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The Resistance of Pathogenesis-Related Protein StPRp27 to Erwinia Carotovora Subsp. Carotovora

Author: ChenWeiDa
Tutor: LiuJun
School: Huazhong Agricultural University
Course: Genetics
Keywords: Pathogenesis-related proteins StPRp27 The prokaryotic expression Konjac Erwinia carotovora subsp.carotovora
CLC: S436.32
Type: Master's thesis
Year: 2009
Downloads: 74
Quote: 2
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Abstract

Amorphophallus as Araceae Konjac is a perennial herb, and is widely distributed in the mountainous areas of southwest China, and has a high value in use because of its the corms rich glucomannan. Soft Rot is a bacterial disease caused by subspecies by carrot carrots soft rot Erwinia soft rot (Erwinia carotovora subsp.carotovora), most of konjac producing areas in China have the disease occurs, resulting in severe konjac devastating disasters. However, konjac not yet found resistance to soft rot germplasm resources. Pathogenesis-related protein (Pathogenesis-related proteins, PR) is a plant pathogen invasion duration process induces the expression of a class of proteins, and has a broad spectrum of disease resistance. Pathogenesis-related protein is a large gene family, the family of the different members of the resistance to pathogens there are some specific. Pathogenesis-related protein gene StPRp27 of potato pathogenesis-related protein gene belongs to the PR-17 family. This topic first study Soft Rot of bacteria inhibition of the gene protein, konjac plant expression vector transformation based on anti-Soft Rot of transgenic lines for resistance to soft rot genetic engineering breeding konjac lay the foundation. The main findings are as follows: bacteria significantly inhibited 1.StPRp27 protein Soft Rot. Potato pathogenesis-related protein gene StPRp27 subcloned into the prokaryotic expression vector pGEX-6p-1 prokaryotic expression plasmid pGEX-StPRp27 and into the prokaryotic expression host strain E.coli strain BL21 (DE3) prokaryotic expression. The results showed that expression of different induction conditions, IPTG concentration of 0.1mM, a temperature of 22 ° C, can be soluble of GST-StPRp27 protein. Further purified to give a molecular size of approximately 51kd the GST-StPRp27 of fusion protein the bacteriostatic test results of the protein, GST-StPRp27 fusion protein compared to glutathione (GST) protein has a stronger inhibition of konjac soft rot bacteria (Erwinia carotovora subsp . carotovora) the effect of the growth, with the increase of the concentration of the bacteriostatic effect enhancing, when the GST-StPrp27 concentration increased to 0.08mM, germs basic stop growth to prove StPRp27 Soft Rot of the growth of bacteria having significantly inhibited. Konjac transformed receptor screening. Tube seedling morphology of the lower part of the petiole cut callus induction rate of 100%, and induces the formation of callus; plantlets morphological upper petiole the cut callus induction rate was only 54.4%, to induce the formation of callus The organization is also smaller; tube seedling morphology central petiole the cut callus induction ability between the upper and lower. The study also found that part of the pre-culture the petiole cut three weeks have been induced to form callus, and petiole cut state there are obvious differences; while the the petiole cut two weeks of pre-culture Amorphophallus konjac plantlets were no obvious callus formation petiole cut state differences are not obvious. Pre-cultured for 2 weeks petiole more appropriate as the transformation receptor. Amorphophallus konjac Transformation System for transgenic plants obtained. Plantlets were pre-incubated for 2 weeks petiole cut into sections as explants for genetic transformation, after co-culture and off the bacteria culture, prior to conversion density is less than water petiole cut part of the density than water, in the subsequent screening culture the petiole cut into sections of this part can not induce the formation of callus. The resistant calli transferred to differentiation culture, early differentiation to form adventitious buds are deformed buds can not develop into normal plants resistant callus culture in differentiation medium to form normal plants. The study is divided into 66 resistant plantlets of which were detected by PCR and 28 positive.

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