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Title: Transformation and evaluation of Anti apoptosis gene in banana cv Rasthali for Fusarium wilt resistance
Researcher: Sunisha C.
Guide(s): Usharani T R
Keywords: Entomology insects
Life Sciences
Plant and Animal Science
University: Jain University
Completed Date: 2020
Abstract: Rasthali (AAB, Silk gp) is one of the choicest dessert banana cultivar of India newlineknown for its flavor and quality. Fusariums wilt disease caused by Fusarium oxysporum f. newlinesp. cubense (Foc) affects the cultivar and has reached severe proportions, almost wiping it newlineout. There are no appropriate control strategies for the disease except the resistant newlinecultivars. Although resistance gene to Foc has been identified in wild bananas but newlinebreeding is a limitation due to its sterile, polyploidy, parthenocarpic nature and long newlineregeneration time. Due to the advancement in technology for transformation and transgene newlineexpression in banana, transfer of fungicidal genes into the genomes of susceptible banana newlinecultivars has become a reality. In the present study, the banana cultivar Rasthali newlineembryogenic cell suspension was stably transformed with dual construct of two newlineantimicrobial genes viz., Ace-AMP1 and pflp, and two anti-apoptosis genes namely Ced-9 newlineand AtBag4 using Agrobacterium mediated transformation. A total of 101 putative newlinetransformants from dual genes, 96 from Ced-9 gene and 89 from AtBag4 gene were newlinedeveloped and acclimatized in net house. The presence of gene in putative transformants newlinewas confirmed by PCR using gene specific primers. A total of 150 independent newlinetransformed lines were randomly subjected for the root challenge bioassay with Foc race1 newlinein pot condition. Based on the bioassay performance, 10 transgenic lines from dual newlineantimicrobial gene construct pCAMBIA1305.2-pflp-Ace-AMP1 gene, 19 transgenic lines newlinefrom antiapoptosis pAJF 422-Ced-9 gene construct and 11 transgenic lines from pAJF newline131-AtBag4 gene were selected to determine the T-DNA integration and copy number of newlinetransgene. Out of shortlisted transgenic lines, 8, 10 and 10 transgenic lines of dual gene, newlineCed-9 and AtBag4 respectively showed the T-DNA integration. Expression analysis of newlinetransgenic line with dual anti -microbial gene (line 42 and line 43) showed high mRNA newlineaccumulation of Ace-AMP1 gene and pflp gene in a Northern blot analysis. Further, the newlinetransgenic lines expressing Ced-9 gene (line 33, 38, 40 and 41) and AtBag4 gene (line 34, newline20, newline33, 35, 36, 37, 40, 42, 43 and 45) showed significant relative expression fold values as newline newlinecompared to the untransformed control plants. The disease progression in transgenic, non- newlinetransgenic and resistant controls was analyzed as Vascular Discoloration Index (VDI) for newline newlinesix months post inoculation of Foc. The lines 42 and 43 expressing dual anti-microbial newline newlinexiv newline newlinegenes showed significantly least VDI (10 and 28 per cent) compared to other transgenic newlinelines (30-45 per cent). The Ced-9 gene overexpressing transgenic line 31 resulted in lower newlineVDI (17 per cent) where as other transgenic lines exhibited 30 51 per cent discoloration. newlineThe AtBag4 expressing transgenic line 36 showed 15 per cent of VDI whereas, other newlinetransgenic lines showed 26-61 per cent of VDI. newline newlineThis is the first report in India on the use of dual anti-microbial genes and anti- newlineapoptosis genes of heterologous origin in banana plant for resistance to Fusarium wilt. In newline newlineaddition, the strategy may be applied in other agronomically essential crops to generate newlinedisease resistance against specific pathogen. newline
Pagination: 189 p
Appears in Departments:Department of Biotechnology

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certificates-2-6.pdf1.47 MBAdobe PDFView/Open
chapter 1. introduction.pdf193.17 kBAdobe PDFView/Open
chapter 2. review.pdf1.03 MBAdobe PDFView/Open
chapter 3. material and methods.pdf545.16 kBAdobe PDFView/Open
chapter 4. results.pdf2.57 MBAdobe PDFView/Open
chapter 5. discussion .pdf166.44 kBAdobe PDFView/Open
chapter 6. summary.pdf146.36 kBAdobe PDFView/Open
cover papge.pdf118.42 kBAdobe PDFView/Open
table of content.pdf139.17 kBAdobe PDFView/Open

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