【作者】 张俊；

【导师】 刘学群；

【作者基本信息】 中南民族大学 ， 生物化学与分子生物学， 2019， 硕士

【摘要】 sm-Nvas是来自烟草的受水杨酸(SA)和茉莉酸甲酯(MeJA)双重诱导的植物维管束特异表达的新基因,本文主要研究转sm-Nvas基因水稻的子代遗传稳定性、在转基因水稻中的整合位点和转基因株系对稻瘟病的抗病性。取得如下研究结果:1.连续两年种植15个转sm-Nvas基因水稻家系,提取两代家系的基因组DNA用于sm-Nvas基因和转基因标记检测。有4个家系无sm-Nvas基因和潮霉素抗性基因,但有转基因载体部分原件。10个家系中有sm-Nvas基因及载体原件,它们在2代中都能稳定遗传,1个家系还处于分离状态。2.在转sm-Nvas基因阳性家系中,使用TAIL-PCR方法进行T-DNA侧翼序列分析以确定sm-Nvas基因在水稻基因组的插入位点。确定有8个转基因家系sm-Nvas定位在水稻第6号染色体中,在这些株系中,有一个株系有转基因杂合子株,其余株系是转基因纯合子。3.对已经定位的转sm-Nvas基因阳性植株的农艺性状的对比研究,发现转sm-Nvas基因植株较转基因受体品种除具有更好的水稻稻瘟病抗性外,且分蘖能力也更强。更多还原

【Abstract】 sm-Nvas is a novel gene from tobacco,dually induced by salicylic acid(SA)and methyl jasminate(MeJA)and specifically expressed in plant vascular bundles.In this paper the genetic stability of the progeny of transgenic rice,the integration site in transgenic rice and the resistance of transgenic plants to blast were studied.The following results have been obtained:1.Fifteen transgenic sm-Nvas gene rice families were planted for 2 consecutive years,and the genomic DNA of the 2 generations of these families was extracted for detection of sm-Nvas and related transgenic markers.There were 4 families without carrying sm-Nvas gene and hygromycin resistance gene,but with carrying some component of transgenic vector.There were 10 families which carried both sm-Nvas gene and some components of transgenic vector.All of them can stably inherited in the 2 generations.2.To analyze the insertion sites of the sm-Nvas gene in the rice genome in the sm-Nvas transgenic positive families,the flank DNA sequences of T-DNA inserting sites were detected by TAIL-PCR method.the DNA sequence alignments showed that the sm-Nvas was located on the chromsome 6 in those 8 transgenic families,of which 7 familys were homozygotes and 1 family was still with some heterozygotes.3.It was found that the positve sm-Nvas transgenic plants has not only the characteristics of rice blast resistance,but also with stronger tiller ability compared with the transgenic receptor varieties.更多还原