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-uaM (SoilMicroorganisms)
No. 44, pp. 53N68(1994} 53

VA
S- >t"{ S;'i Plant-Growth-Promoting Fungi from Turfgrass
Rhizospherewith Potentialfor Disease Suppression*
Mitsuro HyAKuMAcHi
of Agriculttfre, Gdu Universily,Gipt 501-1 1, kPan
Fkecitgt.v

Plant-growth-promotingfungi (PGPF)were isolatedfrorn the rhizosphere of

turfgrass and cultivated crops. Frequencies of eccurrence of I'GPF thetotal
out of
fungi isolatecl
from turfgrass, wheat, corn and eggplant were 46.0%, 47.0%, 37.9%
and 10.0%, respectively. The most efficient PGPF were isolatedfrom turfgrass.
Among the 32 PGPF isolates screened, 14 belonged to the sterile group and 9 were
Tn'choderma,5 Fletsan'um, 3 R]nicilliunz ancl 1 Mztcor isolates, In pot experiments
where barley grain inoculum of PGPF was used, the increase in plant height and
weight varied dependlng upon the PGI'F.The increaseindry weight forthe treated
compured with untreated plants was as fo]lows:bentgrass,9,1times;rye-grass, 4.
4 time$: wheat, 1.5times; tomato, 5.1 times; cucumber, 75.2 times;radjsh, 1.9times.
Promotion effect was detected both in steriiized and non-sterilized soil amended
with PGPF inoculum and most conspicueusly in nutrient-deficient one.
Experiments were conducted to determinewhether the control of plant path-
ogens was associated with PGPF. Each representative isolateof the main groups,
i.e., sterile, Tim'choderma, Fletsan'um, and PeniciUir{m, was selected. On the ether
hancl,1?hizoctoniasolani AGI(IC), R. solani AG2-2,R. solani AG4, 15,thium
ttltimum, R mpanidewwatum, R Paroecandnfm, Sclerotiblne iw4fsii, Flersan'ztm o.opor
um f.sp. melonis, F. ompt,f.sp. cuct{merimtm and Gaettmannomyces g7aminis var.
trzttctwere used as target pathegens. PGPF and the pathogen were co-inoculated
and the disease-suppressive effect was cvaluated. Although, the degree of disease
suppression depended Qn the inoculum leveland the cembination of PGPF and

pathogen, PGPF afforded a potentia]protectionto the hosts from infection with

the pathogen in sterilized and non-sterilized soil. Above all, sterile and 7'richoder-
ma isolatesshowed $table and conspicuous suppressive effects.

KeyWords:Plant-growth-promoting fungi,Diseasesuppression

that some of the PGPR could induce resistance

Introduction
in plants has been revealed, and more

Recently, some rhizosphere bacteria have attenti()n24,25,2S) isbeing paid to the mechanisms
been reported to be endowed with the ability of of incluction of plant protectiun.

both plant growth promotion and suppression Although, many studies on PGPR havc been
of soil-borne diseases5'i3'i6''7).
These bacteria conducted, only a few have dealt with plant-
are designated as "plant-growth-promoting

growth-promoting fungi(PGPF). So far, T)'i-

rhizobacteria(PGPR)". The mechanisms of choderma ha7zianttm2,3,",i527},TL koningi27), non-

plant growth promotion havebeenattributed to pathogenic Rhizoctoniasolanii`2i), Rhytophthoiu

plant hormone production'3) and clisease sup- Paiusiticag}, sterile black fungusZ2), sterile red

pression due to the production of antibiotics or fungus7'S) and sterile dark fungus20) have been
siderophoresi'a,ii,iT,iS,'3). More recently, the fact reported as PGPF. Interestingly, all thesePGPF
were also found to be endowed with the ability
*199dilisH13Hi-teoblfift <ltl.)ICtsLif of diseasesuppression.
th?fi In the course of our study for the identifica-
1994 {f 5 H l3 H 9ew.
tion of biocontrol agents, we observed that

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many soi] fungi from the rhizosphere of turf- growing, plant heightand wet and dry weight of

grass and rhizoplane of varieus crops displayed the

the ability of bothplant growth promotion and

disease
suppression. A part of our current study

is reported here.

Materials and Methods

Isolation of rhizosphere and rhizoplane

fungi(RF)
RF iselatedfrom the rhi2osphere
were of

turfgrass (Zbysia tenuijblia WMd. ex Thiele)

and rhizoplane of whcat (Triticum aestivum L.),
corn (Zeainays L.),cggplant (Solanztmmelon-
gena L.) and green pepper (Cmpsiczfm annuum

L.) by using the soil dilutionp]ate and root

washing methodsi2), respectively. As for turf-
grass, after therough mixing of the root-soil
zone, the samples of the root-soil mixture were

used for the soil dilution plate method.

Waksman's medium acidified with lacticacid
(plI4.5)was used for the isolation of the rhizos-
phere fungi. Te ebtain the rhizoplane fungi,
roots of each crop were washed twenty times
serially using sterile distilled
water, Root seg-
ments 5mm long were placed on Czapek-Dox
medium containing streptomyein C30ppm) and

rose bengal(10eppm). RF were randomly iso-

lated from the colonies growing on each

rnedium, 7 to 10 day after incubation.There-

after RF were classified into several groups
according to the cultural characteristics and

spore merphology.

Assay for plant growth promotion of RF

Randomly selected RF from each group were
inoculatedintobarley grains and incubatedfer
]O days at 25 C. Barley grains infested with RF
were amended intononsterilized and sterilized
Yanagido soil, and potting soil nained Gransol
at the rate of 2% (wlw).Yariagido soil is a
nutrient-deficient soil sampled from the fieldof
Gifu UniversityFarm and potting soil is a
commercial nutrient.-rich soil. Plant seeds were

sown in soil previouslyamended with infested

barley grain inoculum and four weeks after

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Plant-Growth-Prornoting Fungi 55

Four weeks aftcr sowing, wheat seedlings eggplant and green peppcr).The occurrence of

-,ith roots were intactfrom the soil

removed genera and their frequency differeddepending
which had been infestedwith GZItalone or on the type of p]ant species, sampling time and
PGPF alone or det +PGPF, The roots were locations. Total number of RF isolatedfrom 6
washed thoreughly in running tap water and differentplants was 1399, and the order of the
finallvwashed distilled
with water, The roots frequency of the main genera from these plants
were surface-sterilized with 2% NaOCI for 30 was Fbusarium> 7'richodernea> steri]e >
seconds and cut into segments 5 mm in length. knicillittm> 1:3,tlzittm > Rhieoctonia> Alter
These segments were placed on PDA(pH 4.5) nan'a > Mucor (Tab]e O.
acidified with 10% lacticacid, ancl incubated

fer3 to 10 days. The fungal colonies growing Table 1 Frectuencyof isolation of PGPF frorn
from the root segments were compared with the several plants.
original culture and the colony-forming seg-
ments enumerated. Genus No. of PGPF!tested isolates(%)

Tn'chodem2a 1541 187(82,4)

Results fi'ztsa}'iztm 114,/'261(43.7)
SterilePvthiztinR3nicillium
51! 128{39.8)
Isolation of plant-growth-promoting fungi 47! 63(74.6)
(PGPF) 441 64(68,8)
Altemaan'a I21 l9(63.2)
Isolationfrequenciesof RF belonging to main 3,, 8(37,5)
A4ntcorRlzi:-octonia
fungalgroups from the rhizoplane of seyeral o! 22co.e)
crops and rhizosphere of turfgrass Vnknown 194! 647(30.0)

Rhizosphere fungi (RF)were isolatedfrom

Total 619f1399i[44.211
turfgrassand cultivatecl crops (wheat,corn,

1oo
90A5
so'a
7og"-
60igo
sooge
40v8
3oots
20pt
10
oTtrtgrasswneatCbrnEggplantGreenpePPer

Plantroots forisolation
of fungi

Fig.1, Percentageof fungithat induced a significantlncreaseor deqrcase of the dry weight of bentgrass.

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Table 2 Dry bentgrass after inoculation

weight of Table3 Distribution of the 32 selective PGPF
with PGPF isolated
from several plants. isolated
frem the rhizosphere of turfgrass.

IsolateNo. Plantsforisolatton Dry weight(mg) Genus Ne, of selective PGPF % distribution

GT21GF1913A-1GSP82GSI)1223A-7ITB-74FI-t4ITB--
turfgrass Sterile1>ichoderma
130I29121109106le2 I49531 43,828.115.6
turfgrass
cornturfgra$s Fhrsarium
Penicitlittm 94
turfgrass Mucor 3.1
cornwheatcggplant
98 Total 32 100.0
87
1IE3K5TA3FB-l3ED3PY-9Control
Ns,heatwheatcornwheatcorncorncorn 83
81
67 promoting isolates, from individualcrop rhizos-
58 phere, several representative isolateswere
54 selected - 5 isolates of 7)'ichodefma,4 of Iile{sar-
52 izama, 2 of the sterile greup, 2 of Agterna}ia,1
52 each of I5)thiitm and unknown greuP, and their
45 effect was compared. The results indicated that
the 4 most efficient PGPF out of the top 5
belonged to RF from the turfgrass
obtained

rhizosphere (Table 2). Among the turfgrass RF,

Frequency of iselationof PGPF from seyeral 32 isolateswere selected as efficient PGPF in 11
plants- ,' individualexperiments using bentgrass based
Six hundred nineteen isolatesout of 1399 (44. on the results of the actual weight of dry mass
2%) showed a pronounced p]ant
much more er the value of the increaseof weight over the

growth promotion effect in bentgrass,and they control. Among these 32 isolates, 14 were ster-
werc considered to be PGPF (Table1). Fre- ile,9belongedto 7lrichoderma, 5to fa. sarium, 3
quency of isolationof PGPF from each plant to Renicilliuin and 1 to Mucor (Table'3). The 13
was: turfgrass, 46.0%; wheat, 47.0%; corn, 37. sterile isolatesseemed to exhibit similar cul-

9%; eggplant, 10.0%; and green pepper, 67.2% tural characteristics and were considered to
(Fig.1).Plant-growth-inhibiting fungi (PGIF) belongto one group.
were also isolatedirom these plants except for The isolatewhich induced a considerable
green pepper in the range from 2.7 to 28.4%. plant growth promotion effect in one plant also
PGIF were isolatedmainly from corn rhizo- showed the same effect inother plants <datanot

plane.
i
shown). Isolatesbelonging to the sterile group
Frequency of PGPF from each genus was: showed the most constant effect in bentgrass,
Trichodemaa, 82.4%; Jb;thium,74.6%; Rrnicil- cucumber, wheat and by the
toinato, followed
liz{m,68.8%; Alternan'a, 63.2%; Fttsmizapn,
43. isolatesof Fttsarium,T)'ichoderma
was less
7%; sterile, 39.8%; ancl Mucoz 37,5% (Table1). effective as itsperformance was low in tomato.
Among the isolatesof Rhizoctoniaspp., none of Some examples of the remarkable effect of

them promoted plant growth. PGPF in bentgrass,rye-grass, tomato, cucum-

ber and radish are shown in Table 4 and Fig.2.
Seletion of PGPF showing conspicuous plant The increase in dry weight in the treated
'
growth promotion effect compared to untreated plants was: bentgrass,9.1
Among the top perferming plant-growth- times; rye-grass, 4.4 times; tomato, 5.1 times;

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In the case of cucuinber, there was a remark-

Duration of plant growth prornotioneffect of able plant growth promotion effect with values

PGPF as high as 75 times in the PGPF-treated soil

Increased growth response of wheat plants compared te the nonsterilizcd Yanagido (con-
as$ociated with PGPF isolatesas compared to troD This phenomenon
soil, was mainly

contrul during variuus growth stages such as attributed to the death of cucumber seedlings in

seedling stage (2weeks after sowing), vegeta- the control soil due to severe damping-off.In
tive stage (4 weeks), pre-flowering stage (6 the PGPF-treated soil, no disease occurred, and
weeks), flowering stage (10weeks) and seed the cucurnber seedlings "iere bigger and quite
maturation stage (14weeks)
was observed. All healthy.Therefore experiments were c;onducted

the isolateswhich increased plant height also tu determine whether such control of damping-

significantly increasedthe ear-head length and off diseasewas associated with PGPF. First]y,

biomass(Fig, 4).The ear-head lengthincreased damping-off pathogens were iso]atedfrom the

two times, while, the fresh and dry weight of nonsterilized Yanagido soil. Many R),tlaium
spp.

ear-heads increased 15-20 time$, and the sced wcre predeminantly isolatedfrom cucumber
no. 10-15times (datanot shown). seedlings with damping eff and some of them
were found to be pathogenic to cucumber (data
not shown). Isolatesof Rhi.,.o(/lonia and Ettsar-
Suppressienef soil-borne di$eases
by PGPF
izfi-n were also isolated,but the frequency of
Suppression of damping-off diseaseof cucum- isolation ss'as v ry lew and they were not path-
'by
ber seedlings PGPF in nonsterilized ogenic te cucumber. These results indicated
Yanagido soil that the inain causal pathogens of damping-off
of cucurnber in the Yanagido soil were B'thium
1 ) Isolationof causal pathogen of damping- spp. They wcre nainly divided into 3 groups,
off of cucumber seedlings Two groups of Fl}'thiptmwere identifiedas

y lcontrol
4oo
agx.s8N.oo-a8L,O-'
350

300
250
200
150
GptgritA
1oo
50
o
Non- SteriIized Pottingsoil
sterilized Yanagido soil
Yanagido soil
Fig. 3, Effect of PGPF on plant growth in sterilized and non-sterMzed Yanagiclo soil, ai'id in nutrient-rich
potting soil,

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Table5. Protectionof cucumber seedlings from b/li?.ium' dttmping-off disease (R),thittne

i7Tngttla}e,
R
paroeeandn{m, Rsp, R aPhanidermatztm) by plant-growth-prometing fungi (PGPF).

Treatment I)isease
severity index Protection(%)

P. irragttinre 2.67a -
P. ir,ugulare+SterileGSPI02 O.58c 78.1
Rirragulare+TrichodermaGTII2 0.93c 65.U
I'.irragulare+Fusarium
GF191 1.73b 35.0
P. irragulare+-Peniciltium GP172 l.80b 32.5

llytltiumsp. 1.60a -
Il)Vhittmsp,+Sterile GSPI02 O.40b 75.C)
Ib,tJziuin GT32
sp,+ 71,'ichocienna O.60b 62.5

P. Paroecandiwm 3.93a -
P. Paroecaizdn{nr+Sterile
GSPI02 ?.87a 1.7
P. Paroecandmm+Tn'chodervnaGT32 1.93b 50.9

P. mphanidern2atum 2.93a -
P. ophanidermatzt,n+Sterile
GSPI02 1.93b 34.1
7)'ichodervnaGT32
P. mphanicte"natttml L.07b 29.5

'These
Rytkiumspp, except forR aPhanidermditttm were isolatedfrom nonsterMzed Yanagido soil.

low and high inoculum levels of GT32 and

pathogen, respectively (datanot shown).

Suppressionof Pythium fo}iar blight dis-

ease of bentgrassby PGPF
AII the 4 selected PGPF also showcd a pro-
nounced suppressive effect on foliar blightdis-
ease of bentgrasscaused by P. aPa72idermatztm
except for FzrsariumGF191 (Table6, Fig. 6).

Suppression Rhizoctonia diseaseby PGPF

of

Suppression of Rhizoctonia disease by soil

amendment with PGPF was evaluated by using
cucumber, bentgrass and rye-grass. All the
selected PGPF signMcanlty suppressed
Fig, Effect of PGPF (7beichodenna
5, GT32 and damping-off disease of cucumber caused by
sterile GSPI02) on the suppre$sion of B'thit{m Rhigoctonia sola;ti ACll(IC),AG2-2 and AG4
clamping-off disease of cucumber. From leftto right (Table7, Fig.7).A!1 these isolates alsu showed
: (A) non-inoculated contrel, R}!thittm i)'nguime significant suppressien against brown patch
a]one, P. irngttlare 1 Tn'chode7?na GT32, Tn'choder- diseaseof bentgrassand rye-grass caused by R.
ma GT32 alone. (B) non-inoculated control, P.
solani AG4 except for I7letsai'ium
GF191 which
irnggztlare alone, P. irrqgulare-i sterile GSPI02, ster-

ile GSPI02 alone. was ineffectiveagainst brown patch of rye-

grass (Table8),SterileGSP82, GSPI02 and

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Fungi 61

Fig, 6. Effect of PGPF (7'n'choderma

GT32) on the suppression of bthium
foliar
blightin bentgrass. From leftto right : non-inoculated control, Rs,thium
ophanidermatum alone, R ciphanide"natum+ Tri(rhoderma GT32.

Table 6. Protection of bentgrass Ry/thiumfoliar by plant-

blightdisease (B,thizeinophanide77nattim)
growth-promoting fungi (PGI)F),from

Treatmeiit Diseaseseverit}, index Protection(%)

-Exp.1'-
P.mphanidewwattfm 4.0aL7b1.7b3.0a1.7b
P.ophanidermatum+Sterile GSP82 57.557.525.057.5
P.ophanidewnatttm + Trichodeitma
GT32
GF191
Rmphanide}vnatt{m+Fbes'an'um
GP172
Rmpizanidemaatum-Pei2icilliz{m

-Exp.2'-
Rmphanidennatttm 3.3a1.7b1
Rmphanidef}natum+Sterile GSP82 48.569.769.739.4
P,ophaniden7iatum+ 7}iichoderma GT32 Oe1.0c2.0b
GF191
P,mpItanidermatttm+Fbtsariztpmz
P,mphanidevmatum-t- Penicilgittm
GP172

'Pottingsoil
Yanagidosoilwere
(Granso])and used for exp.1 and exp.2, respectively.

T) ichoderma GT32 showed stable and conspicu- Suppression of other soil-borne diseasesby
ous suppressive effects, In the case of the combi- PGPF
nation of TVichoderma GT32 ancl Rhigoctonia PGPF isolatesalso suppressed Sclerotiuma
solani AG4 (l272), in contrast to the suppression damping-off and Fintsan'um wilt diseases of

results ef Trichoderma and Il,thiztm ir7agutare, cucumber, and take-all diseaseof wheat caused
disease suppression was conspicueus for co- by (;aeumannompces g7uminis var. trz'tici. As for
inoculationwith lo"r and high inoculum levels Sclerotiztmdarnping-offof cucumber, sterile

of the pathegen and PGPIi respectively (data GSP 102 did not show a conspicuous $uppres-

not shown). sive effect, unlike ]Fbasan'um GF191 (Table

9).
Fusarium GF191 also showed
a strong suppres-

sive effect against Fletsarium wilt of cucumber

caused by F, oxl,, f,sp. cucttmerinum, but not

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Table7. Protectionof cucumber seedlings from Rhizoctonia damping-offdisease (R.soinniAGUICL

AG2-2<IV)by plant-growth-prometing fungi (PGPF).

Treatment Diseaseseverity index Protection(%)

AG2-2AG2-2tSterile 2.36aO,36dO,94c1.42b1.00bc
GSPI02 84.760.239.857.6
AG2-2t- 7'ric,hodennaGT32
AG2-2+ FletsaritfmGF191
AG2-2+feniciUittmGP172

AGI{IC) 3.S7a2.93bO.96c
AGI<IC)+SterileGPS]02 24,275.2
AGI(IC) + Ti7'choderma
GT32

AG4AG4+Sterile 4.00a2.73b1.80c
GI'SI02 31.755.0
AG4+ 7'n'choderma
GT32

Table 8. Protection of bentgrassand by

plant-growth-prometing fungi(PGPF).ryegrassfrombrownpatchdisease(RhixoctoniaselaniAG4)

Treatment Disease severity inde'x Protection(%)

Bentgrass -exp.1'-
AG4AG4+Sterile 3.3aL7b1.0bcI.ebcO.7c
GSI'82 48.569.769.778.8
AG4-- T}ic'hodervna
GT32
AG4+Fusarium GF19I
AG4+ Penicilliu-nt
GPI72

Bentgrass -exp.2"-
AG4AG1+Stcrile 3.3ae.obO.ObO.
GSP82 10010090.9100
AG4+ 7)'ichoderma GT:32
AG4 F-Fztsan'umGF191 be.ob
f,)
AG4TPeniciUiu,n GP172

Ryegrass'
AG4AG4+Sterile 3.7aL5b
GSP82 c1.0c:S.2a2.lb 59.573.013.543.2
AG4 t Trichoderma GT32
AG4tthtsan'ttmGF191
AG4+Pe"icillittmGP]72

*Seeding
was performed at the same time uf the inoculatien of the pathogen and PGPF.
*'Seedjng
was performed 1 week after the fnoculation
of the pathogen and PGI'F,

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Fig. 7. Effect of PGPF (Tric]?oderma GT32) on the disease suppres-

sion ol Rhigoctonia damping-off of cucumber. From left to right :
non-inoculated control, Ritiuactonia sotani AG2-2 alone, 1?. solani
AG2-2+ 7VichodermaGT32, Tiichoderma GT32 alone.

Table 9 Protection of seedlings from Sclerotitt'hnblight

disease(.S/clerotiztm
,v(lsii) by plant-

growth-promoting (PGPF),
fungicucuinber

Treatment Disease severitv index Protection{%)

S, ]'Oly'Sill 2 53aL60bO,07de.soc3
4tsii+Sterile
S. IVs,S. GSI'102 36 8
reftsii+ V'n'choderma GT32 97.4
,oijisii -tFlets[pizt?n
GF191 68,,1-18.6
s. ro GP172
tfsii+PirniciUitem
'
Ooa

against Fletsarittm wilt of cucumber caused by Ilv,thibmz

diseases,it also showed pronounced
a

F. oi;y. f,sp. ;nelonis {Table le).As far as the suppressive effect against R irre,g.ulare
and P.

suppression of take-all disease caused by sp. but not against P. Paiwecandrum and P.
C;aeztmanno17zyces gnznnnzs var. tritici is con- ztttimttm. The suppressive effect of I'GPF
cerned, sterile GSP122 and 7'rt'choderina GT21 varied depending on the genera or species of

induced pronounced suppressive effects (Table pathogens.

11, Fig. 8).Fig. 9 shows a summary of the
diseasesuppression PGPF against
of several Competitive root colonization of PGPF
soilborne pathogens. Among the 4 representa- and pathogen
tive PGPF, TVtichoderma GT32 and sterile Reisolation frequenciesof Glgtand PGPF
GSPI02 showed conspicuous suppressive {sterile GSP122 and TviclzodennaGT21) from
Trz'chodeiTna GT32 afforded
effects. Especially, wheat roots were compared {'I'able12).
a stable protection against all the pathogens, Althoughboth colonized roots fairly welt, the
SterileGPSI02 showedprcmounced suppres-
a reisolation frequency of Cig'twas markedly

sive effect against Rhiffoctoiiia sogani, P. reduced when itwas co-inoculated with PGPF,
irragttlaiv, F, ox),. f. sp. c/ttctcmerinum and

melonis but not against S. ro4fi;ii. As for

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Table 10, Protection of eucumber seed]ings from F)tsan'um

wirt disease {E oxysPontm f.sp. citcztiTterinunt. F,
o. f.sp. melonis) by plant-growth-promoting fungi(PGPF),

'
Treatment Dtsease severity inclex Protection{%)

F.EEEEf.spf.cucumen7zztm
o.o.o.o.o. 1 67aO
sp.f.cttcumen'nu?n -iSterileGSPI02 40bO.60b0 75.963.968.036.0
sp-f,cucumerixztm + 7>ichodervvaaG'l'32
spf.ctict{men'num GF191
--1;latsan'um 53b1
sP cucttmen'nt{m+kificiUium GP172 07ab

EF,F,p',E
f.$pf.melonismelonis+Sterile
o.ao.o.o. 2.93aU.47c1.00c.2.33
spf. GSPI02 84.165.920.512.7
spf.ntelonis -f7)'iclzoderma
GT32
-- Fitsav'iumGF191
sp.f.melonis b2.60
sp melomlg. +ReniciUittnz GP172 a

Table 11, Protection of wheat seedlings froin take-all disease(GaeumannomvcesgreTnzmsvar.tn'tici)

by
plant-growth-premoting fungi (PGPF).

1'reatment Diseas severit}. index Protection(%)

Exp.1Ggt(G2-91)
2,73aO.48cO.27c1,33b1.33b
Cigt+Sterile
GSP122 82.49U.151.35I.3
(nygt+
Ttichode,ena
GT21
(;{gt+1;besa,iu7'n
GF191
opt-PenicilliblinGI'151

Exp,ZCigt(Gl-1)
391aO44bO.32b
dwt+Sterile GSP122 S8.891.8
( kt+ Ttrichode"na GT21

Table 12. Reisolationfrequencies of Caeumannom.vces gmminis var, triticiand PGPF(stcr{le GSP122and

Trichodeniia GT21) from wheat roets.

Traatment
ClgrtReisolation(96)PGPF

Non-inoculated
control o90oe10o o
GgtSterile o
GSP122 85
T)ichodeT7iiaGT21 95
Ggt+SterileGSPI22 t)5100
Gke't
+Trichoderma GT21

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Plant-Growth-Prornoting Fungi 65

Fig8 Effect of PGPF (sterile GSP122) on the suppression of take-all disease in wheat. From leftto r{ght
: non-inoculated centrol, Clgtalone, sterile GSP122 alone, Ggt+sterile GSP122.

1oo
- GSPI02
m GT32
80 pmGF191
waGP172
9v
60g-ri86

4ocS

20

o
R.solani PLirregulare S ro1fsi.i E o. f.sp. E o. f.sp.
AG2-2 cucumerinum melonis
Fig. 9 Comparison of the cross-protection effect of certain PGPF isolates.
'Disease
was more severe than that of control,

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66 HyAKulfAcHi

Discussion These results suggest thatthe mineralization of

the substrate of orgar]ic matter in soil by PGPF

There are many reports on the isolationoi may bc related to the plant growth promoting
fungi from the rhizosphere or roots of several effect of PGPF. PGPF may heTp the plant to
crops which were not considered to bc path- derive necessary mineral nutrients in an easily
ogenic, but showed a deleterious effect on plant available form. Based on the results of cucum-
'reported
growth. Gamieland Katani") that none ber grown in nonsterilized Yanagido soil, we
of the fungi isolated from the rhizosphere or observed that, the remarkable p]ant growth
roots of prometed plant growth, but
tomato promotion effect C75times) coinpared to the
insteadinhibitedit.Yuen and Schroth2S' also control <Table4) was croselv relatecl to the
reported that several species of Renicilliitmand suppression by PGPF of pathogenic 1]3,thiuni
EmpeniciUittntisolated from roots of zinnia spp, which xN'ere indigenousin that soil. Thus

plants inhibitedplant growth by 23 to 57%. On the suppres$ion of deleteriousmicroorganisms

the contrary, recently, several reports have by PGPF may be one of the mechanisms of
indicated that fungi of the rhizosphere can plant growth promotion.
induce both plant grewth promotion and dis- PGPF showed a pronounced suppressive
ease supperssion effect$, Such effects had main- effect against several soil-borne diseases.The
ly been observed in bacteria designaged as disease-suppressive effect of PGPF, however,
plant-gruwth-promoting rhizobacteria, PGPR. varied depending on the genera or species or
In our invcstigation, many isolatescollected types of pathogens. The degree of suppressioT)
from the rhizoplane of various cultivated crops of the discasesalso varied with the levelsof
and the rhizosphere of turfgrass were found tu inoculum of the pathogen and PGPF. In case of
promote plant growth in a variety of crops, supperssion of take-all disease by I'GPF, root-
whereas those isolated from the turfgrass colonizing ability of the isolatesof the sterile
rhizosphere highly effective
were as plant and 1'richoderma groups played a definiterole.
growth promoters. We designated these as They also showed a pronounced suppressive
plant-growth-promoting fungi, c)r in short effect (Table11).It is interesting to note that
PGPF, in analogy with the terminology of some isolatesof the sterile PGPF recovered
PGPR. Among the PGPF isolated from turf- from the turfgrassrhizosphere induced a resis-
grass rhizosphere soil, many isolatesof the tance against anthracnose diseaseof cucumber
sterile group induced stable growth promotion caused by Colletotrichttm
ldigenan'ttmiY).
Such
and di$easesuppression effects in severa] crops. activity, however, was not always rclated to
As fer the mechanisms of plant growth pro- their colonizing activity on the host roots. Sev-
metion of 1'GPF, several hypotheses have been hypotheses about the mechanisms
eral of dis-
put forward, including hormone production, ease suppression have been proposed as follews.
substrate degradation(mineralization) and sup- (O PIant growth promotion activity by PGPF
pression of deleterious microorganisms. So far, may induce the plant to become tolerant to
we have not been able to demmonstrate that various diseases.{2)Direct suppression against
these PGPF produce hormones.There ",ere, pathogens by the production of antibiotic sub-
however, close relationships among the reduc- stances, hyperparasitism or cempetition for
tion of barley grain weight due to PGPF, their infectioncourt or nutritional substances. (3)
subsequent growth promotion effect, cellulase lnductionof systemic resistance in plat by colo-
and starch degradation activity of L'GPF (data nization by and infectionwith PGPF or by the
not shown). Production of NH,`-N and N02-N in changes in the metabolism of the hostplant
soil was also accelerated by amendment with triggered by certain substances secreted from
PGPF-infested barley grains (datanot shown). PGPF. Although, the mechanisms ef growth

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Plant-Growth-Promoting
Fungi 67

promotion er diseasesuppression by PGPF steriie fungus from "Jheat the effect of soi]
and

have not temperature itssuppression

yet been deterrnined,
PGPF and moitsture on of
seem to
take-all. ildlL)coL Res. 93, 156-160
offer a great potentialas biocontrol agents.
9 ) Do]an,T. E,,Cohen, Y, and Coffey, )'I, D, (1986)
Plant growth promotion by fungi has only been
Protection of bean against anthracnose by
demonstrated in a few cases in comparison to Colgetotrichum on bean.
species non pathogenic
the largenumber of reports on rhizobacteria. A RhytoPathoLZ, 86, 117-126
detailed investigatjon is being conducted to 10) Gamiel,A. and Katan, J.(l991)
Invulvementof
analyse the precise mechanisms of plant growth fluorescentpseudumenads and other micro-
promotion and diseasesuppression. erganisms in increased growth response of

p]ants in solarized so{ls. en)'topatholQgry.

81,
494.5e2
LITERATURE CITED 11) Gardner, J,A,I,, Chandner,J.L. and Feldman,
A. E. (IY84} Growth prornotionand inhibition
1) Ahl, P., Voisard, C. and Defago,D. (]986)Iron
by antibiotie-producing fluorescent
bound-siderophores, cyanic acid, and antibi-
pseudomonads on citrus roots. PkJnt and Soil
otics involved in suppression of 7Vzietaviopsis
77, 103-113
btzsicoia by a A"eudomonas /luorescens strain, 1.
12) IIarley,J.L. and "'aid,J.S. {1955)A method
RhJ,topathol. 116,121-134 of studying active mycelia on living roots and
Z > Ahmad, I.and Baker,R. (l988) Imp]icatiens of
other surfaces inthe soil. Tiuns.Bx mvcol. Soc.
rhizosphere competence of T}ichodenna har-
38, 104-].18
zianum. Ctin.1.Microbiol. 34,229-234
13> Hussain, A,, Arshacl, M., Hussain,A. and
3) Baker, R, {1991) Inductionof rhizosphere com-
IIussain,F. (1987) Re$ponse of maize CZba
petence in the biocontror fungus 7beichodeT7na.
map,s) to Azotobacter inoculation under fertil-
b?.The Rhizosphere and Plant Growth, Ed. D.
ized and unfertilized conditions. Bial. Eextilily
L. Keisterand P. B, Cregan,p,221-228.Kluwer Siols 4, 73-77
Academic I'ublishers,Netherlands 1,l) Ichielevich-Auster,M,, Sneh,B., Koltin,Y. and
4) Chang, Y. -C.,Chang, Y. -C. and Baker, R.
Barash, I, (19S5)Suppressionof damping-off
(1986) Increasedgrowth of plants in the pres- cau$ed by Rhiaoctonia st)ecies b.va nonPath-
ence of the biological control agent T7ichoder-
age?zitr isolate o.f R. sota7ti, RliJ,toPatholag)J 75,
ma haxzianzam. PlaniDisease70,l45-148 1080-le84
5> Chanway, C. P. and Nelson,L. rV{.C1991) Char-
15) Kleifeld, O, and Chet, I. {1992) Tn'chodervna
acterization of cultivar specific growth promo- -interaction
Javmeianptm with plants and effect
tion of spring wheat by Bacilltts sp. in The
on growth response, Pgant a"d Soig 144,267-272
Rhizosphere and Plant Growth, Ed. D. L. Keis-
16) Kloepper,J.W'.,Schroth,M. N. and Miller,T.
ter and P. B. Cregan, p.365,Kluwer Academic
D. (1980) Effects of rhizosphere co]onization by
Publishers,Netherlands
plant-growth-prornoting rhizobacteria on
6 ) Defago,G.,Berling, C. H,, Berger, U., Haas, D.,
potato plant developrnent and yield. PhJ'toPath-
Kahr, G.,Keel,C.,Voisard,C.,XNrirthner, P. II.
ogQg" 70, 1078-1082
and Wuthrich,B, (I989> Suppressionof black 17) Kloepper, J. W., Zablotowicz, R. M,, Tipping,
rot of tobacco by a i]lseudomonas strain: poten-
E, M. and Lifshitz, R. (1991)Plant growth
tial applications and mechanisms, In Biolegi- by bacterialrhizosphere
promotion mediated
eal Controlof Soil-Borne PlantPathogens,Ed.
colonizenq.. In The Rhizosphere ancl Plant
D. Hornby, R. J,Cook and Y. Henjs, p.93-108, Growth, Ed, L. Keister and P. B. Cregan,p.
CAB International,UK 315-326,Kluwer Academic Publishers, The
7 ) Dewan, M. M, ancl Sivasithamparam, K. C1988)
Netherlands
A plant-growth-promoting sterile fungus froin 18) LifshitT, R., Kloepper, J.W., Kozlowski, M.,
wheat and rye-grass roots with potential for Simonson,C.,Karlson,J.,Tipping, E, M. and
suppressing take-all,T7ans.Bn nz.vcol. Soc. 91,
Za]eska, 1. (19S7} Plant growth promotion of
687-717
canola (rapeseed}seed]ings by strain of
8 ) Dewan, M, M, and Sivasithamparam,K. 0989)
ilseudomonas Ptitida under gnotobic conditions.
Growth promotion of rotation crop species by a

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68 HyAKu"{AcHI

Qinf Microbiol., 33, 390-395 mulatien inbio]ogical

control ef F)tsaiit{mwilt

19) Meera, M. S.,Shivanna, M. B., Kageyama, K. of carnations by ltseudomonas sp. strain

and Hyakumachi, M. (1993)Induction of sys- VLrCS 417r. Phyiopathoictgy,81, 728-734
temic resistance in cucumber plants using turf- 25) Van Peer, R. and Schippers, B. (1992)
grass rhizosphere fungi. Ann. RPzytopathol.Soc. Lipopolysaccharides of plant-growth-
Iapan59, 279 promoting Rseudomonas sp. strain MrCS417r
20) Narita, Y. and Suzui,T. (1991) Influence of a induce resistance in carnations to F}usan'um
sterile dark
mycelial fungus on taka-all of wilt. Neth.).
Plant ththol.98,129-139
wheat, Ann. Ilh.ytopath.Soc. JaPan57, 301-305 26) "'ei,G.,Kloeppcr,J. IV. ancl Tuzun, S. (199I)
21) Sneh,B.,Ichie]evich-Auster, M., Barash,I.and Inductionof svstemic resistance of eucumber
Koltin, Y, (1986) Increaseclgrowth responses by to CoUetotrichumorbiculare by select strains
a nonpathogenic Rhieectonia sogani thn. f ofplant-growth-premotingrhizobacteria.pla,to-
Bot. 64, 2372-2378 Patholcng},81,1508-1512
22) Speakman, J.B. and Kruger, W. (1984) Cc/mtrol 27) XVindham, "{.T.,Elad,Y. ancl Baker, R. (1986)
ofGaeztmanno7?ayces gvanzinis var. tn'tictl
by a A mechanism for iT)creasedplant growth in-
black n)yceliai fungus. ! Plant Dis.
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Prot.91,391-395 518-521
Z3) Stutz,E.,Kahr, G, and Defago, G, {1989) Clays 2S) Yuen, G. Y. ancl Schroth, M, N. (1986) Interac-
involvedin suppression of tobacco black root tions of llseudovnonasfZuo?escensstrain E6 with
rot by strain of Rseudomonas .fiuoTescens. Soil itseffect
ornamental plants and on the compo-
Biog. Biochem., 21,361-366 sition of root-colonzing microflora. Pdytopatlr-
24) Van Peer, R., Neimann, G. J.and Schippers, B. otogy 76,176-180
(1991)
Inclucedresistance and phytoalexin accu-

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