Determination of Biodiversity Using ITS-PCR-CAPS Markers For Trichoderma sp. From Rhizospheric Soils of Different Locations in India.
Prasad Lakshman*, Sagar Sushma, Chaudhary Sorabh, Kumar Nitin, Tomar Akash1
Department of Plant Pathology, S.V. Patel University of Agriculture and Technology, Meerut-250 110, India
1Department of Recombinant DNA Technology, S.V. Patel University of Agriculture and Technology, Meerut-250 110, India
*Corresponding author Email: firstname.lastname@example.org
The occurrence of Trichoderma in the various rhizospheric soils can be represented a population dynamic for studying a distribution of Trichoderma between different crop rhizospheric soils. Forty-nine Trichoderma isolates were obtained and identified at the genus level by analysis of morphological characters with help of manuals. However, these Trichoderma spp biodiversity were also be analyzed using the internal transcribed spacer (ITS) of the rDNA region and Cleavage Amplified Polymorphic Site (CAPS) markers. The ITS region was first amplified by polymerase chain reaction (PCR) with specific primers and then cleaved with two restriction enzymes i.e. HaeIII and Cla I. Amplification of product showed extensive length polymorphism and restriction analysis generated bands ranging from 100-500bp. The restriction profiles of PCR products showed that HaeIII enzyme has more discriminatory power than Cla I. A high degree of polymorphism was detected among Trichoderma isolates obtained from various rhizospheric soils. Thus, the CAPS analysis produced different DNA profiles on the gels denoting significant intraspecific genetic variation. Our results confirm that ITS region PCR-CAPS is a potential and valid tool for the study of polymorphism among Trichoderma populations at every locations of each rhizosphere.