[ 205 1

Tram.lJ,it. 1/1)",,1. SOr..o{7 (2), '20j-'.!l3 (1964)'

Printed in Great 8rilaill
USE OF THE HONEYDEW STATE IN THE
IDENTIFICATION OF ERGOT SPECIES
By A. R. LOVELESS
Department of lloiany, Universio' College of Rfwtlesia alld J{)'flsaland, Salisbury
(Willl 31"'xl-('gurcs)
Ahllough the stromal;!, of a species of Clav;ccps must be oll'crvcd bd,'n: a final
identification can be nl<'ldc, the size and shape of the conidia of the honey.kw
state, considrrnl in conjunction with the range, aT'- here n>. ans
of 'I'hifl<""l gn,ups of conidia have b''''ll rc'-ogn;7.(,([ :l""I1'g
"I' l111,,,;ydn, Ii-"", i1,irty-four S. Rho.Jcsiall gras"'. Six of d'esc
conidial gn,ops 'urn"sllOlId to till; six species of Chwiu/'.1 known to OCl"lIr in
S. Rhodl",;a.tl,c rnll"ininl!:seven groups would seem to I<-present undescrihed
Claoiups spp. If conidial charae('rs arc used (0 presume dilT"erentiation,
iris (0 all of (he recorded host ranges of already
crgOIS and also \l) predict hosts of und<:scribcrl oncs.
There afC thirty dcscribed species of Claviceps (Lwgdon, '954; Biblio
of Systematic '953 onwards) which are distinguished from
one another by their host ranges ,,-nd by morphological fC'\lures. Lmgclcl11
('942a), who h:-ls drst:riblt\ eight of the thirty, has noted tlll"
which be has found useful ill separating species. These characters include
the type of sckrotiu1l1, the shape and size of the conidia, thc method of
germination 'lr the scleroli;>., the colour or the stipes ;'llld capitula, the
presence or ab"l'llCC of \('OSC hyphae on the stroma, and the shape and
size of the pcrithcci:'l. The size of the :'Isci is considered an unreliable
criterion, beGl1lse :lsci at dillcrelll stages of maturity may occur in a single
perithecium. As there is no rC:ldy means of determining the tkgree of
their maturity, this COli Id t'xpbill the wide vari:'ltion in ascus length
several workt.:rs have nT,mlecl within single species.
As a prelilllinary to identifying the species of C1avicrps found in S.
Rhodesi;l, the author made a detailed analvsis of the data contained in the
type desaiptions, ;llld in ;"lllY subsequellt descriptions, oral! the described
'species. A kw dcseriptiollS give very full information, some lack data
that arc essclllial Illl" ,omparillg species, while others arc so in3dequate
that the spet:ics to which they refer arc scarcely distinguishable. A com-
plete: description of a Claviccps sp. must include detailed information on
hoth the imperfect and perfect states. Nevertheless, the practical difficulties
in observing the perfect state for making determillations of already
<cribed species cannot be minimized. there is an inc,,itable delay
ITom the tinw llle sclerotia are collected from the host until the stromata
arc produced. This delay may be anything lip to 9 months if the sclerotia
. require a period ofaftcr-ripl'ning. different species oftcn require
different germination conditions. 'J he sclerotia orG. sulcata, after a resting
l'eriod of about Gmonths, are easy to germinate on damo filter-o:lnrr ;n
206 Trarzsactions British Mycological Society Identification of ergot. A. R. Loveless 20
7
the bboratory, whereas sclerotia or olher cnmislclllly fail 10
germinate under the same conditions. For an unkllown species the
requisite germination conditions can be determined only by lrying a
number of different experimental treatments, and this may rCfjllin' many
sclerotia. Thirdly, even when stromata have been produced, wnical
sections through the capitulum must be prepared to determine the shape
and size of the pcrithccia.
In view of the above diO"iculties the possibility or using the honeydew
or SpfJacelia state as an easy means of identiftcation has been explored.
Conidial mounts were prepared from ergotized innorcsCCIlCcs of as many
S. Rhodesian grasses as it has been possible to collect. S. Rhodesi<l is
particularly suitable for such a study becJ.llse, being predominantly a
SaV;lIllla region, it is an area in which an intensive search for crgols is
likely to reve;ll new host rccords and the presence or tlndescribed species,
Since Ig61, wben this investigation bcgan, conidia have been collected
(Ill th irt y-four spo;ies ofgrassl's of seven tecII geller;l. '1'llI're ;\rc 1111( 1\ 11 Ibtedly
rurtlll'r hoslS of C!m'icr{!J to be l'ITorded rrom S. 1{l!o\ksia,
hOIll')'dew has nol yet beell \'ollct;\cd [i'olll gl :ISSCS Oil which
Ctr,'brlla andro{!ogollis Cesali (or f,jJ;co({//TI/ fllldm{!o.!:'I/I;,\ (Cl's.) Schol-
Schwa1"7:) has been recorded in this eOLlntry. From the \lork of LlIlgdon
(Ig.pb) there is littlc doubt that this fungus is a lloncydl'w s:lprophyte,
:lnd that the finding of it on a grass is a safc indic:ltion or [1n::vioLls infection
11 ilh a species of ClllviC1'l's. It is worth mentioning tlwrd'lre th:lt the
conspicuous, black, convoluted mY\'clium of Cachet/a (/Ilrlmj;ov()IIis 011 gr:1SS
infloresccnees Ins proved:l most USdll1 fidd indic:ltlll" III l'l1.('ltiz('(1 plants.
.\!though the growth of C. (/Iu/mjlOgol/;s all an illfi.:ul'd suppresses
Ibc den:lupmell t of ClariujJs in Iha t spi kclet, a Cl ]'IfuI se;\ rdl Ileal' Cacbella-
inli.:etet! plants often revcals all uncontaminated llollcydel\' state thal
l\lighl othcr\\'ise ba\"C nOlice.
:\IETHOO OF RECORD1"G TilE SIZE A",D SIlAI't:: OF HONEYDEW CONIDIA
The conidi;\ of C{oi"iccjJs arc known In sIll IW eonsider:l bll' \'a rbl ion
in size and sklJw bOlh bel\\"cCll :md within SIW( ies. Tl) record lhc degree
of v:lri:ltion wilhin anyone conidial mounl. lhe outliucs of t\\'enty-live
random spores,' x 300) were traced on 'Ethuloll' 1' . .\[, Tr:lcing Film
alt:lchcd 10 the screen of a Rcidlcrt L:lnametn. Thl' dimension" of each
Sp'IlT \ITrC measlIl'I'd and the r;lllge in sll;IJ)(' of tIll' s;llIll'k W;I.S illdicateu
bv placing: thc OUlliues side hy side ill on!t;r or sp"re length.
'For each species of ergolized grass lelllr sep;\I';\le conidial mounts were
examined. The [our mounls were invariably laken frOl\\ Jillcrellt plants
and where possible tbey WCl'e selected so as to represel1l widely sep:lratrd
localities in S. Rhodesia and different seasons of collection.
RESULTS
The conidial mounts, as a. whole, differed very noticeabl\' in appearance,
but they could readily be separated into twelve groups.
Nevertheless, thineen groups (Figs. I, 2) ha\'\.: been n:eogni:l.ed because,
in deciding how to classify the conidial mounts, it was considered necessary
10 take host range into account as well as spore morphology. Except for
purjJurc{/ (Fr.) Tu!., all other known species of Clav;ceps are recorded as
having a relatively narrow host range restricted to grasses belonging to
..ne panicular tribe. species have a monogenerie host range, whilst
others are found on scveral genera restricted to a single tribe. C. purpurea
is unique in that it is recorded as having a very wide host range, spread
over members of several tribes, but even so it is apparently restricted
(Langdon, to grasses of the subfamily Pooide;'e (sensu Stapf, 1'.)19).
Wben the host relations of the twelve morphological groups of conidia
were considned, one group did not conlorm to the general restriction of
elavice/is species to particular host t:n::a. This morphological group was
recorded fl'om both Sorghum caiJrorulIl and Loudet;a simplex, two grasses
which arc Ilut only of different tribes but also of different subfamilies of
the Gratllill(';\C. It is extremely unlikely that the same species of Ciaviceps
could infq:1 wid,-Iy different grasses. This morphological group is
therdore I'q llTsell ted t\\ ice ill the ebssi [ication or conidial IIlOllll ts, once
in tile l'o.. it!l-ae :ll\llllllcC in the l'anicoidc<le, lhereby incre:lsing the total
number of groups rccognizcd !i'om twelve to thirtl"\.'n (Table I).
III aClIJrdanee with expectation, the four conidial mounts from anyone
host species all fitted with the same group. TIlc conidia of certain groups
are so distinctive that they can be e:\sily recognized at a glance. The small
reniform conidia of group tOll R/!1'IIchl'/)'II"IIJII /"fjims and the large' fa\catc
or fusiform conidia of .<..:H)IIP 7 Oll Cl'llc!lrus (if/(/ris and Penllisclum
:Ire examples of sllch di.stilll'ti\"{' cOllidia. ,\ dct:lilcd cxalllinatioll of the
size and sll:l]>e of the \I'ilhilllhc groups \\'il\ sllow, h'l\\'ever,
that cadI dislinct from mher groups on the same lribe. In fact
IllOSt grOllp, can be srpar:l.\"d solely on the basis of mean spore length
{Fig. 3;. \\'here the conidia nf t\\'o groups arc very similar in lenglh and
thus indislinguishable on this b:lsis, they call nevertheless be separ:\lcd by
their respn:tive shapes. Thus, whcreas the eOllidia of C. digitariae (group 5)
"nd C. S1/{Olta (group G) cannot be distinguished by spore measurements
alone, the [;ll'lller h:ls pred\lIllinamly ellipsoid and the latter predominantly
:lllalltoid spores. In view of the inevitable variation in me;lll of
diO'crent s;lI11plcs of the same species, the author has !ound the to
breadth r:1tio a most useful accessory character to spore length in cla: ;i!"ying
conidial sallIples. This r;llio b:lS been lI"idely used lO gi"" an il'dex of
shape :l11\1 in lhis \l"I\rk tl'IHkd 10 be nmsl;\l\t 101' a gi\'cll species, evcll
though the IIIC:llI dimcllsiollS of different samples v;\ried sOlllewh:n,
CONCLUSIONS
On the assulllption that e3ch group of conidia represents a. distinct
taxon, the honeydew stales fWIll the lhirty-four grass species can be
assigned to Ihirtecn species of Claviceps. Before Ig61 only four named
species of C!rw;cejJs (c. C)'lIodont;J. C. digiltlliae, C. paspali and C. sulca/a)
and two undetermined species (onc on Pal/hum maximum and another 011
PmniSdllm l)'jJhoides) were definitely knmnl to occur in S. Rhodesia.
Recelltly the aUlhor (1963) bas rcported in addition C. maximlllsis and
TramQctions British Myc%gical Society
20
10 s
H
Tribe: Panleue
I
.-

----I
...
,
---
-- ;
108
I .. ---- --;
11-8
I
.-
----oj
U8
,
.. ---

------1
13'8
;
1------------
,
--------1
1S8 ,
l-------
-------oj
16,0
l
l--- _____

-------1
1011
Tribe: Andropogonue- ,
.--

-----.;
13-2
)
.--
-----.;
16'4
I
--;
13-S Tribe: Chlorideae
I
--,
13-8 Tribe: Arundinelleac
l
1----------

-- --.;
,
.-
15
Spore length (#)
Fig. 3 The ranges and means of 'pore length for the thirteen group5 of honeyd,'"
conidia. The dotted portion, of the range lines indicate the tail 5-10 %of the frequcnC)"
distribution at each end of the range.
Ide1llification of ergot. A. R. Loveless 21 I
C. pusilla. Both are believed Lo be new r{;cords for southern Africa, though
the former merely represents the identification of the ergot on Panicum
onaximum that was known but had not been identified.
The conidia of the six named species of Claviceps correspond to six of
the thirteen conidial groups recognized above. There is no available
d,scription of the conidia of C. digitariae, but the ergot associated in S.
Rhodesia with Digitaria diagonfllis and D. ga<.tnsis has been confirmed as
( digitariae by successful germination of the sclerotia. For the other five
a;nhenticated species, the conidial characters (Table I) agree reasonably
well with those given eidler in the type or in subsequent descriptions.
It has not been possible to equ<lte the remaining seven groups of conidia
with any known species (of (lavio!ps. From considerations of host range
il would seem that most, if n, ,t all, of them belong to undescribed species,
but this can be established only by observing the stromata produced by
germination of the sclerotia. Attempts are being made to germinate
sclerotia corresponding to these unidentified groups of conidia but, as
j
i

i
1
,
t

,
,
i

;
o
i
o
Cl
1
!

"
"
"

i
o
L
,
.

n
,

'-
"
,
-
,
<
,
i

,
,
<
.'
,

" I . ,
, I
"u ..
-,
" '
.g
$
.,
- "

,

"

"
,
,
I
,

f
'- ,
" ,
, -
" ,
-i'l
b .!.
.. !i
cc':
c co.:
- -.
88
!

00

.
" "
" . .,
o 0
"
,

i
o
j

!;

b
- .,
. '.
- ..
1
g '" :I:
E := :.i
U,; ... i :: .

-'C.
Cl Q.
l
i
< < ..,.- __ ';;_"
'"
J!!;;;";;!:': t "..:l::,."

.

...
""'.1l:::::'::::::::::::':::':
,
,

'", " , -
- .
r :0-
.
;. -:ri
"
" "
,
, -
'
;g
" 0
. o
-;; ==
" .
,
,
!n 'f'
"

"
, ,. ,

-
"
00

.

"

,
" .

, , ... ..,

]
.- .
, ,
c '!;

, 0
i
" "
l:
o .

"
t

"
, -
- ,
- .,

,
.

.,
, 0
0 o
" .
: j
.' , ,
.::
. "

ti ti
! I

:: ::
] "'g
"

o

_0
..
N ;.,

r..!.-
':'; r
,!.,.!<
,
f:
- -.
-
.
- ,
,-
-.
Fb
.
I!
.
.

0
;., ;.,
:,. ;.,
"




'!''!'
;.. '"
0

,.
, 0
._ 0
0-
" .
<0
i

i
i
, .
-u

It
.
).,
I' ;,

.:! ..
-
... ..:..
;, :
f!
:e
"
i! "E .2
2
'. .

:I: ::: __ ." "!
I.l "! li-S 'Q.
<.i ...... <ii-" Cl !)
:;; ...
.== 'co""' -:;;- ... E
;::; :l-B t: ... IJI ';'::::rii
j
Co ""=
...

;
't,i1.
g <i: e';;:';0 <5ci c::::::<:i::i::,; wO:: e
""
o 0
C ... t,
;, ;,

..'
r
3
.,
]
]
"j
i
]

,

"
,

E"
o.

,.


-,.
,,2 ..
]11
"
210

eo
il

"
"
.0

"
-
'"

.,

-;:

0;

.'i!
"
"
o
""
u
"
0-
V>

."
-

"
"-

"
-
'" '.,
20ts "J'ransactions British Myc%gical SopelY
SOa-FAMILY; PAN1COlOEAf
Tribe; P,nicue
Group 1 C, rhynchelytri
aOOaOOOOOGOOaOaOOGGOOOaGG
Group 2 (C.
UOOOODOOOOOOOOOOOOOOOOOOO
Group J
0000000000000000000000000
\
Group 4 f

Group 5 (c. ditiurlu)

Group 6 (C. JUkIU)

Group 7 C, fusiformis


Fig_ I. Spore umples or the eighl conidial groups 011 S. Rhodesian .
grasses belonging to the tribe P:r.niecae, E."lch sample CQllsisll or ralldolll spores
()C 'Cl),
Idemifi<;alion of ergot. A. R. Qveless 209
SU8-FAHILy : PANICOlDEAE (eone/nlltd)
Tribe: Andropocone1e
Group 9 (C, puSWI)
O()()()() OOO()() 0l]O()()OOO(]OOOOOG
G I.
C. africana
0000000000000000000000000
G 11
Claviceps Sp. HYP
oooooooOOOOOOOOODOOO
00000
SUB-r \HILY : POOIOEAl:
TrIbe: Chloridele
Group 12 (C. cynooOl'ltls)
ooOGOGGGGeuOGOGOGddOGGddG
Tribe: Arundlnellue
Group 13 Claviceps Sp. LOU
0000000000000000000000000
Fig", Spore samples of the five conidial groups on Rhodesian
ITauet belonging to the tribes ,\ndropl!gonc:ac. ChlQrlde:l.e and ,\rundmelloe. The
samples are drawn at Ihe same magnifies-don (le JO) :is those in Fig. I.
(AtUpltdfOT p1/blicaJion 8 August (963)
REFERENCES
LA/'tGDO/'t, R. F. (1942a). Ergot ornativr grasses in Quunsland. PTtX. ro:!. Soc. Qd 54.
23-32. '
WGDO/'t, R. F. (19420). The genus CTrbtlla Cesati-its biologic status and we. Phylo-
/NJI!toloD, 32, 613-017.
I..vtODO/'t, R. F. (195i). The origin and differentiation of Clauiups species. VI/iD. Qd
Pap. (1101.),3, fil-68.
Lonuss.. A. R..(19"4). Studies of Rhodesian ergots. I. Cltwitrps ma.nmrlUU Theis and
ClaDIUPS ",mUD <eati. KiTkia, 4, 35-40.
Sn.p" O. (19'9) (;ramineae (incomplete). In Flora of Tropuaf "friea g. Ashrord
Kent: L. Reevc and Co. ' ,
2I2 Transactions British Myc%gical Sodety
indicated above, this is likely to prove a lengthy process the
appropriate germination conditions are unknown.
Besides the general conclusion that there are probably several un-
described species of Clauiups in S. Rhodesia, the following additional
points arising out of this investigation deserve mention:
(I) On the basis of the occurrence of the very characteristic triangular
conidia of C. pUJilla, it is safe to extend the recorded host range of this
species to include the genus HyporThLnia. A similar conclusion may also
be tentatively drawn for C. sultata. This ergot has hitherto been recorded
only on BrlUhiaria spp. but allantoid conidia similar to those of C. sulcala
occur also on Urodtloa, closely related to BraclliaTia. It seems likely there-
fore that Uroddoa is also a host genus of C. micala, though this can be con-
firmed only when sclcrotia from Ur()thioa have been germinated.
(2) The occurrence of conidia of the same rather unusual f,,\catf' Jlhape
on both Ctnc!lrus ciii"Tis and tJ/1!loitlrs would indie.lle tllOll these
are both hosts of the same Ciauiceps, which is almost certainly ;111 un-
described species. It may be possible, therefore, to predict the host range of
an ergot species not yet dc...'scribed.
(3) Although most CinvicejJs Jlpp. have a narrow host r.mge,
it cannot be nssumed that the dilferent species of onc host genus will
always be attacked by the same ergot. From the present study it seems
a Imost certain that the ergot ofPnlliCll1ll roloTalul/I is not, as mi)..:"ht be expected
un a Iiost basis, the same as that on P. maximum. Simil:uly, J-(I'flarrlltl/ia
appears to be parasitized by two species, C. pusilla alld another (with
conidia in Group 11) that is probably undescribed.
(4) Judging from the host records it would seem that Ciauiup.r has
cvolved along ...ery different lines in the two subfamilies of thc Gramine;1e.
In thc Panicoideae it has apparently evolved into a number of distinct
species, each with a restricted host range. The results of the prCJlcnt in-
vestigation endorse this view. In the Pooidcac, by contrast, physiological
l:volution does not seem to have been accompanied by a concurrent
morphological evolution. Only six Ciaviups species are known on grasses
of the Pooidene, and four of these arc reported to have a monogeneric
host range: n fiflh, C. iiforalis Kawatani, is recorded 011 El.pm'.f and
If/m/mm in .Jnpan. ,.\[1 othcr crgots rccorded liom the Pooid,ae, covcring
nearly 200 host spccies of li tribes and 50 gCllera (Langdol1, arc
ascribed to Clavictps pflTpuTta. The author wonders to what extent the
tremendous width of this reputed host range is the result of arbitrary
identifications rather than genuine physiologic specialization. It is
suggested that a critical examination of honeydew conidia from recorded
hosts of C. pflTpflTea, of widely differing taxonomic status, would be well
worthwhile.
I am grateful to t\[rJ. B. Phipps and 1,,[rs Luey Crook (nee Chippindall
of the Federal Herbarium, Salisbury, for invaluable help in identifyin:: ,
ergotized grasses, to Mr A. R. A. Nod lill reading the manuscript, and
to my wife for continual assistance throughollt the work.
Identificatiqn of ergot. A. R. Loveless
21
3