BIOCONTROL POTENTIAL OF SOIL HYPHOMYCETES FOR ROOT-KNOT NEMATODES
Dissertation submitted for the Degree of
ifWaittr of ^l)ilo$(opI)p
\ ^ AGRICULTURE (PLANT PATHOLOGY) ^ ^
BY
MQflD. AY£1QB MA?4TQQ
INSTITUTE OF AGRICULTURE ALIGARH MUSLIM UNIVERSITY
ALIGARH (INDIA)
1993
DS2146
He (A1I2*') tdught man th<a.t which he Knevif T»ot CThe Qurc»\96:5)
In memory
Of my Late
Grandmother
O/ io/. ,yfl. //aau/ '-yo/ic^'^^-I.Sc. (Ban.) Ph.D. (Alig.), F.L.S.. F.P.S.I. rijjtessor of Botany
n a m r a m w i w y v a"*^ ' •""» i»wiii"*^"»-
Laboratories
DEPARTMENT OF BOTANY ALIGARH MUSLIM UNIVERSITY AUGARH-202002 INDIA Tel: (0571) 25676 (Office)
D a t e d . 1 1 . 1 2 . 9 3
C E R T I F I C f i T E
This is to csY'tify that Mr*. Mohd. Ayoob Mantoo has
prepav'ed this dissertation as required for M. Phil.
(Agricultuj^e) degree of the ftligarh Muslim University, ftligarh
in Plant Pathology, under rny supervision and guidance. He is
allowed to submit this dissertation in partial fulfilment of the
requirements for the degree of M. Phil, in Agriculture (Plant
Pathology).
(M. Wajid Khan)
Research Supervisor
flCKNOWLEDGEMENTS
From the taegmriing to the end of my M. Phil,
programme, suggestions, assistance and encouragement came from
several sources. Foremost among them was that from my supervisor
Prof- M. Wajid Khan, Depai-tment of Botany, A.M.U. , filigarh, who
went all out to help me. I am conscious of rny debb to hirn and
take this ooporbunity to opine my sense of reverence and
gratitude in all earnestness to him for his indefatigable
efforts, scient 11 lat m g suggestions, constant encouragement and
constructive criticism in the conduct of thus programme.
I owe a great deal of unquantiflable gratitude to
Prof. M. Shamim Jairajpuri, Director, Institute of ftgricultur-e
and Prof. Uazahat Husain, Chairman, Department of Botany,
A. M. U. , Oligarh, for providing necessary facilit3E>s.
It 15 a great pleasure to acknowledge the most
generous help from my seniors, Dr. N. A. Onsari, Dr. fl. ft. Khan,
Dr. M. R. Khan, Dr. K. S. Yadav, Messers S. T. Nabi, N. H. Shah,
F. ft. Lone and Z. ft. Makdoomi,
I obediently and dutifully offer my gratitude to
Messers M. H. Siddiqui, B. Khan and ft. H. Wani m particular <\rid
to Ms. L. Khalil, Mr. N. ft. Baba in general for their kind help.
Words are inadequate to expross rny gv^atitude towards
my parents, brothers and the sister Parveena Shaheen, for their
everlasting love and affection, who remained the source of
inspiration to my work.
l i
ftllah a l o n e i s t o be s o u g h t foi" h e l p and on Hirn
a l o n e we d e p e n d .
Mohd. ftyoob Mantoo I n s t i t u t e o f ftgricultur'B A.M. U. , flligarh.
C O N T E N T S
Page No.
1. INTRODUCTION 1
£. REVIEW OF LITERATURE e
3. MflTERIftLS fiND METHODS 43
1. Survey and collection 43
a. Isolation of fungi from soil samples 44
3. Isolation and identification of fungi
associabed with egg masses 44
4. In vitro inoculations ^5
5. In VIVO inoculations 47
(i) Nematode inoculation 47
(ii) Fungus inoculation 48
6. Root penetration 49
7. Soil drench and spray treatment 50
8. Recording of data 50
9. Root-knot disease parameters 51
10. Experimental design and statistical analysis 51
4. LITERATURE CITED 53
INTRODUCTION
Plant parasitic nematodes ar^e heterotrophic
organisms and occur worldwide in all the kinds of agricultural
soils. Depending upon their mode of parasitism and host-parasite
relationships, they are variously classified into different
groups like ecto-, semi-endo-, and endo-parasites. Endoparasitic
nematodes are more damaging and agriculturally important than
other groups. Omong the endo-parasities, root-knot nematodes
belonging to the genus Meloidogyne constitute the major group of
plant-pav^asit ic nematodes of outstanding economic importance.
The root-knot nematodes, known for more than a century have
attracted the attention of most nematologists and plant
pathologists all over the world, fi vast amount of literature is
testimony of their importance as parasites of crop. They at^e
ubiquitous and most widespread causing losses of many crops.
Though they attack almost every type of crop, causing
considerable losses of yield or affecting the quality of the
produce, vegetable crops mostly suffer greatest damage (Sasser,
1980).
The losses caused by nematodes both in quatity of
the yield and quality of the produce are enormous, fl loss of
$£50 million was estimated by Hutchinson e^ al. (1961) to be
due to nematodes. United States Department of Agriculture
(USDfi) estimated an annual crop loss of 37£,335000 dollars to 16
crops (Taylor ig&7). The society of Nernatolog ists (SON)
committee on crop losses led by Dr. Feldrnesser, estimated anriual
losses in U. S. Pi. in the fov m values of the order of
$1,036,374,300 in field crops; $££5,£45900 in fruit crops;
$£66,989, 100 in vegetable crops and *59,ai7,&34 in ornamental
crops due to nematodes <ftnon,, 1971). finnual-crop losses due to
nematodes on worldwide basis was estimated to the tune of *100
billion (Sasser and Frechman, 1987). The crop losses caused by
nematodes has given the grim picture of agricultural economy in
the United States (finon. , 1987).
There ar^e also some reports of crop losses from
India due to plant-parasitic nematodes like ftnquina t r i t i c i. the
seed gall nematode causing a loss of *10 million to wheat crop;
Heterodera a.ver\ae, cereal cyst nematode causing $8 million to
wheat and barley in Rajasthan State alone and Pratylenchus
coffeae. root lesion nematode causing *3 million to coffee.
However, (^&r\ Berkum and Seshadri, 1970). However, above figures
are not adequate to highlight the crop losses caused by
nematodes as only little information is available about crop
losses in the developing countries.,) The situation warrants
application of management measures in order to minimize the crop
losses. Various rneasuv-es that have been in use for management of
nematodes are classified as physical, chemical, cultural,
regulatory, use of host resistance and biological methods.
Hot watev^ trearnent, radiation treatmerit, ultrasonic,
washing processes, seed cleaning, heat treatment viz. steam
sterilization, solarisation and pasturization of soil etc.
<3outhey, 1965, 1978a; Mass, 1987) are physical measures applied
for management of certain categories of plant -parasitic
nematodes. Soil solarization is a recent technique to kill
nematodes but it has certain lirnitst ions and can be effective in
summer season in tropical and subtropical regions of the world
only.
Several reviews have provided detailed and valuable
information on the efficiency of chemical measures for
management of plant parasitci nematodes. (Van Gundy and Mc
Kinny, 1977; Whitehead, 1978; Winfield, 1978; Haq et. al.. , 1983).
fi number of chemicals both soil fumigants and
systemic were successfully used for management of plant-
parasitic nematodes including root—knot nematodes. But some
problems are associated with their application, ft number of them
have been banned, therefore, only a few are marketed. These
chemicals also create pollution causing toxic effects on
beneficial flora and fauna and on men involved in their
manufacturing and use. Their influence or\ the non-target
organisms has caused ecological imbalances which are sometimes
difficult to be restored. Furthermore, the nematode population
increases several months after the use of chemical. The
persistence in the soil, and contamination of ground water are
their otlTev characteristics that reject their usefulness.
Some cultural practices like fallowing, flooding,
ploughing, crop rotation etc. (Br-own, 197S; Noe, ISSG) are also
used for management of plant—parasitic nematodes. Fallowing of
the field results in economic loss. Flooding causes adverse
effect on soil structure and monetary losses to the farmers.
Cj- op rotation though effective in controlling nematodes, yet it
sometimes increases the population of other plant-par^asit ic
species. fls majov" species of root-knot nematodes are
polyphagous, it is difficult to design effective rotation
schemes for their management.
Regulatory methods aim at checking of
dissemination of nematodes (Southey, ig7Qb; Maas, 1987). The
method involves passage and enforcement of quarantine laws
desinged to prevent the spread of particular nematodes into
areas Known to be free. It is essentially not an approach to
manage the nematodes in infested fields. Successful employment
of nematodes resistance requires the manipulation of genetic
systems to transfer resistant genes form a resistant plant to
susceptible and acceptable type. Use of host resistance can be
most simple, practical and economical method for management of
planty parasitic nematodes but development of r^esistant
cultivars is itself a very time consuming and costly affair.
Some cultivates found resistant become susceptible subsequently
due to continuous variatioon in pathogens caused by host
selection pressure and erivirorirnental fluctuations. Bornetirnes,
new races of the nematodes may evolve due to selection pressures
caused by rnono-culturing of the resistant varieties in a
particular area (Jalala, 1986). Though some plants and
cultivars were claimed to be resistant to root-knot nematodes
(Fassuliotis, 1979), yet use of host resistance will require
still more efforts and is still a far dream for plant breeders.
Currently biocontrol measures have become most
significant methods for the management of plant-parasitic
nematodes. The biocontrol of pathogens have become a fascinating
dream and ultimate goal of many concerned with the health of
plants. In the present time due to greater awareness of the
essentility of pollution free environment, biocontrol seems to
be the most relevant and practically demanding approach. (Khan,
1990).
Biological control means different things to
different people. In the broadest sense, it is the natural
phenomenon of population regulation created by the interaction
of the biotic components of an ecosystem. The biological control
encompasses all biologically causes of mortality, including
competition between individuals of the same and the different
species, the effect of host defenses and host resistance, and
the direct or indirect results of attcaks by organisms belonging
to higher trophic levels. More applied biological control
involves the manipulation, conservation and augmentation to
sroecific kinds of orgariisms 3 n order to regulate populations of
undesirable species and thus to prevent or reduce their negative
impacts on human well-being.
Biocontrol is considered skilful rnani Dulat ion of the
biosphere against nematode pests for achieving maximum benefits
but m some cases, it has many difficulties m practical
applications. Other drawback of biocontrol is impatience of
scientists as well as farmer who alkways desire for instant
results.
Nematode egg-parasitizmg fungi have been reviewed
by Stirling et. a_l.. <197a), Nigh et. a^, (1979), Jatala (1986),
Cabanillas et. al.. (1388,1989), Khan (1990), fll-Ha = mi and Raz:ik
(1991), Marban et_ B1^. (199S). Some of these fungi were reported
to attack more than one develeomenta1 stages of nematodes.
The literature cited showed that much attention has
been given to the biocontrol of plant-parasitic nematodes
particularly root-knot nematodes. In biocontrol studies m
India, mdeginous soil hyphomycetes have not been evaluated for
their efficacy as biocontrol agents of root-knot nematodes and
other endoparasitic nematodes. Therefore in the proposed study
for Ph.D., investigations on biocontrol potential of soil
hyphomycetes for root-knot nematodes will be undertaken. It is
planned to conduct in the following experiments.
1. Sui- vey and collection of soil and root samples from fields
infested wibh root—knot nematodes from different parts of the
country.
2. Isolation, identification and multiplication of fungi
parasitic to root—knot nematodes from the collected samoles^.
3. In vitro studies of selected (identified) fungi
(hyphornycetes) for their efficacy as biocontrol agents
under:—
a) Effect of culture filtrates of selected fungi on
hatchability of juveniles of root-knot nematodes.
b) Larvicidal effect of culture filtrates of selected fungi C'rt
root-knot nematode juveniles.
c) Efficacv of the selected fungi as parasites of eggs/egg
masses of root-knot nematodes,
4. Effect of the selected fungi ori the development of root-knot
nematodes and root-galling in cowpea and tomato.
5. Effect of the selected fungi and root-knot nemabode
Meloidogyne .lavanica alone and in combination on plant
growth characters of cowpea and tomato.
6. Effect of the selected fungi (biocontrol agents) on
Meloidogyne .i avanica juvenile's penetration m the host roots.
7. Management trials of selected fungi as biocontrol arients
through soil drenching and spv^aying experiments in pots or
microplots.
REVIEW OF LITERftTURE
Root—l^not nematodes, Meloidogyne spp. , constitute a
major group pf plant-parasitic nematodes affecting crop
production. Thoy are world- wide in distribution and show
extensive host range. Their association with other plant
pathogenic organisms causes disease complexes m a number of
plants. They account for significant losses in yields of food,
feed and fibre? crops. The most frequent species encountered
world- wide Ar^e Me loodogyne incognita, M. .lavanica, M. arenaria
and M. hapla (Sasser, 1977). Pi recently concluded International
Meloidogyne Project (IMP) showed that these four species Br-e
major species of Meloidogyne. fibout 95/. of the populations
identified, through the efforts of IMP were represented by the
aboye mentioned four species of Meloidogyne (Taylor et_ al. ,
1982). The frequency of the species encountered was as follows:
M. inconnita 5i2/4; M. j ay an ica 3 IS; M. hapla BY- and M. arenaria
7%. Four races were differentiated in M. incognita. Race 1
comprised of 7c:"/.; Race £, 13%; Race 3, 13"/. and Race 4, cl'A of 472
populations studied. Within the species M. arenaria, two races
were identified and designated as Race 1 and Race £ (Sasser,
igS£).
Root—knot disease was first recorded by Berkeley
(1355) on glasshouse cucumbers i»i England who named the nematode
= s Vibrios. The prc- sent day name Meloidogyne was giyen by Goeldi
(1887, 189a) and Chitrwood (1949) confirmed it. On the basis of
morphological d i f f ey^ences particularly in the? perineal patterns
of adult females, Chitwood (1949) described four species vis.
Meloidogyne incognita (Kofoid and White, 1919) Chitwood, 19A9;
M. .lavanica (Treub, 1885) Chitwood, 1949; M, arenaria (Neal,
1889) Chitwood, 1949; M. hapla Chitwood, 1949 and orxB sub
species M. incognita acrita. Since then fv orn tirne to time new
species were discovered, described and added to the species list
of the genus Meloidoqyne. Till 1990, there Are as many as 73
species and L2 subspecies under the genus Meloidoqyne (Eisenback
et al., 1985; Hirschmann, 1986; Golden and Kaplan, 1986; ftbdel
Rahman and Maggenti, 1987; Rammah and Hirschmann, 1988; Golden,
1989; Rarnmah and Hirschmann, 1990).
Sasser (1977) summarized the distribution of
Meloidoqyne species in different parts of the world. Occurrence
of IS species in the United States, 11 species in fifrica, 11
species in Europe and the Mediterranean region, 10 species in
India and Sri Lanka, 9 species in Central and South America, 5
species in South East fisia, Australia and Fiji Islands and 3
species in Canada was included in the summary. In India, root-
knot nematodes were first reported by Barber (1901) or\ tea in
Kerala and named them as Heterodera rad icicola. Out of Ic!
species so far reported in India, Meloidogyne incoqnita and M.
.lavanica are the most widely distributed and attack a wide range
of crops (Khan and Khan, 1985; Haider and Khan, 1988; Khan,
1988).
10
Biocontrol of Nematodes
Plant-parasit ic riernatodes including root-knot
nematodes are managed by different methods which ar-e categorized
as chemical, cultural, biological etc. Biological or biocontrol
in the present time, seems to be practically demanding and the
most relevant approach due to greater awareness of the
essentiality of pollution-free environment (Khan, 1990).
Biocontrol is a natural phenomenon. Plant—parasitic nematodes
share the habitat with other organisms in the soil. The
relationship that is developed m common habitat is a continuous
process and maintains a natural balance m soil biotic
cornmi. nity. However, the efficiency of the organisms varies under
different ecological conditions or as such in the soil (Khan,
1990). Biocontrol has been defined in various ways by a number
of workers. Almost any process occuring naturally or done
artificially by man, which affects the relationship between
organisms in such a way that the natural biological balance is
restored, can be regarded as biocontrol. Recording to DeBach
(1964), biocontrol is defined as "any condition under which or
practice whereby survival or activity of pathogen is reduced
through the agency of any other living organism (except man
himself), with the result that there is a reduction m incidence
of the disease caused by the pathogen. Sewell (1965) defined
biocontrol as "natural or induced, direct or indirect,
limitations of a harmful organism or its effects by another
11
organism or a group of orgarnsrns"-
Swarup and Gokte (1986) defined biocontrol "as the
exploitation of a living organism for reducing the pest
population, with essential approach of trying to restore that
balance of nature that has been upset to the advantage of some
destructive organisms". Barker and Cook (1974) defined
biological control as "the reduction of inoculum density or
disease producing activities of a pathogen or parasite m its
active or doi-mant state, by oriB or more organisms, accomplished
naturally or through manipulation of the environment, host or
antagonist, or by mass introduction of one or more antagonists".
On average fertile field soil contains approximately
10 bacteria, 10 -10 act inomycet es, 10 -10 fungi and 10"^
proto::oa reproductive units per gram. In addition a large number
of fungivorouB nematodes, tardigrades, collembola, mites and
other assorted micro- and meiofauna are also found. Biological
antagonisms are common in soil, and biocontrol has often
occurred under natural conditions (Baker and Cook, 1974). Thus
in general, biocontrol with reference to plant - parasitic
nematodes is a means of promoting organisms that are harmless to
the plants but attack and destroy those ov^ganisms which are
harmful to the plants.
The possibilty of biocontrol of nematodes was first
suggested by Lohde (1874). This study generated interest in
12
many eav^ly scieritists (Kuhn, 1677; Zopf, 1888; Thome, 1927;
Linford, 1937; Linford and Oliveira, 1938; Daddington et_ al. ,
1956; Loewenbev'g et_ al_. , 1959), but organized and detailed
studies on this subject commenced in the I960's only. Now
biocontrol of nematodes has been the subject of several reviews
(Sayre, 1971, 1980; Mankau, 1980; Kerry, 1980; Jatala, 1986;
Morgan-Jones and Rodrigues-Kabana, 1987; Nordbring-Hertz, 1988;
Khan, 1990; Sundei-land, 1990; Galper et. a_l.. , 1991; Leij et. al. ,
1992, Stirling, 1992; Zaki and Maqbool, 199£).
Plant-parasit ic nematodes including i-oot-knot
nematodes a\^B controlled by a number of different kinds of
enemies. These enemies include predacious fungi, predacious
nematodes, and predacious animals; parasites like viruses,
bacteria, r'lckettsias, and fungi (Sayre, 1971; Manaku, 1980).
Organic amendments, use of trap crops and antagonistic plants
also reduce parasitic nematodes biologically.
The literature ctn predacious fungi, endozoic fungi
and fungi parasitic on eggs/cysts is reviewed.
Predacious Fungi
Predacious fungi are also called as nematophagous or
nematode trapping fungi. More than 100 such species have been
reported now (Mankau, 1980; Jatala, 1986). Nematode trapping
fungi were considered as potential biocontrol agents for the
control of nematodes (Duddington, 1957). Nematode trapping habit
15 found in several fungi belonging to order Zoopagales and
13
Moniliales of the class Zygomycetes arid Deuterornycetes
respectively. Predacious fungi have developed various dt^vices to
capture their prey. These devices are grouped into sticky traps
and mechanical traps. The sticky traps are of three types:
(i) Sticky branches e.g. Pact ylel la lobat a
(ii) Sticky rret works e.g. Pirthrobotrys ol i qospora
(ill) Sticky knobs e.g. Pactylella ellipsospora
The mechanical traps B.re of two kinds:
(i) Non-constricting rings e.g. Dactylaria Candida
(ii) Constricting rings e.g. Dactvlel la bernbicodes
(Duddmgton, 1960)
The mechanical traps were found to be more efficient
in reducing nematodes than sticky traps (Cooke, 196c:). Based on
germination of conidia m 16 fungi, predacious fungi were
divided into two ecological groups by Cooke (196A). ft sensitive
group in which conidia quickly give rise to traps, and
insensitive group in which scant trap formation occurs.
Sensitive group are efficient predators and poor saprophytes but
insensitive group are better saprophytes and poor predators.
The existence of fungi that trap and prey on
nematodes was first reported by Zopf (1888). Nematode trapping
fungi were the first entities with which efforts were made in
the beginning to control plant-parasitic nematodes (Drechsler,
1937). He pointed out that some hyphomycetes prey on free living
terricolous nematodes. When nematodes pass through the trapping
14
rings, are captured by the sticky hyphal networks, killed by the
constrict ion of globose structures and body content consumed by
the hyphae originating from these globose stv^uctures. The root-
knot nematode, Meloidoqyne sp. was claimed to be controlled by
adding chopped pineapple tops to the infested soil (Linford,
1937) but disappointing results were obtained when soil was
inoculated with predacious fungi to control Meloidoqyne sp. in
pineapple (Linford and Yap, 1939). Dactylella bembicodes and
flrthrobotrys oliqospora protected the begonias from infection of
root-knot nematodes (Deschiens et_ al. , 1943).
Experiments conducted under indoor conditions by
the application of predatory fungi to control root-knot nematode
Meloidoqyne marioni demonstrated the possibiity of reducing
nematode infection of plants. The increase in the dosage of the
fungi resulted in greater control of the nematodes (Gorlenko,
1956). Various kinds of fungi were reported to infect free
living nematodes isolated directly from forest litter. The
predacious fungi were found to be active in the natural
population of nematodes from a soil habitat (Capstick et_ al • ,
1957). They reported that some nematodes have been trapped by
hyphal snares of the constricting ring type, and some were
infected by spore forming fungi including Harposporium
oxycoracum but more infections were by hyphae or by small
spherical bodies, often associated with hyphae.
15
The constricting ring mechanism of predacious
hyphomycetes that trap nematodes was studied by Muller (1958),
but the chesrnical factors that induce the formation of traps were
demonstrated by Pramer and Stoll (1959) and Feder et_ al. (1963).
The use of nematode—trapping fungi to control root-knot nematode
diseases of tomato and okra was studied by Mankau (1961). He
pointed out that the immediate protection of crops highly
susceptible to root-knot nematodes by the application of
predacious fungi does not seem possible.
The ecology of nematode trapping fungi in the soil
was studied by Cooke (19G£). He found that the decomposition of
sucrose in the soil stimulates AVI increase in both the
population of free living nematodes and the actvity of
indigenous nematode trapping fungi. He further reported that the
fungi were incapable of v^emaining in a predaciously active state
in the absence of 3.n organic energy source other than the
nematodes. Predacious actvity of different species of nematode
trapping fungi in soil has been found to vary in both intensity
and duv^ation, and that constricting ring type of traps are more
efficient in reducing soil nematode populations than fungi
forming adhesive v^eticulate traps (Cooke, 1963). The
significance of ecological characteristics of nematode-trapping
hyphomycetes in relation to biocontrol of nematodes was
discussed by Cooke (1964).
16
Activity of nematode trappers is greatly influenced
by soil conditions such as pH, availablity of nutrients,
temperature and moisture (Mankau, 1968). The morphology of traps
of about 100 species of nematode trapping fungi used for
capturing nematodes have been studied. They vary from simple,
undifferentiated hyphae to -very highly specialized devices
(Sayre, 1971).
Pactylei la ov i paras i t i ca was found naturally
controlling root—knot nematodes even in the presence of a highly
susceptible variety "Lovell" of peaches in Califor-^riia. (Stirling
and Mankau, 1977). Even though the fungus was able to survive
without the riernatode, yet it control led the Meloidoqvne species
(Stirling et_ al. , 1979). Dactylel la arcuata. a nematophagous
hyaline hyphomycete, forms simple, one-celled, lateral, globose
sticky knobs at first but later on multicellular arches, rings
and networks are formed by branching and anastomosis, and then
trap the nematodes (Scheuer and Webstev^, 1990). The biology of
D. meqalospora was studied by Esser et_ al. (1991). The fungus
trapped and assimilated 18 genera of plant-parasitic nematodes;
9 non-parasitic nematodes were also trapped and assimilated by
the fungus (Esser et_ al. , 1991). Two predacious fungi, an
isolate named "Royal 300", of ftrthrobotrys robusta for the
control of Pitylenchus myeeliophaqus on mushrooms (Cayrol et
al.. 1978) and another isolate of flrthrobotrys named "Royal 350"
for the control of Weloidoqyne on tomato (Cayrol and Frankawski,
17
1379) are marketed for commercial use iri France.
In a study on attraction, induction of trap
formation and capturing of plant parasitic- nematodes by 13
nernatophagous fungi, it was observed that the ability of
different fungi to attract tended to increase with increasing
dependence of the fungi on nematodes for nutrition. Induction of
trap formation depended on their motility and on the
concentration of nutrients in the culture medium, fill nematodes
were rapidly captured when traps were present (Jansson and
Nordbring-Hertz, 1980).
The diversity, adaptations and distribution of
nematode destroying fungi, and taxonomic problems encountered in
their study are reviewed by Mankau (1980). He suggested that
fungi may be utilized as alternatives to chemical control after
a more thorough and expanded study of their biology and ecology.
In microplot and greenhouse experiments, it was
found that the reproduction of Meloidoqyne incognita and the
incidence of root galling in corn, were reduced by the addition
of flrthrobotrys conoides and/or green alfalfa. No interaction
was found between the fungus and green alfalfa in the reduction
of the nematode population <fll-Hazmi et_ al. , 198£). Further, the
effect of Q. conoides on M. incognita population densities as
affected by soil ten^peratut^e, inoculum density and green alfalfa
was determined in greehouse experiments. Q. conoides was most
effective in the nematode control, when the fungus was
18
iritroduced into the soil £ weeks before the nematode inoculation
and planting of c o m (Pll-Haarni et_ al. , 19a£) . Culture filtrates
of Rrt hrobot rys oli qospora, Dactylaria brochopaqa and Curvulan a
pallescence inhibited juvenile hatching of Weloidogyne incognita
and Hetev'odera zeae (Walia et_ al. , 1985).
The efficiency of nernatophagous fungus P)rt hr obot rys
dasquptae attacking a range of plant-parasitic nematodes was
studied by Bong et al. (1988). Predatory fungus ftrthrobotrys
irregularis protected against light and moderate infestation of
Meloidoqyne spp. (Cayrol, 1988). Use of nematode-trapping
fungus, ftrthrobotrys tortor against Meloidoqyne spp. on
different substrates in greenhouses was reported by Jawich and
Bochow (1989).
Entraping and assimilation of nematodes by fungi
like flrthrobotrys, Monacrospori um and Dactylaria spp. by
employing constricting rings was briefly reviewed by Sunderland
(1990). The nature of their nematode entrapment and their
biological control potential were also studied (Esser and
Shubert, 1991). The formation of three dimensional sticky
netwoks in flrthrobotrys oli qospora and Monacrospori um cionopaqum
and the capture of nematodes in soil was observed by using
fluorescence microscopy (Jensen and Lysek, 1991). In vitro
studies were carried by Nakasono and Gaspard (1991) to determine
the effectiveness of two nernatophagous fungi, ftrthrobotrys
dactyloides and Dactylei la haptotyle for Meloidoqyne incognita
19
and free living nematodes in soil.
In samples collected from forest, annual and
perennial crops in Minas Gerais (Brazil), the occut^r^Bnce^,
predatory capacity and growth of flrthrobotrys musiformis and 0.
conoides was studied by Naves and Campos (1991). Pria et. al.
(1991) also isolated and identified the predacious fungi from
soil samples collected from various regions of Brazil. In India,
Saxena and Mukerji (1991) surveyed 165 soil 3.rtd vegetation
samples from various sites of Varanasi and found that
nematophagous fungi were distributed in a variety of habitats.
ftrthrobotrvs oliqospora produce extra-cellular
proteases when grown in a liquid culture. The involvement of
protease in the infection and immobilation of nematodes by the
nematophagous fungus was investigated (Tunlid and Jansson,
1991). fl lectin- carbohydrate interaction leads to adhesion of
nematodes to Q. olioospora, which is the first step in the
infection of the nematodes (Tunlid, 199£).
In a study on the formation of conidial traps in
flrthrobotrys oligospora. it was found that the fungus developed
the traps in response to diffusing substances from cow faeces
rather than in response to living nematodes (Dackman and
Nordbring-Hertz, 199£). Trap productoon by nematophagous fungi
(0. dactyloides. ft. olioospora, Monacrospori urn ellipsosporium
and M. cionopaqum) growing from parasitized nematodes was
investigated by Jaffee et. al. (199S).
20
Endozoic -Fungi
In addition to nematode trappers, some endozoic
fungi s^re natural enemies of nematodes. fin endozoic fungus,
Harposporium anqui1lulae was first to be found effective in
checking nematode populations (Lohde, 1874). About 50 endozoic
fungi mostly belonging to Phycomycetes and Deuteromycetes srs
now known to be parasitic on nematodes. Ingestion of spores of
H. anqui11ulae prior to their germination, by plant-parasitic
nematodes was found to be effective for their control (Aschner
and Kohn, 1958).H. arthrosporum was reported to parasitize
nematodes only when its arthroconidla become lodged in the
nematode's oesophagous (Barron, 1979). H. arcuatum was observed
attacking nematodes in farmyard soils collected in Ontario,
Canada. The conidia were readily ingested by the nematodes and
conidiophores emerged through the nematode cuticle at the tail
end (Barron, 1980). H. oxycoraeum, H. cycloides and H. sicyodes
isolated from soil in Varanasi, India were found to be
parasitizing the plant -parasitic nematodes (Srivastava and
Dayal, 1983). The electron microscope study of initiation of
infection by conidia of H. subuliforme was carried out by
Saikawa and Morikawa (1985).
M e n a coniospora, Ar\ endoparasit ic nematophagous
fungus infects nematodes specially at the chemosensory organs
(Jansson and Nordbring-Hert z, 1933). Jansson et. §2,. (1984) gave
a additional information on M. coniospora at different stages of
21
nematode infection using LSM and TEM. M. coniospora conidia
showed diffE?rent patterns of adhesion to the cuticles of
nematodes (Jansson et_ al. . 1985).
Jansson et_ al • (1985) evaluated potential of M e n a
coniospora in reducing root galling caused by root-knot nematode
iri tomato. Experiments conducted to determine whether M.
coniospora, ftrthrobotrys oligospora and ft. flaqrans control the
root-knot nematode, Meloidogyne hapla on alfalfa and tomato,
showed that there was reduction in root galling without any
improvement in plant growth (Townshend et. al. . 1989).
Hirsutella rhossi1lensis produces spores that adhere
to and penetrate the nematode cuticle and assimilate the body
content prior to its emergence and sporulation (Jaffee and Zehr,
1985). The adhesion and infection of Drechmerla coniospora to
the plant-parasitic nematodes was studied and protein was found
to be involved in the adhesion process (Jansson et. al . , 1987).
Mycelial growth and rate of sporulation of Dj;_ coniospora.
Vert 1c1111um balanoides and Harpospori um angui1lulae were tested
in different liquid cultures. Conidia produced in liquid culture
retained their parasitic capabilities (Lohmann and Sikora,
1989). The ultrastructural study of adhesion and initial stages
of infection of nematodes by conidia of D. coniospora was
reviewed by Dijksterhuis et_ al. (1990). The colonization and
digestion of nematodes by D. coniospora was investigated by
Dijksterhuis et. al.. (1991).
22
Catenaria aux i laris arid Nernatophthor£^ qynoph i la are
two important zoosponc fungi involved in biocontrol of plarit-
parasitic nematodes. C. auxi 1 l a n s was observed on females of
Heterodera schachtii (Kuhn, 1877). It also parasitizes females
of H. avenae and Globodera rostochiensis (Kerry, 1S75; Kerry et
al_. , 1976). N. qynoph ilia parasitises H. avenae, H. trif ol ii, \±.
cruci ferae. H. gott mqiana, H. schacht 11 but not G.
rostoch1ensIS (Kerry and Crump, 1977). The ability of N.
qynophila to infect nematodes is dependent upon adequate soil
moisture (Kerry et_ al. . 1980). Roy (1984) reviewed the effects
of nitrogenous soil amendments on the parasitic actvity of C.
anqui 1 lulae c<ri root-knot nematodes. Jaffee (1986) examined the
parasitism of Xi phinema riven and X.. amer icanum by C.
anqui 11 ulae^, Laqenid lum caudaturw. ftphanomyces and Leptol eqnia.
Both nematodes were infected by all fungi. Parasitism by
flphancmyces and Leptoleqnia was imcreased when nematodes were
incubated m BY- soil extract for 4 days before exposure to
fungi.
Voss and Wyss (1990) determined the potential of
Catenaria anqui11ulae as a control agent against several plant-
parasitic nematodes. Variation between strains of the fungus and
its attack to Heterodera schachtii was also studied (Voss et
al. , 199i2). Two of the three isolates (C^, C J Q ) reduced
significantly the number of cysts produced while one isolate
(C^) showed no effect on H. schacht 11 population. In vitro
23
observation on the infection of Meloidoqyne incognita eggs by
the zoosponc fungus C. anqui 1 lulae was reported by Wyss et_ al_.
il33S.) . The embryo was killed withen a few rninutes following
mass aggregation and encystment of the zoospores. Jansson and
Thiman (139£) studied chemotxis of zoospores of C. angui1lulae.
The characterization, entrapment, habitat, hosts,
life cycle and biological control potential of fungi that
utilize zoospores to parasitize the nematodes were briefly
outlined by Esser and Schubert (1963). Three species of
Myzocytiurn viz. M. papi1latum, M. q1utinosporum and M. anomalum
isolated from Ontario, Canada produce zoospores which encysted
directly on the host cuticle before penetration and produce
zoospores (Barron, 1976a), whereas M. hurnicola produced
zoospores that did not attack the nematodes directly but
encysted and produced adhesive buds that atacked and penetrated
nematodes (Barron and Percy, 1975). In Myzocytlum intermedium,
the encysted zoospores germinated by germtubes which penetrated
through the body orifices of the nematodes (Barron, 1976b).
Fungal parasites of eggs/cysts
Several fungal species have been found to be
parasitic on eggs and/or cysts of plant parasitic- nematodes.
Vertici11lum chlamydosporlum, V. falcatum, Cy1indrocarpon
destructans. Humicola qrisea, Fusariurn oxysporum and F. so1ani
were parasitic on cysts of Heterodera schachtii (Vinduska, 1979;
Heijbroek, 1983). Chaetomiurn cochllodes. Exophiala pisciphi la.
Fusarium oxysporum. F. solani, Phytophthora cinnamomi. Pythium
sp. and Trichosporon beiqelii were isolated from young cysts and
24
newly exposed females of Heterodera glycines from soybean fields
in Alabama, U.S.ft- (Ownley, 1383),
fl filamentous, non-sporulating fungus, designated as
Arkansas Fungus 18 <fiRF 18), was isolated from Heterodera
q 1 ycines in Arkansas, U.S.A. It parasitised 89^- of H. glycines
eggs in =cysts when tested in Petri dishes. The fungus also
infected eggs of Meloidoqyne incognita and eggs in the cysts of
H. qraminophila. H. lespedezae. H. leuceilyma, H. schachti i, H.
trifoli i and Cactodera betulae (Kim and Riggs, 1991).
Eggs of root-knot and cyst nematodes Bf^e found to be
parasitised by certain fungi. Destruction of nematode eggs by
Fusarium and Cephalospori urn species was first reported by Lysek
(1963). Later on Vert ici11ium chlamydospori um, V. bulbillosum,
Paecilomyces 1ilacinus, Acremonium bacilosporum etc. were found
to perforate egg shells and enter eggs of nematodes (Lysek,
1966).
The eggs of Heterodera schacht i i were found to be
attacked by flcremoni um strict urn and Fusarium oxysporum in
California sugarbeet fields (Nigh, 1979). Of 14 fungal isolates
evaluated as biocontrol agents isolated from the Meloidoqyne
incognita egg masses, Paecilomyces 1ilac inus was most effective
(Villanueva and Davide, 1984). The infection by F. so1ani on and
within the females, eggs and juveniles of Meloidogyne incognita
was observed by Khan and Hussain (1986). Infected juveniles and
adults were defov^med and thickly covered with fungal mycelium
25
arid chlarnydospores. F. solarn as natuY-^ally present m seal and
may be an impov^tant factor m natural biocontrol of root-knot
nematodes M. .lavanica and M. incognita as it infects the eggs
and larvae of these nematodes.
Vertici11lum ch1amydosporium has been reported not
only infecting Meloidogyne species but developing females of
Heterodera avenae prior to their production stages also (Kerry,
19S0). The A species of Vertici11ium and 2 species of
Paecilomvces have been found as parasites of cysts and eggs of
Heterodera glycines^ Meloidogyne arenaria and M. incoqnita in
Alabama U. S.0. glasshouse studies (Morgan-Jones and Rodriguez—
Kabana, 1984). Out of 15 different fungi isolated from the
different stages of H. a\/eriS(B life cycle, the egg parasite V.
chlamydospor1um was common in young cysts (Dackman and
Nordbring-Hertz, 1985).
The infection of Heterodera avenae and H. schachtii
eggs by six strains of Vertici111um chlamydosporium on water
agar was studied. Dead and immature eggs were most readily
colonised by all strains of the fungus. Eggs of H. avenae were
more susceptible to parasitism than those of H. schachtii
(Irumg and Kevn-y, 1986). The average growth rate, optimum
temperature for growth and production of chlamydospores varied
m these strains. They also varied significantly in their
pathogenecity to H. avenae eggs (Kerry et. al. 1986). However,
ore pathogenic strains of V. ch lamydospor i um are beirig m
26
developed at Rotharnsted Ex per-" i mental Station, U.K., to control
cyst and root-knob nematodes (Carlisle, 1387). Two methods seed
inoculation and paper pot, were used by Coosemaus (1991) for
introducing V. ch lamydospori um into the soil to conty'^ol H.
schacht11. The entomopathogenic fungus V. lecani i has been
tested for utilizing as an biocontrol agent against H. schacht i i
cysts (Hanssler and Hermanns, 1981) and against Globodera
pal 1 ida under field conditions (flvia and Sil^ora, 199£; Crump and
Irving, 199£; Meyer, 199£; Uziel and Sikora, 1992).
Some factors in the development of Vert ici11ium
chlamydospori um and Pasteuria penetrans as biocontrol agents of
cysts and root-knot nematodes were studied by Kerry (1988). Leij
et al. (199S) tested potential control of Meloidoqyne incognita
by V. ch 1 amydospi-irium and P. penetrans, alone and in combination
in a pot experiment. V. chlamydospori um was most effective.
Majority of egg masses were colonized by £. penetrans and both
resulted 9£"/4 population control.
The biocontrol potential of the Vert ici11i um
chlamydospori um for root-knot nematodes has been reviewed by
Cabrera et. at.. (1987) and Leij and Kerry (1991). The effect of
temperature on interactions between V. chlamydospori um and
Meloidoqyne spp. , was demonstrated by Leij et. at. (1995). V.
chlamydospori um required some external nutrients for its
establishment in soil. The colonization of nematode eggs and egg
masses depended on fungal inoculum and on galling caused by
27
nematodes <Leij et_ al. , 19S£)-
Vert ici 11 i urn ch 1 amydospori um parasitises females of
Meloidogyne arenaria (Morgsn-Jones et_ al. 1961), and cysts of
Heterodera glycines (Ginitis et. al. , 1983). It reduces the
population of M. arenaria. by adversely effecting its egg
hatching and inducing juvenile mortality. Both egg shell and
larval cu-ticle aY"-B disv^upted by the fv^r^gus. Ul-trastructural
studies have shown that chitin and lipid layers of egg shell and
basal layer of larval cuticle are disorganised (Morgan-Jones et_
al., 1983).
Strains of Cv1indrocarpon radicicola, Monotorpora
deleae, Catenar ia anqui 11 ulae, Fusari um oxysporum s/av^. lonqius
were tested in laboratory and greenhouse experiments for the
control of Heterodera rostoch iens is. Cylindrocarpon radicicola
1320 and Catenaria anqui11ulae £436 gave the best control of the
nematode (Kondakova, 1973).
For assessing the ability of a soil to suppress
multiplication of the sugar beet cyst nematode and for isolating
the fungus, Cylindrocarpen destructans from the infected
females, was devised by Crump (1987). Surveys of tropical
regions indicated that C. destructans and Ulccladium atrum were
the most commom species associated with root-knot and cyst
nematodes. It has been suggested that multi- cropping systems
may be designed to increase the frequency of occurrence of
microbial species antagonistic to phytonematodes in most
28
tropical regions (Rodr iguez-Kabana and Morgc^n-Jones, 1988).
Dadri and Saleh (1990) tested the impact of sevriral
nernatophagous fungi on Heterodena schacht 11 and Meloj doqyne
lavanica by inhibition of the fungi in field soil and by their
addition to nematode cultures on culture media and m pots.
flcremonium sclerotiqnum, Preussia sp., Vertici11lum
ch I anrydospor i urn and Fusari um so I a m inflected 8B-7c:'/- i±. schacht 11
eggs. The latter two fungi similarly infected the eggs of M..
.lavanj ca. Pis per pi 1 lus versicolor, ft. f urn i gat us, Cyl indrocarpon
ol id 1 urn. Fusarium oxysporum, F. equiseti also infected 25-46"/.
eggs of H. schacht 11 and/or M. .lavanica. Out of six fungal
species tested as biocontrol agents, only two species flcremonium
persicinum and ftsperqi11 us ochraceous were found most effective
against M. .lavan ica (fil- Ha=mi and Razik, 1991). Culture
filtrates of ftcremonium strictum, ftcrophia1ophora fusispora,
fllternaria alternata. ftsperai11 us flavus, ft. niqer and
Penici11lum spinulosuw were found to be inhibitory against M.
incognita (Shabana and Khan, 199£) .
Pythium monospermum^ P. aphanidermatum and
Phvtophthora palmivora were found to be capable of destroying
some non-stylet bearing nematodes through endozoic parasitism by
hyphae from ingested zoospores (Tzean and Estey, 1981). Pythium
tracheiphilum. reduced the population of Meloidoqyne hapla and
Pratylenchus penetrans on lettuce (Gracia, 1991).
29
The perforation in nematode egg shell by
Paeci lornyces 1 i lac in us was first reported by Lysek ( t966) .
Later, infection of eggs, and females of Meloidoqyne incognita
acrita by the fungus was recorded by Jatala et. aj^. (1979). P.
1i1acinus produces the antibiotic P-168 which has a wide
antimicrobial activity not only against fungi, yeast and
gram positive bacteria but also on the eggs, juveniles and
females of nematodes (Isogai et. al. , 1380). The efficiency oF P.
1i1acinus in controlling M. incognita and Globodera pal 1ida was
demonstrated by Franco et. al. (1981) and Jatala et_ al. (1981).
Paecilornyces 1i1acinus was found associated with
developmental stages of Heterodera glycines in soybean field in
Alabama (Gintis et. al. 1983). The parasitism of Meloidoqyne
arenaria eggs and juveniles by P. 1i1acinus was studied by
Morgan-Jones et. al. (1984). They found two species of
Paeci1omyces as parasites of cysts and eggs of H. glycines. M.
arenaria and M. incognita.'The eggs of M. incognita are deformed
by the fungus with the help of diffusable toxic metabolites
(Jatala et. aj.. , 1985; Jatala, 1986).^ Effectiveness of £.
1 i 1 acinus against M. .lavanica on tomato was studied by Croshier
et al. (1985). Davide and Zorilla (1985) reported that P.
1ilacinus significantly reduced the M. incognita population when
applied as soil drench or mixed with substrates such as rice
hulls and rice bran. They also found that nematicide treatment
using isozafos at 3Kg a. i./ha gave about 10"/- higher control of
30
the nematode in the soil than the fungus tr'eat rnenb, Roman and
Podi-^iguez-Marcano (1985) examined the effect of £, 11 I acinus on
the larval populations and root-knot formation of M. incognita
in tomato. The fungus controlled the nematodes and reduced root-
knot formation. Significantly fewer larvae were found m roots
and soil of plants inoculated with the fungus 5 days prior to
nematode inoculation.
Paec11omyces 1i1acinus has demonstrated tremendous
potential as biocontrol agent of nematodes. In case of
Meloidoqyne, Tylenchulus and Nacobbus. the fungal hyphae first
grow m the gelatinous matrix, then form a network around the
eggs and finally penetrate them (Jatala, 19S6). In Panama, £.
111acinus protected tomatoes and potatoes in field conditions
from infection of M. incognita. The fungus treated plots had
higher yield than did the control plots and those treated with
carbofuran (Jatala, 1986). Penici11lum anatolicum a related
fungus, also produces noxious compounds which interfere in the
development of Globodera pal1ida and G. rostochiensis without
parasitizing the females (Jatala, 1988).
The possibility of using Paeci1omyces 1ilacinus
along with the bacterial parasite Pasteuria penetrans for
controlling Meloidoqyne mcopnita in field microplots was
investigated. Nematode control was more effectively obtained
when both organisms were applied together (Dube and Smart,
1987). Similar results were obtained m case of M. .lavanica on
31
tomato by Maheshwari and Mani (1986). The effects of application
of P.. li lacrinus on the development and population of Meloidogyne
5pp., on tomato plants was tested by Rohana et. aj... (1987). The
fungus inhibited the population development of the nematode arid
maintained growth of tomato at 0.5 and 1.0 g inoculum/750 g
soil. Shahsad and Ghaffar (1987) reported that P. lilacinus
alone and combined with carbofuran reduced root-knot disease of
okra and mung.
Cabanillas et. al. (1988) studied the histology of
the interactions of Paecilomyces 1i1 acinus with Meloidogyne
incognita on tomato. Root galling and giant cell formation were
absent in tomato roots inoculated with nematode eggs infected by
P. 1 i 1 acinus. Hewlett et_ al.. (1988) evaluated the efficacy of P.
1i1acinus alone and in combination with phenamiphos and
ethoprop, for controlling M. .lavanica on tomato. The fungus did
not control the nematode. Plants with M. ASiva.r^ica alone or in
combination with £. 1i1acinus had galling indices of 5.0. The
latter produced lower yields than all other treatments.
Jimenez and Gallo (1988) found that £. 1i1acinus
under glasshouse conditions infected eggs and sometimes females
of M. incognita, M. lavanica and M. arevaria.
Microplot experiments conducted to evaluate the
effects of inoculum level and time of application of
Paecilomyces 1i1 acinus on Meloidogyne incognita to protect
tomato (Cabanillas and Barker, 1939) and to determine the
32
irifluerice of carrier and storage oF £. 1 i ] acinus on tornat-o
against M. incorjriita (Cabanillas et_ al.. , 1989), showed that best
protection against M. incognita was attained with 10 and £0 g of
fungus infested wheat kernels and fungus alone delivered into
soil 10 days before planting, and that greabest suppression of
egg. development occured in plots treated with £. 11 lacmus jn
pellets, wheat gv ain and granules. The effects oF temperatuY^e
on growth of 13 isolates of £. 1i1acinus and on their efficacy
in controlling M. incognita were investigated by Cabanillas et
al. (1989). MaKimurn fungal growth occoured from 54 to 30"C and
least growth was found at 12 and 36 C. fis soil temperature
increased from 16—28 C, both root—knot damage caused by M.
incognita and percentage of egg masses infected by P. 111 acinus
increased. The culture filtrates of £. 1i1acinus were inhibitory
to juvenile hatching of M. incognita (Khan et_ al. 1988) and
showed nematicidal actvity mostly against Heteroderidae
(Meloidogyne and Heterodera) (Cayrol et_ al. 1989) .
The genus Paecilomyces was first described by Bainer
(1907) as a close relative of Penici11lum but differing in the
absence of green coloured colonies and by short cylindrical
phial ides which taper into long neck. Samson (1975) placed
Penici11lum 1ilacmum and some other fungi m the genus
Paecilomyces and proposed Paecilomyces 1ilacinus.
Paeci lomyces 1<»lacinus has been the subject of many
recent researches for evaluating its biocontrol potential
33
against nernat odes. In Philippines, it is being produced
commercially under the trade name of "BIOCON" (Jatala, 1986).
Khan and Esfahani (1990) showed the efficacy of
Paeci lomuces' 1 i 1 acinus for controlling Meloidoqyne .i av an ica on
tomato in greenhouse. Root galling and egg mass production were
greatly reduced. The fungus was severely effective when both
organisms were inoculated simultaneously or the fungus preceded
the nematode in sequential inoculation . ft high percentage of
eggs were found to be infected. £. 1i1acinus also reduced damage
to cowpea caused by M. incognita and Rotylenchulus reni formi s
(Khan and Hussain, 1990). Combination of £. 1i1acinus and castor
leaves was found to be better for reduction in gall index,
second stage juveniles, eggs/egg masses, egg destruction and egg
mass infection of M. .lavanica than either alone (Zaki and
Bhatti, 1990).
Two applications of isolates of Paecilomyces
marquandi i from suppressive Chinampa soils or P_. 1 i lacinus from
Peru, was reported to give better control of tomato root-knot
due to Meloidoqyne incognita than did a single application
(Marban et_ al. , 199£). Siddiqui and Mahmood (1992) found that
out of P. 1ilacinus. ftcroph ialophora fusisphora. Baci1lus
1icheniformis and ftlcaliqenes faecal is used for the control of
M. incognita race 3 and Macrophomina phased ina on chickpea, the
last one was leess effective and that first one was most
34
effect ive.
Organisms with inhibitory metabolites
Besides predators and parasites of plant-parasitic
nematodes, some other organisms have also the capacity of
biochemically harming them by the way of producing toxicants
which are mostly by products of metabolism. Ori the hand, some
microorganisms act as decomposers of organic residues and
release substances deleterious to nematodes. These metabolites
and decomposition products s^r^e quite comparable to nematicides
and thus their producers may be considered as biocontrol agents.
Fungi are known to produce metabolites with
nematicidal, antibiotic or fungistatic potentialites.
Penicil1ium anatolicuw produces a series of compounds that can
alter the permeablity of the egg shell to cause free movement of
noxious compounds into the eggs, causing abortive embryonic
development and complete vacuolation of eggs (Jatala et. al. ,
1985). The inhibitory effects of Gliocladium spp., Trichocladium
spp., Trichurus spp., UIocladium spp. and Drechsiera spp. were
manifested as embryonic obliteration, alteration in the egg
shell makeup and reduced hatching which were attributed to the
actvities of their enzymatic and exopathic diffusible toxic
metabolites (Jatala et. al. , 1985). The toxic substances produced
by Paecilomyces 1ilacinus, also cause above mentioned effects
when come in contact with eggs of nematodes (Jatala et. al.. ,
1985). fill concentrations of culture filtrates of P. 1i1 acinus
35
were found effective m inhibiting hatching of Me] oidogyi'ie
incognita juveniles (Khan et_ al. , 1988). The nematicidal
efficiency of metabolites from 16 strains of fungi and their
specificity in controlling nematodes viz. Mtiloidogyne incogni ta,
Globodera pal 1 ida and Nacobbus aberrans were tested by Jatala et_
al. (1990).
The current knowledge on the metabolites (toxins)
produced by fungi is summerized under two main headings (Cayr-'ol ,
1989).
1. Toxins produced in cultures ori liquid media
£. Toxins produced in cultures on solid gel media
Withm the first category, metabolites active
against larvae and adults include species of Fusariurn,
Tr ichoderma and Pis per gi 11 us ni ger (effective against
Meloidoqyne and Heterodera) . Filtrates of 0. niqer and P..
11lacmus were also active against Meloidoqyne eggs (Cayrol,
1989).
The effect of culture filtrates of Rhizoctonia
solam and Sclerot i um rolfsii on hatching and mortality of
Meloidoqyne .lavanica was reported by flli (1989). The culture
filtrates of flspergill us niqer and Rhiroctonia solam reduced
larval penetration, suppressed nematode reproduction and gall
formation on tomato roots. The culture filtrates of former were
more effective than latter (Khan et. aj_. , 1984; Mukhta et_ al.. ,
1991). The culture filtrates of fl. mqer. ft. fumiqatus, ft.
36
ochr-aceus, Peru ci 111 urn isc''la>"'dicurn. P.. no bat urn and P. utriae
were found responsible for considerable reduction m root and
soil populations, maturatic^n and reproduction of Meloidoqyne
incognita and Roty lenchul us reniforrnis (01 i, 1990), Psperai 11 us
ochraceous culture filtrates were tested against M. incoqniba,
Roty lenchul us reniforrnis and He 1 i cot y 1 ench us spp. , in the
laboratory. The nematodes showed iOO'/- mortality withen two
minutes <fimeen, 1991). The structure and confirmation of
ophiobolin K and 6 epiophiobolin K from Q. ustus as a
nematicidal agent was reported by Singh et. ai.. (1990).
Some characteristics of culture filtrates of
Fusari urn so1ani toxic to Meloidoqyne incognita were reported by
Mani and Sethi, (19a4a). They also reported (in 1984b) that
culture filtrates of F. oxyspo^'i urn f. sp. c i cer i and F. so 1 ar, i
have a profound effect on the hatching and mortality of
Neloidoqyne incognita.
Pimong bacteria, Clostridium butyricum produces a
mixture of formic, acetic, propionic and butyric acids in the
culture filtrates that can be toxic to Tylenchorhynchus martini
(Johnston, 1959) and Desulfo'>^ibrio desulfurirans releases H^S to
control nematodes in flooded rice fields (Rodriguez-Kabana et.
ai. , 1965).
Roots of tomato ^nd cucumber plants were inoculated
with 3£6 bacterial isolates and £8 actinomycetes and then
infected with Meloidogyne incognita. Serrat i a marcescens was
37
found to produce volatile metabolite that wa<3 nenictt ox ic. firnmorna
was found to be the m a m metabolic product involved m nematode
inactivation in in vitno conditions (Zavaleta-Mejla, 1985;.
Influence of organic additives on soil microorganisms
Several kinds of organic additives have been found
effective in suppressing plant-parasitic nematodes (Sayre, 1971;
Khan et_ al. , 1974; Rodr i guez-Kabana e; aJL- 1983;'Haq et_ al. .
1986; fibu-Laban and Saleh, ISSZ; Borah and Phukan, 1992).
Initially it was the finding of Lmford (1937) who claimed a
degree of control of Meloidoqyne by adding chopped pineapple
tops to the infested soil. The multifold increase in the
rhizosphere population of saprophytic fungi with application of
oil-cakes in soil prior to transpi atat ion was studied by Khan et
al. (1974) and Khan et_ al. (1976). Soil treatment with organic
matter not only increased microbial actvities but also caused
enhanced enzymatic actvities (Rodriguez-Kabana et_ a_l_. , 1983).
Soil treatment with ground oil-cakes, DD, DBCP, phorate,
fensulfothion, aldicarb or carbofuran reduced soil population of
plant parasitic nematodes in presence or absence of tomatoes
(Haq et. al. , 1986). Fresh and composted rice straw, cacawate
leaves, saw dust and chicken dung were found to reduce nematode
populations of root-knot and reniform initially but results were
not lasting long (Duhaylongsod, 1988). Effects of organic
amendments for controlling root-knot nematodes were also studied
by Darekar et. al, (1980) and Saifullah and Gul (1990).
38
fi col lagenolyt ic fungus C urm i n q h ame 11a eleqari'5
reduced root galling, egg hatching and immobilized second stage
juveniles of Meloidoovne ,iavanica in tomato plants when collagen
was used as soil amendment <Galper et_ al. , 1991). Four oil-cakes
VIZ. castor, groundnut, honge and neem, in combination wibh
carbofuran gave least root galling and highest yield of okra
(Reddy and Khan, 1991). Omong different organic amendments,
karang, neem and mahuva cakes supported growth and sporulation
of Paecilomyces 1ilacinus. Pressmud and farm yard manure were
least effective while piludi cake was toxic to fungus growth
(Patel et. al, , 1991).
Different animal manures were evaluated and compared
with wheat g r a m medium for mass production, storage and
application of some nematode egg parasitic fungi (fibu-Laban and
Saleh, 199E:). Mycelial growth indices of flcremonium
sclerot iqnum, Fusari urn solani, F. oxysporum, Microascus
triqanosporus. Paecilomyces 1i1acinus and Phoma level 1 lei on
animal manures were similar o^'^ higher than on wheat grains. The
fungi were effective in reducing root galling on glasshouse
tomato plants caused by Meloidonvne .lavanica. The organic
amendments vin. poultry manure, mustard cake, neem cake, proved
to be effective in reducing galls and egg masses in roots of
green gram (Borah and Phukan, 199£).
The role of microbes associated with chicken litter
in the suppression of Moloidoqyne arenaria in amended soil was
39
investigated. Microbial degradation of the egg shell and deaxtli
of Snd stage juveniles was apparent (Kaplan et_ aj^. , 199S).
Bacteiria
Bacteria attacking the nematodes are pathogenic or
saprophytic is not yet distinct, because in most cases the
bacteria have been observed within the body cavity, gut and
gonads (Dollfus, 1346). Sayre (1971) stated that tv-ue nature of
bacterial parasitism on nematodes could be resolved only after
application of Koch's postulates.
Pasteuria penetrans; a bacterium attacking nematodes
has been effectively used as biocontrol agent against root-knot
nematodes. Biocontrol efficiency of this bacterium has been
observed to be varied under greenhouse and field conditions
(Mankau, 1975; Sayre, 1980; Stirling, 1984).
The spore-forming bacterium Pasteuria penetrans,
was originally described as a protozoan Duboscqia penetrans by
Thorne (1940). Then it was renamed as Baci1lus penetrans by
Mankau (1975) when its prokaryotic nature was established by
electron microscopy. Sayre and Starr (1985) later placed it in
the genus Pasteuria.
Pasteuria penetrans has shown excellent
synchronisation with the devlopmental stages of root-knot
nematode and that is why it has proved very effective. Baci11 us
penetrans inhibited the penetration by Meloidoqyne incognita 2nd
stage juveniles into tomato roots (Brown et_ al. 1985).
40
Population of M. incognita were lower in the presence oF B.
penetrans in tabacco at 59 days after planting and were higher
after 91 days in tobacco and at sowing with soybeans. The
pathogenic effect of Baci11 us penetrans on Meloidoqyne incognita
was investigated in concrete bordered plots of tobacco,
soyabeans and Vicia y i 1 losa (Brown et_ al. , 1985). Treatment of
M. .lavanica infested soil with Pasteuria penetrans spores as
well as aldicarb or carbofuran, reduced root galling of tomato
seedlings (Brown and Nordrneyer, 1985). The bacterium Pseudornonas
denitrificans has been found to play an important role in
reducing the population of X i phinema americanum (fldams and
Eichenmuller, 19&3). The effect of Ozotobacter chroococcum VP—5
on the hatching of egg masses and eggs of M. incognita was
investigated. Eggs were more susceptible as compared to egg
masses (Chahal and Chahal, 1986). In in vivo and in vitro tests,
0. chroococcum inhibited the hatching of egg masses of M.
incognita and did not allow the larvae to penetrate into the
roots of host to form galls (Chahal and Chahal, 1988).
Preliminary studies on the potential of Pasteuria
penetrans to control Meloidoqyne spp., were done by Channer and
Gowen (1988), Jaya Raj and Mani (1988), Maheswari and Mani
(1988) and Daudi et_ al. (1990). During interaction between
bacteria and nematodes in the soil, the residual products
released are toxic, antibiotics or inhibitory to plant nematodes
(Sayre, 1988). £. penetrans parasitized larvae of Heterodera
41
species also. Maxirmjrii reductions in the cys:,t population of H.
averiAB and H. zeae were qibtained when spore infested soil was
incubated at 30'"'c before application (Bhattacharya and Swarup,
19S9).
Of five bacterial isolates tested against larvae of
Meloidoqyne incognita, Hfeterodera ca.iani, H. zeae, H. a venae,
the most effective were Baci 11 us subt 11 is and B. purni lis. B.
cerus and two Pseudornonas species were also larvicidal (Gokte . r-
and Swarup, 1989).
The biology, specificity and invasion to nematodes,
survival and movement of spores in a soil of Pasteuria spp. is
discussed by Dickson and Oostendon (1990) and Gowen and flhmed
(1990). £. penetrans with Paecjlomyces 1i1acinus gave a greatest
suppression of Meloidoqyne .lavanica ori mung bean (Shahzad et
al. , 1990). The attachment of P,. penetrans spores tq the root-
knot nematode M. .lavanica m soil and its effects on infect ivity
was demonstrated by Stirling et. al. (1990).
Out of four biocontrol agents including bacteria
VIZ. Baci 1 lus 11 cheh i form is. Pseudornonas m m d o c m a ,
Rcrophialophora fusispora and ftsperqi11 us flavus, of root-knot
nematodes, individually B. 1icheniformis was found to be the
best (Siddiqui and Hussain, 1991). The effect of temperature on
attachment, development and interactions of £. penetrans on
Meloidoqyne ay^eris^r i a. was traced out by Hatz and Dickson (199c:).
The greatest attachment rate of endospores of £. penetrans
42
occurv^ed on Snd fc>baQe juveniles at 50"C.
The attachment of g. penetran'3. on Me I oi dopvne larvae
was found to vary with the concentration of spores in the
suspension (Zaki and Maqbool, 1992). The undiluted concen-crat ion
was most effective and reduced 50% root—knot ii'ifection.
Significant reduction of root-knot nematodes on brinjal and niung
beans and increased plant ^ro^^th were obtained when P. penetrans
was combined with Paecilomyces 111 acinus (Zaki and Maqbool,
133£).
The literature r&viewed shows that in recent years
more attention has been given to corlt 'o endopaf-asi t ic nematodes
particularly root-knot nem^E^todes bv usmq opportunistic soil
hyphornycetes, especially P. 11 1 acinus and V. ch 1 amyd'"'soon ura.
Some success has been achieved. Efforts are m progress to sbudy
various aspects of their occurrance, growth, culturing,
adaptablity and efficiency under various agro-climatic
conditions. Some studies in this direction have been made m
India as well. However, in most studies made in the country,
imported strains of these fungi particularly P. 1ilacinus have
been used. Evaluation of Iindian soil fungi (hyphornycetes) has
not been done for their efficacy as biocontrol agents of root-
knot nematodes and other er^doparasites.
MftTERIfiLS AND METHODS
Materials to be used and methods to be employed to
carry out the proposed stilidy are generalized as follows:
1. Survey and collection
Survey will b^ conducted to collect and soil samples
from fields infested with the root-knot nematodes and grown
preferrably with vegetable crops, in different parts of the
country. For collection of the root samples, top soil will be
removed at the base of a plant showing poor growth and .roots
will be examined for root-knots. Five to ten root samples will
be collected at random from each locality of the area undev-
survey, in polythene bags. Samples will be properly labelled and
brought to laboratoty for-^ further examination.
Soil samples will be collected from the vicinity of
the root system having root-knots, fi number of sub-samples from
a particular field will be mixed together and from the bulk
about £50 g of soil will be taken in polythene bags. Samples
will be properly labelled and brought to laboratory for further
examinat ion.
Root samples will be throughly washed and examined
for the presence of galls and egg masses. Root-knot nematode
species will be identified by using perineal pattern
characteristics (Eisenbabk et. al_. 1981) and conducting North
Carolina host differential test (Taylor and Sasser, 1973).
44
Meloidoqyrie .lavardca (Treub) Chit wood will be selected for the
proposed study. Single egg mass culture (Taylor and Sasser^,
1378) of the species will be raised and population will be
multiplied and maintainedion tomato in greenhouse for use in the
experiments.
2- Isolation of fungi from 5oil samples
Soil samples collected from different parts of the
country from root-knot infested fields will be used for
isolation of fungi. Warcup method will be employed for isolation
of fungi from the samples, ft small amount of soil will be added
to sterlized petriplates with the help of spatula and cooled
potato dextrose agar (PDA) or Mav^tin's rose bengal agar will be
poured m the plates. The plates will be shaken gently in order
to attain uniform distribution of soil particles in the plates.
The petriplates will then be incubated at c:5"C in 3.ri incubator.
Soil hyphomycetes present in the samples growing in culture
plates will be identified, isolated and stocked on PDfi slants
with proper markings for future use in experiments. Sub-
culturing will be done periodically.
3. Isolation and identification of fungi associated with
egg masses
0 few egg masses selected randomly from the roots of
the infected host plants collected during the survey will be
washed throughly and plated on PDA contained in sterli::ed petri
dishes. Potato dextrose agar (PDO) will contain the follov«jing
45
consb ituents:
Peeled and sliced potato - £00 g
Dextrose - £0 \;i
agar - £0 g
Distilled Water - 1000 ml
The medium will be prepared in the laborator^y and £0
ml of it (heated and cooled at 45 C) will be taken m each
sterlized petv^iplate. The egg masses will be plated under
aseptic conditions on a laminar flow bench. Before it, a small
amount of streptocycline ^ill also be added to each petriplate.
The petriplates will be then incubated at £5~C for a week in an
incubator and fungal colpnies devloping around egg masses will
be examined and identified. The fungi apparently penetrating the
eggs and egg masses will be then isolated, multiplied and
maintained on PDR slants for future studies.
4. In vitro inoculations
To study the effect on hatchability of juveniles of
the nematode Meloidoqyne .lavanica. 5 egg masses of the nematodes
will be placed in 5 ml of different concentrations of culture
selected filtrates of fungi contained in sterlized petriplates
(3 cm dia. >. The plates' will be examined after £4, 43 and 7£
hours for hatching of eggs. The number of juveniles hatched will
be counted.
To study the larvicidal effect of culture filtrate
of selected fungi, 100 second stage juveniles (Jg) of root-knot
46
nematode, Meloidoqyrig JLiv aniES. will be placed in 5 ml of
different concentrations,of culture filtrate of selected fungi
contained in sterlized petriplates. The plates will be examined
after £4, 48 and 7£ hours. The number of dead juveniles will be
counted.
To study thi efficacy of some selected fungi as
biocontrol agent, egg miisses of Meloidoqyne .lavanica will be
taken and treated with mei"curic chloride (0.01"/.) for 1-3 minutes
and washed in disti1 led,water repeatedly to remove mercuric
chloride. The surface sterlized egg masses will be then placed
on PDO in petriplates. The fungus to be tested will be
inoculated over these egg masses. The whole procedure will be
carried out at 1 ami nor -flow bench. In some petriplates, only
surface sterlized egg masses will be plated and rio fungal
inoculum will be added. Tt;ie5e petriplates will serve as control.
The petriplates with egg masses, inoculated or uninoculated with
i-i
the fungus will be then i lcubated at £5 C for a week.
fit the end o f the incubation period, the egg masses
will be taken out from the petriplates and examined for the
penetration of the fungus into the egg masses, subsequently
infecting the eggs. For ^determining the percentage of infected
eggs in each egg mass, the egg masses obtained from both
treatments (inoculated or control), will be stained with cotton
blue in lactophenol and each egg mass will be gently pressed
o\er a glass slide to sbperate the eggs. The number of eggs
47
irjfocted by thf? test fungus will be counted under the
microscopic fields and percentage oF infected eggs will be
calculated. The deForrnity and abnormal development of root-knot
nematode juveniles if afiy in the inoculated egg masses, will
also be examined and noted.
5. In vivo inoculations
For m vivo studies, root—knot nematode, f. .lavanica
and some selected fungi kill serve as test pathogens. Cow pea,
Vi nna sinensis L- and tomato, Lycopersican eoculent um Mill.,
will serve as test plants.
(i) Nematode inoculation
For inoculation, nematode inoculum will be obtained
by either of the following two methods:
(a) A la>-"ge number of egg masses of M_. lavanica
collected from the roots maintaining single egg mass culture of
the species will be kept on a double layer of facial tissue
paper supported by a coarse sieve. It will be placed over a
petriplate (9 cm dia.) having sufficient water to touch the
bottom of the support, ft small amount of water will also be
poured over the egg masses. PIfter £4 h, the hatched juveniles
will be collected from the petriplate and used for inoculation
of the seedlings in the experiments. Their number m measured
quantity of the suspension will be determined.
(b) The roots maintaining single egg mass culture of M_.
.Tavanica will be cut into pieces (£-4 cm) after thoroughly
48
washing with i tap water. The pieces will be placed m a 1000 ml
container with £00 rnl faf 0.5"/. sodium hypochlorite (NaOCl )
solution. The tightly capj^ed container will be shaken vigorously
for three minutes. Shkking will partially dissolve the
gelabmous matrix, thus freeing eggs from the egg masses. The
liquid suspension of eg^s will be poured through a .SOO-mesh
sieve, nested upon a 50O-rnesh sieve- Eggs suspended in the
agitated solution will pass through the £00-mesh sieve and will
be collected on the 500-mesh sieve. Eggs will be washed free of
residual NaOCl solution under a slow stream of tap water. The
concentration of eggs pfer milliliter will be standardised by
counting the eggs from ten, 1 ml samples and the average number
will be used to represent the number of eggs per ml.
For inoculation, depending upon the inoculum
density, volume of the suspension containing eggs or second
stage juveniles will be taken in pipette. Roots of seedlings (3-
4 week old) will be partially exposed carefully removing the top
layer of soil. The suspension will be poured uniformly on the
exposed roots. Then, the roots will be covered with same soil
and light watering will be done.
(ii) Fungus inoculation
Pure cultures of the selected fungi will be
maintained in the culture tubes, containing PDA. The fungi will
be grown on Czapek's liquid medium in Erlenmeyer flasks. The
sterilized medium in the flasks will be inoculated V'jith the
49
desired fungus and the? flasks will be incubated in an incubator
at £5 C for a week.
After the intubation period, the mycelial mat will
be removed and washed in distilled water to remove the traces of
the medium. Then it will be gently pressed between sterile
blotting papers to remove the excess amount of water. Inoculum
will be prepared by shaking 10 g fungal mycelium in 100 ml of
sterilized distilled water and blending it for 30 sec. in a
waring blender (Stemerding, 1963). In this way each 10 ml of
this homogenate will coV-itain 1 g of the fungus. The desired
amount of the suspensibn will be added to roots exposed by
removing the top layer of the soil. The roots will be covered
with the soil again. Cane will be taken to keep the inoculated
pots moist for 3-4 days for the stabilization of the fungus.
6. Root penetration
For studying the effect of the selected fungi on
root penetration by juveniles of M. .lavanica in their presence,
the experiment will be set up by adding suspension of the
biocontrol agents (selected fungi) and M. .lavanica as given
above. One replicate from each treatment will be uprooted after
every four days of inoculation. Th^ roots will be washed,
labelled and taken to laboratory for examination. The roots will
be stained with acid fuchsin in the laboratory and cleaned in
lactophenol. The number of penetrated juveniles or the other
development stages of the? nematode in the roots will be counted
50
under the microscope.
7. Soil dr-ench and spray treatment
Efficacy of the selected soil fungi found to be
effective as biobontrol agents in earlier experiments will be
tested for their application as soil drench and spray
treatments. The fungus inoculum will be applied on the soil
surface in quantities sufficient to wet 10-15 cm of the soil in
the microplots or £'4" clay pots. This treatment will be applied
before or after sowing of cowpea seeds and tomato seedlings. The
nematode suspension will be added in the soil at the primary
seedling stage. Uninoculated plants will serve as control.
To study the efficacy of biocontrol agents by
spraying the fungus, the inoculum will be sprayed on the host
plants in the microplots or £4" clay pots. The nematode
suspension will be added in the soil after one week of spraying.
Uninoculated plants will feerve as control.
8. Recording of data
fit the end of the experiments, plant growth
parameters and parameters related to root-knot disease on the
host will be considered. Plants will be uprooted after 60 days
of inoculation and the roots will be thoroughly and gently
washed. The length (in cm) and fresh and dry weights <in g) of
shoot and root will be determined separately. Before weighing
the plants for weight, excess amount of water will be removed by
putting shoot and root between blotting sheets. For dry weight,
51
o shoot and root will be dried in 3.ri oven at 60 C and weighed,
3. Root—knot disease parameters
Before drying the plants, roots of plants from the
treatments will be examined. The number of galls and egg masses
present will be counted. Root—gall iridex (GI) and egg mass index
(EMI) will be rated on 0-5 scale of Taylor and Sasser (1976) as
given below:
1_ = O No galls, no egg mass
£ = 1-10 galls/egg masses
3 = 11-30 galls/egg masses
4 = 31-100 galls/egg masses
5 = Pibove 100 galls/egg masses
The percentage of egg masses infected with the
fungus added as biocontrol agents will be determined. Similarly,
percentage of eggs in each egg mass infected by the fungus will
also be determined by exafnining the eggs under the microscope.
Root population and soil population of the nematode
will also be determined by the standard methods.
10. Experimental design and statistical analysis
Pots or pet^'i plates in the experiments will be
arranged according to the complete randomized block design
(CRBD). The data will be subjected to Analysis of Variance
(RNOVft) and L.S.D. will be calculated to determine the
significance between the treatments.
52
The materials and methods described above will be suitably modified or changed during the course of investigations, whenever felt necessary.
LITERftTURE CITED
Pbdel Rahman, Fawzia and O. R. Maggenti (1387). Meloidogyr e ca 11 forniensis n. &p. (Nernata: Meloidogyninae) , Parasitic on Bulrush, Scirpus robust us Pursh. Journal of Nernatology 13 <.2.) z 3.07.
flbu—Laban, O.Z. and ft.H. Saleh (1992). Evaluation of animal manures for mass production, storage and application of some nematode egg-parasit ic fungi. Nernatologica 3S(£);£37-244.
fldarns, R. E. and J. J. Eichenmuller (1963). fi bacterial infection of Xi ph inema amen can urn. Phytopathology 53:745.
ftl-Hazrni, ft. S. and ft. T. ftbdul Razik (1991). Evaluation of some fungal species as biocontv^ol agents of Meloidoqyne .lavanica. Pakistan Journal of Nernatology 9(l):39-47.
fil-Hazmi, ft.S., D.P. Schmitt and J.N. Sasser (1982). Population dynamics of Meloidoqyne incognita on corn grown in soil infested with ftrthrobotrys conoides. Journal of Nematology 14(l):44-49.
k
ftl-Hazmi, ft.S., D.P. Schmitt and J.N. Sasser (1982). The effect of Ort hrobot rys conoides on Meloidoqyne incognita population densities in corn as influenced by temperature, fungus inoculum density and time of fungus introduction m the soil. Journal of Nematology 14(2):168-174.
flli, H.H. (1989). The effect of culture filtrates of Rhiroctonia solam and Sclerotlum ro1fsii on hatching and juvenile mortality of Meloidoqyne .lavanica. Japenese Journal of Nematology 18(7):36-3B.
fill, H.H. (1990). Nematicidal action oF fungal culture filtrates. Japenese Journal of Nematology £0:1-7.
Anieen, H. H. (1991). Nematicidal effect of ftsperqi 1 lus ochraceous filtrate. Bulletin of Faculty of Agriculture, University of Cairo 4£(3):963-970.
ftschner, M. and S. Kohn (1958). The biology of Harposporiurn anquillulae. General Microbiology 19:182-189.
ftviva U. and R.ft. Sikora (1992). Use of non-target isolates of the entornopathogen Vert ici 111 urn lecani <Zimm) Vieges to control the potato cyst nematode, Globodera pal 1ida (Stone), Nematologica 38(1):123-130.
54
Baker, K. F. arid R.J. Cook (1974), Biological coritr-ol of Plant Pathogens. W.H. Freeman, San Francisco, pp. 435.
Barr ori, G. L. (1976a>. Nernatophagus fungi: three new species of Myrocyt i urn. Canadian Journal of Botany cic: (5) :752-765.
Barron, G. L. (1976b). Nernatophagus fungi: a new endoparasite intermediate between Myzocyt lurn and Leg en id x um. Canadian Journal of Botany 54(1/£):l-£.
Barron, 6. L. (1979). Nematophagus fungi: a new Harposporiurn producing aerial arthroconidla. Canadian Journal of Botany 57(8):88&-889.
Barron, G. L. (1980). Nematophagus fungi: a new Harposporium from soil. Canadian Journal of Botany 58(4):447-450.
Barron, 6.L. (1982). Nematode-dsstroying fungi, In: Experimental Microbial Ecology (Eds. R.G. Burns and J.H. SlaterO, Blakcwell, Oxford, pp.533-555.
Barron, G. L. and J.G. Percy (1975). Nematophagus fungi: a new Myzocytium. Candian Journal of Botany 53(13):1306-1309.
Barkeley, M.J. (1855). Vibro forming cysts on the roots of cucumber. Gdnr's Chron. 14:£c:0.
Bhattacharya, ft.d. and G. Swarup (1989). Pasteuria penetrans. a pathogen of the genus Heterodera, its effect on nematode biology and control. Indian Journal of Nematology 18(l):61-70.
Bong, B., W. M. Roberton and L.F. ftinsworth (1988). Observation or\ the specificity of the nematophagus fungus ftrthrobotrys dasguptae (Shome and Shome) to plant parasitic nematodes. Nematologica 34 (5) : £'38-545.
Borah, fiparajita and P.N.Phukan (199£). Effect of different organic amendments for the management of Meloidogyne incognita on green gram. Current Nematology 13(1):49-50.
Brown, E.B. (1978). Cultural and biological control methods, In: Plant Nematology (Ed. J. F. Southey), GD, Ministry of fiqriculture, Fisheries and Food, Her Majesty Stationery Service, London, pp. 569-585.
55
Brown, B. M. and D. Nordrneyer (1985). Synergist jc reducb K-ICI in root galling by Me] oidogyne .lavanica with PastRKrj a penotraris and nematicides. Revue ds- Nernatologie 6(3):£85-£a6.
Brown, S.M. and B.C. Jr Smart (1985). Root penetration by Meloidoqyne incogruta juveniles infected with Baci31 us pgnetrans. Journal of Nernatology 17 (2) : lc:3-l£6.
Brown, S.M., J.L. Kepner and B.C. Jr. Smart (1985). Increaced crop yields following application of Baci1lus penetrans bo field plots infested with Meloidogyne incognita. Soil Biology and Biochemistry 17(4):483-486.
Cabanillas, E. and K. R. Barker (1989). Impact of Paeci lornyr'es 111acinus inoculum level and application time on control of Meloidogyne incognita on tomato. Journal of Nernatology £1 (1) : 115-lc:0.
Cabanillas, E. and K.R. Barker and L.ft. Nelson (1989). Survival of Paecilomyces 1i1acinus in selected carriers and related effects on Meloidogyne incognita on tomato. Journal of Nematology £1 (1) : liE:l-130.
Cabinillas, E., K.R. Barker and L.ft. Nelson (1989). Growth of isolates of Paeci1omyces m biocontrol of Meloidogyne
Lilacinus and their on incognita
Journal of Nematology £1(£):164-17£.
efficacy tomato.
Cabinillas, E. , K. R. Barker and M. E. Daykin (1988). Histology of interactions of Paecilomyces 1i1acinus with Meloidogyne incognita on tomato. Journal of Nernatology £0(3) :36c.'-365.
Cabrera, R.I., J.P. Latge, D. Dominguez and Y.G. Gonzalez (1987). The fungus Vertici111um chlamydospora um m biological control of the phytonematodes of the genus Meloidogyne in guava orchards, Psid ium qua.iava. Ciencia Teenica el la Ogricultura, citricos y Otros Erutales 10(4):79-87.
Capstick, C.K-, D.C. Twinn and J.S. Waid (1957). Predation of natural populations of free-living nematodes by fungi. Nematologica 2:193-501.
Carlisle, D. (1987). Farmers fight Scientists 1558:35.
their foes with fungi. Ne->w
56
Cayrol, J. C. (1988). Biological conbrol oT MeJi-i idogyne with predatory fungus flrthrobot rys i rrpqu 1 ar is. EPPO Bulletiri lS(l):73-75.
Cayrol,
Cayrol,
Cayr'ol,
J.C. (1989). r4ematic:idal Horticulture £93:53—57.
toxins of funrj 1. Revue
J . C , C. Djian and L. Pijarowski (1989). Study of the nernaticidal properties of the culture filtrates of the nematophagus fungus Paeci lornyces 111 acinus. Revue de Nematologie IS(4):331-336.
J.C. and J. P. Frankowski (1979). Une rnethode de lutte biologique contreles nematodes a galles des racmes appartenant au genre Meloidoqyrie. Revue Horticulture 193:15-23.
Cayr'ol, J . C , J. P. Frankowski, ft. Laniece, G. d'Hardemare and J. P. Talon (1978). Contre les nematodes methods de lutte biologique a I'aide d'un hyphomycete predateur. ftrthrobotrys robust us souche untipolis (Royal 300). Revue Horticulture 184: £"3-30.
Chahal, P.P.K and V.P.S. Chahal (1986). Effect of flzotobacier chroococcum on the hatching of egg masses and eggs of Meloidoqyne incognita. Plant and Soil 95:589-c:91.
Chahal, P. P.K and V.P.S. Chahal (1988). Biological control of root-knot nematode of brinjal (Solanum melonqena L. ) with Ozotobacter chroococcum. Hematology (Eds. M.Maqbool
In: Advances et a 1. , )
Nematological Research Centre, Karachi, pp. :c:57-c:63.
University of
m Plant Natlonal Karachi,
Channer, ft. G. and S.R. Gowen (1988). Preliminary studies on the potential of Pasteuria penetrans to control Welotdoqyne Goeldi 1887. Proceedings of Helmmthological Society of Washington 16:90-104.
Cooke, R. C (1962). The ecology of nematode-trappmg fungi the soil, finnals of Applied Biology 50(3):507-514.
in
Cooke, R. C. (1963). The predaceous activity of nematode-trappmg fungi added to soil. finnals of Applied Biolorqy 51(£):£g5-£99.
Cooke, R. C. (1964). Ecological trapping Hyphomycetes. 54:375-379.
characteristics of nernatode-AnnuaIs of Applied Biology
57
Cossernans, J. C. (1991). Methcids for introducing Vert i cri L13 urn ch I anrnydospor 1 urn into soil. EoUletin SROP 14 (£): 35-46..
Croshier, R., 6. Montecinos, M. Jimenez and P. Gallo (1985). Effectiveness of Paeci lornyces 11 I acinus (Thom Samson) XYi the control of the root-knot nematode Meloidogyne .Tavanica. International Nernatology Network Newsletter £(3):3.
Crump, D.H. (1987). fl method for assessing the natural control ' of cyst nematode population. Nematologica 33 (£) : £35-
Crump, D.H- and F. Irving (1992). Selection of isolates and methods of culturing Vert ici ill um ch 1 arnydospor 1 urn and its efficacy as a biological control agent of beet and potato cyst nematodes. Nematologica 38(3):367-374.
Dackraan, C and B. Nordbring—Hertz (1985). Biocontrol: Fungal parasites of the cereal cyst nematodes Heterodera avanae in Southern Swedan. Journal of 17(1):50-55.
Nematology
Dackman, C.
Darekar, K.S., N.L. Mhase and S.S. Shelke (1990). Effects of placement of non-edible oilseed cakes on the control of root-knot nematodes on tomato. International Nematology Network Newsletter 7(l):5-7.
Daudi, ft.T., fl.G. Channer, R. fihmad and S.R. Gowen (1990). Pasteuria penetrans as a biocontrol agent of Meloidopyne .lavanica in the field in Malawi and in microplots m Pakistan. Proceeding of the British Crop Protection Conference on Pests and Diseases l:£53-£57.
Davide, R. G. and R.ft. Zorilla (1985). Evaluation of a fungus Paecilornyces 111acinus for the biological control of root-knot nematode M. incoqnita on okra and compared with nematicide isai^ofos. Philippine ftgriculturist 68(4):493-500.
Dickson, D. W. and Ostendon (1990). Biological control of nematodes with Pasteuria spp. Nematology Circular (Gainesville) 175:4.
58
Dijhsterhuis, J., M. Veenhuis and W. Haidei" (iggO) . Ultrastructural study of adhesion and initial stages of infect lori of nematodes by conidia of Drechmc?ri a coniosDova. Mycological Reseai-^ch 94(1) :l-a.
Djksterhuis, J., W. Haidei", U. Wyss and M. Veenuhuis (1991). Colonisation and digestion of nematodes by endoparasitic nematophagus fungus Drechmeria conispora. Mycolgical Research 95(7):B73-878.
Dollfus, R.P. (1946). Parasities des Helminthes. In Encyclopedia Biologique, ed Lechevalier, P. vol. £7. P a n s 48E' pp.
Drechsleir, C. (1937). Some hypomycetes that pr ey on free living t e m c o l o u s nematodes. Mycologia £9:447-45£.
Drechsler, C. (1941). Predacious fungi. Biological Reviews, Cambridge Philosphical Society. 15:£65-£90.
Dube, B. and C. S. Grover Jr. (1987). Biological control of Meloidogyne incognita by Paecilomyces 11lacinus and Pasteuria penetrans. Journal of Nematology 19(£) •.5.2.B.—2.E.7.
Duddington, C.L. (1956). The Predacious fungi: Zoopagales and Moniliales. Biological Reviews, Cambridge Philosphical Society 31 : 15iz:-193.
Duddington, C.L. (1957). The Friendly Fungi: Faber and Faber, London, p. 188.
Duddington, C.L- (1960). Biological control-predacious fungi,In: Nematology (Eds. Sasser, J.N. and W.R. Jenkins) University of North Carolina Press, Chapel Hill, pp.461-465.
Duddington, C.L. (1962). Predacious fungi and the control of nematodes. Viewpoints in Biology l:15l-£00.
Einsenback, J. D. , Baojun Yang and K.M. Hartman (1985). Description of Meloidoqyne pini n.sp., a root—knot nematode parasitic on Sand Pine (Pmus claus) with additional notes on the morphology of M. megat via. Journal of Nematology 17(2):£06.
Eisenback, J.D., H.Hirschmann, J.N. Sasser and fl.C. Triantaphyllou (1981). fi more complete characterization of the four most common specLies of roo-knot nematodes (Meloidoqyne spp.) with pictorial key. IMP Publication, Raleigh, North Carolina, U.S.ft.
59
Esser, R. P. and E. K. Sober-s (19B4). Natural errernies of nematodes. Proceedings of Soil and Crop Scierice Society of Florida. c:3:l£l-13S.
Esser, R. P. and T. S. Schubert (1983). Fungi that entrs^p arid assimilate by employing constricting rings. Nematology Circular (Gainesville) 192:3.
Esser, R. P N. E. El-Gholl and M. Price (1991). Biology of Dactyle]la menalospora Drechsler: a nematophagus fungus. Proceedings of Soil and Crop Science Society of Florida 50:173-180.
Fassuliotis, G. (1979). Plant breeding for root—knot nematode resistance, In: Root-knot nematodes, (Meloidoqyne spp. ) systematic, biology and control (Eds. F„ Lamberti and C.E.Taylor) Ocademic Press, London., pp. 4c:5-45c:.
Feder, W.A., C.O.R. Everard and L.M.O. Wootton (1983). Sensitivity of several soecies of the nematophagus fungus Dactyleila to a morphogenic substance derived from free-lving nematodes. Nematologica 9:49-54.
Franco, J., P. Jatala and M. Bocangel (1981). Efficiency of Paec1]omyces 1ilacinus as a biocontrol agent of Globodera pal 1ida. Journal of Nematology 13(4):438-439.
Galper, S., E. Cohn., Y. Spiegel and I. Chet (1991). fi col lagenolyt ic fungus Cunnmqhamel la elegans for biological control of plant-parasitic nematodes. Journal of Nematology £3 (3) :iE:69-£74.
Gintis, B. O. , G. Morgan—Jones and R. Rodriguez-Kabana (1983). Fungi associated with several developmental stages of Heterodera q1ycines from an Alabama-soybean field soil. Nematropica 13:18i-c:00.
Goetdi, E. fl. (1887/1892). Relative sobra a molestla ds caffeeiro na provmcia de Riode Janeiro flrch. Mus nac. Rio de Janerio 8:7-121.
Gokte, N. and G. Swarup (1989). On the potential of some bacterial biocides against root-knot and cyst nematodes. Indian Journal of Nematology IS(1):152-153.
Golden, ft. M. (1989). Further details and SEM observation on Meloidoqyne marylandi (Nematoda: Meloidogynidae). Journal of Nematology 21(4):453.
60
Golden, ft. M. and David T. K. (1986). Meloidogynf? christ lei n. sp. (NematodarMeloidogyne from oak with 5EM and host range observations. Jc>\.\-rri3.1 of Nernatology lfl<A):533.
Gowen, S. R. and R. Rhmad (1990). Pastsuria perietrans for control of pathogenic nematodes. Aspects of Applied Biology •24: £5-32.
Gracia, J. ft. R. D. Reeleder and G. Belai»~ (1991). Interactions between Pythiurn tracheiphilum, Meloidoqyne hapla and Pratylenchus penetrans on lettuce. Phytoprotection 7£:105-144.
Grandison, G.S. and K.M. Cooper (1986). Interaction of VflM and cultivars of alfalfa susceptible and resistant to Meloidogyne hapla. Journal of Nernatology 18 (c.'): 141-149-
HansBler, G. and M. Hermanns (1981). Vert icill m m lecani i as a parasite on cysts of Heterodera schachtii. Zeitschrift fur Pflanzenkrankheiten and Pflannenschatz 88(11):678-681.
Haq, S., ft.U. Siddiqui, S.K.Saxena and M.W. Khan (1986), Effect of groundnut oilcake and certain nematicides on the population of nematodes and fungi m the presence and absence of tomato. Indian Journal of Nernatology 16(1):109-110.
Haq, S. , S. K. Saxena and M. W. Khan (1983). Toxicity of systemic nematicides in relation to larval hatching and mortality of root-knot nematode (Meloidoqyne incoqnita). Indian Journal of Parasitology 7(£):193-194.
Hatz, B. and D.W. Dickson (1992). Effect of temperature on attachment, development and interactions of Pasteuria penetrans on Melodooyne arenaria. Journal of Nernatology £•4(4) :51£-5£1.
Heejbroek, W. (1983). Some effects of fungal parasites on the population development of the beet cyst nematode (Heterodera schacht 11) . Mededelmgen van de Faculteit Landbouwwetenschappen Rijksuniversitlet Gent 48(£): 433-439.
Hewlett, T.E., D.W. Dickson, D.J. Mitchell and M.E. Kannwischer-Mitchell (1988). Evaluation of Paeci Imycre s 11lacinus as a biocontrol agent of Meloidoqyne lavanica on tobacco. Journal of Nernatology £0(4):578-584.
61
Hirschrnann, H. (198&). Meloidoqyne hisp^tnica n. sp. (Nematoda: Meloidogynidae) the Seville root-knot nematode. Joui-'nal of Nernatology 1B(4):5£:0.
Hussey, R. S. and K.R. Barker <1973). ft comparison of methods of collecting mocula of Meloidogyne spp. including a new technique. Plant Disease Reporter 57: 10c:5-10iE:6.
Ibrahim, I.K. A , ^..x. n. , M. M. Joshi and J. P. Mollis (1973). The swarming virus disease of Tylenchhorvhynchus mart xni. International Congress of Plant Pathology. Minneapolis, Minnesota, Abstract 555.
Iruing, F. , and B. R. Kerry (1986). Variatiion between strains of the nematophagus fungus Vertici11ium chlamydosporlum Goddard II. Factors affecting parasitism of cyst nematodes eggs. Nematologica 3£(4):474-Ae5.
Isogai, ft.fi., S. H. Suzuji and S. tanura (1980). Constituents of a peptidial antibiotic P-168 produced by Paecilomyces 111acinus (Thorn) Samson. Agriculture Biological Chemistry 44(1£):3059-3031.
Jaffee, B. fl. (1986). Parasitism of X i ph i nema riven and X,. americunum by zoosporic fungi. Journal of Nematology 15(1):a7-93.
Jaffee, B-fl., ft.E. Mukdoon and E.G. Tedford (1992). Trap production by nematophagus fungi growing from parasitized nematodes. Phytopathology Sc! (6) :615-6c:0.
Jaffee, B. fl. and E. Zehr (1985). Parasitic and saprophytic abilities of the nematode attacking fungus Hirsutella rhossi11ens15. Journal of Nematology 17:341-345.
Jain, R. K. and C.L. Sethi (1988). Interaction between VftM and MeloidoQvne incognita and Heterodera calani on cowpea as influenced by time of inoculum. Indian Journal of Nematology 18(2):a63-e68.
Jansson, H.B., fl. Jeyaprakash and B.M. Zuckerman (1985). Control of root—knot nematodes on tomato by the endoparasitic fungus M e n a comospora. Journal of Nematology 17(3) :3£7-3£9.
Jansson, H.B., fl. Jeyaprakash and B.M. Zuckerman (1985). Differential adhesion and infection of nematodes by the endoparasit ic fungus M e n a comospora (Deuteromycetes) . Applied and Environment Microbiology 4g:55£-553.
62
Jansson, H.B., ft. Von Hofsten and C. Von Mecklenberg (1984). Life cycle of the endoparasit ic nernatophaqous fungus M e n a cornospora: a light and electron microscopic study. Aritonic Von LeeuwenhocP Journal of Microbiology 50(A):3£l-3£7.
Jansson, H.B. and B. Nordbring—Hertz (1980). Interaction between nernatophagous fungi and plant parasitic nematodes: attraction induction of trap formation and capture. Nernatologica £6 (4) :3a3-389.
Jansson, H. B. and B. Nordbring—Hertz (1983). The endoparasitic nernatophagous fungus M e n a coniospora infects nematode's specifically at the chemosensory organs. Journal of General Microbiology lc:9 (4) : 1121-1 lc:6.
Jansson, H.B., C. Dackman and B.M. Zuckerman (1987). Odhesion and infection of plant parasitic nematodes by the fungus Drechrnena coniospora. Nernatologica. 33(4): 109-11c:.
Jansson, H.B. and L. Thiman (1992). ft preliminary study of chernotaxis of zoospores of the nematode parasitic fungus Catenaria anqui 11 ulae. Mycologia 84 (1): 109-1 liE:.
Jatala, P. (1986). Biological control of plant-parasitic nematodes, ftnnual Review of Phytopathology £4:453-489.
Jatala, P., B. Manrique and L. Gavilano (1990). efficiency of some fungal metabolites specificity in controlling plant parasitic Fitopatologica £5(1):13.
Nernat icidal and their nematodes.
Jatala, P., J. Franco., 0. Gonzalez and C M . O'Hara (1985). Hatching stimulation and inhibition of Globodera pallida eggs by ennymatic and exopathic toxic compounds of some biocontrol fungi. Journal of Nematology 17:501.
Jatala, P., R. Kaltenbach and M. Bocangel (1979). Biological control of Meloidoqyne incognita acrita and Globodera pal 1 Ida on potato. Journal of Nematology 11:303.
Jatala, P., R. Sales., R. Kaltenbach and M. Bocangel (1981). Multiple application and long term effect of Paec-T omuces 11lacinus in controlling Meloidoqyne mcoqnita under field conditions. Journal of Nematology 13(4):445.
53
Jawich, M. and H. Bochow (1989). Us« of the neniatode-trappi ng fungus Orthrobott-'vs tortoi-' against Weioidopyrie spp. on different substrates in green houses. Reihe firjrarwissenschaften 36 (.5) :i215-c:£l.
Jaya Raj, M. fl., and ft. Mani (1988). Biocontrol of Mel .lavanica with the bacterial spore parasite penetrans. International Nernatology Network New 5(1):3-4.
idoqyne
Pasteuria iletter
Jensen, C. and G. Lusek (1991). Direct observation of trapping activities of nernatode-destroymg fungi m the soil using fluorescence microscopy. Microbial Ecology 85 (3) : 507-=: 10.
Jimenez, R.M. and D.D. Gallo (1988). New contribution on the effectiveness of Paeci lornyces 11 1 acinus (Thorn) Samson m the control of root-knot nematodes (Meloidoqyne spp.). IDESIft 10:29-33.
Johnston, T. M. (1959). Effect of fatty acid mixtures or\ the rice stylet nematode (Tylenchorhynchus martini Fielding, 1956). Nature 183:1392.
Kaplan, M J. P. Noe and P. G. Hai-tel (1992). The role of microbes associated with chicken litter in the suppression of Meloidoqyne arenarla. Journal of Nernatology 24 (4) :5c!£:-5c:7.
Kerry, B.R. (1975). Fungi and the decrease of cereal cyst-nematode population in cereal monoculture. EPPO Bulletin 5:353-361.
Kerry, B.R. (1980). Biocontrol: Fungal parasites of female nematodes. Journal of Nernatology 12:253-259.
cyst
Kerry, B.R. (1984). Nematophagous fungi and the regulation of nematode populations in soil. Helmmthological Abstract 53B:1-14,
Kerry, B.R. (198B). Two microorganisms for the biological control of plant parasitic nematodes. British Crop Protection Council 2:603-607.
Kerry, B.R. and D.H. Crump (1977). Observation on fungal parasites of females and eggs of the cereal cyst nematode, Heterodera avanae, and other cyst nematodeEr>. Nematologica 23:193-201.
64
Kerry, B. R. , D. H. Crump and L.ft. Mullen (1980). Parasitic fungi, sell moisture and multiplication of the cereal cyst nematode Heterodera avenae. Nernatologica £6:57-6S.
Kerry, B. R. , F. Irving and J.C. Hornser (1986). Variation between strains of the nematophagous fungus Vert ici 11 lum ch lamydospor i urn Ooddard 1. Factors affecting growbh in vitro. Nernatologica 3£'(4) :461-473.
Kerry, B. R. , B.C. Jenhinson and D. H. Crump (1976). Fungal parasites of cyst nematodes. Rotharnsted Experimental Station Report Part l:c:01-£0c:.
Khan, fl. ft. and M. W. Khan (1985). Root-knot nematodes infecting some common weeds m vegetable fields in western Uttar Pradesh (India;. International Nernatology Network Newsletter £(4):15-16.
Khan, ft.M. (1976). Control of diseases caused by nematodes by the application of oil-cake manures. Final Technical Report PL-480 scheme, Botany Department, flligarh Muslim University, filigarh, India, pp. 94.
Khan, M.W. (1990). Biocontrol of plant nematodes in closer perspective, In: Progress m Plant Nernatology (Eds. B. K, Saxena, M. W- Khan, ft. Rash id and R.M. Khan) Print House (India), Lucknow, pp.367-385.
Khan, M. W. and M.N. Esfahani (1990). Efficacy of Paecilomyces 11 lac in us for controlling Meloidoqyne .lavanica on tomato m greenhouse in India. Pakistan Journal of Nernatology 8(2): 95-100.
Khan, M.W., S.S- fishraf and S.R. Haider (1988). ftn analysis of biocontrol efficiency of Paecilomyces 1ilacinus , in management of root-knot nematodes. (Abstract). International Conference on Research and Plant Sciences and Relevance to the Future, Delhi, India.
Khan, T.ft. , M.F. fizam and S.I. Hussain (1984). Effect of fungal filtrates of Pisperqi 1 lus ni qer and Rhizoctonia so lam on penetration and development of root-knot nematodes and the plant growth of tomato var. Maryglobe. Indian Journal of Nernatology 14 (£): 106-109.
Khan, T. ft. and S.I. Husain (1986). Parasibism of h'ieloidoqyne incognita by Fusari urn so lam. International Nernatology Network Newsletter 3(£):11-13.
65
Khan, T. ft. and S.I. Husian (1990). Biological conbrol oF i-oot-UriCit end reniforrn nematode?:; and roob—t^ot fungus orr cowpea. Bioved l<l):19-iE:4.
Kirn D. G. and R. D. Riggs (1991). Characteristics and eFfnciency of a sterile hyphomycete (ftRFlS) a new biocontrol ager.b for Heterodera qlyemen and other nematodes. Journal of Nernatology £5 (3) :£75-iE:8c:.
Kondakova, E.I. (1973). Use of parasitic fungi m the control df the potato cysb nematode. CfibstractH. In: Materially vsesoyu:::nogo simponiurna po bor'be s kartofel noi nematodoi, Tartu, pp. 53-59.
Kuhn, J. (1S77). Varlaufiger Bev^icht iiber die bisher iijeri Ergebnisi^e der siet dern Jahre 1S75 m fiuftrage des Vereins fCJr Rubenzucker Industrie ausgefuhrben Versuche nur Errnit tenl ung der Ursuche der Rubenrni id igkei t des Bodens Und zur Erforschung der Nabur des Nernatoden. Z. Ver. Rubenzucker Ind. Dtsch. Reiches, pp. 452-457.
LeiJ, F.fi.O.M. De and B. R. Kerry (1991). The nematophagous fungus Vert ici 111 urn chlamydospor i urn as a potential biological control agent for Meloidoqyne arenarj a. Revue de Nernatologie 14 (1): 157-164.
Lei J, F.fl.O. M. De. , B. R. Kerry and J. ft. Dennchy (1992). The effect of fungal application rate and nematode density on the effectiveness of Vert ici 11 i urn ch larnydospor i urn as a biological control agent for Meloidoqyne incognita. Nematologica 38 (1) : 1 Icl-lclc:.
LeiJ, Frans De, G.D. Keith and B.R. Kerry (1992). The use of Vert ici 1 1 lurn ch 1 arnydosporiurn Goddard and Pasteuria penetrans (Thome) Sayre and Starralone and in combination to control Meloidoqyne incognita on tomato plants. Fundamental and Applied Nematolony 15(3):235-£4i2.
Lei J, F.fl.R. M. De., J. ft. Dennchy and B. R. Kerry (1992). The effect of temperature and nematode species on interactions between the nematophagous fungus Vert ici 1 \ lum chl arnydospor i urn and root-knot nematode (Meloidogyne spp.). Nematologica 3S(l):65-69.
Linford, M. B. (1937). Stimulated activity of natural enemies cif nematodes. Science 85:1£3-1£4.
66
Lirifoi-d, M. B. arid F. Yap (1939). Ror.t-knot nerncHtcde injnr-y re<3tricted by a fungus. Phytopathology £'9:5&9-809.
Lohde, G. (1874). Einige neuerr para<3 i.t isrichieri Pil::e. Tegcblatt der 47. versarnrnlung deutscher Nat uraf or^cher urid fierzie iri Bre5lau:£:03-£06.
Lohraann, U. and R.fl. Sikoi^a (1989). Mass production of the endoparasit ic fungi Drechsmer la conispora, Ver-t icl 1 iurn ba lane ides and Harpospon urn anaui 1 lulae in liquid culture. Nernatlogica 35(1):97-104.
Lysek, H. (1963). Effect of certain soil organisms on the eggs of parasite roundworms. Nature 199:9£5.
Lysek, H. (19S6>. Study of biology oF geohelrninths. II. The importance of some soil microrgamisms for the viability of geohelminth eggs in the soil, ficta Univ. Palacki. Olomuc 40:83-90.
Maas, P.W. Th. (1987). Physical methods and quaranatine, In: Principles and Practices of Nematode Control in Crops (Eds. R. H. Brown and B.R. Kerry), Academic Press, Australia, pp. S65-S91.
Maheswari, T., Uma and ft. Mani (1988). Combined efficacy of Pasteuria penetrans and Paecilomyces 111 acinus on the biocontrol of Meloidoqyne lavanica on tomato. International Hematology Network Newsletter 5(3):10-11.
Mani, ft. and C. L. Sethi (19a4a). Some characteristics of culture filtrates of Fusarium solani toxic to t^eloidoqyne incognita. Nematropica 14(£):139-144.
Mankau, R. (1961). The use of nematode-trapping fungi to control root-knot nematodes. Nematologica 6( ):3c:6-33c:.
Mankau, R. (1958). Reduction of root-knot disease witn organic amendments under semi field conditions. Plant Disease Reporter 52:315-319.
Mankau, R. (1975). Bacil lus penetrans n. comb. cau'smg a virulent disease of plant parasitic nematodes. Journal of Invertebrate Pathology £6:333-339.
Mankau, R. (1980). Biocontrol: Fungi as nematode control £\DentE>. Journal of Hematology 1£(4):£44-£5£.
Marban
67
Mendoza, N. , E. Garcia., M.B. Dicklow and B. M. Zuckerrnan (1992), Studies on Pf^eci lornycec rae rqua- idi i frctrn nematode suppressive Chmariipa soils. Jouri'ial of Chemical Ecology 18(5):775-783.
Meyer, S. L. F. (1992). Induction of ivicreased benornyl to ier^^ncs in Verbici11ium lecanii, a fungus antagonistic to plant pai^asitic nematodes. Journal of Helminthological Society of Washington 59(£):537-239.
Morgan—Jones, G. and R. Rodriguez—Kabana (19S4>. Species of Vert ici 11 lum and Paeci lornyces as parasites o-*" cysts arid root-knot nematodes. Phytopabhology 74(7):a3i.
Morgan-Jones, G., J.F. White and R. Rodriguez-Kabana (1983). Phytonematode pathology: Ultrastrucbural studies. 1. Parasitism of Meloidogyne arenarla eggs by Vertici11ium chlamydosporium. Nematropica 13:245-260.
Morgan-Jones, G., J.F. Uhite and R. Rodriguez-Kabana (1984). Phytonematode pathology: Ultrastrucbural studies. 11, Parasitism of Meloidoqyne arenaria eggs and larvae tay Paecilornyces 1ilacinus. Nematropica 14:57-71.
Mukta, S. S. , M.F. fizam. , A.M. Khan and S. K. Saxena (1991), Effc^ct of flsperqi11 us niger and Rhiroctonia sol a m on the development of Meloidoqyne incoqnita on tomato. Current Nematology 2(2):163-166.
Muller, H.G. (1958). The constricting ring mechanism of two predacious hyphomycetes. Transactions of British Mycological Society 41:341-364.
Nakasona, K. and J.T. Gaspard (1991). Effectiveness of two nematophagous fungi added to soil in v3tro. Proceedings of the Association for Plant Protection of Kyushu 37(3):126-129.
Naves, R. L. and V. P. Campos (1991). Dccur-rence of neinatode-trappmg fungi in south of Minas Gerais State (Brazil) and m vitv-o studies of the preadatory capcity and growth of some of their isolates. Nematologica Brasileira 15(2):152-162.
Nigh, E. ft. (1979). flcremoni um strict um and Fusar lum oxyspor i urn two fungal egg parsites of Heterodera schachiii the sugarbeet cyst nematode. Ph.D. Thesis. University oF California, Riverside, California, pp.79.
68
Noe, J. P. (1386). Croppii-ig sy'B>LE?riiB an.-iLys-is For Ijmjting lw-><-.t-z due to plant parasitic nematodes: guide to resear'.lT methodology. North Carolina States University Graphics, Raleigh, 18 pp.
Patel, R. R., D.J. Patel and B. ft. Patel (1331). Influence of organic amendments on growth and sporulation of nematophagous fungus Paeci lomyces 111 acinus (Thorn; Samson. Current Nematology d(1):57-40.
Prarner, D and N. R. Stoll (1353). Nernen: a morphogeriic substarice causing trap formation by predacious fungi. Science 1£9: 366-967.
Pria, M. Dalla., S. Ferraz and J.J. Muchovej (1331). Isolation and identification of nematode-predaceous fungi fr jm soil samples from various regions of Brazil. Nernatologica Brasileira 15 (£): 170-177.
Qadri, ft.N. and H.M. Saleh (1330), Fungi associated with Heterodera schacht 11 (Nematoda) in Jordan II. Effect <Z'ri H, schacht 11 and Mel oidoqyne .lavanica. Nernatologica 36:104-133.
Ramniah, ft. and H. Hii"schmann (1988). Me Lu idoqyne mayaqneriqis n.sp. (Meloidogynidae) root-knot nematode from Pureto Rico. Journal of Nematology £0(l):5a-69.
Rammah, ft. and H. Hirschmann (1330). Mel oidoqyne rnororrn en< >i s n.sp. (Meloidogynidae) in root-knot nematode from Morocco. Journal of Nematology c:i2: i=:73-c:91.
Reddy, P.P. and R.M.Khan (1331). Integrated management of root-knot nematodes infecting okra. Current Nematology a(£:) :115-116.
Rodriguez—Kabana, R., G. Godoy, G.Morgan—jones and R.ft. Shelby (1383). The determination of soil chitinase activity. Conditions for assay and ecological studies. Plant and Soil 75:95-106.
Rodriguez—Kabana, R. and G. Morgan—jones and R.ft. (1388). Potential for nematode control by microfloras endenn c in the tropics. Journal of Nematology £0(£):191-203.
Rodriguez-Kabana, R., J.W. Jordan and J.P. Hollis (1965). Nematodes: biological control m rice fields: role of hydrogen sulphide. Science 148:514-515.
69
Rohana, D. , B. Matwoto arid I. Djatnika (1987). Tha effo(:?bs cf application of Psea'i ] omyces 11 l3r::iriu' and populatiun levels of root-knot nematode (rnel'ji doqyne spp. ) and toarnto plant growth. Bulletin Penelitian Hort i kul tura 15(4):109-111.
Roman, J. and ft. Rodr'iguez—Marcano (1985). E F f tjct of the fungus Paeci 1 omyces 11 1 acinus ori the larval populations and root-knot formation of Meloxdoqyne incognita m tomabo. Journal of Agriculture of the University of Puerto Rico 6g(,2) : 159-167.
Roy, A.K. (1384). Effect of nitrogenous soil amendments on the parasitic activity of Caternarla anqui11ulae on root-knot nematode. Journal of Hematology 14 (c:) : 190-193.
Saifullah, M. Zulfxqar and ft. Gal (1990). Organic amendrnents aE control of root-knot nematodes. International Hematology Network Newsletter 7(l):£7-c:4.
Saikawa, M. and C. Morikawa (1985). An electron microscope stduy oF initiation of infection by conidia of Harpospon i irn subul 1 forme, Br\ endozoic nematophagous fungus. Transactions of the Mycological Society of Japan ^&^^):£i5-ai9.
Sarason, R.O. (1975). Paeci1omyces and some allied hyphomycetes. Studies in Mycology 6:1-119.
Sasser, J.N. (1977). Worldwide dissemination and importance o' the root-knot nematodes, Meloidogyne spp. Journal of Nematology 9:£6-29.
Sasser", J.N. (19SS). Relative importance and frequency of occurrence of various species, pathogenic variation and host races^ In: Proceedings of IMP Research Planning Conference on Root—knot nematode<-», Meloidogyne spp. (Region 111) Brasaillia, Brazil, pp.165-153.
Saxena, G. and K.G. Mukherji (1991). Distribution of nematophagous fungi in Varanasi, India. Nova Hedwigia 55(3/4):487-495.
Sayre, R.M. (1971). Biotic influences in soil environment. In: Plant Parasitic Nematodes, Vol.1 (Eds. B. M. Zuckerman, W. F. Mai and R. Pi. Rohde), Academic Press, New York, pp. £35-c:55.
70
Sayre, R. M. (1980). Promising org<ariJE>ms for bioconfcro] of rrernatodes. Plant Disease 64:526-53iE:.
Sayre, R.M. (19a&), Pathogens for biological control of nematodes. Crop Protection 5(4):£6S-£7£.
Sayre, R. M. (1988). Bacterial diseases of nematodes and their role in controlling nematode populations. flgr icult urt?, Ecosystems and Environment £4 (1-3) s 5&3-c:79.
Sayre, R. M. and M. P. Starr (1985). Pasteui-'ia penetrans <os-Thorne, 1940) nom. rev. comb. n.sp. n., a mycelial and endospore-f orrning bacterium parasitic in plarit parasitic nematodes. Proceedings of Helminthological Society of Washington 5£:149-165.
Scheuer, Ch. and J. Webster (1990). Dactyl e?! la arcuata sp. nov. , a nematode-trapping hyphomycete. Mycological Researcrh 94(5):7ia-7£0.
Shabana, T. Khan and T.fl. Khan (1992). Effect of culture filtrates of soil fungi on the hatching 3.rid mortality root-knot nematode (Meloidoqyne incognita) . Current Nematology 3(l):53-60.
Shahzad, S. and fl. Ghaffar (1987). Field application of Paecilomyces 1i1acinus and furadan for the control of root-knot disease of okra and mung. International Nematology Network Newsletter 4(l):33-34.
Shahzad, S., S. Ehteshamul-Haque and fl. Ghaffar (1990). Efficacy of Pasteuria penetrans and Paecilomyces 11lacinus for the biological control of Meloidoqyne .lavanica or\ mungbean. International Nematology Network Newslette."?r 7:34-35.
Siddiqui, Z.ft. and I. Mahraood (199£). Biological control of root-rot disease complex of chickpea caused by Meloidoqyne incognita race 3 and Macrophomma phaseolina. Nematologia Mediterranea £0(£):ISS-EOS.
Siddiqui, Z.fl. and S.I. Hussain (1991). Studies on the biological control of root-knot nematode. Current Nematology 2(l):5-6.
Singh, S.B., J.L. Smith., G.S. Sabins., ft.W. Dombrowski., J.M. Schaef f aer. , M. ft. Goetz and G. F. Bills (1991).. Structure and covifonnat ion of Ophiobolin K and 6~-epiophiobolin K from ftsperni1lus ustus as a nematicidal
71
agent. Tet rahedrori 47 (54) : 6931-&93a,
Sinha, B. K. , R. P. Nath and M. G. Haider C1977). Eff(:?<.t <:<f interact ion of Meloidcigyne incognita and Ozonj urn texanurn \3.r^. par a =5 it icurn on brinjal. Tndiai-i Journal <-jf Nematology 7(l):l-7,
Smith, G. S. , R. S. Hussey and R. W. Roncadon <1SB&). Penetration and post— infect ion development of Meloidonvne incognita on cotton as affected by Glomus mtraradices and phosphorus. Journal of Nematology 18(4):429-435.
Southey, J. F. (197Ba). Physical methods of control, In: Plant Nematology (Ed. J. F. Southey ). GDI Ministry of ftgriculture, Fisheries and Food, Her Majesty's Sbationery Service, London, pp. c:48-iE:61.
Southey, J.F. (197Sb), Regulatory controls m Plant Nematology (Ed. J. F. Southey ) GDI, Ministry of Ogncultur^e Fisheries and Food, Her Majesty's Stationery, London, pp. 3c.'6—335:.
Snvastava, S.S. and R. Dayal (1983). Fungal parasibes of nematodes Harpospor i urn. Ind i an Phytopathology 36 (i2) : 392-394.
Stenierding, S. (1963). Een mi xer-wal lenf i Iter methods on Vr 1J beweegl 1J ke endoparasit lare nematodem wit woi- lets to Verzamalen. Verls. Pliziekt. Diqnst. 141: 170-175.
Stir-ling, G. R. (1984). Biological control of Meloi dogyrre .lavanica with gaci 1 lus penetrans. Phytopathology 74:55-60.
Stirling, G.R. (igsS). Biological control of plant parasitic nematodes. Progress, Problems and Prospect's. Nematologica 38(3):392-394.
Stirling, G.R., M.V. MeKenny and R. Mankau (1970). Biological control of root-knot nematodes (Meloidoqyne spp.) on peach. Phytopathology 69(8):306-809.
Stirling, G.R., R.D. Sharma and J. Perry (1990). Attachment of Pasteuria penetrans spores to the root-knot nematode Meloidoqyne lavanica in soil and its effects on mfectivity. Nematologica 35 (2) :i246-c:5£.
Sunderland, K.D. (1990). The future for biological control. Professional Horticulture 4(l>:ll-50.
72
Taylor', O. L. and J.N. Sassei" (1978). BioJcigy, 3 dent i f lacb lori arid control of root-knob nematodes, Meloidogyne spp. II'iP Publication. Raleigh, North Carolina.
Taylor, fl.L., J.N. Sasser and L. ft. Nelson (1982). Relationshjp of climate and soil characber i&t ic<3 to geographical distribution of heloidoqyne species in agricultural soils. IMP Publication, Raleigh, North Carolina.
Thome, G. <1939). fl monograph of the nematodes of the superfamily Dorylaimoidea. Capita Zoologica S:l-i261.
Townshend, J.L. , M. Meskme and G. L. Barron (1989). Biological control of Meloidoqyne hapla on alfalfa and tomato with the fungus Mer3 a conispora. Journal of N'ematoloqy £1 K£) :179-183.
Tunlid, ft. and S. Jannson (1991). Protease and their involvement in the infection and imniobol izat ion of nematodes by the nematophagous fungus flrthrobobrys oliqospora. Applied and Environmental Microbiology 57 (10) : SB6a-c:87c:.
Tunlid, ft. and S. Jannson (1992). fl lect m—cvarDohydrate interaction leads to adhesion of nematodes to ftrthrobobrys ol1qospora. The first step in infection of the nematodes. Mycological Research 9&:401-41c:.
Tzean, S.S. and R.H. Estey (1981). Species of Phytophtnora and Pythi um as nematode-destroying fungi. Journal 'nf Nematology 13 (lE:) : 160-163.
Uziel, fl. and R.fl. Sikora (1992). Use of non-target isolates of the entomopathogen Vertici11ium leacni1 to control the potato cyst nematode Globodera pal 1ida. Nematologica 38(1):123-130.
Van Berkum, J.ft. and fl.R. Seshadri (1970). Some important nematode problems in India. Society of Nematoloijy Pascara. International Symposium, Europe.
Van Gundy, S. D. and M. V. McKenry (1977). flction of nematacidcs, In: Plant Disease- an Advanced Treatise Vol.1 (Eds. J. G. Horsfall and E. B. Cowling), Academic Pre5<3, New York pp.£63-283.
Villanueva, L. M. and R.G. Davide (1984). Ev.-iluation of several isolates of soil fungi for biocontrol of root—kn':'t nematodes. Philippine flgricul t ui-'ist 67 (4) :361-371.
73
Vinduska, L. <ig79). Funrji in Heteroderr^ schach-: i i. cysfc 3„ Ce&ka Mykologie 33:170-175.
Voss, B. and U. Wyss (1990). Variation bebween strains of the nernatophagous endoparasit ic fungus Cat ev^riAr 3 a anqui11ulae BoroUm 1. Factors affecting parasitism m vitro. Zeitschrift fur Pflanzenkrankheiten und Pflanzenschutz 97(4):41&-430.
Vos5, B. , P. Ukerrnohl and U. Uyss (199£). Variation between strains of the nernatophagous endoparasit ic fungus Caternarla anqu111u1ae Sorok1n II. Attempts to achieve parasitism of Heterodera schacht11 in pot trials. Zeitschrift fur Pflazenkrankeiten und Pflanzenschutz 99(3):311-31S.
Ualisa, K. and Gopal Swarup (1985). Effe3ct of some fungi on nematode hatching and larval root penetration. Indian Journal of Nematology 15 (i2) : 174-176.
Whitehead, ft. G. (1978). Chermcal control (a) soil treatmei'it. In: Plant Nematology. 3rd Ed. (Ed. J. F. Southey) GDI, Ministry of Agriculture, Fisheries and Food, Har Majesty's Stationery Service, London, pp. c:S3-£96.
Whitehead, ft. G. and fl. L. Wxn-field (1978). Chemical control, In: Plant Nematology (Ed. J. F. Southey), GDI, Ministry of figriculture, Fisheries and Food, Her Majesty's Stationery Service, London.
Wyss, U. , B. Voss and H. B. Jansson (1992). In vitro observation on the infection of Meloidoqyne incognita eggs by the zoospore fungus Catenaria anqui11ulae. Fundamental and Applied Nematology 15 (i2): 133-139.
Zaki, F.O. and D. S. Bhatti (1990). Effect of castor (Ricmus communis and the biocontrol fungus Paecilomyces 111 acinus on Meloidoqyne .lavanica. Nematologica 36(1):114-1 D r»
Zaki, M.J. and M.fl. Maqbool (1992). Effect of spore concentraions of Pasteur 1 a penetrans on the attachmei'it of Meloidoqyne larvae and growth of okra plants. Pakistan Journal of Nematology 10(l):59-73.
Zaki, M.J. and M.ft. Maqbool (1992). Effect of Pasbeuria penetrans and Paeci lomyces penetrants on the control cf root-knot nematodes on brinjal and mung. Pakistan Journal of Nematology 10(l):75-79.
74
Zavaleta-Mejia E. (1985). The effect of soil bacteria on Meloidogyne incoqriita (Kofoid and White) Chit wood infection. Dissertation Plbstracts, A&(4>:10ia.