Isolation, Characterization of Naative Trichoderma Isolates and Bioinformatics Analysis of Pathogenesis Related Genes During Interaction . Of Trichoderma, Fusarium and Tomato

Abstract

Thesis Title: Isolation, characterization of native Trichoderma isolates and Bioinformatics analysis of pathogenesis related genes during interaction of Trichoderma, Fusarium and Tomato. With the advent of. synthetic insecticides in the 1950s, easy control of insect pests appeared at hand. However, It soon became obvious that there were problems associated with the use of insecticides. Sotne insect pests becatne resistant, and son1e non target organis1ns vve re adversely affected, and pest resurgence occutTed. Additionally, enviromnental and health concerns arise now need to develop alternatives to conventional pesticides. The use of micro organisms to manage pests is known as biological control. To manage agriculturally important diseases main focus enlighten on Trichode17na due to its unmatching beneficial activities. Trichoderma species are important biological control agents used in plant disease management. Certain species of the fungal genus Trichoderma are potent mycoparasites and are used for biological control of fungal diseases by mycoparasitism and competition for plant exudates. Trichoderma species are free living fungi that are common in ecosystem and found in nearly all type of soils. They colonized in the roots of plants and releases variety of compounds that produces localized and systemic resistance responses in plants. The fungi Trichoderma species were isolated from soil of distinct regions of Sardhna, Mainpuri and Meerut. The fungal species were identified on the basis of their morphological and molecular characteristics. A total of 10 isolates were used for ITS analysis. Internal transcribed spacer- 1 region (ITS 1) of the ribosomal DNA was amplified by polymerase chain reaction (PCR) by using selected universal primers (ITS 1 and ITS4) for ten Trichoderrna isolates. The size of PCR product amplified with the primer pair ITS l and ITS4 ranged frotn 563-602bp and contained ITSl, ITS2 and 5.8 S rDNA gene and also llbp of3'end of 18S r DNA and 36bp of the 5' end of 28S rDNA. The amplified DNA was sequenced and aligned against ex-type strains sequences from Tricho BLAST /GenBcftlk and established Trichoderma taxonomy. Total ten native Trichoderma isolates were identified at the species level by n1otphological charactetistics and analysis of their ITS 1, ITS2 nucleotide sequences as: nine of thetn belongs to Trichode~.,,~a harzianum and one was Trichoderma brevicompactzon, Isolates had been tested for antagontstlc effects against soil borne plant pathogen Fusarium oxysporum spp lycopersi~ii. The . most effecti·v e I· solates were of harzianum. For the further transcn·p tomt·c s st ud.t es pnm er destgned with the help ofbioinfonnatic tools. Sequences downloaded from databases and then NCB~ bla~t for similar sequences was used. Primer3 and primer blast are the important _softwar~ used 1.n thl~ approach. These primers help in the study of genes which are activated dunng the Interaction ° Trichoderma, Tomato and Fusarium.