【摘要】 通过根癌杆菌介导法 ,将维管束特异启动子 (竹节花黄斑病毒启动子CoYMV)启动的几丁质酶和CaMV3 5S启动的 β 1,3 葡聚糖酶嵌合双价基因 ,导入陆地棉品种冀合 3 2 1和中棉所 3 5。利用卡那霉素涂抹法对转基因株的卡那霉素抗性进行初步鉴定 ,再经PCR和Southern杂交确证 ,结果显示双价抗病基因分别以 1～ 2个拷贝整合到棉花基因组内。对转基因株系T3 幼苗进行无底塑钵菌液浇根法鉴定和大田病圃鉴定 ,结果显示 :7个转化株系均表现不同程度的抗或耐黄萎病性 ,苗期鉴定结果和大田病圃调查结果基本一致 ;其中D9910、D9915和D9919等 3个转基因纯合系的大田病情指数分别为 6 5、9 4和 9 5,均达到高抗水平。对以上 3个高抗黄萎病的纯合系进行遗传分析 ,结果显示这 3个抗病纯合系的卡那霉素抗性符合一对显性基因分离规律 ,结合分子杂交验证结果 ,可以认为这3个转基因株均含有一个拷贝的双抗病基因。更多还原

【Abstract】 The two disease-resistance genes chitinase and glucanase,which were respectively directed by commelina yellow mottle virus promoter （CoYMV,vascular specific） and CaMV35S promoter,were introduced into cotton genome via Agrobacterium tumefaciens.Transgenic plants were obtained from two popularly cultivated varieties Jihe321 and CRIC35.After screening by spraying kanamycin over unfolding leaves,the kanamycin resistance （Km^R） plants were tested by PCR and Southern blot.The results showed that there were one or two inserts of transgenes in cotton genome.Performance test of resistance of T₃ families in field and greenhouse showed that seven lines were resistant or tolerant to Verticillium dahliea.Meanwhile,the resistance at seedling stage in greenhouse was in accordance with that at the boll-setting stage in field.Among the seven lines,D9910,D9915 and D9919 had a disease resistance index of 6.5,9.4 and 9.5,respectively in field,which showed a high resistance level.Genetics analysis of the three lines showed a classical Mendelian pattern of one pair of genes,which meant that each of the three lines contains one copy of transgene.Southern blotting also confirmed the copy number of inserts.更多还原