Vipul Kumar

Optimal Physical Parameters for Growth of Trichoderma Species at Varying pH, Temperature and Agitation

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.

Trichoderma Genome to Genomics: A Review

Trichoderma species are widely used in agriculture as biopesticides. These fungi reproduce asexually by production of conidia and chlamydospores and in wild habitats by ascospores. Trichoderma species are well known for their production of enzymes called Cell Wall Degrading Enzymes (CWDEs). All living organisms are made up of genes that code for a protein which performs the particular function. Some genes that play an important role in the biocontrol process are known as the biocontrol genes. These genes send some signals which help in secretion of proteins and enzymes that degrade the plant pathogens. These biocontrol genes can be cloned in huge amounts and can be used on large scale for commercial production. Some Trichoderma genes are also helpful in providing resistance to the biotic and abiotic stresses such as heat, drought and salt .The major biocontrol processes include antibiosis, mycoparasitism and providing plant nutrition.

To Develop Strain Specific Molecular Marker for Easy and Proper Identification of Fungal Species Based on Molecular Characters: A Review

The sequencing of fungal genomes is advancing at breakneck-speed, producing voluminous amounts of data. Within the next five years, it is possible that over a couple thousandgenomes, representing every major fungal family will be completed and available to the scientific community. In order for this data to have a truly transformative effect on mycological and other research, however, several factors need to be addressed. These include; (1) the establishment of user friendly platforms for examining, sorting, and sifting through the genomes, (2) integration, or at least cross-communication, between the various databases that house the genomic data, and (3) investment in community resources that can act as repositories for and provide materials to researchers, i.e. strains, clones, plasmids, etc. The frameworks for some these needs, e.g. the materials available from the Fungal Genetics Stock Center (FGSC, University of Missouri), are already established and should be reinforced, whereas for others, e.g. data accessibility, the sooner that a plan can be implemented the better. The Fungal Kingdom is considered to contribute greater than 15% of the species richness found in the major groups of organisms .This study is a reflection of the usefulness of sequence analysis of the 28S ribosomal RNA gene in identifying fungal as well as determining fungal diversity. Various techniques that are based on utilizing the 28S rRNA have been discussed. Of critical importance is the manner in which massively parallel sequencing was exploited to correct the under representation of fungal species in compilations of fungal hat were drawn using traditional methods of surveying fungal species from ecosystems.

Genetic Determination of Potential Trichoderma Species Using ISSR(Microsatellite) Marker in Uttar Pradesh, India

Seven Trichoderma sp. were collected from different locations of Uttar Pradesh, India for evaluating their bioefficiency by determining their genetic variations. PCR-based Inter Simple Sequence Regions (ISSR) Marker employing 6 primers produced 30 scorable bands out of which 27 bands were polymorphic. The Unweighted Pair Group Method of Arithmetic Means (UPGMA) dendrogram constructed from Nei’s [14] genetic distance produced 2 main clusters (1 isolates in cluster 1 and 6 isolates in cluster 2). The result indicating their genetic diversity has opened new possibility of using the most efficient and more isolates of Trichoderma in the preparation of effective biopesticide.

Phylogenetic diversity analysis of Trichoderma species based on internal transcribed spacer (ITS) marker

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 Tk-5201/CSAU and Trichoderma virens Tvi4177/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 Tk-5201/CSAU and T. virens Tvi-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.

Trichoderma species Cellulases Produced by Solid State Fermentation

The main aim of this study was to analyze eight species of Trichoderma for cellulase enzyme production by solid state fermentation. Different carbon sources such as wheat bran, corn cob, sucrose, maltose and filter paper were used. Highest celluase enzyme production was achieved with T. harzianum on media supplemented with corn cob. The optimum pH, temperature and thermal stability of isolated enzymes were also analyzed. The best pH for enzyme production was found between 4-6. The optimum temperature range for cellulase production ranged between 30-40°C. Choosing the optimum pH, temperature and best carbon source are essential for the enzyme production. Compare to other fungal genera it has been found that Trichoderma spp. have the greater potential to synthesize cellulase enzyme.

Enhancing Seed Germination and Vigor of Chickpea by Using Potential and Effective Strains of Trichoderma Species

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.

Comparative Study of Biological Agents, Trichoderma harzianum (Th-Azad) and Trichoderma viride (01PP) for Controlling Wilt Disease inPigeon Pea

The paper aims to study the morphological, physiological, molecular characterization and bio-formulation of Trichoderma harzianum (Th Azad) and Trichoderma viride 01PP-8315, an effective fungicide and a biological control agent too that protects the plants and seeds from other pathogenic fungi. The physiologic study is done in an attempt to find the effective management of the disease caused by soil borne pathogens. Trichoderma harzianum (Th Azad) and Trichoderma viride 01PP-8315 isolated from the infected soil samples of Pigeon pea fields and grown at preferable temperatures, pH and different solid and liquid culture media. Most preferable temperature for the growth and sporulation of Trichoderma harzianum (Th Azad) and Trichoderma viride 01PP-8315 has been observed up to 30oC (210.5 mg dry weight of mycelium). A detailed morphology of the strain is done in this study including colony growth rate, colony color, colony edge, mycelial form, conidiation, conidiophore branching, conidial wall, conidial color, etc. The molecular characterization of the strain is carried out using an 18S rRNA gene sequence with the help of a universal Internal Transcribed Spacer marker that gives an amplicon of a total of 1173 base pairs and 546 bp of the 18S rRNA gene was sequenced and used for the identification of isolated fungal strains that is later sequenced and allotted with Gene Bank Accession no. JX119211 and KC800922 respectively. To check the presence of endochitinase gene in two of the potential strains of Trichoderma species viz. T. harzianum (Th Azad) and T. viride 01PP an ech42 primer was used. A talc based bio-formulation has been prepared with this strain where the population of the spores was found to decline after 180 days.

Effect of Biopriming with Biocontrol Agents Trichoderma harzianum (Th.Azad) and Trichoderma viride (01pp) on Chickpea Genotype (Radhey)

Chickpea is an important pulse crop of India. Its productivity is quite low due to several biotic and abiotic stresses. Among the biotic stress disease are the major constraints. Wilt caused by Fusarium oxysporum f. sp. ciceri has been considered as devastating one to cause up to 10 per cent loss in yield every year. For eco-friendly and sustainable management of the disease, two species of antagonists (Trichoderma viride 01PP and Trichoderma harzianum Th azad) and chemical fungicide (Bavstin @ 0.2%) were evaluated against the pathogen. Two bioagents Trichoderma harzianum Th azad and Trichoderma viride 01PP were evaluated for their efficacy on colony growth by dual culture plate method. The results showed that the two bio-agents suppressed the colony growth of Fusarium oxysporum f. sp. ciceri, which ranged between 53.38-57.99 per cent the suppression of the growth of the pathogen was significantly higher with Trichoderma harzianum Th azad. Seed treatment in vivo showed that out of six treatments including control, T1 (5%) followed by T2 (10%), T3 (20%), T4 (2% @ 2.5 kg h-1), T5 (Bavistin @ 0.2%) and T6 (Control) were found superior seed treatments in enhancing quality seed parameters (germination, plant death on different dates, plant survival and yield), which can be finally converted in superior yield even in adverse conditions. T1 treatment (5%) was found to be significantly superior and effective in increasing 79% and 71.67% respectively (in both strains) more germination from control followed by bavistin (T5). Results of the study show that bio-agents significantly reduced the wilt incidence, and increased seed germination and plant growth parameters as compared to chemical fungicides.