Weed Science Society of America

An Evaluation of Hydrophilic Polymers for Formulating the Bioherbicide Agents Alternaria

cassiae and A. eichhorniae
Author(s): Yasser M. Shabana, R. Charudattan, James T. Devalerio and Mohamed A. Elwakil
Source: Weed Technology, Vol. 11, No. 2 (Apr. - Jun., 1997), pp. 212-220
Published by: Weed Science Society of America and Allen Press
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Weed Technology. 1997. Volume 11:212-220

An Evaluation of HydrophilicPolymers for Formulatingthe BioherbicideAgents

Alternariacassiae and A. eichhorniael
YASSER M. SHABANA, R. CHARUDATTAN,JAMES T DEVALERIO,
and MOHAMED A. ELWAKIL2
Abstract: Eight polymers capable of forming aqueous gels were comparedfor their capacityto retain
hydrationover time, to promote spore germination,and to prolong the viability of germinatedspores
(= germlings) of Alternariacassiae, a bioherbicideagent for sicklepod. When comparedat a standard
0.1% w/w (gel/water) concentration,the eight gels retained hydration for 6 d with no significant
differences among them in the rate of dehydration.The best concentrationof each gel that yielded
95 to 100% spore germinationwithin 6 h after hydrationwas then chosen, and the gels were compared at these concentrationsto determinethe durationof effectiveness of the gels. The effectiveness
was rated on the basis of the proportionsof alive germlings versus germinated spores and alive
germlings versus total spores, determinedwith the aid of a fluorescent vital stain. Based on these
two parameters,the most effective gel was Kelzang xanthan gum. However, all gels supported>
50% alive germlings over a period of 1 wk, suggesting that the addition of any of these polymers
to the inoculum suspension should enable the fungal propagules to remain moist for a prolonged
period, benefit from the high ambient moisture to improve germination,and promote disease development. Accordingly, seven of these gels were tested for their ability to enhance pathogenicityof a
mycelial inoculum of A. eichhorniae, a bioherbicide agent for waterhyacinth.Gellan gum and Kelgin?-HV were most effective in promoting disease, followed by Evergreen? 500 polyacrylamide,
were no better than the control
and Kelging-LV. Metamucil?, Kelzang xanthan gum, and N-Gel@3'
inoculum without any gel. Thus, the gels may have differentialeffects on different fungi and inoculum types. Nonetheless, the results confirm the utility and feasibility of hydrophilic gels as formulating materialsfor bioherbicides.
Nomenclature: Sicklepod, Senna obtusifolia L. #3 CASOB; waterhyacinth,Eichhornia crassipes
(Mart.) Solms, # EICCR;Alternaria cassiae Jurair& Khan;A. eichhorniae Nag Raj & Ponnappa.
Additional index words: Adjuvant,alginates, aqueous gel, bacterialpolysaccharides,cellulose, formulation,humectant,moisturerequirement,mycoherbicide,polyacrylamide,psyllium mucilloid, CASOB, EICCR.
Abbreviations: A, alive germlings;DS, disease severity; G, germinatedspores;PDB, potato dextrose
broth; RH, relative humidity; T, total spores.

INTRODUCTION

relativehumidityplay key roles in the infectionand

sporulationprocessesof foliarfungalpathogens(Huber
A majorobstacleto the use of foliarfungalpathogens 1992). The influenceof wateron the infectionprocess
for weed controlis the need for moisturefor the fungal has been well documented(Huber1992; Ingold 1978;
propagulesto germinateand infect and, to an extent, Lacey 1986). The occurrenceof moisturelowers the
colonize the weed. Free moistureon plant surfaceand host'stranspiration
rate,bringsthe waterpotentialclose
to zero,andrendersa greateravailabilityof internalwaIReceived for publication June 3, 1996, and in revised form October 23,
ter
for the pathogen(Huber1992).
1996. Publication R-05182 Florida AgriculturalExperiment Station Journal
The limiting effect of moistureon the efficacy of
Series.
2 First and fourth authors: Assistant Professor and Professor, Plant Pamany weed pathogenshas been well documented,and
thology Department,Faculty of Agriculture, Mansoura University, El-Manmoststudiesdemonstrate
a relationship
betweendewpesoura, Egypt; second and third authors:Professor and Senior Biological Scientist, Plant Pathology Department, University of Florida, Gainesville, FL
riod and the amountof disease (TeBeestet al. 1992).
32611.
Moistureis perhapsthe most constrainingfactorto the
3Letters following this symbol are a WSSA-approvedcomputercode from
Composite List of Weeds, Revised 1989. Available from WSSA.
developmentof mycoherbicides.The successfuluse of
212

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WEED TECHNOLOGY

Collego?4 [Colletotrichumgloeosporioides (Penz.) Sacc.

f.sp. aeschynomene], a commercial mycoherbicideused
to control northernjointvetch [Aeschynomenevirginica
(L.) B.S.P., # AESVI] is attributedto the high moisture
conditions in rice (Oryza sativa L., # ORYSA) and soybean [Glycine max (L.) Merr.]fields in which it is used
(Templetonet al. 1979). Adding materials such as wetting agents, stickers, emulsions, oils, and polymeric gels
that extend the leaf-wetness durationhas been shown to
be necessary to increase the efficacy of the inoculum
(Boyette et al. 1992). However, so far there have been
only limited attempts to explore the utility of the numerous hydrophilic polymers used in agricultural,food,
and pharmaceuticalindustries as formulating materials
to increase the efficacy of bioherbicides.
The objective of this research was to determine the
ability of eight different polymeric gels to hold moisture
for prolongedperiods, promoteviability and germination
of spores, and improve pathogenicity of a mycelial inoculum. Two bioherbicide agents, Alternaria cassiae
(Walkerand Boyette 1985) and A. eichhorniae (Nag Raj
and Ponnappa1970), were used. The former,a pathogen
of sicklepod, was tested as spore inoculum, the inoculum
type developed for possible commercial use (Charudattan et al. 1986). The latter,a pathogen of waterhyacinth,
was used as mycelial inoculum since this fungus is able
to infect from a mycelial inoculum (Shabana 1992).

plastic dish. Five replicateswere used for each of the

polymers.The weightof the dish plus the polymerwas
recordedbeforeaddingwater.Theplateswereplacedon
a laboratorybenchat 55 ? 5% relativehumidity(RH)
and23 ? 2 C in a randomizedcompleteblockarrangement.Eachplatewas weighedat regularintervalsuntil
the weight equalledthe prehydration
weight.The time
elapsedbeforereachingthis pointof completedehydration was calculated.This experimentwas repeatedthree
times. The data were analyzedby repeated-measures
analysisof variance(ANOVA)and by separatingthe
meansby multiplecomparisonsby Fisher'sLSD (SAS
Institute1985).

Effect of Gels on the Germinationof Alternariacassiae Spores and on the Viability of Germlings.The
influenceof eight gels on sporegermination,germling
development,and germlingsurvivalwas testedover a
7-d periodusing A. cassiae spores.9This fungus was
used as a model since an abundantand standardsource
of sporeswas available.Five concentrations
of each gel
were preparedin 70 ml distilledwater(0.0004, 0.004,
0.04, 0.4, and4.3%w/v) in glassbottles(9.5 by 4.5 cm).
Fourreplicatesof eachgel concentration
wereused.The
sporeswere addedat the rateof 0.01 g, and the sporegel suspensionswere kept on a laboratorybenchat 55
? 5% RH and 23 ? 2 C. The numberof spores,germinatedspores,and viable sporeswere countedafter6
h andafterevery 24 h for 7 dayswith a hemacytometer
MATERIALSAND METHODS
mountedon a Zeiss fluorescencemicroscope.Sporeviby usingfluoresceindiacetatedye
Retention of Hydration by the Polymeric Gels. The abilitywas determined
a vitalstainthatstainsonlyviable
(diacetylfluorescein),10
durationthe gels remained hydrated was determinedin
protoplast.
The
stain
preparation
was storedin acetone
order to select a polymer that retainedwater for the lonstock
solution
(1% w/v) at -10 C. At each counting
gest duration. Eight polymers representing different
the
time,
was
dye
addedto 5 ml of the spore-gelsuschemistries and gelling qualities were tested: Kelgin?5
pensions
taken
from
the bottleto give a finalconcentraHV, MV, and LV (sodium alginates of high, medium,
tion
of
0.1%
stain.
After3 to 5 min at roomtemperature
and low viscosities); Kelzan?s xanthan gum (polysac?
2
(23
the
C),
spores
were examinedfor fluorescence.
charide gum, produced by the bacteriumXanthomonas
The
exciter
filter
BG12
and the barrierfilter47 were
campestris, with a cellulose backbone and trisaccharide
used
(for
of
wavelengths
330
to 500 nm and > 460 nm,
side chains having a pentasaccharideas the repeating
respectively).Four sets of spore-gel-stainpreparations
unit); Gellan gum5 (a polysaccharidecomprised of glucwereused as replicates,andat least 85 sporesper treaturonic acid, rhamnose, and glucose; Kelco); N-Gela'6 (a
ment
were observed.
nonionic derivative of cellulose); Metamucil?7(psyllium
The abilityof the dye to stainonly the live protoplast
hydrophilicmucilloid);andEvergreen? 5008 (polyacrylwas
confirmedby adding14 dropsof 37%formaldehyde
amide). One hundredgrams of distilled water were addto
2
ml of sporesuspensionto kill the spores,staining
ed to 0.1 g of a polymer placed in an open 15-cm-diam
after10 minwiththe fluoresceindye (finalconcentration
IUniversity of Arkansas,Fayetteville, AR 72701.
andexaminingthe spores5 min laterfor fluores0.1%),
5The NutraSweetKelco Company,Monsanto Company,8355 Aero Drive,
cence.Twoml of the sporesuspensionwithouttheformSan Diego, CA 92123-1718.
6

Hercules Incorporated,Hercules Plaza, Wilmington, DE 19894.

7 Procter & Gamble, Cincinnati, OH 45202.

8

Chemie Linz AG, A-4021 Linz, Austria.

9Provided to R. Charudattanby Mycogen Corp., San Diego, CA 92121.

'?Esta

Kodak Company, Rochester,NY 14650.

Volume 11, Issue 2 (April-June) 1997

213

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SHABANA ET AL.: HYDROPHILICPOLYMERSFOR FORMULATINGBIOHERBICIDEAGENTS

aldehyde treatmentwas used as a control sample. The

examination showed that 99% of the spores treatedwith
formaldehydedid not fluoresce, confirming that the dye
stained only live spores.
The numbers of total (T) and germinated (G) spores
and alive germlings (A) were recordedand the ratios of
G/T, A/T, and A/G were calculated. Statistical analyses
were performedwith SAS (SAS Institute 1985). Arcsine
of square root transformationswere used for G/T, A/T,
and A/G proportionsto induce homogeneity of variances
among treatments.Plots of residuals and predicted values were used to determine the appropriatetransformation (Steel and Torrie 1980). The square-root transformed data were subjected to ANOVA, and pairwise
contrasts between treatmentswere performed. The best
dilution of each gel with respect to the proportionsA/G
and A/T (since they were considered the most accurate
indicators of gel's efficacy) was selected, and the data
were subjected to ANOVA. Fisher's LSD test was used
to determine the significance of pairwise comparisons
between treatments.

Effect of Gels on the Pathogenicityof Mycelial Inoculum of A. eichhorniae. Seven gels (Gellan gum, Kelgin? HV, Kelgin? LV, Metamucil?, Evergreen? 500
polyacrylamide, Kelzan? xanthan gum, and N-Gela'
were used at the best concentrationsdeterminedfrom the
previous gel-A. cassiae experiment.
The cultureof A. eichhorniae (isolate Ae5) was grown
in stationary Erlenmeyer flasks containing potato dextrose broth (PDB) at 25 ? 3 C and 12-h diurnallight of
36 puE/m2/sfor 4 wk. Mycelium from several flasks was
harvested on cheesecloth, pooled, and blended at 20%
w/v in sterile deionized water in a Waring blender for
10 s at the high-speed setting. The final inoculum consisted of each gel added to the mycelial suspension in
sterile deionized water to give a final concentrationof
0.5% for each gel, except for Gellan gum, which was
tested at 5%. Six individual detached waterhyacinth
leaves, as replicates, placed in six 15-cm-diam plastic
petri dishes lined with moist filter papers, were inoculated with each inoculum. Inoculation was made by
brushingthe gel-inoculum mixture, except in the case of
Gellan-gum-containing inoculum, which was spread
with a spatula. The dishes with inoculated leaves were
placed uncovered in a darkened dew chamber at 28 C
for 7 h, after which they were covered and transferred
to a growth chamber (25 + 3 C and 24 pIE/m2/s)with
a 10-h diurnal light. Control leaves were treated in the
same mannerwith gel suspensions and/orgel paste without the fungus. Seven days later, the leaves were rated

100,

oo

-_

F:
z

60-

ol
0
1

5.5

6.5

7.5

DAY
G-gum EK-HV EK-LV DK-MV
~Metam MN-gel oPoly fX-gum
Figure 1. Retention of hydration over time by eight different hydrophilic
polymers preparedas 0.1% (w/w) aqueous gels. G-gum = Gellan gum; K-HV
= Kelging-HV; K-LV = Kelging-LV; K-MV = Kelgin?-MV; Metam = Me-

tamucil?; Poly = Evergreen? 500 polyacrylamide;X-gum = Kelzang xanthan gum. Water retention was calculated from the weight of water in the
hydratedgels at different intervals. See text for statistical analysis.

for disease with the help of a pictorial disease scale

(Freeman and Charudattan1984). This experiment was
repeatedtwice. The data were analyzedby ANOVA, and
the significantdifferencesbetween treatmentmeans were
determinedwith Duncan's new multiple range test.
RESULTSAND DISCUSSION

Retention of Hydration by the Polymeric Gels. The

gels retained hydration for 6 d (Figure 1) with no significant differences among them with respect to water
retention as determined by weight loss, although there
was a significant time effect (P = 0.0001), a block effect (P = 0.0001) (i.e., due to differences in the physical conditions to which the plates were exposed), and
a time block interaction (P = 0.0001). There was about
18 to 22% water loss per day from each gel until the
fourth day after initiating the experiment. From the
fourth day to the fifth day, the amount of dehydration
decreased to about 14 to 17% (i.e., the rate of dehydration diminished), and from the fifth to the sixth day the
amount of dehydrationdecreased to about 7 to 9%. The

214

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WEED TECHNOLOGY

-.M.NM

C.~

~~~~~~~~~~~

..

__~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

C.,

C._

Figure 2. Comparison of fluorescein diacetate-stainedAlternaria cassiae spores under visible (A and C) and UV lights (B and D). Viable spores and live
germinatinghyphae fluoresce under UV light, dead spores and hyphae do not fluoresce. This technique was used to study the effect of various hydrophilic
polymers on the germinationof Altemnariacassiae spores and on the germling viability.

gels were totallydehydratedonly after7 to 7.5 d (Figure 1). Thus, the gels remainedhydratedfor a length
of time sufficientto promotesporegerminationandinfection (6 d).

total spores (A/T) and alive germlings among germinated spores (A/G). Based on statistical analysis of these
data (ANOVA and contrasts;available from the authors
but not shown here because of the voluminous data outputs), the following summary is drawn about the reEffect of Gels on the Germination of A. cassiae spective gel-concentrationcombinationsthat yielded the
Sporesand on the Viabilityof Germlings.Thedataon highest levels of germination and germling viability
the numberof total spores(T), germinatedspores(G), (G/T, A/G, and A/T values): All gels promotedhigh levand alive germlings(A) (Figure2) were used to deter- els (95 to 100%) of spore germination(data not shown)
mine the abilityof the gels to sustainviable germlings and germling viability (52 to 94%) at certain concentraof alive germlingsamong tions (Table 1); within each gel, the concentrationeffect
by calculatingthe proportions
215

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SHABANA ET AL.: HYDROPHILICPOLYMERSFOR FORMULATINGBIOHERBICIDEAGENTS

Table 1. Levels of viability of Alternaria cassiae spores at the best concentrationsof gels tested.a
A/Tc

Gelb
%

A/Gd

Gel

wlv

%Wlv

Kelzan' xanthan gum (0.4)

Metamucil? (0.4)
Gellan gum (4.3)
Kelgin6-HV (0.04)
Evergreen? 500 polyacrylamide (0.04)
Kelging-MV (0.04)
N-Gella (0.04)
Kelgin0-LV (0.004)

87.13
81.88
77.50
69.25
62.88
58.63
57.00
51.63

ae
a
ab
bc
cd
cd
d
d

Kelzan? xanthangum (0.4)

Kelgin?-LV (0.004)
Evergreen? 500 polyacrylamide(0.04)
Kelgin?-HV (0.04)
Kelgin?-MV (0.04)
N-Gell3 (0.04)
Metamucil? (0.4)
Gellan gum (4.3)

94.00 a
92.25 ab
90.88 ab
89.00 abc
86.50 abc
85.75 bc
84.13 bc
82.63 c

aThe values are the means of eight observations obtained over the experimentalperiod of 1 wk.
bThe best concentrationof each gel that promotedthe highest levels of spore germinationand germling viability.

c The proportionof alive germlings/totalspores.

dThe proportionof alive germlings/germinatedspores.
eValues followed by the same letter(s) within each column are not significantly different according to Fisher's LSD test (P

was significant(P < 0.02) with respectto A/T andA/G

proportions.Therefore,it was possible to separatethe
most effective concentrationfor each gel from others
thatwere less effective.
On thebasisof the abilityto sustaingermlingviability
amongtotal spores(A/T), 0.4%Kelzan?xanthangum,
0.4% Metamucil?,and 4.3% Gellangum were equally
effective(Table1). These gels were followedby 0.04%
Kelgin?-HV,0.04% polyacrylamide,and 0.04% Kelgin?-MV.The least effective were 0.04%N-Gels and
0.004%Kelgin?-LV.Whencomparedon thebasisof the
abilityto sustaingermlingviabilityamonggerminated
sporesonly (A/G),0.4%Kelzan?xanthangum,0.004%
0.04%Kelgin?-HV,
Kelgin?-LV,0.04%polyacrylamide,
and 0.04% Kelgin?-MVwere equallyeffective.These
gels were followed by 0.04%N-GelI?,0.4% Metamucil?, and4.3%Gellangum.
The datafromthe best concentrationsof each of the
gels were plottedtogetherto comparethe gels with respectto the proportionsA/T (Figure3) andA/G (Figure
4) over time (i.e., to estimatetheir ability to sustain
viable germlingsover time). The gels differedin their
ability; in general, the proportionof alive germlings
amongthe total spores(A/T) increasedup to 72 to 120
h from time zero and then decreaseduntil the end of
the experiment(Figure3). On the basis of A/T,the gels
fell into three categories:those that sustained80 to

0.05).

the gels sustained> 60% viability of germlingsover

the 1-wk experimentalperiod(Figure4).

Effect of Gels at Selected Concentrationson the

Pathogenicityof MycelialInoculumof A. eichhorniae.
The highest levels of disease severity(DS) were obtainedfrominoculacontainingGellangum(5%w/v) and
Kelging-HV (0.5% w/v). Evergreen?500 polyacrylamide(0.5%w/v) andKelgin?-LV(0.5%w/v) werenot
significantlydifferentfromGellangumandKelgin?-HV;
treatments
containingthesepolymerswereno betterthan
inocula with no gel or with Metamucil?(0.5% w/v),
Kelzan?xanthangum(0.5 w/v), or N-Gela'(0.5%w/v).
Also, therewere no significantdifferencesamongMetamucil?,Kelzan?xanthangum, N-Gela', andthe control withoutany gel (Table2).
Thus, the gels used can be categorizedinto three
groups on the basis of disease promotion.The first
group of the most effective gels consisted of Gellan
gum and Kelgin?-HV.The second groupincludedthe
moderatelyeffective gels, Evergreen?500 polyacrylamide and Kelging-LV,and the thirdgroupcontained
the least effective gels, Metamucil?,Kelzan?xanthan
gum, and N-GelI?.
Theproblemof the lackof optimumdewperiodsnecessaryfor fungalgerminationand infectionis common
to a numberof potentialmycoherbicides;
researchindi100% alive germlings (Kelzan? xanthan gum and Mecatesthatthis mightbe overcomeby certainformulation
tamucil?);60 to 80% (Gellangum, Kelging-HV,Kel- techniques(Amsellemet al. 1990;Boyetteet al. 1992;
gin?-MV,polyacrylamide,andN-Gel'?;and40 to 60% Heiny and Templeton1991; Quimbyet al. 1989;TemKelgin?-LV)(Figure3). The proportionof alive germl- pleton and Heiny 1989). An extendedperiodof foliar
ings among germinatedspores (AMG)
continuedto be wetnessis requiredfor a fungusto infectthe aerialparts
high until72 to 96 h afterinitiatingthe experimentand of the targetweed. For this reason,the influencesof
then decreaseduntil the end of the experiment(Figure eight hydrophilicgels on sporegerminationand germ4). Withthe exceptionof Metamucil?andKelgin?-MV, ling viabilityweretestedusingA. cassiae andon patho216

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WEED TECHNOLOGY

100

0 100
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200

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Issue 2 (April-June)199721780

60 -

POLY

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040
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AGENTS
BIOHERBICIDE
FORFORMULATING
POLYMERS
SHABANAET AL.:HYDROPHILIC

~''100

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WEED TECHNOLOGY

genicityof a mycelialinoculumusingA. eichhorniae, to

select suitableformulatinggels.
Assuming100%viabilityof sporesandavailabilityof
free moisture,the germination
processin a populationof
sporesis a functionof timein at leasttwo ways(Gottlieb
1978). First,the sequenceof events leadingto the formationof germtubesin any single sporeis time-dependent, and this in itself may vary with manyparameters
such as temperature,nutrition,and osmotic pressure.
Second,the heterogeneityof the populationis suchthat
the sporesarenot synchronizedto begingerminationsimultaneously.Attemptshave been madeto synchronize
the germinationprocessby priorhydrationtreatmentor
cold and heat shocks(Gottlieb1978). In our study,we
used priorhydrationin gel solutionsto synchronizethe
germinationprocess and obtained95 to 100% viable
germlingsfromthebestconcentration
of eachgel in only
6 h afterhydration.
We used the proportionsof alive germlingsvs. the
total spores (A/T) and alive germlingsvs. germinated
spores(A/G) as criteriato evaluatethe effectivenessof
the gels. Wereasonedthatthe abilityof a gel to promote
sporegerminationandto sustaingermlingviabilityover
severaldays would be the most importantand desired
characteristics
in a formulatingmaterial.Basedon these
criteria,all eightgels wereacceptable,althoughKelzan?
xanthangum was clearlythe best.
The secondimportantparameterused to evaluatethe
gels was the durationof hydration.The longera gel remainshydrated,the longerit is likely to supporta conducive environmentfor inoculumgerminationand survival. Hence,this parameterwas determinedfor the differentpolymersfor morethana week. For all gels, the
longevityof hydrationextendedto about6 d, suggesting
thataddinganyof thesepolymersto thefungalinoculum
will enablethe propagulesto benefitfromthe high ambient moistureand improvetheirgerminationand host
infection.
Althoughthe eight hydrophilicgels were similarin
theirabilityto remainhydratedwith no significantdifferencesamongthem (Figure1), they differedin their
abilityto promotegermination
andviabilityof A. cassiae
spores.Thismaybe dueto the differencesin the amount
of free waterthatwas availableto the sporesandthe pH

Table 2. Effect of seven polymeric gels on the pathogenicityfrom a mycelial

inoculum of Alternaria eichhorniae to waterhyacinthleaves.
Gel

DSa
% w/v

0 to 9 scale

Gellan gum (5)

Kelgin?-HV (0.5)
Evergreen? 500 polyacrylamide(0.5)
Kelgin?-LV (0.5)
Metamucil? (0.5)
Kelzan? xanthan gum (0.5)
N-GelS' (0.5)
No gel (control)

5.50 ab
5.50 a
4.00 ab
3.83 ab
2.83 b
2.67 b
2.17 b
2.00 b

a Disease severity (DS) was recorded 7 d after inoculation using a pictorial

scale of leaf area damaged by disease on a scale of 0-9, where 0 = healthy
and 9 = completely dead.
b Values followed by the same letter(s) are not significantly different according to Duncan's new multiple range test (P = 0.05).

and nutritionalcompositionof the gels. The gels also

differedin theireffects on promotingthe infectivityof
the mycelialinoculumof A. eichhorniae;a differentset
of gels from those thatwere best for A. cassiae spores
were effective (Table2). This may be due to different
responsesbetweenthe two Alternariaspp. or between
sporeand mycelialinocula,explainedon the basis that
differentmoisturedurationsfor germinationmay be involved (Gottlieb1978).
Theresultsfromthis studyhavebeenusedto develop
an effectiveformulationof A. eichhorniaefor field use
in Egypt.In greenhousetrials,this formulation
has been
successfully used to control waterhyacinth(Shabana
1992).Thus,thehydrophilicpolymersusedin thisstudy,
whichare readilyavailablein the marketplace,
offer an
effective, easy-to-useoption to formulatebioherbicide
agents.
ACKNOWLEDGMENTS

We thankthe International
Foundationfor Science,
Stockholm,Sweden,for providingmajorfinancialsupportfor this research.Additionalfundingwas provided
by the CulturalandEducationBureau,EgyptianEmbassy, Washington,
DC. We thankSteveLinda,formerlyof
the Universityof Florida,Departmentof Statistics,for
assistancewith statisticalanalyses.

Figure 4. Effect of the best concentrationof each polymer tested (concentrationthat yielded the highest percentages of spore germination)on the proportion
of alive germlings vs. germinatedspores (A/G) of Alternaria cassiae spores over a 1-wk period. The plotted data are: 4.3% for Gellan gum (G-gum); 0.4% for
KelzanS xanthangum (X-gum) and Metamucil? (Metam); 0.04% for Kelging-HV (K-HV), Kelgin?-MV (K-MV), Evergreen? 500 polyacrylamide(Poly), and
N-Gel; 0.004% for Kelging-LV (K-LV). Means from four replicates of each gel.

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SHABANA ET AL.: HYDROPHILICPOLYMERSFOR FORMULATINGBIOHERBICIDEAGENTS

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