Seven different strains of Trichoderma are isolated from wilt infected leguminous crops of an Indian state and tested for their antagonistic activity against Fusarium (soil borne pathogen) which is expressed as a zone of inhibition in the culture plates. The seven strains are identified as Trichoderma viride, T. harzianum, T. asperellum, T. koningii, T. atroviride, T. longibrachiatum, and T. virens. Upon successful identification, morphological description and sequencing of the isolated strains with the help of universal ITS primers, the sequences are submitted to NCBI and allotted with the accession numbers JX119211, KC800922, KC800921, KC800924, KC008065, JX978542 and KC800923, respectively. Genetic variability studies reveal that a percentage of polymorphism in SSRs is obtained within the seven strains of Trichoderma species which is comparatively higher (>77%) than with RAPD primers (~50%). This study aims at selecting the best strain of Trichoderma species (Trichoderma viride 01PP) and then preparing a simple bioformulation that is cheap, easy to apply and readily accessible to the farmers. Shelf life of the prepared bioformulation is even checked for 180 days and it is concluded that the number of propagules start declining from 30 th day onwards when the bioformulation is prepared in talc as a carrier material.

The study was aimed to carry out experiments to determine the optimal parameters for the biomass production of Trichoderma. It is quite essential to determine the physical conditions that are favorable for the growth of Trichoderma species. The seven species under study have been isolated from the rhizospheric soils of chickpea; pigeon pea and lentil crops of different areas of an Indian State (Uttar Pradesh) and these were later tested in vitro at different pH, temperatures and varying agitation speed. A significant difference in the biomass production was recorded among the species at tested pH levels i.e. 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5 and 8.0. The most favorable pH ranges between 5.5 and 7.5 in which total dry weight of mycelium varies between 1.41 and 1.35 g. Although all the species of Trichoderma produced sufficient biomass at different temperatures viz. 20°C, 25°C, 30°C and 35°C but they were found to be best grown at a temperature range of 25°C to 30°C. Aeration by agitation was also checked at different speeds such as 100, 150, 200 and 250 rpm but greatest biomass was recorded at 150 rpm. Preparation of standardized inoculumsSpore suspensions were prepared by adding 15 ml of sterile distilled water to mature (4-5 days) fungal colonies on PDA plates to dislodge the spores from the mycelium. The spores were counted using a heamocytometer (Neubauer, Germany) to obtain a spore concentration

The phylogeny of Trichoderma and the phylogenetic relationships of its species was investigated by maximum parsimony analysis and distance analysis of DNA sequences from multiple genetic loci 18S rDNA sequence analysis suggests that the genus Trichoderma evolved at the same time as Hypomyces and Fusarium and thus about 110 Myr ago 28S rDNA sequence analysis shows that the genus Trichoderma is part of a monophyletic branch within the Hypocreaceae. Most isolates of the genus Trichoderma were found to act as mycoparasites of many economically important aerial and soil-borne plant pathogens. Trichoderma has attained importance as a substitute for chemical pesticides and hence an attempt was intended to corroborate the positive relatedness of molecular and morphological characters. Two fungal strains, Trichoderma koningii T k-5201/CSAU and Trichoderma virens T vi-4177/CSAU were isolated from a soil sample collected from CSA Farm, Kanpur district of Uttar Pradesh, India. The universal primers (internal transcribed spacer, ITS) were used for the amplification of 18S rRNA gene fragment and strains were thus characterized with the help of ITS marker. It is proposed that the identified strains T. koningii T k-5201/CSAU and T. virens T vi-4177/CSAU be assigned as the type strains of a species of genus Trichoderma based on phylogenetic tree analysis together with the 18S rRNA gene sequence search in Ribosomal Database Project, small subunit rRNA and large subunit rRNA databases. The sequence was deposited in GenBank with the accession numbers KC800923 and KC800924, respectively. Thus an integrated approach of morphological and molecular markers can be employed to identify a superior strain of Trichoderma for its commercial exploitation.

In this study, potential and effective strains of Trichoderma such as T. harzianum (Th. Azad) and T. viride (01PP) have been investigated and their effect of pre sowing seed treatment on germination, seedling establishment, seedling dry weight and vigour in chickpea genotype (Radhey) was observed. The different pre sowing seed treatments showed different responses against all seven seed quality parameters. Chickpea seeds were treated with different concentrations of Trichoderma bioformulation such as 5%, 10%, 20% gm/kg seed followed by treatment with 0.2% Bavistin. As a result, the percentage of seed germination was found to be higher in T. harzianum (Th. azad) & Trichoderma viride (01PP) treated seeds with 5% bioformulation as compared to the other concentrations. Various attributes with their observations include seed germination (92% and 90%), root length (12.38 and 12.19 cm) shoot length (4.97 and 4.32 cm) seedling length (17.38 and 16.50 cm), dry weight (1.19 and 1.88 cm), vigour index I (5197.12 and 1485) and vigour index II (109.48 and 169.20). Among all treatments, control showed the poorest performance for all seven seed quality attributes.

Trichoderma species are morphologically very similar and were considered for many years as a single species. A new strain Trichoderma longibrachiatum 21PP was isolated from a rhizospheric soil sample collected from Kaushambi district of an Indian State (Uttar Pradesh). The universal primers (ITS-1& ITS-4) were used for the amplification of 28S rRNA gene fragment that produced a sharp band of about 700 bp on the gel. The amplified gene fragment was then sequenced (664 bp) and then deposited in GenBank with the Accession No. JX978542. Thus, the molecular identification of the specified strain enabled us for further characterization as this strain of Trichoderma species can be used as a biocontrol agent against pathogenic fungi such as Fusarium, Alternaria, Sclerotinia, etc. It is proposed that the identified strain T. longibrachiatum 21PP be assigned as the type strain of a species of the genus together with the 28S rRNA gene sequence search in Ribosomal Database Project, small subunit rRNA and large subunit rRNA databases.