转基因烟草植株的检测  

The Demonstration of Transgenic Tobacco

作　　者：张彦妮[1] 曲敏 卓丽环[1] 徐香玲[3] 杨利平[4] 

机构地区：[1]东北林业大学,哈尔滨150040 [2]黑龙江省旅游学校 [3]哈尔滨师范大学生物系 [4]韶关学院

出　　处：《东北林业大学学报》2003年第6期36-40,共5页Journal of Northeast Forestry University

摘　　要：对采用叶盘法以质粒PBLGC为介导转化烟草的 3个品种 ,经Kan抗性筛选获得的 119株转化的烟草再生植株分别以启动子CaMV3 5S、几丁质酶基因和β -1,3 -葡聚糖酶基因的引物进行PCR检测 ,分别获得 91、72、47株阳性植株。其中 ,同时具有几丁质酶基因和 β -1,3 -葡聚糖酶基因的植株有 3 6株。对部分转化植株进行PCR -Southern杂交实验的结果表明 ,外源基因的转化频率与植物品种、外源基因片段的大小有关。又对 4株转基因植株后代 (T1)进行了PCR、PCR -Southern杂交 ,结果发现 ,几丁质酶基因和 β -1,3 -葡聚糖酶基因已经稳定地遗传给后代 ,但二者的遗传传递率不同 ,β -1,3 -葡聚糖酶基因似乎比几丁质酶基因更易于传递。另外 ,对转化植株T0 代几丁质酶活力检测结果表明 ,转基因植株中几丁质酶活力显著增强 ,含有几丁质酶基因和 β -1,3transformed regeneration plants derived from three varieties of tobacoo whic h were transformed by the leaf-method and Kanamycin selection were tested throug h PCR electrophoresis assay, under the media of plasmoid PBLGC (PBLGC contained chitinase gene and β-1, 3-glucanase gene and NTP-II gene) by using promtor 35S, chitinase gene and β-1, 3-glucanase gene primer separately. 92, 72 and 47 posi tive plants were obtained respectively. There were 36 positive plants containing both chitinase gene and β-1, 3-glucanase gene. Parts of transgenic plants were tested by PCR-Southern. The result showed that the transferring frequency of fo reign gene was related to plant variety and the length of foreign gene. The prog enies of four transgenic tobacco were demonstrated by PCR and PCR-Southern hybri dization analysis of tobacco DNAs. The result indicates that chitinase gene and β-1, 3-glucanase gene had stably inherited to progenies, but their rates of inh eritance were different. The inheritance of β-1, 3-glucanase gene seemed to be more stable than that of chitinase in progenies. On the other hand, the detectiv e result of chitinase activities of T 0 generation for transgenic plants showed that chitinase activities increased in transgenic plants, and chitinase activi ties in the plants containing chitinase and β-1, 3-glucanase gene were the high est.

关 键 词：转基因烟草 植株检测 叶盘法 介导转化 抗性筛选 再生植株 几丁质酶基因 

分 类 号：S572[农业科学—烟草工业]