壳寡糖诱导植物抗性相关基因mRNA差别显示分析

壳寡糖是一种天然的信号分子，可以诱导植物产生各种防御反应。该文利用mRNA显示技术。研究壳寡糖引起的植物抗性相关基因的变化。以50mg/l壳寡糖喷洒的烟草叶片为材料，分别提取处理8h、7d和对照的叶片总RNA，利用一个锚定引物和三个随机引物组合，通过mRNA差别显示技术共获得壳寡糖处理后发生变化的8h差别条带22条，7d后的差别条带为74条。将壳寡糖处理8h得到的17条差别条带克隆到载体上后，进行反向Northern鉴定，有6条可能为真实条带。测序结果表明：其中一条差别片段与烟草热激蛋白90基因具有97％的核苷酸同源性。说明50mg/l壳寡糖处理的烟草叶片在mRNA水平上发生了明显的变化，烟草的热激蛋白90基因可能参与到壳寡糖诱导的抗性信号传导通路中。

mRNA Different Display Analysis of Plant Resistance - related Gene Induced by Oligochitosan

Being a natural signaling jmolecule,oligochitosan can induce various plant defense responses.In this paper,It investigated the variation of resistance-related genes induced by oligochitosan using mRNA Differential Display technology.Tobacco(Nicotiana tabacum var.samsun NN)with 4-5 leaves was treated with 50mg/l oligosaccharides.After 8 hours and 7 days treatment one anchor primer and three arbitrary primers were used in mRNA differential display.Altogether 22 differential bands of 8 hours after treatment and 74 differential bands of 7 days after treatment were observed and recovered from the sequencing gel,among them 17 differential bands were cloned and the false positive bands were excluded by reverse northern blot.Sequencing results of 6 positive bands indicated that one of them has the 97%identity with tobacco heat shock protein 90 gene.Results showed that distinct difference took place in mRNA level after oligochitosan treatment,and heat shock protein 90 gene may involved in the signal pathway of resistance induced by oligochitosan.