatives; however, their primary demand is
as a soil fumigant (and perhaps the readily-detected stench of this
for drop-in replacements that must be equally powerful (Martin,
sulfur-containing compound and its irritant characteristics gives
2003).
the illusion of higher volatility than it has). Like all fumigants,
In terms of the broader capacity for exercising choice in how
it is applied directly as supplied and at high volumes (c. 500 L
best to manage soilborne pests and diseases, arguably it is the
ha−1 ) rather than the small doses of agents typically thought of
broad sweep of what might for convenience be termed “inter-
as pesticides. Because of the dependence on moisture in the soil
mediate” production systems that have over the years most com-
for realization of this effect (El Hadiri et al., 2003), it is perhaps
monly been able to do so. These are systems with an intensity of
more accurately described as a fumigant-like pesticide. Never-
production beyond those of broad scale where specific interven-
theless, the inclusion of methyl isothiocyanate amongst the soil
tion for soilborne pest management is generally impractical, yet
fumigants has long been in common use and for all intents and
less than those where such management is a core (and invariably
purposes it is regarded as such.
highly costly) element of a rigid and highly evolved production
Methyl isothiocyanate is one of an array of isothiocyanates,
process, or is legislated or market-mandated for phytosanitary
naturally-occurring compounds characterized by a common
reasons. They are systems where there is economic and practi-
structural entity with a variable side group that distinguishes
cal scope to intervene at varying levels and with some degree
one from another (Kjaer et al., 1963). The toxic effects of pure
of flexibility to manage the threats posed by noxious soilborne
isothiocyanates have been known for many years (Walker et al.,
organisms, but still within the constraints of the prophylactic
1937). Methyl isothiocyanate decomposes relatively quickly to
approach imposed by the soil-based system. A good general ex-
innocuous compounds (Lloyd, 1961; Turner and Corden, 1963)
ample of such a system is potato production. An example of an
and so does not present the residue threats that can be of concern
extensive system is broad-scale cereal growing; strawberry fruit
with xenobiotic pesticides. Despite the broad-spectrum efficacy
production is an example of a highly intensive system, whereas
of methyl isothiocyanate, however, metam sodium is too costly
strawberry nursery (runner) production is an example of an in-
for use in many production systems. Also, many producers seek
tensive system bound by strict phytosanitary (certification) con-
biologically-based alternatives because they would prefer not
trols that provide no leeway for partial solutions. The context of
to use broad-spectrum synthetic agents if possible. However,
these examples and this discussion is that of western-style agri-
unlike the demands for identical performance inherently char-
culture; small-scale or tropical agriculture may present unique
acteristic of the search for methyl bromide replacements, these
opportunities and issues.
producers have more flexibility in their choices. This logically
provides richer opportunities for researchers seeking to develop
D. The Biofumigation Concept non-pesticide-based management options than do highly inten-
1. True Fumigants and Synthetic Isothiocyanate sive systems.
Broad-spectrum true gaseous fumigants of high vapor pres-
sure that diffuse and penetrate rapidly and far through soil such as 2. Naturally Occurring Isothiocyanates
methyl bromide have historically not found a place in intermedi- Fortuitously, a characteristic of isothiocyanates is that they
ate production systems because of their high cost, including the occur conveniently in nature, notably deriving from the glucosi-
need for plastic tarping of the soil to retain the gas while it acts. nolates common as secondary plant compounds in the diverse
However, the less potent and less costly broad-spectrum pes- Brassicaceae (Rosa et al., 1997; Fahey et al., 2001). The pest
240 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
suppression potential of isothiocyanates released from Bras-
sica residues has long been generally recognized (Grainge and
Ahmed, 1988). However, the use of brassicas in an attempt to
suppress noxious soilborne organisms has had sporadic atten-
tion with generally mixed results, ranging from pest, disease, or
weed suppression (Blank et al., 1982; Chan and Close, 1987;
Muehlchen et al., 1990; Mojtaheedi et al., 1993; Davis et al.,
1996; Akiew and Trevorrow, 1999; McLeod and Steele, 1999)
to no suppression (Johnson et al., 1992) to in some instances
pathogen stimulation (Stephens et al., 1999).
The term biofumigation was coined by J. A. Kirkegaard
(Kirkegaard et al., 1993). As originally conceptualized, it was
a crystallizing term for that part of the suppressive effects of
Brassica species on noxious soilborne organisms that arose
quite specifically through liberation of isothiocyanates from hy-
drolysis of the glucosinolates that characterize the Brassicaceae
(Kirkegaard and Matthiessen, 2004). Since isothiocyanates are
biologically active, and the methyl isothiocyanate generator
metam sodium is widely used as a broad-spectrum fumigant-
like pesticide, there existed a logical link and an impetus to
the notion of harnessing, and further developing, a biological
source of isothiocyanates for suppression of soilborne pests and
diseases (Brown and Morra, 1997). The term biofumigation at-
tempted to ascribe, in a simplified way, a mechanistic name
to this particular part of a general phenomenon of allelopathic
(Whittaker and Feeny, 1971) effects that have been observed in
the Brassicaceae for centuries and given them a reputation as
poor companion plants (Bell and Muller, 1973; Chew, 1988).
Since being coined, the initially adjectival term biofumiga-
tion has morphed into a noun and has rapidly entered the pest
management lexicon in a much broader and more popularized
way to encompass any beneficial effect arising from green ma-
nure or rotation crops, and even composts. This is far beyond
what was originally conceived or intended when the single-
word label was given to the putative process. This extension or
perhaps embellishment of the expression has probably largely
come about because of its all-embracing pest suppression over-
tones, and has very likely been amplified by the euphony of the
word. It has probably also been influenced by the high level
of attention being given to management of noxious soilborne
organisms associated with the phase-out of methyl bromide,
and the global-wide search for alternatives. Nevertheless, the
notion of emulating the well-known pesticidal (fumigant) ef-
fect of a synthetic isothiocyanate through biological sources
of isothiocyanates remains the central theme of the concept,
notwithstanding the recognition of a broad range of other agro-
nomic and pest suppression benefits from Brassica, and other,
rotation and green manure crops (Kirkegaard and Matthiessen,
2004).
E. Enhanced Biodegradation
Before addressing some of the details and recent develop-
ments in biofumigation it is pertinent to bring into the discussion
an issue that can bear heavily on the success of methods used
to manage soilborne noxious organisms. Methyl isothiocyanate,
like many of the more modern nonresidual soil-applied pesti-
cides, is also susceptible to the phenomenon of enhanced (or
accelerated) biodegradation, whereby the efficacy of a pesticide
can be seriously compromised by its unusually rapid breakdown
in soil. In summary, this phenomenon is caused by adaptation of
microbes (usually bacteria) and subsequent stimulation of their
populations by natural selection to use the pesticide as a nutri-
ent and energy source. It is induced by repeated applications of
a susceptible pesticide to the same area of soil (Kearney and
Kellogg, 1985; Felsot, 1989; Cork and Krueger, 1991; Aislabie
and Lloyd-Jones, 1995). Strictly, in the soil pest management
context, biodegradation only reaches the level of “enhanced”
when it becomes so rapid that it compromises management of
the target pest. That in itself presents a problem, as analyti-
cal diagnosis may show an increased rate of disappearance of
a pesticide from soil, but its efficacy may not at that point be
compromised. Predicting where that threshold falls is virtually
impossible in practice.
Halogenated pesticides (e.g., DDT, dieldrin, heptachlor) are
highly recalcitrant to biological breakdown and have now been
widely banned because of their capacity to persist in the envi-
ronment and concentrate up food chains (MacRae, 1989; Ais-
labie and Lloyd-Jones, 1995). Methyl bromide is also recalci-
trant, and although some biotic degradation is possible (Dungan
and Yates, 2003), for all practical purposes it is not subject
to enhanced biodegradation. Given the high vapor pressure of
methyl bromide (1380 mm Hg at 20◦ C, compared to 21.0 mm
for methyl isothiocyanate (Lembright, 1990)), it would seem
likely to be nonpersistent in soil more because of its volatil-
ity than recalcitrance, hence the need for plastic tarping in its
use.
It is probably fair to say that the concept of enhanced
biodegradation of a soil-applied pesticide reducing its pesti-
cidal effect is alien to users of methyl bromide. That may
be about to change. Several of the existing registered fumi-
gant pesticides that are now being drawn into use as sub-
stitutes for methyl bromide, notably metam sodium and 1,3-
dichloropropene (Duniway, 2002; Ajwa et al., 2003) have long
been known to be highly susceptible to enhanced biodegrada-
tion (Smelt et al., 1989; Warton et al., 2001a; Dungan et al.,
2003). Chloropicrin, which is commonly mixed with 1,3-D in
particular to broaden its spectrum of activity breaks down much
faster than methyl bromide and mainly by microbial degrada-
tion (Gan et al., 2000a). Enrichment of chloropicrin-degradative
microbial populations is possible (Ibekwe et al., 2004). While
no control failures attributable to enhanced biodegradation of
chloropicrin have been recorded, its greater susceptibility to mi-
crobial degradation than methyl bromide suggests that it could
occur if conditions were favorable.
Given that no highly recalcitrant pesticide is ever again likely
to gain regulatory approval, any prolonged use of labile sub-
stitutes for methyl bromide would seem to be at considerable
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
risk of pest management failures attributable to enhanced
biodegradation because of the characteristic highly intensive
use pattern in such systems. It would be remiss when consider-
ing the use as soil-applied pesticides of a family of compounds
known to be susceptible to the phenomenon, particularly in
the context of integrated pest management principles, not to
incorporate discussion of enhanced biodegradation—much as
ignoring insecticide resistance management in a discussion
of IPM in, say, cotton production would be unusual. It is a
concept that often seems poorly understood and confused
with resistance. The notion of organisms using an apparently
toxic compound for metabolic advantage, as opposed to
the metabolic cost of detoxification in pesticide-resistant
organisms is counterintuitive. Yet it is a practical issue that has
well-documented examples of causing economically significant
pest management failures, wasteful overuse of pesticides, and
difficulties overcoming the problem (Felsot, 1989; Racke,
1990; Felsot and Shelton, 1993; Suett et al., 1996).
F. Scope and Purpose
It is within the broad framework of IPM or “clean and green”
ideals, flexibility, choice, producer desires, economic and bio-
logical limitations in soilborne pest management solutions that
biofumigation has been driven out from its initially descriptive
meaning and empirical roots to have a more targeted devel-
opment in recent years. It has become a name for a tool that
provides an opportunity for management of noxious soilborne
organisms in a way that meets many pest management ideals
(Brown and Morra, 1997; Kirkegaard et al., 1998; Kirkegaard
and Matthiessen, 1999; Matthiessen et al., 2001; Kirkegaard
and Matthiessen, 2004). In effect, the specific underlying intent
of this thrust of research has been to systematically determine
how close it is possible to come to emulating the efficacy of
metam sodium using biofumigation, and what is required to do
so.
It is the purpose of this review to describe that evolution,
and in a broad practically-orientated context critically and prag-
matically assess progress against potential, suggest needs, and
highlight impediments for the further development of the pro-
cess, weigh its inherent advantages and limitations, and make a
realistic assessment of feasible applications. A similar contex-
tual objective holds for the linked topic of enhanced biodegra-
dation. Our viewpoint is that a substantial knowledge base al-
ready exists on the history and background fundamentals of these
topics and that it is readily available and well-reviewed, thus
needing only overarching reiteration. We believe that what re-
quires attention is the more holistic integration and development
of that knowledge aimed at practical applications, progressing
development and management within and between these top-
ics, together with integration of new information particularly
pertinent to those aspects. Those aspects are the theme of this
review.
241
III. DEVELOPMENT OF THE BIOFUMIGATION
CONCEPT
A. Chemistry and Biological Effects of Glucosinolates
and Derivatives
Glucosinolates are a class of sulfur compounds occurring
as secondary plant products almost exclusively in families of
the Order Capparales: Tovariaceae, Resedaceae, Capparaceae,
Moringaceae and Brassicaceae (Fenwick et al., 1983; Brown
and Morra, 1997; Rosa et al., 1997). They are particularly com-
mon in the Brassicaceae, plants that are widely cultivated as im-
portant vegetable, condiment, oilseed and forage crops (Fenwick
et al., 1983). Comprising some 350 genera and 2500 species
(Rosa et al., 1997), the Brassicaceae therefore present a rich
potential pre-adapted source of material for investigation and
application for beneficial uses in cropping systems.
Brassica species glucosinolates in particular have been the
subject of many extensive reviews over the last half-century. This
would seem to be principally because of the effects of various
derivatives in human and livestock foods, such as initially flavor
and antinutritional characteristics but more recently on thera-
peutic and prophylactic properties. Such aspects are outside the
scope of this review but recent reviews by Fahey et al. (2001)
and Holst and Williamson (2004) provide a lead-in to those top-
ics. Increasingly, from a plant sciences perspective, attention has
been directed at brassicas in relation to their long-recognized al-
lelopathic properties and hence their potential to be harnessed
for use in the management of pests in agriculture (Chew, 1988;
Brown and Morra, 1997; Rosa and Rodrigues, 1999; Chitwood,
2002). However, despite a large body of evidence indicating pos-
itive effects and considerable potential, there has until recently
been a lack of significant adoption of brassicas for soilborne pest
and disease management in cropping systems.
1. The Glucosinolate-Myrosinase System
Glucosinolates are characterized by a common core chemi-
cal entity, with a side-chain group of variable chemical structure
that distinguishes individual types, of which more than 120 have
been identified in three groups: aliphatic, aromatic, and indolyl
(Fenwick et al., 1983; Brown and Morra, 1997; Fahey et al.,
2001). Only the aliphatic and aromatic glucosinolates liber-
ate isothiocyanates upon hydrolysis (Bones and Rossiter, 1996;
Rask et al., 2000). Fahey et al. (2001) provide a comprehensive
survey of the chemical structure of all glucosinolates known to
that time, and an outline of their synthesis and biosynthesis. Glu-
cosinolates consistently occur in plants in conjunction with a hy-
drolytic enzyme thioglucosidase hydrolase, known commonly
as myrosinase (Rosa et al., 1997). Of themselves, glucosino-
lates have no or very limited biological activity. But they are
important because of the wide variety of active products that
derive from them as a result of myrosinase action, a dynamic
link that has led to the interaction commonly being dubbed the
“glucosinolate-myrosinase system” (Bones and Rossiter, 1996;
Rask et al., 2000).
242 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
It is generally believed that the glucosinolate-myrosinase
system is part of plant defense against insects and possibly
pathogens and that it evolved from the more prevalent system
of cyanogenic glucosides (Rask et al., 2000). Glucosinolates
are very stable water-soluble compounds, while myrosinase is
sequestered in vacuoles within myrosin cells, with both com-
pounds separately coexisting distributed throughout the plant
(Rosa et al., 1997; Fahey et al., 2001). It is only upon phys-
ical disruption of the plant tissue that the physical separation
of the myrosinase from its glucosinolate substrate is overcome,
triggering in the presence of water the hydrolysis reactions that
release a variety of biologically-active products such as isothio-
cyanates, organic cyanides, oxazolidinethiones and ionic thio-
cyanates (Brown and Morra, 1997). Among the degradation
products of glucosinolates, the isothiocyanates have been gen-
erally reported as the most biologically active, being recognized
since early in the twentieth century as broad-spectrum biocides
(Brown and Morra, 1997; Rosa and Rodrigues, 1999).
Reaction conditions that favor the production of isothio-
cyanates are pH around neutral, warm temperature and high
dilution with water; at more acid pH, lower temperature and
drier conditions the less biologically active nitriles predominate
(van Etten and Tookey, 1979; Rosa et al., 1997). The type of
isothiocyanate generated corresponds to the type of glucosino-
late substrate, as characterized by its side-chain (Chew, 1988).
Interestingly, and perhaps a little ironic given that the biofu-
migation concept was partly founded on the basis that the syn-
FIG. 1. Mechanisms by which Brassica green manures can influence the growth and yield of following crops. The pathway by which biofumigation related to
glucosinolates (GSL) and isothiocyanates (ITC) is shown by the large arrow. (From Kirkegaard and Matthiessen, 2004)
thetic methyl isothiocyanate generator metam sodium has found
widespread commercial use as a pesticide, methyl glucosinolate
does not occur in the Brassicaceae, although it is a characteris-
tic component of the closely related Capparaceae (Fahey et al.,
2001).
B. Green Manuring and Biofumigation
Green manuring is a traditional technology for providing gen-
eral benefits in terms of nutrients, soil properties, organic matter
and break-crop effects, but it has been largely supplanted in mod-
ern agriculture by the advent of inorganic fertilizers. Figure 1 is
a diagrammatic summary of the many ways in which an incorpo-
rated Brassica green manure can influence the yield and quality
of a subsequent commercial crop. While the primary focus of
this review will be on the pathway shown for biofumigation as an
isothiocyanate-based concept for suppression of soilborne pests
and diseases, indicated by the large arrow, it is appropriate to
briefly mention some of the related aspects.
The pathways shown in Figure 1 include several that are un-
related to soilborne pest and disease control and which are com-
mon to most green manures, including beneficial impacts on or-
ganic matter, nutrition, soil structure and erosion control. Those
related to pest and disease suppression include non-host or trap
crop effects, and indirect effects on the pest or pathogen associ-
ated with changes in the populations of antagonistic organisms,
as well as the more direct suppressive effects of compounds
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
released from the tissues that may not be related to glucosi-
nolates or isothiocyanates. It is clear from Figure 1 that the
isothiocyanate-related biofumigation pathway is only one of
many ways in which Brassica green manures can influence
the growth of a crop. It is worth highlighting some of the no-
table, previously reported non-biofumigation impacts of green
manures before a more detailed consideration of the ways in
which the biofumigation pathway has, particularly in the last
5–10 years, been refined for pest suppression.
1. Green Manures
Green manures, tillage practices, and organic amendments
can provide benefits to following crops and to farming systems
in general associated with maintenance of soil cover and soil in-
tegrity, soil sanitization effects, reduced erosion, greater soil or-
ganic matter and soil structural improvements that improve wa-
ter penetration (Bailey and Lazarovits, 2003; Thorup-Kristensen
et al., 2003). Brassica green manures are no exception, and the
combination of a large taproot with a dense network of fine sur-
face roots appears to make some Brassica species particularly
useful in this regard. For example, on the sandy irrigated soils in
the western United States the inclusion of mustard green manure
crops has reduced the level of wind erosion and improved the
water infiltration of the soils, with a steady improvement in soil
structure where they are included in the system for some years
(Gies, 2004; McGuire, 2004). These impacts of brassicas on
soil structure and water penetration have also been observed in
hard-setting soils in the Australian wheat belt following canola
rotation crops (Chan and Heenan, 1996), where they appear to
be related to the influence of the canola roots on soil aggregation.
Brassica green manure crops are effective at capturing soil
mineral nitrogen that may otherwise leach from the soil, and
when incorporated into the soil can provide a source of organic
nitrogen that can become available to subsequent crops. The re-
cent review by Thorup-Kristensen et al. (2003) provides a very
comprehensive account of the use of green manures and catch
crops in this regard, and the significant body of literature re-
viewed includes many studies involving Brassicaceous species
such as radish (Raphanus sativus), white mustard (Sinapis alba),
and winter rape (Brassica napus) in comparison with legumi-
nous and grass species.
2. Trap Crops and Non-Hosting
Brassicaceous green manures can also act as trap crops
(Thorup-Kristensen et al., 2003). Perhaps the most well doc-
umented case involving their use for this purpose is for the
control of sugar beet nematodes (Heterodera schactii) in north-
ern Europe (Muller, 1999; Schlathoelter, 2004). Fodder radish
(Raphanus sativus) and white mustard (Sinapis alba) cover crops
have been selected and developed to be grown as green manures
preceding sugar beet crops. The brassicas are invaded by the ne-
matodes, which develop within the roots, but have their sexual
differentiation disrupted. This results in very low numbers of fe-
males in the subsequent generation, causing significant decline
243
in the population and reducing the infection of subsequent sugar
beet crops. It is a unique example related to specific nematode-
resistant brassicas, rather than a general non-hosting effect.
For a green manure to be effective in disease control it is gen-
erally desirable that it not host the pathogen in question so that
a decline in population or inoculum occurs during its growth.
Indeed the fact that Brassica species are generally moderate
hosts to some other important plant parasitic nematodes (e.g.,
Meloidogyne spp.), has reduced their applicability as biofumi-
gant green manures as very careful management is required to
avoid population increases on the biofumigant crop, particularly
in warmer climates (McLeod et al., 2001; Stirling and Stirling,
2003). In such cases, it is possible that the suppressive effects
of the incorporated tissues could still reduce the population to
manageable levels, but it is generally desirable to select biofu-
migants that are poor hosts, or to grow them at times of the year
when pathogens do not build up.
3. Non-Glucosinolate or Isothiocyanate-Related Effects
There are several examples in the literature of significant
suppression by incorporated Brassica amendments that are not
associated with the glucosinolate concentration of the tissue.
The most consistent of these are those related to nematode sup-
pression. A study by Potter et al. (1998) showed clearly that
the significant suppressive effects (60 to 95%) of six diverse
Brassica species were unrelated to either the total glucosino-
late content of the tissues, or to the concentration of propenyl
glucosinolate, the major glucosinolate found in mustard which
is known to produce the biologically active propenyl isothio-
cyanate upon hydrolysis. Similar results have also been shown
for Meloidogyne javanica by McLeod and Steele (1999), and
in glasshouse studies of the microbial complex associated with
apple replant disease (Mazzola et al., 2001). The most likely
explanation for these observations are either that the incorpo-
ration of organic matter itself increased the populations of an-
tagonistic organisms in the soil, or that non-glucosinolate com-
pounds released by brassicas are toxic to nematodes. Indeed
many potentially biologically active, non-glucosinolate sulfur-
containing compounds are released from Brassica amendments
(Bending and Lincoln, 1999), and other products of microbial
decomposition of tissues including fatty acids can also be biolog-
ically active. Irrespective of the mechanism responsible, these
levels of suppression that are unrelated to glucosinolates can sig-
nificantly confound the interpretation of biofumigation studies
if appropriate controls are not included in the experiments.
In addition to isothiocyanates, hydrolysis of glucosinolates
in incorporated biofumigants can release a range of other po-
tentially toxic products including nitriles, epinitriles, and ionic
thiocyanates (Brown and Morra, 1997; Morra, 2004; Rollin and
Palmieri, 2004; Palmieri, 2004). Although isothiocyanates are
generally considered the most toxic of these hydrolysis products,
they may not be released in large concentrations in all circum-
stances following incorporation (Bending and Lincoln, 1999).
Morra (2004) has shown that much of the weed suppression
244 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
noted following incorporation of Brassicaceous seed meals is
likely to arise from the ionic thiocyanate rather than the isothio-
cyanates. Often experimental protocols used to determine the
toxicity of different amendments can favour different com-
pounds depending on volatility (e.g., sealed containers), water
solubility (irrigation/soil water content) and reactivity (soil or-
ganic matter content). To resolve the question of isothiocyanate-
related suppression it is generally desirable to correlate the level
of pest suppression with measured levels of isothiocyanate re-
leased in soil, a task that is not trivial given the relatively rapid
loss from soils as a result of many different processes (Brown
and Morra, 1997).
4. Processed Brassica Amendments
Another mode for achieving biofumigation effects is to
use Brassica-derived isothiocyanate-rich materials such as seed
meals or oils as soil amendments to achieve pesticidal effects
(Lazzeri et al., 2004). There may be a niche for such products,
but they are likely to be regarded as pesticides by regulatory au-
thorities and will therefore potentially face significant hurdles in
implementation that the growing of a Brassica crop rich in ap-
propriate isothiocyanates and manipulating it to maximize their
release would not face (Askew, 2004). While there is other liter-
ature on this aspect, and it may well be a viable option in some
niches, we regard it as outside the scope of this review which is
focused on the use of isothiocyanate-releasing plants grown in
rotation with the crop for which protection from soilborne pests
and diseases is sought.
C. Biofumigation Potential—Beyond Empiricism
The type, concentration and distribution of glucosinolates in
different plant parts vary between Brassica species and culti-
vars (Josefsson, 1967; Sang et al., 1984). Hence the capacity to
generate isothiocyanates varies accordingly. Although the bio-
cidal activity of various isothiocyanates released by Brassica
tissues in vitro is well established (Brown and Morra, 1997),
difficulty in interpreting the role of isothiocyanates in the varied
suppressive effects reported has characterized most field stud-
ies of green manuring with brassicas. A significant number of
previous studies have reported suppression of a range of pests
and pathogens in the field using a range of different Brassica-
ceous green manures (Table 1). While the levels of suppression
reported in some experiments are encouraging, their variability,
even for the same pathogen is problematic for the wider adoption
of the approach. In many cases there is insufficient information
on key aspects of the approaches used to evaluate the likely role
of isothiocyanates in the responses observed, or to assess the
likelihood that the level of suppression could be enhanced with
better management.
The great difficulty with such empirical studies that char-
acterized earlier work in the area of soilborne pest and disease
suppression by brassicas is that they provide limited scope to as-
sess the basis of observed effects. More tellingly, it restricted the
capacity to pursue opportunities to improve the positive effects
that were implicit in the broad general knowledge that existed.
In highlighting the need for a systematic approach to research in
order to confirm and, if warranted, establish and further develop
biofumigation as a harnessed, workable process, Kirkegaard and
Matthiessen (1999) pointed out that the risks of empiricism are
twofold: firstly, without information on glucosinolate types and
concentration, biofumigation may be disregarded as an option
when different species or cultivars may be suppressive; secondly,
biofumigation may be pursued as an option when the observed
impacts are unrelated to glucosinolate hydrolysis products
(Potter et al., 1998; Kirkegaard and Matthiessen, 2004).
Kirkegaard and Matthiessen (1999) contended that results
existing to that time made effective and consistent adoption
of the concept too problematic. It was feared that a failure to
adopt a systematic approach to biofumigation research may re-
sult in both lost opportunities and inappropriate applications.
They went on to propose a systematic approach with two basic
steps: establishing the potential for suppression and linking sup-
pression to glucosinolate hydrolysis products, and to field test
efficacy in relation to other practices within the farming sys-
tem. Several international research groups have adopted such a
systematic and collaborative approach to assessing and develop-
ing the biofumigation potential of Brassicaceous amendments,
most notably groups in Australia (Kirkegaard et al., 1998), Italy
(Lazzeri et al., 2004) and the United States (Brown and Morra,
1997). In the light of recent findings, a third element, that of
the actual biofumigation related to release efficiency of isoth-
iocyanates and their fate in soil, which was initially implic-
itly part of the biofumigation potential component, has been
teased out and placed separately into the systematic approach
schema. Figure 2 summarizes the components of the system-
atic strategy adopted in order to maximize the isothiocyanate-
related suppression of pest organisms outlined in Figure 1 and,
through that, to achieve adoption of biofumigation for manage-
ment of soilborne pests and diseases in appropriate agricultural
systems.
1. Glucosinolate Profiles
Elements in the determination of biofumigation potential
were to establish the glucosinolate profile of a diverse range
of field-grown brassicas (Kirkegaard and Sarwar, 1998), to de-
termine the effect of environment and ontogeny of the plants on
glucosinolate production and the implications of that for screen-
ing (Sarwar and Kirkegaard, 1998), and the in vitro testing of
the toxicity of some of the more common isothiocyanates occur-
ring in brassicas to soilborne fungal pathogens (Sarwar et al.,
1998). The results of this systematic set of studies revealed a
range of results relevant to the development of biofumigation
as a pest management tool. In a slightly different, but concep-
tually similar approach, tests of the biocidal activity of various
purified glucosinolates hydrolyzed with myrosinase were used
to determine the most active hydrolysis products (Manici et al.,
1997). The results were then used to find Brassicaceae with high
 TABLE 1
A summary of pest suppression in field studies directed at Brassica-based biofumigation, including information relevant to assessing the role of
isothiocyanate-related suppression in the observed responses. Rates of incorporated green manure are based on % w/w of fresh material assuming soil bulk
density of 1.0 g cm−3 and published incorporation depths (or 20 cm if not provided). Level of suppression is calculated as the change in the level of plant
infection/damage or soil populations compared with a non-Brassica green manure control where possible, or fallow. (na: information not provided in reference).
Suppression BiomassC Soil D Rate Glucosinolate Growth Incorporation
Pest Brassica (%) (t ha−1 ) type (% w/w) (µmole g−1 ) stage strategy Reference
Nematodes
Tylenchus semipenetrans B. juncea 92 na na na na Late flower Chopped with mower, Daugovish et al. (2004)
surface irrigated within
30m (30 mm)
Tylenchus semipenetrans B. napus 0 55–89 SL 0.5–6.8 na na Rotary hoe, no irrigation Walker and Morey (1999)
B. juncea 0 11–137
Meloidogyne chitwoodi B. napus 78 20–44 LS, 6.9, 0.9, 2.0 1.3–2.9 na na Rototiller, no irrigation Mojtahedi et al. (1993)
Meloidogyne javanica B. juncea 4–14 fold 90–115 SL, 7.5 7.2–9.3 na 25% flower Slashed, disc incorporated Rahman and Somers (2005)
Meloidogyne javanica B. napus 0 50 na 3.3 na na Chopped (2 cm), soil dry Stirling and Stirling (2003)
B. juncea
Meloidogyne incognita B. napus 0 30–61 LS, 6.7, 0.5 1.0–2.0 2–4 na Flail chopped, disc harrow, Johnson et al. (1992)
Meloidogyne javanica 0 (15–20 cm), moldboard
Criconemella ornate 0 plough (25–30 cm)
Fungi
Sclerotinia minor B. juncea 68 na na na na Late flower Chopped with mower, Daugovish et al. (2004)
S. alba 91 surface irrigated (30 mm)
Aphanomyces euteiches S. alba 32 91.6 SiL, 6.9 4.6 na Start flower Chopped by hand to 15 cm, Muehlchen et al. (1990)
spade incorporated
Rhizoctonia solani B. juncea 25 5.6 S, 7.0, 1.0; 0.2 na Flowering Chopped to 2 cm, Van Os et al. (2004)
incorporated
Pyrenochaeta lycopersici B. rapa 12–52 45 na 2.3 na na Rototiller (assume to Amenduni et al. (2004)
B. oleracea 23–43 30 1.5 20 cm)
Verticillium dahliae B. oleracea 35 56 SL, 6.9, 2.4, 6.8 2.0 na Post harvest Post-harvest residues Subbarao and Hubbard (1999)
chopped and disc
incorporated
(Continued on next page)

245
246
TABLE 1
A summary of pest suppression in field studies directed at Brassica-based biofumigation, including information relevant to assessing the role of
isothiocyanate-related suppression in the observed responses. Rates of incorporated green manure are based on % w/w of fresh material assuming soil bulk
density of 1.0 g cm−3 and published incorporation depths (or 20 cm if not provided). Level of suppression is calculated as the change in the level of plant
infection/damage or soil populations compared with a non-Brassica green manure control where possible, or fallow. (na: information not provided
in reference). (Continued)
Suppression BiomassC Soil D Rate Glucosinolate Growth Incorporation
Pest Brassica (%) (t ha−1 ) type (% w/w) (µmole g−1 ) stage strategy Reference
Verticillium dahliae B. napus 0 100 L, 8.2 na na na Disking or rotovating Davis et al. (1996)
Didymella bryoniae B. oleracea 16 na LFS na na Post-harvest Pots harvest residue Keinath (1996)
chopped, dried and disc
incorporated
Pythium spp. B. napus 0 98 SL, 7.9 4.9 na 50% flower Disked and then rotary Stephens et al. (1999)
B. juncea 0A 122 6.1 na hoed
Bacteria
Ralstonia solanacearum B. juncea 62 45 na 1.2 na Flowering Cut to 3–5 cm lengths and Akiew and Trevorrow (1999)
incorporated by hand
Streptomyces scabiei B. oleracea 90 - na 0.33 B na Post harvest Post harvest residues, dried Gouws and Wehner (2004)
and ground and
incorporated
Insects
Inopus rubriceps B. oleracea 76–86 80 na — na Vegetative Cut and grazed but not Blank et al. (1982)
R. sativus incorporated
Weeds
Various species B. napus 50–96 41–58 LS, 6.7, 0.9 4.1–5.8 na Start flower Rototilled Boydston and Hang (1995)
A
Some evidence for pest stimulation by brassicas.
B
Calculated on dry weight basis.
C
Wet biomass.
D
Texture (S=sand, L=loam, Si=silt, F=fine), pH, OM%, clay%.
Notes: wide range in suppression reported, even for the same pests; some suppression levels reported are promising, but results are variable with some pathogens even
increased; wide range in amounts incorporated and incorporation strategies; only one study gives glucosinolate content, and other data necessary to evaluate the approach for
isothiocyanate-based suppression is mostly incomplete; most studies have not adopted all of the ‘best-bet’ approaches, at least for optimizing the isothiocyanate approach (as
below); some studies reveal other likely causes unrelated to glucosinolates/isothiocyanates and often these are not adequately separated.
“Best-bet” checklist: high glucosinolate variety; 5% W/W of fresh material/soil; pulverize thoroughly and incorporate; moist soil and add water; cover; sandy soils.
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
FIG. 2. Diagrammatic representation of the systematic approach adopted to optimize the isothiocyanate-related biofumigation potential of incorporated Brassica
green manures. (From Kirkegaard and Matthiessen, 2004)
levels of those glucosinolates and implement them in field use
(Lazzeri et al., 2004).
Key findings of Kirkegaard and Sarwar (1998) were that the
variation in biomass, glucosinolate profiles and concentrations
in both shoots and roots provide significant scope to select or
develop brassicas with enhanced biofumigation potential. Total
glucosinolate production on a ground area basis from almost
eighty brassicas sampled at mid-flowering showed wide varia-
tion (0.8–45.3 mmol m−2 ) that derived equally from differences
in biomass and glucosinolate concentration, and that were not
correlated in either root or shoot tissues. Roots contributed an
average of 23.6% (2 to 81%) of the total plant glucosinolates,
with their contribution limited more by low biomass than glu-
cosinolate concentration, which was usually similar or higher
than that of shoots. The glucosinolate concentration in root and
shoot tissue did not correlate with seed levels, ruling out seed
glucosinolate levels as a selection parameter for higher biofu-
migation potential.
The types of glucosinolates present in the tissues varied con-
siderably between species but were consistent within species.
Shoots contained predominantly aliphatic glucosinolates while
aromatic forms predominated in roots. Some species (e.g., B. na-
pus) had predominately non-isothiocyanate liberating glucosi-
nolates in the tissues while others (e.g., B. juncea) produced
mostly isothiocyanate-liberating glucosinolates. Ultimately,
what these results showed was that it is not possible to randomly
use any Brassica (extending even to cultivars within a species)
for biofumigation purposes. It is essential to be selective.
247
Furthermore, the clear implication was that it is not helpful,
and may even be counter productive, to report studies imput-
ing biofumigation effects without describing in some way the
biofumigation potential of the species or cultivar used, as their
capacity in that regard varies greatly. In this regard, only one of
the seventeen field studies in Table 1 reported the glucosinolate
concentration of the Brassica tissues.
Given that biofumigation effects by the definition used
here are related to isothiocyanates, the use of glucosinolate
concentration gives an indirect measure of the biofumigation
potential of plants and is inherently predicated on a high
level of conversion to isothiocyanates. To validate this as-
sumption, and to develop a more direct and simpler analyt-
ical procedure, Warton et al. (2001b) analyzed the glucosi-
nolate concentration and isothiocyanate release by hydrolyz-
ing tissue of matched samples of Brassica shoot and root tis-
sues. In around 80 percent of samples, the molar concentra-
tion of glucosinolates in tissues was greater than that of the
isothiocyanates released upon hydrolysis. The reasons for the
observed trends were obscure and remain speculative with-
out further study, yet are nevertheless noteworthy in terms
of realizing biofumigation potential. Not only was the direct
measurement of isothiocyanate evolution by gas chromatog-
raphy considered a desirable method because it was directed
at the primary bioactive principles, but also it was a simpler
analytical procedure that avoided several steps involving wet
chemistry (Warton et al., 2001b). As will be seen later, isothio-
cyanate release from incorporated fresh tissues, rather than from
248 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
ground, freeze-dried tissues is influenced by even more complex
processes.
2. Effects of Plant Growth and Environment
In a study using a representative selection of brassicas from
five species, the total glucosinolate concentration in root and
shoot tissue varied considerably (3- to 10-fold) with environment
(Sarwar and Kirkegaard, 1998). Interactions between species
and environments meant that the ranking of the brassicas for total
shoot and root glucosinolates changed with environment, while
the added effect of environment on phenological development
and biomass production further influenced overall glucosinolate
production by a crop on a ground area basis. Total glucosinolate
concentration generally declined with the onset of flowering and
was lowest at maturity, whereas spring-sown plants generally
had greater concentration than those sown in autumn.
These results suggested that there will be an optimum time
when the balance of biomass and glucosinolate concentra-
tion yields the greatest glucosinolate production per unit of
ground area. Usefully, the study was conducted simultaneously
in the field and glasshouse, producing results suggesting that a
glasshouse environment can be used to determine the types and
proportions of glucosinolates present in a Brassica and to rank
various types within, but not between, species for the total con-
centration in the tissues. Overall, the results indicated that both
environment and ontogeny can influence all of the plant-related
components of the biofumigation potential. Thus, the glucosino-
late production and biofumigation potential has the capacity to
vary significantly. An accurate estimate of biofumigation poten-
tial therefore requires measurement in the target environment.
Again, few of the studies in Table 1 included more than super-
ficial information on growing conditions or development stage
of the plants, thus nullifying comparisons and assessments of
mechanisms.
3. Biological Activity of Various Isothiocyanates
Sarwar et al. (1998) tested the in vitro toxicity of six
pure isothiocyanates (four aliphatic and two aromatic) to
the mycelial growth of five root pathogens by adding them
to the headspace above, or dissolving them in, the agar
growing medium. Aromatic isothiocyanates were less toxic
in the headspace tests because their lower volatility less-
ened headspace concentration, but were more toxic than the
aliphatic isothiocyanates when dissolved in the agar. In both
the vapor exposure and direct contact experimental methods,
the toxicity of the aliphatic isothiocyanates decreased with
increasing length of the side chain, although there was little
difference between methyl and propenyl isothiocyanate (the
latter occurring commonly in mustard, B. juncea and B. ni-
gra (Kirkegaard and Sarwar, 1998)) in the headspace experi-
ment. Similarly, the shorter chain of the two aromatic isoth-
iocyanates was more toxic in the agar. Thus, shorter-chained
isothiocyanates within each molecular structure class are inher-
ently more toxic, irrespective of volatility; this supported sim-
ilar contact-test findings using an insect by Borek et al. (1995,
1998).
In other studies where only the volatiles released from hydrol-
ysis of Brassica tissues contacted test fungi, tissues dominant in
more volatile types of isothiocyanates such as propenyl isothio-
cyanate were more toxic (Angus et al., 1994; Kirkegaard et al.,
1996; Mayton et al., 1996) while in experiments using isoth-
iocyanates dissolved in agar, aromatic isothiocyanates were up
to 20 times more toxic (Drobnica et al., 1967). In an in vitro
study using an insect as the bioassay subject that compared va-
por exposure of pure methyl (aliphatic) and pure 2-phenylethyl
(aromatic) isothiocyanates, Matthiessen and Shackleton (2000)
found little difference in toxicity at warmer temperature, but tox-
icity of the less volatile aromatic isothiocyanate fell away partic-
ularly sharply as temperature was reduced across the lower part
of the range 20, 15, 10 to 5◦ C. It was concluded that the equiv-
alent toxicities at warmer temperature were a consequence of
the more than 70-fold higher contact toxicity of 2-phenylethyl
than methyl isothiocyanate (Borek et al., 1995) overriding its
much lower volatility. Similarly, when the insects were exposed
to vapors from hydrolyzing plant tissues rich in either propenyl
or 2-phenylethyl glucosinolates at 15◦ C, the absolute amounts
of dry tissue required were similar for given mortality levels,
indicating that the effect was consistent when plant tissue sub-
stituted for pure isothiocyanates (Matthiessen and Shackleton,
2000).
Sarwar et al. (1998) found up to a seven-fold variation in
susceptibility of the individual fungi they tested. Smith and
Kirkegaard (2002), seeking to explain whether the beneficial
break-crop effects of canola in canola-wheat rotations in the
Australian cereal belt (Angus et al., 1991) may be associated
with biofumigation effects, recorded considerable variation in
the sensitivity of different fungi to 2-phenylethyl isothiocyanate,
which occurs in the roots of canola (Kirkegaard and Sarwar,
1998). This is consistent with differences in sensitivity of fun-
gal species to pure isothiocyanates that have been reviewed by
Brown and Morra (1997), Fenwick et al. (1983) and Rosa et al.
(1997), where fungicidal concentrations of isothiocyanates for
given fungi may differ by an order of magnitude. The varia-
tion in sensitivity to isothiocyanates provides opportunities to
select Brassica species that produce the largest quantities of the
isothiocyanates most toxic to the target organism.
Tests carried out using hydrolysis of pure glucosinolates
in vitro revealed that sulfur-substituted aliphatic types were the
most toxic to Fusarium (Manici et al., 1997), and to Pythium and
Rhizoctonia (Manici et al., 2000). Subsequent wide screening of
several glucosinolate-containing plant species has led to the se-
lection and recent commercialization of several biofumigants
that produce high quantities of the most potent glucosinolates
(Lazzeri et al., 2004). Using this systematic approach to match
the most potent brassicas with target organisms known to be
sensitive to particular isothiocyanates provides the best possi-
ble chance of achieving successful suppression in the field. Not
only do these results indicate that the glucosinolate profile of
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
Brassica biofumigants will differentially affect pathogens, but
they possibly help explain why the efficacy of metam sodium is
widely regarded as variable. Again, the paucity of information
in the studies listed Table 1 make it difficult to determine the role
of isothiocyanates in suppressive effects of brassicas or suggest
methods to improve observed beneficial effects.
4. Implications for Adoption
The effectiveness of brassicas for biofumigation will ulti-
mately depend on many factors beside the biofumigation poten-
tial of the particular Brassica used. The timing of incorporation
or exudation of the tissue needs to coincide with the suscep-
tible stage of the pest organism and suppression must be of
a sufficient level and persistence to provide protection for the
crop of interest. Additionally, the effectiveness of biofumiga-
tion will be influenced by the effectiveness of incorporation,
activity of the hydrolyzing myrosinase enzyme, efficiency of
the hydrolysis and losses resulting from volatilization, sorption
onto soil components, leaching and microbial degradation. Nev-
ertheless, the concept of biofumigation potential has provided a
foundation from which to consider strategies to manipulate its
components to enhance suppression for particular management
scenarios.
Some of the more speculative early conclusions of Kirkegaard
and Sarwar (1998) (and others over the years), in particular
those based on extrapolation of in vitro determinations of isoth-
iocyanate toxicity to the field, have proven inapplicable once
assessed in vivo (Matthiessen and Shackleton, 2005 and see fol-
lowing). In general, however, subsequent findings by a range
of researchers, particularly on the most appropriate Brassica
species to use, the plant parts and types of isothiocyanates on
which to focus and the conditions required to achieve maximum
release of isothiocyanates from Brassica tissue in soil, have ulti-
mately shown the prescience of this integrative approach. It has
been valuable in taking research beyond the limitations and even
misdirections stemming from what was arguably hitherto over-
extrapolation from in vitro laboratory and empirical field studies,
to systematically go much closer to achieving field-applicable
outcomes (see following). Previously, few field studies reported
this crucial information necessary to assess potential to improve
field efficacy (Table 1).
It is relevant to comment that optimistic extrapolation from in
vitro results is commonplace in many areas of biological science
that propose potential solutions to a problem. There are a variety
of reasons for this. Unfortunately, extrapolation or overstatement
particularly from in vitro laboratory studies (which are much
simpler to do than in vivo or field studies), that is not carefully
qualified can slow progress or even be counter-productive to the
development of a concept because its proponents can be seen
by practitioners as unrealistic toward real-world situations and
needs. We believe that the concept of biofumigation has had
its share of this and there is a need for care in presentation of
results to avoid any proliferation of the issue. Similarly, poor
or inconsistent results from empirical field studies that have not
249
adequately represented crucial factors (Table 1) may stultify
progress by being given inappropriate levels of credence.
D. Realizing Biofumigation Potential
To add further complexity to the contrasts and variations in
biological activity of the different types of isothiocyanates un-
der simplified in vitro test systems (Borek et al., 1995; Borek
et al., 1998; Sarwar et al., 1998), methyl isothiocyanate was
strongly (relative to methyl bromide) sorbed by soil, particu-
larly that containing a high level of organic matter (Matthiessen
et al., 1996). The ultimate test of the use of metam sodium
or isothiocyanate-based biofumigation is the biological activity
achieved by the isothiocyanates in soil. Despite its propensity
to be sorbed, practical field use over the last 50 years shows
that methyl isothiocyanate from metam sodium can achieve its
potential by effectively controlling many soilborne noxious or-
ganisms, although the variability of results from even the typi-
cally high commercial application rates required to achieve that
potential is well-recognized (van Berkum and Hoestra, 1979).
Given that a wide range of other isothiocyanate precursor glu-
cosinolates, both aliphatic and aromatic, occur in brassicas, the
question arises as to what is the biological activity of other isoth-
iocyanates (see Kjaer et al. (1963) for details of the structural
chemistry of the isothiocyanates) relative to that of methyl isoth-
iocyanate in the soil environment and what may be the effect of
different environmental conditions such as temperature on the
expression of that activity.
1. Beyond In Vitro Studies—Activity of Isothiocyanates in Soil
Soil-dwelling organisms are notoriously difficult subjects for
dose-mortality testing of biological activity under in vivo con-
ditions. Cryptic, resting and hidden stages, dependence on the
presence of soil for even short-term survival which potentially
causes high mortality in untreated controls, and extraneous mor-
tality that can be caused during extraction from the soil can
confound mortality caused by test agents. Emulating the soil
environment presents a major challenge for realistically deter-
mining the likely practical biological activity of isothiocyanates
and the biofumigation potential of various putative biofumigant
brassicas against relevant soilborne pests or diseases. It is there-
fore not surprising that there has been a strong reliance on in
vitro studies for gauging biological activity.
Recently, examination of the isolated biological effects of
pure and plant-derived isothiocyanates was extended to emu-
late their use in soil (Matthiessen and Shackleton, 2005). Their
relative biological activity was determined both in vitro and in
the presence of contrasting soils and over a range of tempera-
tures. Vapor exposure of contrasting pure isothiocyanates (the
aliphatics methyl and propenyl and the aromatics benzyl and
2-phenylethyl), metam sodium and Brassica tissues generating
contrasting isothiocyanates (propenyl and 2-phenylethyl) was
tested. The in vivo tests gave results that contrasted sharply
with those from various in vitro tests that have appeared
250 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
in the literature over the years (Matthiessen and Shackleton,
2005).
Low effective doses and steep responses of methyl isothio-
cyanate, structurally the simplest of the organic isothiocyanate
molecules and the most volatile, showed it to be the most bi-
ologically active isothiocyanate under all test conditions. The
results obtained using metam sodium closely reflected those
from pure methyl isothiocyanate. The equivalence of the re-
sults was indicative of high conversion to methyl isothiocyanate
in soil, despite the capacity of metam sodium to also produce
other degradation products, and supports the conventional inter-
pretation from quantitative analytical studies that the pesticidal
efficacy of metam-sodium is largely, if not entirely, the result of
its decomposition to methyl isothiocyanate (Turner and Corden,
1963).
Biological activity of methyl isothiocyanate was maintained
across the broadest range of temperatures. It only declined to any
appreciable extent at 5◦ C, which is below or at the lower end of
the range of soil temperatures where agricultural activities in-
volving the use of fumigant-like pesticides would commonly be
undertaken. The factor having the greatest dampening effect on
the biological activity of all compounds and tissues was the pres-
ence of soil, particularly that with high organic matter content.
Compared to in vitro, 10- to 20-fold more methyl isothiocyanate
was needed for the same mortality in the presence of sand and
loam, while in the presence of a peaty soil the extra requirement
was 25- to 50-fold (Matthiessen and Shackleton, 2005).
In comparison to methyl isothiocyanate, the activity of the
other pure isothiocyanates tested was 2 to 4-fold less in vitro.
This result was in striking contrast to the substantially greater
toxicity of propenyl, benzyl and 2-phenylethyl isothiocyanates
(3-, 56-, and 72-fold, respectively) compared to methyl isothio-
cyanate in direct contact tests against another soil insect (Borek
et al., 1995). At 25◦ C propenyl, benzyl and 2-phenyethyl isoth-
iocyanate are 5, 1700 and 2300 times less volatile than methyl
isothiocyanate, respectively (Boublik et al., 1973; Howard and
Meylan, 1996). The higher ratios for the biological activity of
the aromatic isothiocyanates relative to methyl isothiocyanate in
the vapor-exposure in vitro tests of Matthiessen and Shackleton
(2005) compared with the direct-contact in vitro tests of Borek
et al. (1995) suggests that the inherently high contact activity of
aromatic isothiocyanates is generally outweighed by their low
volatility tending to depress activity in non contact tests.
The vapor-exposure activity of the other isothiocyanates com-
pared to methyl isothiocyanate was somewhat lower in vitro, but
it was under the influence of the environmental factors tested,
particularly soil type, that much more striking and relevant
contrasts emerged. The biological activity of propenyl isoth-
iocyanate declined 2- to 5-fold in the presence of soil, although
it was only at the lowest temperature and in organic matter-rich
peat soil that activity fell away more sharply. Activity of the
aromatic isothiocyanates was suppressed to a much greater de-
gree, most evident for 2-phenylethyl isothiocyanate at 5◦ C in
peat soil where activity was many thousand-fold reduced. The
bioassay results of Matthiessen and Shackleton (2005) suggest
that aromatic isothiocyanates in particular are sorbed by soil to
an even greater degree than what, from direct analytical mea-
surement and comparison with methyl bromide was interpreted
as highly active sorption of methyl isothiocyanate by the same
soils (Matthiessen et al., 1996). It would, however, still be un-
realistic to extrapolate results derived in laboratory microcosms
to directly attempt to quantify amounts of pure isothiocyanates
or biofumigants to achieve suppression of soilborne pests and
diseases under field conditions.
2. Implications for Biofumigation
A consequence of revealing these sharply contrasting pat-
terns of biological activity of the pure isothiocyanates in soil
is that it raises questions about the nature of the link between
intrinsic activity measured under simplified in vitro conditions
and the activity that may be realized under field conditions. It
seems clear that initial optimism derived from in vitro labo-
ratory studies about the greater potential of aromatic isothio-
cyanates for suppression of soilborne pests and diseases (Borek
et al., 1995) may have been misplaced. The dramatically re-
duced biological activity and extreme flattening of the slope
of the response for the aromatic isothiocyanates in peat soil at
lower temperatures shows that aromatic isothiocyanates are in-
fluenced much more by the environmental conditions than are
aliphatic isothiocyanates (Matthiessen and Shackleton, 2005).
It is possible that the very much greater contact toxicity of
the aromatic isothiocyanates, which was attributed to charac-
teristics of their molecular structure that influence their re-
activity with essential enzymes (Borek et al., 1995), also en-
hanced their reactivity with other substrates such as the or-
ganic components of soil. If so, their low volatility would be
expected to interact with this imputed heavy sorption to very
much further reduce their biological activity as temperature
is lowered, as was observed by Matthiessen and Shackleton
(2005).
Similar responses of the test organism to hydrolyzing plant
tissues supported the conclusions from the pure isothiocyanates
that aliphatic isothiocyanates are likely to have greater bio-
logical activity than aromatic isothiocyanates in natural condi-
tions, despite the opposite effect by contact in vitro (Matthiessen
and Shackleton, 2005). Mustard tissue produced almost entirely
propenyl isothiocyanate; fodder rape tissue, while not so narrow
in its isothiocyanate-producing spectrum, produced more aro-
matic isothiocyanate than aliphatic isothiocyanate. Plant tissue–
derived isothiocyanates responded in a similar manner to soil
and temperature as the pure form of isothiocyanates they con-
tained. Laboratory tests of the effects of cellular-disrupted In-
dian mustard tissue on proliferation of the bacteria Ralstonia
solanacearum in three soils varying in clay content showed that
larger quantities of tissue were required to suppress the organism
as clay content increased (Wang et al., 2006).
The activity of different types of isothiocyanates is expected
to be related to the amount available to readily contact a target
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
organism relative to the rate of deactivation by sorption onto soil.
Consequently it is conceivable that aromatic isothiocyanates that
have the opportunity to readily contact the target (e.g., in solu-
tion) may achieve their greater potential for biological activity.
This is an aspect that is speculative and requires further work
to elucidate. Given present information, however, the indica-
tions remain that structurally simpler aliphatic isothiocyanates
are more likely to achieve biological activity under field condi-
tions than their structurally more complex counterparts or aro-
matic isothiocyanates.
In terms of practical implementation of biofumigation, it
therefore now appears clear that brassicas producing high
amounts of short-chain aliphatic isothiocyanates are likely to
have the greatest potential for suppression of noxious soil organ-
isms. Conversely, those producing long-chain aromatic isothio-
cyanates are likely to have the lowest potential for a role as
biofumigants, despite such isothiocyanates being intrinsically
more biologically active in contact tests (Borek et al., 1995;
Smith and Kirkegaard, 2002). Although in the diverse and com-
plex soil environment there are likely to be major influences
and interactions of many factors on realizing potential such as
greater sorption of the isothiocyanates by organic matter- and
clay-rich soils (Matthiessen and Shackleton, 2005; Wang et al.,
2006), the results indicate that the better starting point for se-
lection of brassicas for isothiocyanate-based biofumigation is
those producing simple aliphatic rather than those dominant in
complex aromatic isothiocyanates.
Generally, shoots of brassicas (which represent >90% of
total plant biomass for most brassicas) contain predominantly
aliphatic, and roots predominantly aromatic, glucosinolate pre-
cursors of isothiocyanates (Kirkegaard and Sarwar, 1998). Thor-
ough pulverization of plant tissue is only readily achievable
on above-ground parts of plants using mechanical implements;
attempts to do so for below-ground plant parts will result in
counter-productive destruction of soil structure and would, in
any event, be unrealistic as a practical farm operation. These
various aspects come together to strongly suggest that the focus
for achieving practical benefits from isothiocyanate-based bio-
fumigation should be on agronomically appropriate brassicas
with high above-ground biomass rich in precursors of aliphatic
isothiocyanates.
Under some circumstances there may be soil structure ef-
fects that override sorption effects. Ben-Yephet and Frank (1985)
found that the penetration and efficacy of metam sodium could
be higher in clay soil than a more sandy soil provided that the
aggregate structure allowed rapid percolation of the aqueous so-
lution. Conceivably, this effect could apply to biofumigation so
long as the soil was not disaggregated inappropriately by incor-
poration of plant material. Most field studies of brassicas used
as biofumigant green manures have not provided sufficiently
detailed information on soil characteristics to enable adequate
assessment of likely effects (Table 1). In general, it seems most
likely that sandier soil with low organic matter content will allow
better expression of biofumigant effects.
251
3. Achieving High Release of Isothiocyanates
It is possible to select and grow biofumigant brassicas that
contain enough glucosinolate to potentially produce levels of
isothiocyanate equal to or greater than the levels applied as com-
mercial synthetic methyl isothiocyanate-generating pesticides
(Kirkegaard and Sarwar, 1998). However, it is the efficiency with
which the glucosinolates can be converted into isothiocyanates,
and their fate in the soil that ultimately dictates their effective-
ness as pesticides. Logically, some form of incorporation of
above ground biomass was recognized early as being necessary
to promote maximum whole-plant isothiocyanate release into
soil. Initial attempts tended to involve ad hoc use of cultivat-
ing implements such as rotary tillers to chop and bury the plant
material (of unspecified biofumigation potential) into the soil in
the one operation (Table 1). Few, if any, attempts were made to
measure the results of such approaches in the field, particularly
the levels of isothiocyanates present in soil. But in laboratory
studies simulating such field operations, isothiocyanate forma-
tion from incorporated fresh tissues and measured in soil, was
less than 5 percent of the potential amount present in the stand-
ing biomass, assuming complete conversion of glucosinolates to
isothiocyanates (Gardiner et al., 1999; Morra and Kirkegaard,
2002). Such low efficiency of this conversion gives absolute
amounts of isothiocyanate markedly less than achievable with
recommended doses of metam sodium (Brown et al., 1991).
Farmers often have concerns about the use of rotary cultiva-
tors, as they tend to destroy soil structure. For this reason, there
is a general preference to avoid unnecessary use of rotary tillage,
except possibly where soils are structureless coarse sands. Ide-
ally, this preference needs to be factored into the development of
biologically-based alternative methods of soilborne pest and dis-
ease suppression. This is to ensure that advantageous attributes
are not downgraded by other characteristics that may be dis-
advantageous and that could lead to premature rejection of the
technique by farmers.
A recent watershed in the understanding of isothiocyanate re-
lease from glucosinolate-rich plant tissue came with the finding
in laboratory experiments that disruption of Brassica leaf tissue
at the cellular level, demonstrated by freezing and thawing, could
increase isothiocyanate concentration in soil by around two or-
ders of magnitude compared to cut fresh leaf tissue (Morra and
Kirkegaard, 2002). Moreover, the addition of sufficient water to
the soil samples to waterlog them doubled the already high level
of isothiocyanate detected in the soil (Morra and Kirkegaard,
2002). These revelations inspired field studies (Matthiessen
et al., 2004a) aimed at determining whether similarly high levels
of isothiocyanates could be achieved in soil under field condi-
tions, and whether there may be scope to reduce the use of rotary
tillage in so doing.
By thoroughly pulverizing the above-ground tissue of mus-
tard (B. juncea), leaving it on the surface of the ground and soon
after watering it heavily, Matthiessen et al. (2004a) achieved
propenyl isothiocyanate concentration in field soil of about
100 nmol g−1 . This compared with the 200 nmol g−1 of methyl
252 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
isothiocyanate measured in the same soil fumigated with com-
mercial rates of metam sodium (B. Warton and J. N. Matthiessen,
unpublished data). Initially, based on the knowledge from in vitro
laboratory studies that propenyl isothiocyanate is intrinsically
twice as toxic as methyl isothiocyanate (Borek et al., 1995), it
was conceivable that 100 nmol g−1 propenyl isothiocyanate in
soil may therefore be the equivalent of 200 nmole g−1 methyl
isothiocyanate. However, the in vivo bioassay of Matthiessen and
Shackleton (2005) described above demonstrated that a greater,
rather than lower, propenyl isothiocyanate concentration in soil
is likely to be required. This was because of the greater suppres-
sive effect of soil and temperature on its biological activity than
on that of methyl isothiocyanate. Gratifyingly, on the other hand,
the concentration of ex-Brassica isothiocyanates measured in
the field equaled that achieved in the laboratory experiments of
Morra and Kirkegaard (2002) using thawed frozen leaf tissue
and excess water.
The field results therefore emphatically emphasized the prin-
ciples of cell-level tissue disruption and need for excess water
to maximize isothiocyanate production and release that were
vividly demonstrated by Morra and Kirkegaard (2002). They
also suggest that the experimental system used in the field was
optimal and so have identified the maximum benchmark against
which to develop robust practical methods usable by farmers.
These findings also suggest it may be necessary to re-visit the
conclusions drawn from studies on biofumigation where tissues
have been rotavated into relatively dry soil, or where no detailed
information on either the incorporation process or soil water
content was reported (Table 1).
E. Field Implementation
1. Key Factors in Field Implementation
Pulverization of the above ground plant material can be read-
ily achieved, so long as it is recognized that the mulching im-
plement must have hammers, rather than blades and must be
operating at high speed. This ensures that the plant tissue is effec-
tively broken down at the cellular level, rather than chopped into
fragments (Matthiessen et al., 2004a). However, even with such
treatment of the plant tissue, it is clear from both the laboratory
(Morra and Kirkegaard, 2002) and field studies (Matthiessen
et al., 2004a) that its intrinsic moisture content is insufficient
to maximize the hydrolysis of glucosinolates to isothiocyanates.
The experimental system used in the field studies of Matthiessen
et al. (2004a) simulated flood irrigation and used an unrealisti-
cally large amount of water, in order to demonstrate key prin-
ciples. Further work is necessary to determine the effectiveness
of alternative irrigation methods such as sprinklers, and require-
ments for the amount of water applied, in order to work this new
knowledge into an adoptable system. It seems very likely that
soil type will have a substantial effect on the amount of water
required and rate of transport of the isothiocyanates in solution
into the soil. A question that remains is whether the very high
concentration of isothiocyanates measured in the soil under pul-
verized mustard tissue left on the soil surface and watered could
also have been achieved had the plants been pulverized and in-
corporated into the soil and then watered. However, heavy wa-
tering of recently rotavated soil is probably a practice that many
farmers would prefer to avoid.
The preliminary results outlined above primarily illustrate
the important principles of tissue disruption and water addition
in maximizing isothiocyanate production, and that they can be
achieved in the field. They point the way very clearly to the type
of further work that is necessary to build robust practical sys-
tems to achieve the same results, and to evaluate their efficacy
in suppressing soilborne pests and diseases. The requirement
for considerable extra water possibly presents a larger logisti-
cal challenge than pulverizing the plants. The crucial practical
issue would now appear to be to determine the minimum wa-
ter requirement for effective isothiocyanate release, and how
to achieve the high levels of water that are seemingly required
to both enhance isothiocyanate release and to rapidly carry the
isothiocyanates so formed into the soil. It is this area of research
that rates as the greatest priority in bringing about the adoption
of biofumigation, particularly at the more intensive end of the
production system spectrum where the sole aim is pest and dis-
ease management. That is to say in production systems where
biofumigation sensu stricto (the emulation of metam sodium)
is the key objective, as compared to systems where agronomic
benefits are also highly valued in their own right such as potato
production in the Pacific Northwest of the U.S.A. (Gies, 2004;
McGuire, 2004).
2. Farming Systems Fit
Following the establishment of a conceptual framework and
methodology to assess and express biofumigation potential, it
is then possible to undertake a systematic process to implement
biofumigation in the field and assess its efficacy. The selection
of plant ecotypes based on their qualitative (type of dominant
glucosinolate) and quantitative glucosinolate content needs to
be complemented by selection for agronomic performance that
makes their cultivation practical (Lazzeri et al., 2004). Although
seemingly self-evident, it is important to consider at the outset
the potential fit of a biofumigant green manure in the farm-
ing system of interest. In some cases, the green manure may
fit easily into the system, replacing an existing green manure
(e.g., grass or legume) or periods of fallow or pasture leys. An
example of the latter is the use of mustard green manures in
the potato farming system in the irrigated areas of the Pacific
Northwest of the U.S., where it was utilized on over 16,000 ha
in 2003 (Gies, 2004; McGuire, 2004). In other circumstances
adjustments may be necessary to the timing of other system
components in order to accommodate a period of green ma-
nure, or indeed it might be immediately evident that a Brassica
rotation will not be possible to fit into the cropping system be-
cause of such factors as an insufficient temporal or inappropri-
ate seasonal window, and so other alternatives will need to be
sought.
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
The soil type, climate and time of year in which the crop is to
be grown will influence the choice of species. For example, most
mustard (B. juncea) varieties flower in response to long days and
if sown in spring will bolt into flowering and produce limited
biomass, while winter-adapted brassicas with a vernalization
requirement will remain vegetative for long periods when sown
at the same time. The crops may be susceptible to local pests
and diseases and pathogens for which control is prohibitively
expensive, may harbor or promote organisms noxious to nearby
or following crops, or may be sensitive to frost. Management of
the crop needs to be compatible with the equipment available
on-farm to contain costs—establishment of small-seeded crops
such as brassicas can be problematic, particularly for broad-scale
horticultural producers such as potato growers unaccustomed to
their management.
The wide variety of phenological and morphological diver-
sity within the Brassicaceae provides good scope to select those
most appropriate for different environments and circumstances.
However, the system will need to allow enough time, at the ap-
propriate time of the year, to allow soil preparation, sowing,
sufficient growing period to obtain required biomass, pulverisa-
tion, incorporation and breakdown of the biofumigant crop be-
fore preparations begin for growing the production crop. Clearly
a biofumigant Brassica that needs to be grown at a time that does
not work in a complementary way with the growth cycle of the
crop for which pest or disease suppression is sought will not be
a suitable option.
3. Case Study 1—Commercial Implementation in Potatoes
This case study (McGuire, 2004) was an examination in com-
mercial potato crops comparing the pest and disease manage-
ment efficacy of a mustard green manure with the conventional
application of metam sodium. It also examined the effects of
the green manure on soil quality (measured by water infiltra-
tion, which was both meaningful to the farmers and because
it integrates several soil quality attributes such as bulk density,
aggregation and pore space). In that regard alone it was valu-
able and comprehensive, but it was unique in also including a
comparative economic analysis of the costs. These studies did
not include isothiocyanate analysis in the soil, so it is not pos-
sible to discriminate to what degree the beneficial effects were
based on biofumigation effects sensu stricto or other attributes
of the mustard green manure. However, in the particular potato
production system investigated, there were no statistically sig-
nificant differences in potato yields between the two treatments,
over three trials. Nor were there any significant differences in
tuber size classes, an important quality parameter.
The economic analysis of direct costs, even without conduct-
ing a detailed nitrogen budget that may capture some additional
benefits through cycling between that applied to the mustard
and a likely possible reduction in that needing to be applied to
the subsequent potatoes, showed that the mustard green manure
substituted for metam sodium increased net return by USD169
ha−1 (McGuire, 2004). This case study demonstrates that in-
253
volvement of growers and agronomists with research teams at
the outset is much more likely to provide the most expeditious
development of the technology. It also points up the importance
of growers conducting their own trials to ensure that such rota-
tions will be appropriate to their production system, and to tune
agronomic details appropriately.
4. Case Study 2—Bacterial Wilt
This case study supports the conclusion that the incorpora-
tion strategy is critical to ensuring the biofumigation potential
of brassicas is realized in the field. Bacterial wilt (Ralstonia
solanacearum) is one of the most important diseases of bacte-
rial origin in the world, particularly in tropical areas (Haywood,
1991). Studies in Australia demonstrated for the first time that
biofumigation using Indian mustard (B. juncea) green manure
was effective in reducing the level of bacterial wilt in the soil and
reducing the severity of the disease in a following tobacco crop
(Akiew and Trevorrow, 1999). Using the systematic approach
outlined in Figure 2, a project currently being undertaken by S.
E. Akiew and J. A. Kirkegaard first sought to identify brassi-
cas that would grow in tropical northern Australia in the period
from July to September, preceding a tomato crop. A wide range
of the species tested including B. juncea, B. napus, S. alba, R.
sativus, B. nigra grew well during this cooler winter period,
providing flexibility to choose from a range of potential bio-
fumigants based on the toxicity of their isothiocyanate to the
bacterial wilt pathogen. The general sensitivity of the pathogen
to isothiocyanates was confirmed in laboratory assays, where
bacterial wilt was found to be the most sensitive of a range of
bacteria tested to 2-phenylethyl isothiocyanate in vitro (Smith
and Kirkegaard, 2002).
A number of different approaches were used to assess the
suppressiveness of incorporated tissues in soil. Preliminary test-
ing of chopped Brassica leaf tissues incorporated into soil in
muslin bags and buried in the field showed that most Brassica
tissues were more suppressive than non-Brassica controls, but
no difference in the toxicity of different Brassica tissues, and
therefore isothiocyanate types, was identified. In other experi-
ments, non-Brassica tissues or brassicas low in glucosinolates
were also shown to have some suppressive impacts, suggesting
a more general long-term impact of organic matter on bacterial
wilt rather than only a biofumigation effect related to isothio-
cyanates. The experimental approach was modified based on
the results of Morra and Kirkegaard (2002) and Matthiessen
et al. (2004a), that showed the high and rapid release of isothio-
cyanates following cellular-level tissue disruption and the need
for considerable water. A novel bioassay technique was devel-
oped to separate the short-term impacts of isothiocyanates from
other longer-term impacts of incorporated tissues (Akiew et al.,
2005). In this approach, bacterial wilt-inoculated soil was placed
above frozen leaf tissue in small tubes, which were then incu-
bated at room temperature for three days before the soil above the
tissue was tested to determine bacterial wilt survival. Using this
technique, only volatile compounds released from the thawing
254 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
tissue contacted the soil above during the relatively short incuba-
tion period. These experiments showed that brassicas containing
higher levels of more volatile isothiocyanate precursor glucosi-
nolates such as the propenyl form (e.g., mustards) were more
suppressive than those with lower concentrations or less volatile
types such as 2-phenylethyl (e.g., rape).
The information from the laboratory experiments supported
the findings of Matthiessen and Shackleton (2005) suggesting
that mustards high in propenyl glucosinolate would be the most
suppressive biofumigants if conditions conducive to isothio-
cyanate release could be satisfied in the field. The effective-
ness of a range of biofumigants for suppression of bacterial wilt
was then evaluated in a highly bacterial wilt-infected field us-
ing a non-Brassica control (soybean) and a range of different
Brassica species including mustard and rape. The biofumigants
were grown to produce around 5 to 6 kg m−2 of fresh biomass.
They were then pulverized and incorporated, and immediately
irrigated, before beds were formed and covered with reflective
plastic for weed control, as is common in commercial practice.
During crop growth all of the biofumigants delayed the onset
and reduced the incidence and severity of wilting in the subse-
quently planted tomatoes compared with the soybean control.
At harvest the mustard treatments were more effective than the
rape treatments in reducing wilt severity and increasing tomato
yield (S. E. Akiew and J. A. Kirkegaard, unpublished). A second
growing season of more comprehensive field studies to confirm
results across years is currently in progress.
This case study demonstrates that for pathogens sensitive to
isothiocyanates, careful selection of brassicas high in the most
toxic isothiocyanates that are least sorbed by soil, and man-
agement of the incorporation process to maximize their release
in soil can result in significant levels of biofumigation-based
control in the field. It is also useful to note that significant non-
glucosinolate related disease suppression has been observed dur-
ing the course of the work. This non-glucosinolate related sup-
pression can confound attempts to interpret experimental results
if adequate controls are not included in the experiments and this
is a priority of the field experiments currently in progress. Al-
though the exact mechanism of those other effects is at present
uncertain, it is likely that similar mechanisms have been operat-
ing in historical studies where pest and disease suppressiveness
was variable and inconsistent.
IV. FUTURE DIRECTIONS
A. Capitalizing on Results and Maintaining Momentum
Recent research and applied use has substantially changed the
conceptual basis and applied aspects of biofumigation. It should
signal that the concept provides an opportunity for soilborne
pest and disease suppression, and other benefits, but it must be
applied in appropriate agricultural systems. It should also signal
the end of the usefulness of isolated in vitro laboratory studies
aimed at demonstrating potential, but that fail to address key
practical issues, and in the process risk compromising the cred-
ibility of the concept. Sufficient information now exists to make
judgments about the best approach to implement biofumigation.
It is only with testing of biofumigation within such a context that
real judgment about its applicability to individual situations will
be possible. We suggest some relevant issues to consider.
1. Revisit Previous Work and Adopt a “Best Bet” Approach
The recent advances in strategies to enhance the isothio-
cyanate release from Brassica biofumigants suggest that there is
a need to revisit the previous studies such as those summarized
in Table 1. The level of suppression achieved in those studies
has clearly been limited by the lack of understanding of some
of the key elements important to maximize success including:
(a) selection of Brassica varieties high in isothiocyanates that
are most toxic to pest organisms and less subject to losses in soil,
(b) growing critical amounts of material for incorporation (e.g.,
5% w/w fresh plant material/soil), (c) thorough pulverization of
above-ground tissue and rapid incorporation, (d) watering and/or
covering to increase isothiocyanate release and reduce volatile
loss, (e) targeting light textured, low organic matter soils for best
effects. Adopting these elements, or as many as appropriate to
the agricualtural system, as “best bet” approaches in both future
research studies and practical applications will enhance chances
of success and further identify characteristics of situations where
biofumigation has a role in the management of soilborne pests
and diseases. A very useful adjunct to such studies would be the
inclusion of relevant controls to separate isothiocyanate-related
effects from other green manure impacts on pest suppression.
2. New Research to Increase Use of Biofumigation
Further immediate progress in increasing the isothiocyanate-
related suppression could arise from continued attention to selec-
tion or purposeful breeding of brassicas high in the most effective
isothiocyanate-liberating glucosinolates. Although there is some
specific development of appropriate varieties at present (e.g., as
used in Italy and the United States (Lazzeri et al., 2004; Gies,
2004)) this is not known to have progressed to breeding that
is targeted specifically at increasing glucosinolate concentra-
tion. Rather, it has been directed at identifying existing material
within selections with high glucosinolate and biomass poten-
tial (Gies, 2004; Patalano, 2004). The elucidation of the genes
involved in the biosynthesis of glucosinolates may in future pro-
vide opportunities to engineer specific biofumigant types to raise
levels above those likely to be achieved in conventional crossing
programs (Halkier and Du, 1997; Mithen, 2001). However, such
investment in traditional or molecular breeding approaches will
necessarily be predicated on a clear market potential for such
lines. Those market signals will only come with farm-scale ap-
plications initially flowing from implementation of the “best
bet” approach, followed by positive results and the user-driven
evolving improvements that would inevitably follow successful
beginnings.
The other most immediate increase in isothiocyanate-related
suppression is likely to arise from building upon the focus on
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
techniques of pulverization and incorporation that maximize
isothiocyanate release and minimize losses in soil. At present
existing machinery is being utilized for this purpose. There are
no doubt opportunities to innovate in the engineering and design
of machinery designed to simultaneously pulverize and incor-
porate green manures into the soil with rapid covering. This
could occur in much the same way that specific machinery for
synthetic fumigant application has evolved, often borne from
end-users building their own. Again, it is market signals that
will ultimately drive such innovations. Equipment used in Eu-
rope to incorporate farmyard manures and slurries into the soil
to prevent undesirable odours near urban centers already incor-
porate some of the necessary features. Such machinery would
have applications beyond those specifically related to biofumi-
gation, as most green manures could be incorporated in this
way.
3. Environmental Issues—Fate of Isothiocyanates
and Glucosinolates in Soil
The appeal of biofumigation is in its “clean and green” ap-
proach, although technically it attempts to emulate the same
process achieved with methyl isothiocyanate through the use
of metam sodium. The fate of isothiocyanates and glucosino-
lates will come under the spotlight and the general environ-
mental implications will need to be identified. In first moves
addressing this theme, Gimsing and Kirkegaard (2006) have re-
cently found that both isothiocyanates and glucosinolates can
remain in soil for several days after incorporation and both
can be leached below the level of incorporation by irrigation.
The potential non-target and off-site impacts of these com-
pounds will be of interest especially given the other mecha-
nisms of biological suppression operating with Brassica green
manures in some cases. This information will be necessary
to provide confidence in the process and avoid unintended
consequences.
V. ENHANCED BIODEGRADATION OF SOIL-APPLIED
PESTICIDES
A. Background and History
1. Environmental Fate
The fate of xenobiotics in the environment is determined by
both abiotic and biotic factors, but it is now recognized that
the mineralization, or complete degradation, of a compound in
the environment is almost always a consequence of microbial
activity (Alexander, 1980). In the early Post–World War II era,
longevity of soil-applied pesticides was seen as one of their most
useful properties. However, Audus (1949) noted that the ultimate
recovery of soil from the toxic action of the herbicide 2,4-D was
important in practice and demonstrated with repeated soil per-
fusion in the laboratory that its detoxification resulted almost
entirely from the activity of microorganisms. Prophetically, the
temporal dynamics of the degradation indicated that microor-
ganisms had proliferated in response to the applied metabolite.
255
But it appears that such processes, if they occurred in the field,
were regarded as having little consequence and Audus’s (1949)
study seems to have been relegated to the realm of laboratory
curiosity for some time. Subsequently, during the 1950s and
1960s great concern arose about the accumulation of the chlori-
nated hydrocarbon and cyclodiene insecticides in the biosphere,
coming sharply into the public consciousness with publication
of Silent Spring (Carson, 1962). Ultimately, during the 1970s
and 1980s these particularly persistent pesticides were phased
out and replaced by highly toxic but biodegradable compounds
such as the carbamates and organophosphates.
Paradoxically, biodegradation of these newer pesticides,
which had been generally seen as a positive process, is a double-
edged sword in relation to management of many soilborne pests
and diseases because of the requirement for prophylactic treat-
ment and the need for longevity into the crop growth cycle. The
history of observation that biodegradation of soil-applied pes-
ticides can be so rapid that it adversely affects control of pest
insects, the recognition that the process could become enhanced
or accelerated by repeated application of the pesticide, general
characterization of the mechanisms underlying the phenomenon
and many specific early examples have been comprehensively
reviewed by Felsot (1989), in Racke and Coats (1990), Suett
(1991), Felsot and Shelton (1993), and Suett et al. (1996). As
the broad scope of the phenomenon’s history and basic char-
acteristics has remained unchanged, no detailed description is
needed here. A general summary will suffice to give context to
recent findings relevant to contemporary issues and the theme
of this review.
2. Microbially-Mediated Transformations
Examples of control failures associated with rapid degrada-
tion of pesticides were recorded in the literature, but they were
isolated. While microbial action was known to be the major
cause of xenobiotic degradation, it would be safe to say that
this was not widely appreciated. Therefore it appears clear that
the adverse implications for pest management of such processes
were initially not recognized. The major issue that brought the
phenomenon to the fore was large-scale control failures of corn
rootworms (Diabrotica spp.) that require season-long control in
the US cornbelt where carbamates had replaced cyclodiene in-
secticides by the mid-1970s. Initially, by logical inference, resis-
tance was suspected to be the cause, as the insects had developed
resistance to the cyclodienes, but no resistance could be demon-
strated. It also became apparent that the failures were specific to
fields within a region rather than area-wide, further discounting
resistance as the cause. The epithet “problem soils” emerged to
describe the unexplained phenomenon and it became apparent
that fields that had been treated multiple times with the same
insecticide fell into that group whereas control was acceptable
in previously untreated fields. Furthermore, greater persistence
of pesticides in samples of soil that had been sterilized com-
pared with the parent soil indicated that rapid degradation was
microbiologically-mediated.
256 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
Eventually, it became clear that microbial (generally bacte-
rial) activity was the cause of the control failures through dis-
sipation of pesticides and that the repeated application of the
pesticide (or sometimes structurally similar compounds) accel-
erated the rate of dissipation through natural selection and pro-
liferation of adapted microorganisms. In this way, initially suc-
cessful pesticides could become ineffective. The phenomenon
was labeled enhanced or accelerated biodegradation to at once
describe the basis of the phenomenon and to indicate when a
production system was adversely impacted by pesticidal effi-
cacy falling below a required level. To reiterate, biodegradation
that can be shown to have been increased in its rate by such
a natural selection/proliferation mechanism only truly becomes
described as enhanced when there are negative consequences for
pest control in a particular cropping situation, which may be a
single field. Indeed, the impact of enhanced biodegradation will
be much more site-specific than resistance because it is not mo-
bile pests acquiring the capacity to metabolize the pesticide, but
rather sedentary and otherwise innocuous organisms in the soil.
3. Determination of Enhanced Biodegradation
Analytical chemistry can determine the rate of degradation
of a pesticide in soil and show large variation between loca-
tions depending on, inter alia, treatment history (e.g., Warton
et al., 2001a). However, whether the observed rates of break-
down constitute a meaningful case of enhanced biodegradation,
and whether it may vary between the targets in a multiple-species
suite of pests or diseases is, unlike resistance, not readily defined
by such direct measurement. Unfortunately, it is often only the
effects of pest control failure that lead to the eventual realization
that the effect is caused by enhanced biodegradation.
Another practical aspect that is likely to be overlooked
by those not familiar with soilborne pest and disease issues
is that very often the pest or disease may not be present,
giving the illusion that the prophylactic pesticide application,
targeted at the perceived risk rather than a confirmed reality,
has been effective. In such instances, the pesticide application
is nurturing enhanced biodegradation, to be manifest as control
failure if and when a pest appears in the system. It is probably
the highly localized aspect of the phenomenon, the indirect
action of otherwise innocuous organisms, the counter-intuitive
notion that such secondary organisms could be utilizing a
pesticide for metabolic advantage rather than the metabolic
cost required of resistance, inability to measure the problem
directly, and its often cryptic nature that has resulted in the
lower profile of enhanced biodegradation as a potential problem
for pest management compared to resistance. For individual
producers impacted by enhanced biodegradation, however,
the consequences are very real, and extrication from what can
become an impasse presents an often formidable challenge.
B. Mechanisms and Modifying Factors
1. Soil pH
There is an extensive literature on enhanced biodegradation
of numerous soil-applied pesticides correlating the speed of on-
set, the severity and the persistence of the phenomenon on var-
ious factors, including raised soil pH, frequency of application,
and soil type (Read, 1983, 1986; Smelt et al., 1996; Suett et al.,
1996; Smelt et al., 1989; Walker and Welch, 1990). It is clear
that soils can differ greatly in their ability to develop enhanced
biodegradation (Smelt et al., 1996), and there is no clear rela-
tionship between the number of applications of a pesticide and
the occurrence of enhanced biodegradation (Smelt et al., 1989).
Over time, soil pH was identified as the most significant correlate
(Houot et al., 2000; Bending et al., 2003; Singh et al., 2003a,
2003b), seemingly to the point of an implied direct causative
role. However, the possibility of covariates either confounded
or interactively associated with elevated soil pH in the devel-
opment, severity and persistence of enhanced biodegradation of
soil-applied pesticides did not receive specific attention. A possi-
ble reason for the observed correlation between the degradation
rate of pesticides in soil and soil pH is that bacterial adaptation
occurs more readily at higher pH (Suett et al., 1996). With only
one exception (Read, 1986), early studies compared different
soils, so consequently the observed differences could depend on
factors associated with soil type as well as pH.
When a non-degrading field soil (pH 5.4) was amended with
calcium carbonate (lime) to increase pH to either 5.8 or 6.4,
aldicarb was degraded after a prolonged lag period, suggesting
a biological rather than a physico-chemical mechanism (Read,
1986). The degradation rate was decreased when the soil pH was
lowered from 6.4 to either 5.8 or 5.4 with sulfuric acid, although
not to the same extent as with soil sterilization. More recent
studies that utilized soil from a single field with nominally uni-
form soil characteristics showed that the dissipation rate of the
herbicide isoproturon, and proliferation of microbial biomass
and isoproturon-degrading microorganisms, was positively cor-
related with a marked natural gradient in soil pH (Walker et al.,
2001; Bending et al., 2003).
It should be noted that most of these pH measurements were
carried out in water, a few in calcium chloride solution, and some
were not specified. Soil pH is commonly measured in either
water or CaCl2 , as there are benefits in using both extractants
(Slattery et al., 1999), but it can make comparison of results
difficult. Soil pH measured in CaCl2 gives a lower value than in
water, but fortunately the comparative values can be determined
in a look-up table (Henderson and Bui, 2002).
The specific effect of soil pH on the enhanced biodegrada-
tion of metam sodium had not been studied until recently. Of
seven soils where nematode control was problematic with metam
sodium, six with pH values between 4.7 and 5.3 had methyl
isothiocyanate half life (DT50 ) values of 3.4 to 19 days, while
the seventh soil, with a pH value of 7.3, had a much lower DT50
of 0.5 day (Smelt et al., 1989). There were, however, a number
of other differences between the soils including organic matter
content, clay content and the number of prior metham sodium
treatments which meant that the observed variation in metam
sodium degradation rate could not be conclusively ascribed to
soil pH alone.
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
2. Soil Calcium Level
Particularly severe enhanced biodegradation of metam
sodium, attributable to a suite of spore-forming bacteria that
degrade the methyl isothiocyanate that is produced almost as
rapidly as it is generated, occurs in a region of Australia (Warton
et al., 2001a). The region has coarse sandy soils that have an in-
trinsic pH ranging from acidic to alkaline, but a common practice
is to add lime (calcium carbonate) to the more acid soils to in-
crease their pH for agronomic reasons (Russell, 1961), and to
suppress Pythium spp. fungi (El-Tarabily et al., 1997). Addi-
tion of lime to a soil not only increases pH, but obviously also
increases calcium levels. Calcium is an important nutrient for
bacteria and calcium ion is commonly a metal co-factor crucial
in enzymatic reactions (Brock, 1970). In a study of acceler-
ated degradation of the herbicide atrazine in 47 different soils,
Houot et al. (2000) gave a tabulation of their various physical
and chemical characteristics, including pH and calcium carbon-
ate concentration. When these calcium carbonate values were
taken and plotted against the soil pH, the relationship was de-
scribed by an exponential function that had an inflection around
pH 6.5–7.5 (Warton and Matthiessen, 2005). In soils with pH
below this inflection point, dissipation of atrazine was negligi-
ble, even after repeated applications to the same soil; however,
it was notable that the concentration of calcium carbonate in-
creased by almost 200-fold over the narrow range of pH from 7
to 8 (Houot et al., 2000).
Until recently, studies on the biodegradation of soil-applied
pesticides have correlated the influence of elevated soil pH with
the phenomenon, but had not considered the possible effect
of increased calcium, which is confounded with pH increase
at a very high quantitative level in what is a particularly rel-
evant soil pH range. By modifying the pH and the calcium
levels of a single sandy soil separately and in unison, Warton
and Matthiessen (2005) induced enhanced biodegradation of
methyl isothiocyanate only when both factors were together ele-
vated. Alone, neither elevated pH (using magnesium carbonate)
nor elevated calcium level (using calcium chloride) promoted
biodegradation. It is now clear that the confounding with soil
pH has masked a key role for calcium in the development of
enhanced biodegradation.
Soil pH in the range 5.8 to 6.8 was determined to be near a
threshold value at which the biodegradation of methyl isothio-
cyanate can quickly change from minimal to very rapid (Warton
and Matthiessen, 2005). To raise pH, an increasing amount of
lime had to be added for each pH increment, with the rise becom-
ing exponential around pH 5.8 to 6.8 (Warton and Matthiessen,
2005). Those pH values, measured in calcium chloride, are
equivalent to values of 6.5–7.4 measured in water (Henderson
and Bui, 2002), the inflection point plotted from the field data
of Houot et al. (2000). The similarity of empirical data used to
predict lime amendment required for desired soil pH, and its
derivation from diverse field soils, adds strong support to the
conclusion that a pH/calcium interaction is a major causative
factor in enhanced biodegradation, further indicates its general
257
applicability to pesticides other than metam sodium, and sug-
gests that it was masked within the high pH correlations observed
in earlier studies (Warton and Matthiessen, 2005).
Calcium is a particularly important nutrient for bacteria such
as Bacillus spp. and actinomycetes that form spores or resis-
tant resting stages, being implicated in their capacity for envi-
ronmental tolerance (Brock, 1970). It was such organisms that
were identified as responsible for severe enhanced biodegrada-
tion of metham sodium in sandy soils in Australia (Warton et al.,
2001a). The application of lime to such soil markedly increased
populations of a wide variety of bacteria, notably actinomycetes,
as well as overall microbial activity (El-Tarabily et al., 1996).
In the light of such principles and observations, and given the
extensive literature showing soil pH to be a major biodegrada-
tion correlate, it seems very likely that interdependence of soil
pH and calcium concentration is a generally applicable mech-
anism underlying the phenomenon of enhanced biodegradation
of soil-applied pesticides.
Liming of agricultural soils is often carried out to overcome
natural acidity because it improves plant growth (Russell, 1961).
It is also employed as a means of increasing suppression of plant
fungal disease because elevated pH beneficially changes the bi-
otic and abiotic characteristics of the soil (El-Tarabily et al.,
1996). Notably, raising soil pH greatly favors dominance of
bacteria in the soil microbial population (Alexander, 1977), and
bacteria are the most common causative organisms of enhanced
biodegradation of soil-applied pesticides (Racke, 1990). It has
also been observed that elevating soil calcium using calcium car-
bonate and using calcium sulfate (to control for the coincident
increase in soil pH by the former) were both equally effective
in suppressing fungal disease (Kao and Ko, 1986), further em-
phasizing a diverse or complex role of calcium in soil microbial
functions.
3. Soil Type
Despite inducing rapid degradation of methyl isothiocyanate
in sand, the same effect could not be achieved in loam of similar
pH and calcium content, indicating that soil type is a crucial
factor that can override the otherwise powerful pH/calcium in-
teraction (Warton and Matthiessen, 2005). While this is hardly
surprising in view of the immense complexity of soils, the lim-
ited soil type comparison so far undertaken makes further work
on the influence of soil type necessary to elucidate the mecha-
nisms responsible and to elaborate the risk factors in production
systems. Apart from many mineralogical and physical differ-
ences between soil types, microbiological properties also vary
widely between different types of soils (Houot et al., 2000). It
was clear from the results of Warton and Matthiessen (2005)
that biodegradation of methyl isothiocyanate is fundamentally
driven by an interdependence of pH and calcium concentration,
in complex interplay with the frequency and number of applica-
tions of the pesticide, and soil type.
The results of Warton and Matthiessen (2005) demonstrate
that it is necessary to elevate both soil pH and calcium in a sandy
258 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
soil to substantially increase the risk of enhanced biodegradation
of methyl isothiocyanate, whereas increasing either soil pH or
calcium levels alone does not. With the likelihood that this is a
general phenomenon, producers who use soil-applied pesticides
and add lime to sandy soil to raise the soil pH or calcium for
agronomic reasons or soil pathogen suppression, risk decreasing
the pesticide’s efficacy by triggering enhanced biodegradation.
Unfortunately, sandy and light-textured soils tend to offer agro-
nomic advantages for intensive horticultural production and they
are common where pesticides intrinsically much more suscep-
tible to enhanced biodegradation such as metam sodium and
1,3-D must now substitute for methyl bromide.
The combination of high levels of calcium, high soil pH
and sandy soil is a recipe for enhanced biodegradation disaster
for metam sodium: following an application of metam sodium
less than half the concentration of methyl isothiocyanate was
achieved in degrading soil compared to previously untreated
soil, and the methyl isothiocyanate was present in the soil for
less than 5 percent of the time of that in untreated soil (Warton
et al., 2001a). More optimistically, the findings offer key infor-
mation that provides some opportunity to better manage soil
pH and calcium concentration with other amendments, such as
gypsum which has the effect of raising calcium levels but of
not changing pH appreciably (El-Tarabily et al., 1996), to re-
duce that risk and optimize soil pH and calcium for best overall
benefit in a production system.
4. Overcoming Enhanced Biodegradation
Once a soil develops microbial adaptation to a pesticide and
becomes a case of enhanced biodegradation, there is little or
nothing that can be done to “cure” the problem. In the laboratory,
it can be stopped by sterilizing the soil or reducing the soil pH to
around 4.5 or lower (Read, 1983), but such procedures are obvi-
ously impracticable for field use. Recovery involves leaving the
soil untreated with that compound, often for an extended period.
However, because many of the bacteria commonly responsible
for enhanced biodegradation are types with resistant stages, such
as actinomycetes (Warton et al., 2001a), they can persist quies-
cently for a very long time to quickly resurge when the pesticide
is again applied. Thus it is likely that the re-initiation of en-
hanced biodegradation after a soil seemingly “recovers” would
return very rapidly to the maximum level previously observed,
without going through a more gradual period of build-up that
may have characterized the original onset of the problem.
Since the basis of enhanced biodegradation is microbial, not
unexpectedly the ‘infection’ can be transferred to non-affected
soil by movement of degrading soil; this has been demonstrated
in the laboratory (Harris et al., 1984; Walker, 1987; Walker and
Welch, 1990; Walker et al., 1996) and in the field (Suett and
Jukes, 1993; Walker et al., 1996). However, it seems improbable
that this is a major cause of induction of enhanced biodegrada-
tion under field conditions. Within a soil, there also exists the
possibility of transfer of degradative function between bacteria
through gene transfer on mobile extrachromosomal elements
like plasmids and transposons, a topic beyond the scope and
relevance of this review but which, with other microbiologi-
cal aspects, is well covered elsewhere (Kearney and Kellogg,
1985; Sayler et al., 1990; Cork and Krueger, 1991; Soulas and
Lagacherie, 2001; Top et al., 2002; Top and Springael, 2003;
Springael and Top, 2004). High levels of spatial heterogeneity
in the expression of enhanced biodegradation on small scales
within a field related to variation in the microbial community
activity with soil pH can occur (Bending et al., 2001, 2003), but
this seems unlikely to be a factor that could readily be accounted
for in day-to-day pest management decisions.
C. Link to Biofumigation and Methyl Bromide
1. Enhanced Biodegradation of Metam Sodium
The first reports of inadequate pest control by metam sodium
in areas where efficacy had previously been high began to ap-
pear in the Netherlands in the 1980s. While measurement of
transformation of the metam sodium to methyl isothiocyanate
showed it consistently occurred within one day, the subsequent
time to 50% transformation of the methyl isothiocyanate varied
from 0.5 to 50 days and the fastest transformations occurred in
those soils that had been treated frequently (Smelt et al., 1989;
Verhagen et al., 1996). These analytical determinations indi-
rectly indicated the existence of enhanced biodegradation, and
that it was the methyl isothiocyanate being degraded. Subse-
quently, degradation of methyl isothiocyanate by a bacterial con-
sortium was confirmed as the cause of biodegradation in field-
collected soil where metam sodium had been used regularly, with
its disappearance being a mere seven hours compared to around
18 days in nearby untreated soil or treated soil that was ster-
ilized by autoclaving (Warton et al., 2001a). Matthiessen et al.
(2004b) then demonstrated that the level of enhanced biodegra-
dation in the previously treated soil was sufficient to reduce
the efficacy of metam sodium applied at label rate to control
soil pests. Enhanced biodegradation of methyl isothiocyanate
causing significant reduction in control of soilborne pathogens
with metam sodium and dazomet (tetrahydro-3,5-dimethyl-2H-
1,3,5thiadiazine-thione), a granular methyl isothiocyanate gen-
erator, has also been reported from Israel (Di Primo et al., 2003).
2. Cross-Degradation
When propenyl, benzyl and 2-phenylethyl isothiocyanates,
all of which are commonly derivable from brassicas (Kirkegaard
and Sarwar, 1998), were added to the strongly methyl
isothiocyanate-degrading soil (Warton et al., 2001a) their rate of
degradation was markedly greater than in samples of the same
soil that had been sterilized, despite the soil never having been
exposed to those compounds (Warton et al., 2003). This was
a clear case of cross-degradation, or cross-enhancement, which
had been previously observed between other structurally-related
pesticides, notably within the carbamates (Suett, 1987; Racke
and Coats, 1988; Felsot, 1989; Morel-Chevillet et al., 1996).
Its occurrence suggested that the microorganisms responsible
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
for the breakdown specifically target the isothiocyanate func-
tional group. Cross-degradation of a sort was also demonstrated
to metam sodium or dazomet in reciprocally-challenged soils
whose enhanced degradation origins were linked to the other
of those two compounds (Di Primo et al., 2003), although it
seems likely that in both cases it was the methyl isothiocyanate
that was being degraded. Soil insect bioassays also showed that
substantially more propenyl isothiocyanate-liberating Brassica
plant tissue was required to achieve given levels of mortality in
the presence of the methyl isothiocyanate-degrading soil than
non-degrading soil (Warton et al., 2003). These findings indi-
cate that the effectiveness of biofumigation is highly likely to
be compromised in soil with a history of prior use of metam
sodium, and further suggest that it is likely to occur regardless
of the glucosinolate profile of the Brassica used.
In the light of these findings, a question that arises relates
to the risk of onset of enhanced biodegradation of an isothio-
cyanate that derives from a biofumigant Brassica used often on
the same area of land. There are no known studies specifically
aimed at addressing this question. Purely speculatively, it would
seem that applying a large dose of a single pure isothiocyanate,
such as via metam sodium or dazomet, is more of a selection
pressure risk on the microbial biomass of the soil than plant
tissue that is decomposing into a wide variety of microbial sub-
strates. This may be an aspect that some may consider worthy
of investigation but, pragmatically, it does not seem likely to be
a major issue. Endeavoring to utilize biofumigation as a substi-
tute for failed or sub-optimal metam sodium use brought about
by enhanced biodegradation is much more likely to present a
practical problem.
3. Methyl Bromide Alternatives
With the phasing-out of methyl bromide for use in soil fu-
migation under the international Montreal Protocol, there has
for some years been an active search for alternatives. While
many of the chemical alternatives currently registered and avail-
able have good activity against fungi, nematodes or weeds,
they lack the full spectrum of activity and versatility of methyl
bromide as a pre-plant soil fumigant. For this reason, mix-
tures are common. In most regions there are currently only
three alternatives that are presently registered for use in many
previously methyl bromide-using crops such as strawberries:
chloropicrin, 1,3-D and methyl isothiocyanate generators such
as metam sodium and dazomet (Ajwa et al., 2003). Of these
the most promising alternative is generally considered to be a
mixture of 1,3-D and chloropicrin (Ajwa et al., 2003; Martin,
2003). Relative to methyl bromide, inconsistent control is the
primary problem encountered with metam sodium. Those short-
comings derive from its poorer soil penetration, narrower spec-
trum of activity, potential for enhanced biodegradation to cause
control failures, crucial dependence on application method,
greater potential for unpredictable phytoxicity of the follow-
ing crop through greater effects of environmental conditions on
dissipation rates, and large quantities of carrier water to en-
259
sure uniform distribution in soil (Ajwa et al., 2003; Martin,
2003).
In a twist on inadvertent initiation of enhanced biodegrada-
tion by repeated applications of pesticides to soil, several studies
over the past few years have been undertaken to deliberately ac-
celerate the rate of degradation of soil fumigants being used as
substitutes for methyl bromide. This work has occurred in Cal-
ifornia where there are particular concerns about atmospheric
emissions near urban centers. Dungan et al. (2003) deliberately
increased the degradation rate of metam sodium in soil by adding
organic matter-rich amendments to soil. With a similar objec-
tive, Ibekwe et al. (2004) induced enhanced biodegradation of
chloropicrin and metam sodium in the laboratory by repeated ap-
plication to soils, with those amended with composted manure
showing faster degradation. They also confirmed that bacterial
enrichment was an underlying mechanism for the observed en-
hancement of degradation, but it was not clear to what extent
the addition of organic matter directly or indirectly boosted the
bacterially-mediated biodegradation. 1,3-D, particularly in mix-
ture with chloropicrin, is presently the most commonly used
alternative to methyl bromide. Gan et al. (1998) used organic
amendments to significantly enhance degradation of 1,3-D and
found the mechanism to be a combined result of enhanced chem-
ical and microbial degradation. In a study of the effect of en-
vironmental factors on degradation of 1,3-D, Guo et al. (2004)
found that organic matter promoted degradation directly and that
microbial contributions to degradation, after initially being in-
significant, became important as soil microorganisms adapted
to the fumigant. Similarly, the addition of thiosulfate fertilizers
to soil caused 1,3-D to be rapidly transformed to non-volatile
products but, as might be expected, the mechanism in this case
was chemically based (Gan et al., 2000b).
While reduction of atmospheric emissions is a laudable ob-
jective, it is conceivable that to do it deliberately by inducing
enhanced biodegradation has the potential to be counterproduc-
tive to pest and disease control efficacy, especially in highly
intensive production systems requiring frequent fumigant appli-
cation. Control efficacy could be diminished either by transient
boosting of degradation such as by addition of fertilizer, or per-
manently if enhanced biodegradation were induced. None of the
studies addressed the residence time needed by the fumigants
to achieve efficacy against specific organisms, nor the longer
term implications of perhaps promoting the permanence of en-
hanced biodegradation. The nearest to an exception was Gan
et al. (1998) who suggested that organic matter amendments
would be in the upper layers of the soil and may act as a filter
to reduce emissions but not lower concentration sufficiently to
reduce efficacy in the soil below. Indeed, the general implication
throughout these studies appeared to be that the more rapid the
degradation of the pesticide, the better.
Clearly, there needs to be a balance between ensuring efficacy
and dissipating the xenobiotic. Otherwise, the emissions reduc-
tion objective could eventually be achieved at the expense of ef-
ficacy by eventual cessation of the pesticide’s use if its efficacy
260 J. N. MATTHIESSEN AND J. A. KIRKEGAARD
has been diminished below an acceptable economic threshold
for the given production system. Less intensive systems may
provide more scope for organic matter amendment to provide
a perhaps more transient boost to degradation rates, but atmo-
spheric emissions are probably less likely to be an issue under
such circumstances. There would appear to be a fine line be-
tween addition of organic matter with the intention of boosting
biodegradation rates and the deliberate initiation of entrenched
enhanced biodegradation that could progressively worsen and
permanently interfere with the efficacy of pest control. This
would seem to be an issue that producers would need to be
well advised about before undertaking such actions.
VI. CONCLUSIONS
Biologically-based methods for control of soilborne pests and
diseases have long been touted as an environmentally benign al-
ternative to synthetic chemical pesticides. But the stark reality
is that they have yet to widely meet expectations for consistent
and robust efficacy under field conditions (Lazarovits, 2001).
For biologically-based control methods to succeed, the stan-
dards for efficacy must be those used for chemical pesticides,
or they must be advocated for systems that have no economi-
cally viable chemical control option or which prefer to avoid use
of chemicals in order to meet other objectives. There must be
recognition at the outset of the realities of how to cost-effectively
build in to agricultural systems such methods, and of intrinsic
limitations of biologically-based methods of pest control when
they are set against synthetic chemical benchmarks that were
developed and pre-tested to work robustly under a wide variety
of circumstances.
It is evident that development of biologically-based meth-
ods of soilborne noxious organism control has not reached ma-
turity and that it is often contentious. Practitioners frequently
are skeptical of scientists’ efficacy claims, especially those de-
rived solely from laboratory studies, or dismissive on the basis
of economics or the logistics of implementation. Most (some
may argue perhaps too much) of the effort has consisted of ba-
sic, descriptive research. Much less attention has been given
to systematic implementation based on the results of the ba-
sic research and an understanding of the milieu in which that
implementation must occur. There is a very long history of in-
vestigation of many biologically-based alternatives, which can
often easily be shown to have potential in in vitro laboratory
assays. In many cases, however, these are with concentrations
so great that physiological activity is doubtful and maybe even
contraindicated (Chitwood, 2002). Simple calculations, assum-
ing a soil bulk density of 1 g cm−3 and a depth of incorporation
of an amendment of 25 cm, show that for a 1% w/w amend-
ment of soil 25 metric tons ha−1 of the amendment would be
required. Quantities well above that level are readily achiev-
able in situ with well-grown brassicas (Table 1), but propos-
als to ship in Brassica-based amendments need more careful
examination.
Without listing specific examples, there are published studies
recording suppressive effects at 2–5% w/w of amendments such
as Brassica seed meal in laboratory and glasshouse studies. But
too infrequently is the calculation of the scale-up or “loading”
requirement for field scale application, both quantity and cost,
included. The economic production, shipping, application and
incorporation of such quantities is likely to present insurmount-
able barriers to adoption of such an approach in any more than
niche markets such as nursery beds or in pot culture. These real-
ities need to be more clearly acknowledged in scientific studies.
Inadequate attention has been given to complementing in vitro
experiments with in vivo, soil-based, tests with appropriate field-
achievable loadings in the first instance, and then following with
carefully designed field tests and methods to implement and val-
idate the principles elucidated in the controlled environment. It
is little wonder that touted biologically-based methods often
cannot match highly extrapolated prediction or the efficacy of
chemical pesticides under field conditions, leaving field trials
often floundering in empiricism.
Biofumigation should be seen as an option for suppression,
not a panacea for elimination, of soilborne pests and diseases and
it should be advocated for selective application in appropriate
production systems. It has been demonstrated to have efficacy,
secondary soil benefits and, notably, economic benefits in an
appropriate production system of moderate intensity that tradi-
tionally uses metam sodium, and uptake by producers in at least
one regional production system has been high (McGuire, 2004).
Furthermore, and in contrast to other phytochemically-based ap-
proaches, biofumigation using isothiocyanate-generating brassi-
cas has a strong positive precedent in the commercialization and
use of the methyl isothiocyanate-generating synthetic pesticide
metam sodium. Arguably, the fact that there is such a generally
efficacious, well-understood methodology to strive at emulating
(with the caveat that it be targeted at appropriate systems) is
a distinct advantage because it provides an established bench-
mark based on the same type of active compound against which
to measure performance. Interestingly, and illustrative of the
need to be cognizant of the target production system, biofumi-
gation and like approaches that seek to utilize naturally-derived
biologically active compounds are permitted to be used in or-
ganic production but are regarded as methods of last resort (van
Bruggen and Termorshuizen, 2003).
Alone, biofumigation is unlikely to provide the high-impact
and broad-spectrum control achieved by large doses of synthetic
fumigants in highly intensive production systems with extremely
high pest management benchmarks. If metam sodium or 1,3-D
alone, and particularly in combination with chloropicrin, cannot
under a wide range of circumstances consistently achieve levels
of pest and disease control equivalent to methyl bromide, then
biofumigation cannot hope to reliably emulate methyl bromide.
That is a reality that should be recognized and acknowledged to
avoid over-statement and over-expectation that ultimately risks
leading to dismissal of this, and like, concepts. However, this
is not to say that a useful niche role cannot be achieved for
 BIOFUMIGATION AND ENHANCED BIODEGRADATION
biofumigation in suitable situations, or with some form of sup-
plementation such as solarization (Katan, 1981; Gamliel et al.,
2000) in more intensive production systems.
Grower acceptance of new plant rotation strategies must be
both economically and logistically compatible with the target
farming system’s basic practices. The need to grow the biolog-
ically active green manures is an extreme manifestation of the
need for a prophylactic approach to soilborne pest and disease
management. While some modifications may be possible to ac-
commodate and maximize biofumigation, efficacy alone will be
insufficient to ensure adoption, and this must be recognized.
Many laboratory, glasshouse and microplot studies have shown
that allelopathic rotation crops or green manures can suppress
soilborne pests and diseases, but there are few reports of suc-
cessful application in commercial agriculture. That may in part
reflect the general absence of such reports in the scientific lit-
erature and the scattered uptake and on-farm development by
keen individual producers who source information and apply it
without formal reporting of their experiences.
Despite the toxicity of some Brassica-derived isothio-
cyanates being inherently greater than methyl isothiocyanate,
biofumigation is not likely to be as effective in soils with high
levels of organic matter or clay as sandy soils because of dimin-
ished bioavailability of isothiocyanates resulting from sorption
(Matthiessen and Shackleton, 2005). To some extent, applica-
tions of metam sodium overcome such limitations because field
trials for establishing efficacy and attaining registration as a pes-
ticide have been conducted in a range of contrasting environmen-
tal conditions. The result is that the registered application rates
are set at a level that gives greatest overall efficacy. This means
that many metam sodium applications are likely to be higher
than necessary in sandy soils, yet probably barely adequate in
heavy or high organic matter soils.
While the methyl bromide withdrawal issue can be used as a
particularly strong contemporary impetus to justify the general
search for alternative strategies at all levels, caution and realism
must be exercised in what options are promoted as replacements
for methyl bromide. Biofumigation is far more likely to achieve
acceptance and adoption more broadly if it is demonstrated to be
effective in appropriate less intensive production systems first.
This will provide the impetus to further investigate the conditions
and requirements for achieving consistent performance. To have
any chance of significant positive impact on adoption for routine
application, such studies need to be systematic and field-focused
towards understanding the conditions that need to be met to
achieve consistent results. In that way, evolved improvements in
performance and expansion in use are likely to steadily gather
pace.
It is only with some momentum in adoption achieved by
practical results and evident prospects for improvements that it
will become increasingly financially attractive for seed suppli-
ers to develop and market Brassica lines specifically directed at
biofumigation. This is already beginning (Gies, 2004; Lazzeri
et al., 2004). It is that step of moving out of the laboratory and
261
microcosms into existing agricultural systems that is crucial.
Once markets are initiated and the methodology gains credibil-
ity in a system appropriate to its immaturity, there is a much
greater chance that further research and development will be
well-targeted towards enhancing efficacy and expanding capa-
bility, thereby further improving biofumigation as an option for
soilborne pest and disease management.
With the phasing out of methyl bromide and its general sub-
stitution with 1,3-D, chloropicrin and metam sodium, the risk
of enhanced biodegradation of soil-applied pesticides seems
destined to increase in systems that have up to now not ex-
perienced the phenomenon. Unfortunately, substituting these
generally inferior control methods for methyl bromide is more
likely to result in reduced efficacy which then may be con-
founded with that caused by enhanced biodegradation. Unwit-
tingly, this would cause entrenchment of enhanced biodegra-
dation. Meeting the general challenge of greater risk of en-
hanced biodegradation in what were once not-at-risk methyl
bromide-dependent systems is likely to present significant
difficulties.
Prevention of enhanced biodegradation by careful manage-
ment of pesticide application frequency, allied with knowledge
of degree of risk, is the best strategy but something of an ideal-
ization in the real world. It is comparatively straightforward to
measure the problem; preventing its onset or recovering from it
has no useful precedents, particularly in highly intensive produc-
tion systems where there is little scope for lengthy breaks from
production. However, recognizing risk factors, particularly the
recently-revealed crucial risk combination of sandy soil of high
pH and with a high calcium level, should allow scope for some
management intervention to reduce the risk.
ACKNOWLEDGMENT
We thank Ben Warton for comments on a draft.
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