Morphological, Molecular Identification and SSR Marker Analysis of a Potential Strain of Trichoderma/Hypocrea for Production of a Bioformulation

Abstract

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.

Antipathy of <b><i>Trichoderma</i></b> against <b><i>Sclerotium rolfsii </i></b>Sacc.: Evaluation of Cell Wall-Degrading Enzymatic Activities and Molecular Diversity Analysis of Antagonists

“…So far, Shahid et al [2013] used SSR marker analysis of a potential strain of antagonists of Trichoderma . A set of 20 SSR primers was used, and genetic variability studies reveal that a percentage of polymorphisms in SSRs is obtained within the 7 strains of Trichoderma species which is comparatively higher (>77%) than with random amplified polymorphic DNA primers (approx.…”

“…The SSR was used for molecular characterization of Trichoderma for studying genetic diversity and similarity across 11 Trichoderma strains which differed in their antagonistic activity. A total of 20 SSR primers were selected for the study, and PCR amplifications were carried out according to Shahid et al [2013]. The amplification products were separated in 2% (w/v) agarose gel in 1× TBE buffer and visualized by staining with 0.1% (w/v) ethidium bromide [Sambrook and Russell, 2001].…”

“…There are various well-established PCR-based molecular marker techniques that had been widely used in Trichoderma and other fungi [Joshi, 1999]. The simple sequence repeat (SSR) markers are reliable but rarely reported for Trichoderma characterization [Shahid et al, 2013].…”

Influence of Protoplast Fusion in Trichoderma Spp. on Controlling Some Soil Borne Diseases

“…However, other author found a relationship between RAPD polymorphisms of Trichoderma strains and their antagonism against Aspergillus niger [29]. Moreover, the others found a high relationship between some molecular markers of Trichoderma strains and their antagonism against many plant pathogens as Fusarium oxysporium, Pythium altimum and Sclerotium rolfsii [5,8,24,30]. Moreover, here and in a previous study, T. harzianum was the predominant taxon, T. harzianum is the most commonly reported species in the genus, occurring in diverse ecosystems and ecological niches.…”

Rapid identification of Trichoderma koningiopsis and Trichoderma longibrachiatum using sequence-characterized amplified region markers

“…Therefore, correct identification of Trichoderma at the species level is important. Morphological identification of species has proven unreliable (Sharma et al 2009 andShahid et al 2013) because of their high degrees of similarity.…”