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Authors: Bodade, Shrikant Raghunath
Advisor: Patil, H. B.
Title: Identification and differentiation of strains of xanthomonas axonopodis PV punicae at molecular level
Publisher: Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani
Language: en
Type: Thesis
Pages: 64
Agrotags: null
Keywords: Genotypic, Phenotypic, Cluster, Chickpea, Correlation
Abstract: Pomegranate (Punica granatum L.) is an important fruit crop of India and other subtropical countries. Bacterial blight caused by Xanthomonas axonopodis pv. punicae (Hingorani and Singh, 1959), is becoming a major threat to pomegranate production in Maharashtra as well as other pomegranate growing states of India. It causes yield loss to the extent of 80 - 90 %. Considering importance of this disease, study regarding assessment identification and differentiation of strains of X. a. pv. punicae was undertaken. Based on genetic diversity study of isolates of Xanthomonas axonopodis pv. Punicae, conducted earlier by kale (2012) five representative samples of each region have been selected towards assessment of aggressiveness and nucleotide sequences variation at pathogenicity gene fragment sequence level. A Rifampicin resistant mutant were developed by continuous growing of bacterial cell on Rifampicin (200 pg/ml) supplemented YGCA medium. During syringe-infiltration of known concentration of each of five isolates on pomegranate cv. Bhagwa produced confluent water soaking spots. Based upon appearance of symptoms, there were grouped in to three categories as highly virulent (Xap5), moderately virulent (Xapl, Xap2 and Xap4) and less virulent (Xap3). Further aggressiveness was assessed through Inplanta growth assay. The highly aggressive isolate Xap5 produced 3.1 X 108 cfu/ml bacterial colonies while, moderately and lesser virulent isolates were produced bacterial count of 2 X 107 cfu/ral and 4 X 106 cfu/ml respectively after 5 days of inoculation. The pathogenicity gene fragment of size 460-490 bp were also exploited for restriction digestion by enzyme BamHl, Pstl and Smal. The Restriction enzyme analysis categorized all of them into four groups. Further pathogenicity gene fragment (460- 490 bp) were cloned and sequenced. The BLAST analysis confirmed the identity of the sequences by showing 93 % homology with pathogenicity gene sequences available in public domain. Also the multiple sequence analysis revealed the variation in the sequences of the isolates collected from different regions. Thus based upon the results comprising variation in aggressiveness test and nucleotide sequences of pathogenicity gene fragment, is an indicative of existence of at least three strains among pathogen Xanthomonas axonopodis pv. punicae causing oily spot disease in pomegranate.
Subject: Agricultural Biotechnology
Theme: Agriculture
These Type: M.Sc
Issue Date: 2015-07-09
Appears in Collections:Thesis

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