Isolation, Molecular and In-silico characterization of Trichoderma spp. from Rhizospheric Soil Sample

Author: Vishwajeet Yadav, Mukesh Kumar, R.S. Sengar, Pushpendra Kumar, M.K. Yadav and Vyankatesh Dhanraj Bagul

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Abstract

Trichoderma, soil-born filamentous fungi are capable of parasitizing several plant pathogenic fungi, it is known as biocontrol agents. Five isolates of Trichoderma spp. isolated from various crop rhizospheric soil in Uttar Pradesh districts were characterized by their cultural, morphological, and molecular level. The isolates differed significantly in terms of colony traits, sporulation, branching of conidiophores, and the colour and shape of phialospores. Molecular analysis of the isolates was done by sequencing the ITS region of ribosomal DNA using specific universal primers ITS 1 and ITS 4. Multiple nucleotide alignment of ITS 1, ITS 4 and 5.8s region depicted intra-specific and inter-specific variations in the ITS sequences among the different Trichoderma species. The result of this research obtained at morphological and ITS-based rDNA region sequencing. The sequencing of these five isolates revealed that the Trichoderma were characterized into Trichoderma lixii (Acc. No. OP031646), three Trichoderma harzianum (Acc. No. OP104445, OP104449, OP104451) and Trichoderma sp. (Acc. No. OP104454). However, significant percentage identity between the known isolate of T. lixii from the database and T. lixii (TBT-13) isolates from wheat is 97% whereas the percentage between the known isolate of T. harzianum and three species of T. harzianum (TBT-14, TBT-15, TBT-16) from potato, wheat, and mustard was 97-98% respectively. Similarly, Trichoderma spp. (TBT-17) are 98% identical to Trichoderma spp. result after NCBI-BLAST. As a result, the study was found to be helpful in identifying Trichoderma spp. from the rhizospheric soil of various crops.

Keywords

Trichoderma, Biocontrol, Glucanase, Antagonistic, Pathogen, ITS- region

Conclusion

In this study, different species of Trichoderma have been isolated from rhizospheric soil of various crops of different districts in Uttar Pradesh, such as Trichoderma lixii, three species of Trichoderma harzianum and one of Trichoderma sp. The isolated species of Trichoderma were identified on the basis of cultural, morphological and molecular approaches using Internal Transcribed Spacer (ITS-PCR) region amplification. Further, it may be concluded that the Trichoderma isolates are classified into different species after sequencing of ITS region. The obtained sequenced are in-silico characterized through bioinformatics tools such as NCBI- BLAST, MEGA 11.0, Bioedit, Phyre2 etc. Furthermore, it is beneficial that the Trichoderma species acts as biocontrol agents against several pathogenic fungi and produces some lytic enzymes such as glucanase, chitinase, proteases, tubulises etc. These lytic enzymes helped in the management of fungal diseases in crops pand lants. So, Trichoderma is used as a bio-fertilizer and for seed treatment in various crops.

References

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How to cite this article

Vishwajeet Yadav, Mukesh Kumar, R.S. Sengar, Pushpendra Kumar, M.K. Yadav, Vyankatesh Dhanraj Bagul (2022). Isolation, Molecular and In-silico characterization of Trichoderma spp. from Rhizospheric Soil Sample. Biological Forum – An International Journal, 14(4): 648-652.