trends in plant science
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38 Babakov, A.V. et al. (1995) Culture of transformed horseradish roots as a
source of fusicoccin-like ligands, J. Plant Growth Regul. 14, 163–167
39 Qin, M.B. et al. (1994) Induction of hairy root from Artemisia annua with
Agrobacterium rhizogenes and its culture in vitro, Acta Bot. Sinica 36
(Suppl.), 165–170
40 Trypsteen, M. et al. (1991) Agrobacterium rhizogenes-mediated
transformation of Echinaceae purpurea, Plant Cell Rep. 10, 85–89
41 Yamanaka, M. et al. (1996) Polyacetylene glucosides in hairy root cultures of
Lobelia cardinalis, Phytochemistry 41, 183–185
42 Benjamin, B.D., Roja, G. and Heble, M.R. (1994) Alkaloid synthesis by root
cultures of Rauwolfia serpentina transformend with Agrobacterium
rhizogenes, Phytochemistry 35, 381–383
43 Sato, K. et al. (1991) Anthraquinone production by transformed root cultures
of Rubia tinctorum: influence of phytohormones and sucrose concentration,
Phytochemistry 30, 1507–1510
44 Hu, Z-B. and Alfermann, A.W. (1993) Diterpenoid production in hairy root
cultures of Salvia miltiorrhiza, Phytochemistry 32, 699–703
45 Delbecque, J.P. et al. (1995) In vitro incorporation of radiolabelled cholesterol
and mevalonic acid into ecdysteroid by hairy root cultures of a plant, Serratula
tictoria, Eur. J. Entomol. 92, 301–307
Recent advances in the transformation
of plants Genevi•ve Hansen and Martha S. Wright
Plant transformation technology has become a versatile platform for cultivar improvement as
well as for studying gene function in plants. This success represents the culmination of many
years of effort in tissue culture improvement, in transformation techniques and in genetic
engineering. The next challenge is to develop technology that minimizes or eliminates the
tissue culture steps, and provides predictable transgene expression.
G
enetic engineering has opened new avenues to modify
crops, and provided new solutions to solve specific needs1.
In the future, the proportion of acreage planted with trans-
genic crops, and the range of transgenic crops, is sure to increase.
For example, in the USA, the proportion of acreage planted with
commercial transgenic cotton, soybean and corn was ~25, 18 and
10, respectively, in 1997 (Ref. 2). Forty-eight transgenic crop
products had been approved for commercialization in various
countries by the end of 1997.
The powerful combination of genetic engineering and conven-
tional breeding programs permits useful traits encoded by trans-
genes to be introduced into commercial crops within an
economically viable time frame. There is great potential for
genetic manipulation of crops to enhance productivity through
increasing resistance to diseases, pests and environmental stress
and by qualitatively changing the seed composition. Plant ‘fac-
tories’ are also being designed for high volume production of
pharmaceuticals, nutraceuticals and other beneficial chemicals.
Transgenic plants might become drug-delivery devices, with both
HIV and rabies vaccines being synthesized in plants3, and bananas
have been engineered to produce edible vaccines4. Moreover, with
the establishment and expansion of genomics programs, a much
broader range of genes with potential for crop improvement are
being identified and, in some cases, tailored and/or re-designed for
further enhancement of their properties within specific crops1.
This has further intensified the interest in developing efficient
plant transformation technologies to be able to concurrently test
and capture the value of these genes.
226 June 1999, Vol. 4, No. 6 1360 - 1385/99/$ Ð see front matter © 1999 Elsevier Science. All rights reserved.
46 Takeda, T. et al. (1994) Bryonolic acid production in hairy roots of
Trichosanthes kirilowii Max. var. japonica Kitam. transformed with
Agrobacterium rhizogenes and its cytotoxic activity, Chem. Pharm. Bull.
(Tokyo) 42, 730–732
47 Gränicher, F. et al. (1995) An iridoid diester from Valeriana officinalis var.
sambucifolia hairy roots, Phytochemistry 38, 103–105
Hector E. Flores* and Jorge M. Vivanco are at the Dept of Plant
Pathology and Life Sciences Consortium, 315 Wartik Building, The
Pennsylvania State University, University Park, PA 16802, USA;
Victor M. Loyola-Vargas is at the Centro de Investigacion
Cientifica de Yucatan, Merida, Mexico (tel 152 99 81 3961;
fax 152 99 81 3900;
e-mail vmloyola@cicy.cicy.mx).
*Author for correspondence (tel 11 814 865 2955;
fax 11 814 863 7217; e-mail hef1@psu.edu).
Beyond crop improvement, the ability to engineer transgenic
plants is also a powerful and informative means for studying
gene function and the regulation of physiological and develop-
mental processes. Transgenic plants are being used as an assay
system for the modification of endogenous metabolism or gene
inactivation. Advances in tissue culture, combined with improve-
ments in transformation technology, have resulted in increased
transformation efficiencies. In recent years, many crops, previ-
ously classified as recalcitrant because they were stubbornly re-
sistant to the overtures of genetic engineering, have now been
transformed. In many instances, the technology has been pushed
even further to introduce the gene of interest directly into
elite cultivars5.
Tissue culture prerequisite
A tissue culture stage is required in most current transformation
protocols to ultimately recover plants. Indeed, it is the totipotency
of plant cells that underlies most plant transformation systems.
Plants are regenerated from cell culture via two methods, somatic
embryogenesis and organogenesis. Both are controlled by plant
hormones and other factors added to the culture medium. Some
species, such as soybeans6, banana4,7 and sugar beet8,9 can be
regenerated via either method so the choice is dependent upon
which gives the best yield or the easiest outcome.
Somatic embryogenesis is the generation of embryos from
somatic tissues, such as embryos, microspores or leaves. Prolifer-
ating somatic embryos in liquid culture or on solid medium are
suitable targets for transformation because the origin of proliferating
PII: S1360-1385(99)01412-0

Fig. 1. Examples of cultured tissues for transformation experiments. (a) Type I and (b) type II callus from maize, (c) primordial shoots from
maize. (d) and (e) Callus and shoots from soybean. (f) Cotton shoot apex from four-day-old seedling. M indicates the apical meristem and P the
primordial leaf. Magnification: (a), (b), (d) and (e) 133; (c) 403; (f) 83.
embryogenic tissues is at or near the surface of the older embryos
and, thus, readily accessible to DNA delivery. Embryogenic tissues
are, in general, very prolific and allow recovery of many transfor-
mants that are, in most cases, non-chimeric because of the
assumed single cell origin of somatic embryos10. It is the tissue
culture approach generally chosen for monocots because callus is
easily initiated from the scutellum of immature embryos after
exposure to auxin. The concentration and the choice of auxin (i.e.
2,4-dichlorophenoxyacetic acid, chloramben or dicamba) is to
some extent dictated by the genotype and the species. Auxin is
then reduced or withdrawn to allow the generation of shoots from
callus for the subsequent recovery of plants.
Maize can form two different types of somatic embryos11: type I
somatic embryos occur as a group of fused embryos and type II
occur as discrete, single embryos in a cluster (Fig. 1). Depending
on the transformation method elected, it is possible to favor the
initiation of one type of callus over another. Type I callus is con-
venient for biolistic transformation5,12 whereas a certain type II
callus is preferentially chosen to produce maize protoplasts13.
However, the use of embryogenic tissue can have some limi-
tations. It can be labor intensive to establish and maintain the cul-
ture and the recovery of plants can be a long process, with the risk
of encountering morphological abnormalities and sterility. This
system also requires a constant source of material to initiate new
embryogenic cultures. This inconvenience has prompted the
use of mature embryos from seeds10 as an alternate source of
embryogenic tissues, even though their response might not be
very prolific.
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Organogenesis is the generation of organs, usually shoots, from
a variety of tissues. Except for monocot leaf explants that contain
meristems only at the leaf base10, cotyledons, leaf fragments,
hypocotyls and scutella from embryos generally have the poten-
tial to generate shoots when placed on medium containing a
shoot-inducing hormone, usually a cytokinin, such as 6-benzyl-
aminopurine. For instance, bananas are generally propagated by
shoot tip culture4. Transgenic cassava is obtained only after the
development of shoots14,15. The advantage of this system is that
shoots can usually form roots readily. If roots fail to appear, graft-
ing can be the solution16. Sometimes, tissues can acquire a trans-
lucent and brittle appearance with a high water content. This
abnormal morphology, called hyperhydricity17, is often observed
with woody plants or sugar beet shoots and can compromise plant
recovery. The occurrence of this hyperhydric state can be avoided
or reduced17 by modifying the sugar source, the calcium concen-
tration or by the use of antivitrifying agents, such as phloridzin.
Transformation systems
Successful transformation of plants demands that certain criteria
be met. Among the requirements for transformation are:
• Target tissues competent for propagation or regeneration.
• An efficient DNA delivery method.
• Agents to select for transgenic tissues.
• The ability to recover fertile transgenic plants at a reasonable
frequency.
• A simple, efficient, reproducible, genotype-independent and
cost-effective process.
June 1999, Vol. 4, No. 6 227
trends in plant science
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• A tight timeframe in culture to avoid somaclonal variation and
possible sterility.
At present, three techniques appear to fulfill these criteria:
• Protoplast transformation.
• Biolistics or microprojectile bombardment.
• Agrobacterium-mediated transformation.
How does one decide on the technique to use? In the fortunate
occurrence that several systems are available for a crop, the choice
is based on convenience: tissue accessibility, patent clearance and
equipment availability.
Although rapid commercialization requires the introduction of
the new trait directly into the germplasm of elite varieties elected by
plant breeders, the technique might first be developed using
‘model’ systems. The method is then extended to elite genotypes,
which are sometimes less amenable to in vitro culture. The trait can
also be introduced into the gene pool of the species by conventional
breeding techniques. For instance, the recent transformation suc-
cess of the diploid wild relative Fragaria vesca will allow transfer
of important traits into the octoploid edible strawberry18, which is
less amenable to transformation.
Protoplast transformation
Of the different transformation systems, the protoplast method is
probably the one that necessitates the most finesse. Protoplasts are
isolated either by a mechanical or by an enzymatic process to
remove the cell wall. This results in the production of a suspen-
sion containing millions of individual cells and therefore offers
the advantage of probable single cell targets. Protoplasts are fre-
quently obtained from an established suspension cell line of callus
initiated from immature embryos, immature inflorescences,
mesocotyls, immature leaf bases and anthers10. Interestingly,
many cell suspensions can be cryopreserved and still maintain the
ability to form callus and regenerate plants19. Protoplasts can
either be transformed by Agrobacterium or by direct DNA uptake
methods, facilitated by polyethylene glycol treatment, electropo-
ration or liposomes20. Elegant regeneration systems developed for
protoplasts, once considered the most efficient route to transgen-
ics for monocots, were eventually incorporated into all transfor-
mation approaches10,13.
In some species, protoplasts can be isolated from leaf mesophyll.
This technology was pushed further in its sophistication when
stomatal guard cells from sugar beet leaf protoplasts were identi-
fied as being the cells of choice for transformation because of
their totipotence9. The isolation of such cells follows the standard
protocol of protoplast isolation, with an enrichment of guard cells
by centrifugation. This method appears to be genotype-
independent in contrast to the Agrobacterium-mediated transfor-
mation of sugar beet friable callus or cotyledon explants21. A
similar strategy could thus be attempted with other important
recalcitrant crop species.
Biolistics transformation
Biolistics22 is the delivery of microprojectiles, usually of tungsten
or gold coated with DNA and propelled into the target cells by
acceleration. The acceleration can be provided by gun powder,
by gases, such as helium or CO2, or by an electric discharge. This
method can introduce DNA into virtually any tissue from any
cultivar – success depends critically upon the ability of the target
tissue to proliferate and give rise to a fertile plant.
Minor alterations in the standard biolistic protocol have yielded
tremendous improvements22. These include:
• Proper pre-culture of the explant material.
• Use of baffling screens.
• Use of small size microprojectiles12.
228 June 1999, Vol. 4, No. 6
• Subjecting the tissue to an osmotic pretreatment by either
partial drying in a laminar flow hood or culturing in a medium
containing an osmotic agent22.
For example, the transformation efficiency in the maize elite line
CG0056 was dramatically increased under these conditions12.
Molecular analysis of plants obtained by biolistic transfor-
mation generally reveals a complex pattern of transgene inte-
gration. In addition, delivery of long fragment DNA is challeng-
ing because breaks can occur in the delivered DNA. Although the
fate of introduced DNA is not clear, ligation of the transgenic DNA
fragments before integration is proposed to account for the obser-
vation of arrays of transgenic DNA integrated at the same site into
the plant genome23. This can result in the reduction of transgene
expression by co-suppression. Some of the pitfalls of biolistics are
circumvented by the delivery of fragment DNAs or by using the
‘agrolistic’ approach24. This novel method permits generation of a
simple insert even after biolistic or protoplast transformation.
Agrobacterium-mediated transformation
The natural ability of the soil microorganism Agrobacterium to
transform plants is exploited in the Agrobacterium-mediated
transformation method. During the process of transformation, a
specific segment of the vector, T-DNA, which can be engineered
to contain a selectable marker and/or genes of interest, is trans-
ferred from the bacterium to the host plant cells and inserted into
the nuclear genome. These functions are mediated by a set of viru-
lence genes with optimal expression occurring at acidic pH and in
the presence of phenolic inducers, such as acetosyringone, that are
released by wounded plant cells25. The Agrobacterium system is
attractive because of the ease of the protocol coupled with minimal
equipment costs. Moreover, transgenic plants obtained by this
method often contain simple copy insertions. These advantages were
a driving force to adapt this system to many different crops includ-
ing monocots. To this end, highly efficient vectors25 were designed
with extra copies of the virulence genes (superbinary vector), or with
mutations that enhance virulence gene expression. Already, it is poss-
ible to transfer large fragments (150 kb) into plant nuclear genomes26.
Typically, Agrobacterium-mediated transformation of dicots is
performed using sterile leaf pieces, cotyledons, stem segments, callus
suspension cultures and germinating seeds. Because of having to
deal with two different biological elements, many parameters should
be tested to satisfy both partners and guarantee a successful out-
come. These variables25 include the use of feeder cells, alternative
Agrobacterium strains, infiltration of the bacteria, the duration
and temperature of co-cultivation. Agrobacterium was reported to
penetrate tobacco tissue through stomata cells after spraying with
the bacteria27, so that wounding before inoculation should not be
necessary, at least in green tissues. However, it was necessary to
substitute wounding by pre-induction of the bacterial vir genes.
Pre-wounding25 sunflower shoot apices or banana meristematic tis-
sue, excised from corm tissues or from shoot tips, with microprojec-
tiles or glass beads16 has proven to be beneficial4. This phenomenon
could be explained by the attraction of Agrobacterium to wounded
sites and the increased access of bacteria to plant cells. Subjecting
plant tissues to brief periods of ultrasound by sonication, which
allows Agrobacterium entry throughout the tissues, is yet another
method of increasing transformation efficiency6.
Some crops appear to react, or be hypersensitive, to Agrobacterium
inoculation by forming necrotic barriers25. In grapes, this reaction
has been remedied by the addition of antioxidants, such as
polyvinylpyrrolidone and dithiothreitol. In another example,
shoot tip necrosis of both aspen (Populus alba) and poplar
(Populus trichocarpa) could be overcome by buffering the medium
with 2-(N-morpholino)ethanesulfonic acid and Ca-gluconate.

Interestingly, it is suggested that a block in T-DNA integration
underlies the recalcitrance of some monocots to Agrobacterium
infection28. However, Agrobacterium can transform monocots29 even
if they are generally not their natural host. Transformation of wheat
immature embryos and embryogenic calli is reported30. This success
is attributed to the addition of surfactant to the inoculation medium.
Barley transgenic plants were recovered after transformation of
immature embryos, from which the embryonic axes had been re-
moved, and were wounded subsequently with microprojectiles31.
Transformation of maize immature embryos from the inbred line
A188 and its derived hybrids has also been reported32. This achieve-
ment might be explained by the use of the superbinary vector. For
sugarcane, the use of this vector, as well as the pre-induction of
organogenesis, appears to be crucial for recovering transformants33.
Alternative methods of transformation
There is a perpetual quest to find more efficacious and economical
methods for plant transformation. Some of these are described here,
but are not often used because of their impracticability at present.
Plants can be recovered after DNA delivery using silicone carbide
whiskers34 but the use of these fibers requires caution because of
their potential carcinogenic effects. The electroporation method35
is not often used because of its low reproducibility. Microin-
jecting DNA into zyogtes36 is an alternative method that could
eventually lead to recovery of transgenic plants. Uptake of naked
DNA by plant cells can also be facilitated by laser microbeam37.
However, these options, which are still at the early stages of
development, can be tedious and require sophisticated equipment.
In planta transformation
There is considerable interest in developing plant transformation
methods that exclude the tissue culture steps and rely on simple
protocols. These methods are called in planta transformation
because transgenes are generally delivered into intact plants in the
form of naked DNA or from Agrobacterium. The stage of plant de-
velopment elected is variable but for many species it is around the
time of zygote formation. This choice stems from the underlying
assumption that at fertilization the egg cell accepts the donation of
an entire genome from the sperm cell and it might thus be the appro-
priate stage to integrate transgenes. This concept is exemplified by
the successful transformation method developed for Arabidopsis.
In this method, Arabidopsis flowers are infiltrated or dipped into
an Agrobacterium suspension and subsequently some of the har-
vested seeds are transgenic38,39. Although the overall efficiency is
low, the sheer number of seeds recovered for screening and the ease
of the method makes it an extremely attractive alternative.
A similar approach utilizing naked DNA has been attempted.
Cotton transformants were recovered following injection of DNA
into the axil placenta about a day after self-pollination40. Simi-
larly, a mixture of DNA and pollen was either applied to receptive
stigmatic surfaces or DNA was injected directly into rice floral
tillers41, or soybean seeds were imbibed with DNA (Ref. 6). These
procedures, intriguing as they are, are impractical at present
because of their low reproducibility.
Direct access to the male gamete is an option that can be
achieved by biolistic delivery of DNA into tobacco pollen in cul-
ture. Subsequent pollination with the bombarded pollen led to the
recovery of transgenic plants42. Similarly, this method could most
certainly be applied to other crops43. However, this apparently
clear strategy might, in fact, be complicated by a variety of factors
including the presence of nucleases and methylases in pollen44,
pollen survival and polyploidy45.
In another approach, meristematic cells located in the apical
dome of nodal buds have also been considered ideal recipients for
trends in plant science
reviews
transgenes because growth and development occur in this area.
Furthermore, some cell layers of the apex will eventually con-
tribute to the germline and are thus transmitted to the sexual
offspring. DNA and lipofectin injection followed by electropo-
ration into pea apical meristems allowed the recovery of transgenic
offspring6. Excised apical meristems from embryonic axes and
shoot tips46 have also been used as targets for transformation by
particle bombardment or by Agrobacterium inoculation. After
DNA delivery, these tissues are induced to form multiple shoots
prior to whole plant regeneration. This method is, however, very
labor-intensive because each regenerated plant must be screened
for transgenic sectors that will potentially contribute to the germline.
In addition, this method requires some tissue culture steps. Shoot
apices are far less efficient as the target for transformation com-
pared with embryogenic tissues, with the additional risk of obtain-
ing chimeric plants46. However, the efficiency can be improved by
multiplication of shoot meristems before DNA delivery47.
Scorable markers
For each of the transformation methods, transient expression ex-
periments, generally performed with a reporter gene, are a prelimi-
nary step used to identify conditions that will allow efficient DNA
delivery. With so many variables and no evidence of combinations
that are broadly applicable across plant species, this simple test pro-
vides some guidance for answering several initial questions, by
providing the most convenient way of measuring the quantity of
DNA introduced into the target cells. As the conversion rate from
transient expression to stable integration in the plant genome is low
and estimated to be ,1% to 9% for DNA delivered by bombard-
ment22, it is worthwhile achieving a high frequency of transiently
expressing cells following DNA delivery. Indeed, most transgenic
plants are generated from transformation systems capable of pro-
ducing transient expression in regenerable tissues. However, fas-
tidious research to maximize transient expression is in vain if it is at
the expense of tissue health. The empirical rule for success is to iden-
tify an efficient gene delivery method in the largest possible number
of regenerable cells without negatively affecting tissue survival.
Several reporter genes are used in plants, including b-
glucuronidase, luciferase and genes involved in anthocyanin bio-
synthesis48. More recently, the gene for green fluorescent protein
(GFP) has become an important in vivo reporter in plants. When
expressed in cells and illuminated with blue light, GFP yields stable
bright-green fluorescence, which is easily monitored nondestruc-
tively (Fig. 2)49. It can thus be used as a means to visualize the fate of
transformed cells over time and rapidly test the influence of various
factors through the successive steps of the transformation protocol.
Selection
Selection is an important part of the transformation process. In
general, the gene of interest is co-delivered with a selectable marker
to identify and encourage the growth of recipient cells. Selectable
markers usually confer resistance to chemical agents, such as antibi-
otics or herbicides that inhibit various cellular functions48. Scorable
markers, such as GFP (Ref. 49), are also used for selection. In most
cases, some tailoring of the selection process is required for each crop.
Mannose-6-phosphate isomerase (MPI) is a recently developed
selectable marker. The enzyme is encoded by manA from E. coli,
which converts the unusable carbon source mannose-6P to
fructose-6P. Thus, transformants containing manA can grow on
mannose as a sole carbon source. This selection has a positive
mode of action that encourages the growth of transformed tissues
rather than just permitting it. The MPI marker is extremely effective
for the selection of transformed sugar beet50 (0.94%), maize (.50%)
and wheat (25%) (G. Hansen et al., unpublished).
June 1999, Vol. 4, No. 6 229
trends in plant science
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Fig. 2. Green fluorescent protein expression in oat seeds on either
side of a non-transgenic oat seed (magnification, 653).
Because of concerns raised about the use of antibiotic genes for
selection, several approaches to eliminate selectable markers have
been envisioned51. The selectable marker might be used in the first
generation and eliminated later by conventional breeding. One
possibility is to deliver two separate T-DNAs; one with the genes
of interest and the other with the selectable marker gene, from the
same or two different Agrobacterium cells. The two T-DNAs are
often inserted into unlinked sites in the host plant genome, allowing
later genetic segregation51. Similar segregation can be achieved if
the experiment is performed using the ‘agrolistic’ approach24.
Transgene integration and expression improvement
The perfect transformant would contain a single copy of the trans-
gene that would segregate as a mendelian trait, with uniform ex-
pression from one generation to the next. Ideal transformants can
be found with difficulty, depending upon the plant material to be
transformed and to some extent on the nature and the transgene
complexity. As gene integrations are essentially random in the
genome, variability is often observed from one transgenic plant to
another, a phenomenon ascribed to ‘position effect variation’52–54.
The general strategy to ‘fix’ this situation is to generate, probably
at a high cost, enough transgenic plants to find some with the
desired level of expression.
Efforts are being directed toward achieving stable expression
of the transgene with an expected level of expression rather than
that imparted by the random site of integration. Scaffold attach-
ment regions could potentially eliminate such variability by
shielding the transgene from surrounding influence. Some guid-
ance might come from genome sequencing, which might provide
the necessary DNA ingredients to control gene expression. The
ability to target integration could also lead to some control of
transgene expression55. It is foreseen that site-specific recombi-
nases could assist in this endeavor56. All these areas of research,
which are primed for breakthroughs, should be carefully moni-
tored for immediate implementation in the design of suitable vectors
for use in transformation. In the longer term, it is less expensive
and ultimately more desirable to produce higher quality and fewer
quantities of transgenic plants.
230 June 1999, Vol. 4, No. 6
Prospects
Plant transformation remains an art because of the unique culture
conditions required for each crop species. To accommodate a
genotype or species that has not been manipulated in culture pre-
viously, one must either adapt an established protocol or create a
new one, bearing in mind the efficiency imperatives. There is also
a practical need for a method of transformation that will decrease
the complexity of the pattern of transgene integration and expres-
sion. Presently, most commercial transgenics are altered in single
gene traits. The challenge for the genetic engineers is to introduce
large pieces of DNA-encoding pathways and to have these multi-
gene traits function beneficially in the transgenic plants.
Although a clearer understanding of the events surrounding the
integration and expression of foreign DNA is emerging, there
are many questions that remain unanswered. Are there target cells
or tissues not previously attempted that are more amenable
to transformation? Is there a physiological stage that allows
greater transformation? Can it be manipulated to achieve higher
transformation efficiency? Does the tissue chosen as a target
affect the level of expression? It is becoming increasingly clear
that plants transformed by Agrobacterium express their transgene
more frequently. Can this be partly attributed to the fact that
T-DNAs frequently integrate in telomeric regions57?
Transformation technologies have advanced to the point of
commercialization of transgenic crops. The introduction of trans-
genic varieties in the market is a multi-step process that begins
with registration of the new varieties, followed by field trials and
ultimately delivery of the seed to the farmer. Technical improve-
ments that have the greatest opportunities for new approaches,
probably in the realm of in planta transformation, will further
increase transformation efficiency, extend transformation to elite
commercial germplasm and lower transgenic production costs,
ultimately leading to lower costs for the consumer.
Acknowledgements
Our thanks to the members of the Transformation Technology
Group of Novartis, whose work has contributed to the content
of this review. We thank Heidi Kaeppler, Heng Zhong and
Roberta Smith for providing photographs of oat seeds, maize
shoots and cotton shoot, respectively.
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Genevi•ve Hansen* and Martha S. Wright are at Transformation
Technology, Novartis Agribusiness Biotechnology Research, Inc.,
3054 Cornwallis Road, Research Triangle Park, Durham,
NC 27709, USA.
*Author for correspondence (tel 11 919 541 8685;
fax 11 919 541 8585;
e-mail genevieve.hansen@nabri.novartis.com).
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