The Microbiology and the Integrated Pest Management (IPM) group focuses on research, development and dissemination of ecofriendly technologies through innovative delivery
models to promote sustainable agricultural practices. The research focus is technology
specific (eg. biofertilisers, biopesticides, and bio control agents) and need based to address problems encountered at farm hold level. The group also focuses on extensive demonstrations and trainings in the areas ofbio-pesticides, biofertilisers, soil testing, and
integrated nutrient and disease management to support to the farmers. Some of these
demystified technologies have also been set up as community managed enterprise. Soil
analysis is also undertaken as part of the soil health assessment by analyzing the chemical
and biological properties of the soil and a soil health card has been developed.
Screening for novel microbes and their beneficial functions
Bioprospecting for saline tolerant, nitrogen fixing, phosphate solubilising and plant
growth promoting microbes from the pristine agriculture and mangrove ecosystems to
be used as potential biofertilisers and biocontrol agents in agriculture practice in hear,
normal and stressed soils.
Diversity of Pseudomonas from the saline Agroecosystems: The diversity of Pseudomonas populations isolated from three different plant rhizospheres, namely paddy,
pearl millet and cotton grown in saline soils along the coastline of Southern India was
studied. The genetic diversity was analyzed using molecular tools, Random Amplified
Polymorphic DNA (RAPD) and Polymerase chain reaction Restriction Fragment Length polymorphism (PCR-RFLP). The biochemical characterization, antibiotic resistance
assay and RAPD profiles revealed the presence of a largely homogenous group and
the PCR-RFLP restriction showed two groups predominantly present in all the three
rhizospheres, while the minor group consisted of salt-sensitive isolates restricted to the paddy rhizosphere alone. Increasing salinity caused a predominant selection
of salt tolerant species, in particular Pseudomonas pseudoalcaligenes and Pseudomonas alcaligenes, irrespective of the host rhizosphere. The diversity of Pseudomonas associated
with the finger millet rhizosphere was studied by amplification of the 16S rDNA using
Pseudomonas genus-specific and GTG(S)primers. Free tree V1.6.6. using Nei and Li/Dice
similarity matrix calculation and clustering was done by UPGMA algorithm.
Diversity of Azospirillum from the saline rice ecosystems: The diversity of the
Azospirillum associated with rice fields of the coastal agro ecosystem was estimated by PCR RFLP of the 16S rONA. Among the 302 isolates tested 19 restriction pattern were observed. The frequency of occurrence of salt tolerant spp. of A. brasilense was high than A. lipojernum, which was sensitive to salinity, thereby indicating high association between the salinity and distribution of the Azospirillum in the rice ecoysytems.
Genetic Diversity of Bradyrhizobia isolated from different crops: The genetic diversity
of Bradyrhizobial strains associated with blackgram and cowpea grown in two different agricultural soils (non-saline and saline) along the coastline of Tamil Nadu has been
analyzed. DNA polymorphism of phenotypically indistinguishable isolates was analyzed using RAPD of 16S rONA and nif D, which showed
considerable genetic diversity. Soil pH and salinity
had an effect on the selection of natural populations
as revealed by PCR-RFLP of 16S rONA. A group of
strains and a type strain sharing three different 16S
PCR-RFLP types were shown to have the same set
of symbiotic genes as inferred from the PCR-RFLP
pattern of nif D. The diversity of bradyrhizobia
associated with the Soybean (Glycine max Merrill.) was studied using restriction fragment length
polymorphism (RFLP)and phylogenetic analysis. Cluster analysis of16S rONA restriction
patterns with seven tetrameric endonucleases grouped these isolates with Bradyrhizobium spp. resolving two genotypes within these Bradyrhizobia and the intergenic spacers (IGS)
and RFLPs with three restriction enzymes formed six genotypes. The IGS sequences
of the strains of Indian origin have very low similarity to those of the type strains of
validly described species of Bradyrhizobiwn from other legumes and also from soybean.
The RFLPs of symbiotic genes (nif H and nod C) and phylogeny based on the nif H DNA
sequence delineated all isolates into two biovarieties. It clearly showed the existence
of the potential new biovariety among soybean nodulating bradyrhizobia. One strain
designated M6 can be proposed as a novel sp. in the genus Bradyrhizobium.
Novel genus Swaminathania salitolerans gen. nov. spp. nov. from the mangroves: Screening for microbial strains, from mangrove-associated wild rice (Porteresia coarctata- Tateoka), resulted in the isolation of a number of strains belonging to the taxa Swaminathania, Vibrio, Bacillus, Enterobacter
and Azospirillum. Novel salt tolerant strain Swaminathania salitolerans gen. nov., sp. nov.
isolated from P. coaractata were found to fix
nitrogen and solubilize phosphate and showed
intensive colonization in Pokkali than in P.
caroctata and Ponni. Phylogenetic analysis based
on their 16S rRNA genes showed homology
to the genus Acidomonas, Asaia, Acetobacter, Gluconacetobacter, Gluconobacter and
Kozakia in the Acetobacteraceae. Like
wise a diazotrophic strain MSSRF40T
was identified as new genus in the
family Enterobacteriaceae for which Mangroveibacter plantisponsor gen. nov.
sp. nov., was assigned.
Novel Vibrios associated with
Four novel species of the genus Vibrio sola ted from the mangrove rhizosphere
were named as Vibrio rhizosphaerae sp. nov., Vibrio porteresiae sp.nov., Vibrio mangrovi sp.nov and Vibrio plantisponsor sp.nov is proposed. Following the polyphasic taxonomic approach. Multilocus sequence and phylogenetic
analysis using 168 rRNA, recA, pyrH, 'poA and nifH gene sequences, PCR based genomic
fingerprinting using PCR-RFLP of 168 rRNA,
ribotyping and repetitive extragenic palindrome
(rep)-PCR using GTGs and BOX primers. Fatty
acid analysis, biochemical characterization,
G+C content analysis and whole genomic DNADNA
hybridization, has identified these Vibrio strains
and also defined a new ecological niche for vibrios as a
rhizosphere-associated heterotrophic diazotroph.
Diversity of actinomycetes from different ecological niches
The diversity of the actinomycetes from different
ecosystems have been isolated and efficient
phosphate solubilising strains have been identified.
The isolation of DNA has been standardized and the
amplification of the 16 S rDNA was carried using
fD1 (5'-AGTTTGATCCTGGC TCAG-3') position7-
26 and r P2 (5'ACGGCTACCTTGTTACGAC TT-3')-
position 1513 to 1494 primers. The initial genetic diversity analysis of antagonistic actinomycetes was performed using GTGs primer, which showed
4 different genotypes of actinomycetes. A further study on the characterization of these
genotypes is under progress.
Metagenomic of the unculturables from the mangroves: The diversity of unculturable microbes associated with the rhizosphere of five different mangrove spp. viz., P.coarctata, Rhizophora mucornata, Avicennia marina, Sonnerratia apetla and Excoecaria agallocha collected from Pichavaram mangroves, South India was analyzed using molecular tools like RFLP, Amplified Ribosomal DNA Restriction Analysis (ARDRA), and Single-Strand Conformation Polymorphisms (SSCP). The DNA from the soil was extracted using Soil Master™ and the 16S rRNA was amplified using Coml and Com2-Ph primers. Single stranded DNA was obtained by lambda exonuclease digestion. The SSCP pattern revealed that majority of the dominant populations were closely related to previously
described soil bacteria (i.e., Planococcus, Planomicrobium, Bacillus, and low G+ C Grampositive
bacterium, and glacial ice bacterium).
Biological control of Phytopathogens using PGPRs: Different group of organisms viz., Pseudomonas, Bacillus and actinomycetes were
screened for their antagonistic activity against
Rhizoctonia solani; Macrophomina phaseolina,
Pyricularia oryzae and Xanthomonas oryzae etc. to isolate efficient strains with antifungal and antibacterial activity has been identified.
Antagonistic Pseudomonas was selected using Pseudomonas primers. The diversity of the
antibiotic coding genes for DAPG, PLT and PRNT were determined using specific primers.
About 48% of the Pseudomonas strains showed positive amplification for DAPG gene of size
with DAPG primers Phl2a and Phl2b and 16% of the strains show positive amplification
for HeN primers PM2 and PM7..
Bioprospecting for metabolites from microbes collected from the
The demand for new effective antibiotics to combat the development of resistance by microbes to the existing drugs. Discovering novel biomolecules using the untapped
resources has gained momentum and the modern biodiversity prospecting integrates the systematic search for the new sources of biomolecules, genes and other economically
valuable natural products. The project involves nine institute and Nicholas Piramal
India Limited. This project envisages a mega scale screening for microbes from different ecological niches. In the present study screening for metabolites from microbes isolated
from soil samples collected from different ecological regions of the Eastern Ghats and
the mangrove ecosystem will be in 30 different media and a total of 1000 cultures/ month
(total of 13, 000 cultures) were isolated to purity and the colony characteristics viz., colony
size, configuration, margin, elevation, colour, mucilage, biofilm formation and pigment
production were recorded. The cultures were stab inoculated and dispatched to Nicholas
Piramal Limited (NPL) Mumbai where the isolates were screened for novel biomolecules
with anti-cancer, anti-diabetes, anti-inflammatory and anti- infectivity activity by HITS
(High infectivity throughput screening tests). Two sets of glycerol vials were stored in
cry preservation vials at -80°C. And another set of stab was sent to National Centre for
Cell Sciences (NCCS) for storage.
Development of biological software for good agricultural practices
Trials were conducted in the coastal agriecosystems in farmer's field in Chidambaram
to study the performance of the phosphate solubilizers and nitrogen fixers under field
conditions. The phosphobacterial strains, viz., PS-5 and PS-10, nitrogen fixers MSA- 274 and MSA-160 and S. salitolerans strains, PA-6 and PA-51 were used as these strains were
proved efficient in the green house conditions. The trial was conducted in four different
locations covering 207 acres. Biofertilizers formulated in vermiculite were used. Three
treatments viz., seed bacterization, root dipping and foliar application (in intervals of 30 days) were given. These treatments resulted in an increase in the yield parameters
of 20-30% compared to the chemical control. The Pseudomonas strains MSP-393, MSP27;
MSP147, MSP573 were tested for the control of bacterial blight disease and sheath blight
disease. Toxicological analysis of the strain MSP 393 proved it to be non toxic by HBAT.
Talc and Liquid based formulations: Talc and liquid formulation of the biofertiliser
and the biocontrol agents were standardized. Third party testing of the talc formulations
in the field was carried by Elbitech. The liquid formulations were standardized using
ifferent cell protectants and preservatives and the shelf life of the product was stable up
to six months of storage and the CFU was maintained upto 4x109 and hence proved to be
superior to the talc based formulations. The performance of the liquid formulations in
the field is being evaluated by a trial at the Biocentre in Puducherry.
One of the innovation areas is the
demystification of technologies and
establishing low cost units for the
production of the biological software's
for the promotion of good agriculture
practices as part of promotion of ecoagriculture
and organic farming. The ecoenterprises like Trichoderma viride, Trichogramma and Pseudomonas fluorescens and biofertilisers like Azospirillum and phosphobacteria are successfully management by the women SHGs. To improve their technical skills
representatives from the Jhansi Rani WSHG and Manimegalai WSHG were given hands
on training in the production of Azospirillum and phosphobacteria in the microbiology
lab at MSSRF. Two decentralized units have been
set up one at Kannivadi, Dindigul district and
the other a Kezhsathamangalam in Puducherry. The project interventions helped to enhance the awareness among men and women farmers of the
region on different kind of biofertilizers and its crop
specificity over a period of time
Integrated Pest Management
IPM group is involved in bioprospecting, mass
multiplication of promising biopesticides, field
testing and establishing micro enterprises and
demystification of technologies.
Biological control of Helicoverpa armigera using Cipadessa baccifera, Clausena dentata, Dodonaea angustifolia and Melia dubia (at laboratory &field level) was carried out. Different
solvent as well as aqueous extracts of the traditional plants were tested against different
stages of H. armigera. The adult longevity, eggs laid and hatchability was recorded in the adult, while the pupation, larval duration, pupal weight and adult emergence were
recorded in the larvae. The adult life span of H. armigera was reduced by petroleum ether and chloroform extracts of D. angustifolia, acetone extract of M. dubia and chloroform
extract of Clausena dentata. Among the test plants, D. angustifolia reduced the fecundity
compared to others. The egg hatchability was suppressed by many of the treatments,
despite the induction in egg laying.
Pupation was least and malformed adults were high in petroleum ether extract of
Cipadessa baccifera, acetone extract of Clausena dentata, hexane extract of D. angustifolia and chloroform extract of M. ubia. Life span was less in adults emerged from acetone
and petroleum ether extracts and solvents (Cipadessa baccifera), hexane extract further
fed with normal diet (Clausena dcntata), acetone extract (D. angustifolia) and chloroform
extract (M. dubia).
Efficacy of D. angustifolia extract against H. annigera on rose and
Earias vitella in cotton under field conditions
There was a substantial decrease in the eggs, number of larvae and percent damage of
buds and flowers after 4th spray of extract. Hexane and acetone extracts were highly
effective, and on par with NPV. The compound responsible for insecticidal activity
against H. armigera in leaf extract of Argemone mexicana was identified as 3-hydroxy-2-
(15-methylhexadecyl)-1-cyclopentanone and from Clausena dentata as ((23E)-23, 24,27-
trimeth yl-octocos- 23-enoicacid -[(2E)-4-methyl-pen t-2-enoil-oxy -methyl ester).
Mass multiplication of biopesticide
The multiplication of three species of Trichogramma namely T. chilonis, T. japonicum and T.
brassicae has been standardised for crop specific problems. The group has developed low
cost technology for mass multiplication
of entomopathogens, Beauveria bassiana,
Metarhizium anisopliae, Verticillium lecanii,
Nomureae rileyi and fungus Paecilomyces
lilacinus. Different substrates viz., rice,
sorghum, barley, groundnut, potato, beetroot, carrot, sawdust and paddy
straw were used for the mass production
of the entomopathogens. The addition
of 1% (w/ w) yeast extract (YE) in all
the substrates enhanced the sporulation
of B. bassiana as against the control.
Among the grains tested, the maximum
sporulation of B. bassiana was observed
on rice supplemented with 1%YE (117.0
x 108spores/ g).
Field testing and Establishment of micro enterprises (Extension)
Trichogramma units at Kannivadi (2 nos.), Puducherry (1 no.) and Pudukottai (3 nos.) and
mushroom units at Puducherry (2 nos.) has been established.
Mass production technology of Trichogramma has been demistified and transferred the
technology to WSHG. The crop calendar based production coupled with quality has
been a positive step to manage these notorious pests. The parasitoid production centers
are successfully run by seven groups.
The trainees have now become trainers and the horizontal transfer of knowledge and technology through well-planned objectives and execution has promoted capacity building, leadership quality, employment generation, reduced pesticide usage, higher cost benefit ratio and sustainable agriculture in the villages.
MSSRF has imparted training to one
woman SHG at Kannivadi, Dindigul
District, in the different technologies
involved in rPM, based on the cropping
pattern and the pest incidence in their area.
After one year of initiation, the concept
has spread into other places and the
pioneer group members started training
six groups and now each group run this
as a micro enterprise through which
they reach the farmers in and around
their area and create awareness about
this particular parasitoid and its control.
They have proved this as an economically
and environmentally viable enterprise,
indirectly reduced the usage of chemical
pesticides by 35% in their area.
Edible mushroom is a healthy food and
its production is promoted as a micro
enterprise. Six women and three youth
SHGs benefited from the training imparted by the IPM group. Other groups who were
restricting to savings were also attracted towards such training and expressed their
willingness to participate in the future.
Technologies taken to the field from us and adopted as practiced by others: Use of
Trichogramma chilonis in the management of a wide range of pests; Mushroom production
(oyster and milky); Integrated pest management packages such as multiple cropping,
crop rotation, use of light traps, mechanical removal of pests etc; Spraying botanicals as a
preventive tool against sucking pests, defoliators and borers and use of entomopathogens
in the management of field problems at Thonimalai are among the technologies taken
to the field.
Demonstration of the efficacy of entomopathogens against various insect pests at Thonimalai under organic farming.
Support to organic farming at Thonimalai by way of field survey, identification of pests
and diseases were provided with guidance to application of IPM packages suitable to the cropping system. The major pests and diseases observed and the IPM practices followed
at Thonimalai are given below.
Banana pseudostem weevil: Among the 245 trees in the 2 field, 85 trees were
infested with banana pseudostem weevil. Prophylactic pseudostem injection of neem oil
coupled with soil application of neem cake was carried out to prevent the incidence of
banana pseudo stem weevil. 146 pseudo stem traps + 2.6 Kg of B. bassiana pasted traps
Surveillance of insect pests of Morinda citrifolia in the west coast of
Periodical visits to west coast of Kerala and Karnataka are made to conduct survey in Morinda citrifolia and M. tinctoria (natural habitat as well as farms). The pest population
if any on the above species in different seasons is recorded and documented. The major pests recorded till date are Membracid bugs, Coccinellid, Valley carpenter beetle, Plant hopper, Grasshopper, White fly, Green plant bug, Predatory bug, Mantis, Fruit fly,
Weaver ants, leaf hopper.
Demystification of technology:
The mass multiplication
procedure of parasitoids,
Trichogramma chilonis, and Tetrastichus sp.; predators, green lacewing, Chrysoperla carnea, and aphid predator,Cryptolaemus montrouzieri were demystified. The objective is to reduce the cost, simplify
the methodology and develop a user friendly model so that it can be converted into a
Soil testing helps to diagnose soil health and evolve soil specific and crop specific solution. Soil testing is the basis for scientific management of fertilizer requirements.
The basic objective of the soil-testing programme is to educate farmers on judicious
and economic use of fertilizers and better soil management practices for increasing agricultural production based on the soil nature. Soil health cards have been developed and are being promoted in all sites especially in the watershed sites and places where good agriculture practices are being promoted. GIS based tools are being used to map the
soil profiles and as monitoring tools. So far @ 5000 soil samples from different sites have
been analysed for pH, EC, organic carbon, macro nutrients (N, P and K), exchangeable cations (Ca, Mg, Na & K) and micro nutrients (Cu, Zn, Fe & Mn). 67 water samples from
Jeypore and Puducherry were analyzed for pH, Electrical conductivity, anions, cations,
SAR and RSC. Developed a soil health card for the benefit of farmers.
Biological properties of the soil
The microbial population plays an important role in the maintaining the soil health. The
microbial population and the different enzyme activity related to the microbial activity
has also been estimated. 691 samples from project sites were analysed for dehydrogenase
and phosphatase activity. 180 soil samples from Pudukottai, Puducherry and Jeypore
were analyzed for microbial load. Six phosphate solubilizers and 12 nitrogen fixing
positive strains were isolated and sub cultured for future studies.