84 International Journal of Biotechnology for Wellness Industries, 2013, 2, 84-90 ISSN: 1927-3037/13 © 2013 Lifescience Global Sequencing of 28SrRNA Gene for Identification of Trichoderma longibrachiatum 28CP/7444 Species in Soil Sample Mohammad Shahid 1,* , Anuradha Singh 1 , Mukesh Srivastava 1 , Smita Rastogi 2 and Neelam Pathak 2 1 Department of Plant Pathology, C.S.A. University of Agriculture & Technology, Kanpur, U.P., India 2 Department of Biosciences, Integral University, Lucknow, U.P., India Abstract: Most of the Trichoderma species are morphologically very similar and were considered for many years as a single species. Since new species were discovered, a consolidated taxonomical scheme was needed and proposed and defined nine morphological species aggregates. DNA methods brought additional valuable criteria to the taxonomy of Trichoderma which are being used today for studies that include identification and phylogenetic classification. Most isolates of the genus Trichoderma that were found to act as mycoparasites of many economically important aerial and soil-borne plant pathogens. Trichoderma has attained importance for substitute of chemical pesticides and hence an attempt was intended to corroborate the positive relatedness of molecular and morphological characters. A fungal strain of Trichoderma longibrachiatum 28CP/7444 was isolated from a soil sample collected from Barabanki district of Uttar Pradesh, India. The universal primers were used for amplification of the 28S rRNA gene fragment and strain characterized by using 28S rRNA gene sequence with the help of ITS marker. It is proposed that the identified strain Trichoderma longibrachiatum 28CP be assigned as the type strain of a species of the genus Trichoderma based on phylogenetic tree analysis together with the 28S 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 number JX978541. Thus an integrated approach of morphological and molecular markers can be employed to identify a superior strain of Trichoderma for its commercial exploitation. Keywords: 28S ribosomal RNA gene, Trichoderma, phylogenetic analysis, ITS, DNA sequencing, GenBank. 1. INTRODUCTION Trichoderma commonly available in soil and root ecosystem has gained immense importance since last few decades due to its biological control ability against several plant pathogens. Some strains of Trichoderma like Trichoderma harzianum, T. longibrachiatum, T. atroviride, T. viride, T. virens and T. koningii are efficient bio control agent which have the ability to inhibit pathogen growth in the soil, hence improving the overall health of the plant Antagonistic microorganism, such as Trichoderma reduce growth, survival of pathogen by different mechanism like competition antibiosis, mycoparasitism, hyphal interactions and enzyme secretion. Such micro organisms are now available commercially and are used in crop management and practices. The use of Trichoderma species as biological control agents has been investigated for over 70 years but it is only relatively recently that strains have become available commercially. Biocontrol agents are widely regarded by the general public as “natural” and therefore non- threatening products, although risk assessments must clearly be carried out on their effects on non-target organisms. Moreover, knowledge concerning the *Address correspondence to this author at the Department of Plant Pathology, C.S.A. University of Agriculture & Technology, Kanpur, U.P., India; Tel: 0512-2534113; Fax: 0512-2534113; E-mail: shahid.biotech@rediffmail.com behaviour of such antagonists is essential for their effective use. The morphological and microscopic Figures 1 & 2 of Trichoderma longibrachiatum 28 CP/ 7444 is given below. Figure 1: Trichoderma longibrachiatum 28CP/7444 strain in PDA medium. Figure 2: Trichoderma longibrachiatum 28CP/7444 strain under Microscopic observation.