Institutional Repository
Thesis Issued 2026-09-08 EN

Identification of phomopsis resistance gene in eggplant

Author: Md. Ibrahim Khalil
Eggplant-Genetic aspects

Abstract

The F4, Fs and F, lines of eggplant derived from the crosses of Dohazari G x BAU Begun-1 and Laffa S x BAU Begun-1 were evaluated for resistance to phomopsis blight and fruit rot. The three parents and seven lines were inoculated with Phomopsis vexans under field conditions in winter seasons of 2006-07, 2007-08 and 2008-09. The inoculated plants exhibited differential disease reactions. Among the parents, BAU Begun-1 was resistant whereas Dohazari G and Laffa S were susceptible to P. vexans. All the lines of three generations showed resistant reaction to the disease. Significant differences were observed among the phenotypes in all the yield components. High genotypic and phenotypic coefficient of variation, heritability and per cent genetic advance were recorded for number of fruits per plant, number of secondary branch per plant, fruit length and fruit breadth in all the generations. Significant positive correlation was observed between yield contributing characters. Random amplified polymorphic DNA (RAPD) technique was used for assessing genetic variation and relationship among three parent cultivars and their F4, Fs and Fo progenies of eggplant. Amplification with five decamer primers generated 69.0, 66.1 and 65.2% polymorphic bands for F4, Fs and F6 lines respectively. The dendogram constructed from Nei's genetic distance produced parent cultivars merged into one subgroup with resistant cultivar BAU Begun-1 and formed a separate sub-group in all the generations studied. The lines through F4 to Fo grouped into sub-cluster together, that indicates some common trait was being shared by all of these advanced lines of both the crosses. This might be the resistance trait inherited from BAU Begun-1 as supported by other characters like fruit colour and fruit shape also inherited from the parents. A PCR-based molecular cloning technique was used to identify Nucleotide binding site-Leucine rich repeat (NBS-LRR) type resistance gene sequence in eggplant. Primers were designed based on previously used in sugar beet and the resultant amplicons were cloned. Nine clones were sequenced and found to be about 243 nucleotides in length. Most of the sequences have similarity with TIR and non-TIR types of NBS-LRR gene of Solanaceous crops. These sequences also have similarity with 12 gene sequence which is a non-TIR type resistance gene in tomato- that confers resistances to the tomato wilt pathogen. Another experiment was conducted to determine if an osmotin gene in eggplants correlates with phomopsis resistance. Primers were designed against homologous regions in osmotins, and made to be most similar to sequences in Solanaceous crops. These primers were used to PCR amplify and clone~700 bp sequence from both the resistant BAU Begun-1 and the susceptible Laffa S eggplant cultivars. Analysis of the sequences revealed six different types osmotin like sequence (Genes or alleles) for the phomopsis susceptible eggplant cultivar Laffa S and resistant cultivar BAU Begun-1 which encodes some conserved osmotin amino acids including 16 of the characteristic Cysteine residues and five acidic cleft amino acid residue. This means that there were at least 3 different osmotinlike genes in eggplants. Four types of sequences were amplified from both the cultivars. One type of sequence (C) was found only in BAU Begun-1 and another (E) only in Laffa. PCR products were successfully digested with restriction enzymes in presence of PCR buffer without any purification and addition of any reagents to find out the allele or gene responsible for resistance in eggplant. Further analysis of the C and E type sequence uncommon in the resistant and susceptible eggplant cultivars would reveal the presence or absence of any other resistance trait in BAU Begun-1.