花椰菜花叶病毒(CaMV)35S启动子在转基因棉花中的表达  

Expression of CaMV 35S Promoter in Transgenic Cotton

作　　者：焦改丽[1] 孟钊红[2] 聂安全[1] 南芝润[1] 张换样[1] 李俊峰[3] 王娇娟[1] 赵俊侠[1] 李燕娥[1] 郭三堆[4] 

机构地区：[1]山西省农业科学院棉花研究所,山西运城044000 [2]山西大学生物技术研究所,山西太原030031 [3]中国海洋大学海洋生命学院,山东青岛266000 [4]中国农业科学院生物技术研究中心,北京100081

出　　处：《作物学报》2004年第11期1135-1139,共5页Acta Agronomica Sinica

基　　金：山西省"十五"重点攻关项目 ( 0 3 10 0 5 1);山西省留学回国人员科技资助项目;国家"十五 86 3"计划项目的一部分。

摘　　要：利用GUS作为报告基因 ,通过GUS组织化学定位法检测棉花转化愈伤组织、体细胞胚 ,R0 代棉花根、茎、叶、花器官以及正在发育的胚GUS基因表达情况 ,详细阐述了CaMV 3 5S启动子在棉花细胞中的表达轮廓。结果表明 ,在愈伤组织细胞有丝分裂和增殖过程中GUS基因能稳定表达并遗传给后代细胞 ;在根、茎、叶细胞中检测到GUS表达活性。在胚胎发育过程中 ,最早检测到GUS表达活性的为开花 1 1d的鱼雷胚和胚乳。随着胚胎的发育 ,GUS表达活性逐渐增强并扩展到子叶微管组织 ,表明CaMV 3 5S启动子是随着胚胎发育进程被逐渐调节的。在花粉和特殊的纤维细胞中 ,也检测到GUS的表达活性。资料证明 ,该启动子在棉花不同发育时期的大部分组织和细胞中都是表达的。The CaMV 35S promoter is the most widely used promoter for driving transgene expression in plants. It is effective driving transgene expression in both dicots and monocots. It is usually classified as constitutive promoter. However, some reports suggest that it is not expressed in all cell and tissue types. In addition, the available information on its expression profile in all cell and during the early stages of development is incomplete. In this paper, the expression profile of CaMV 35S promoter was detailedly described using the GUS as a reporter gene in the transgenic cotton. The results of histochemical assay showed that GUS gene could express steadily and transmit to next generation in the course of callus mitotic division and proliferation. Dark blue staining of GUS was observed at various stages of somatic embryos (globular, heart, topedo, cotyledon) and the roots, stems and leaves of R 0. GUS positive expression was first detected during the early stages of torpedo embryo and endosperm at around 11 days after anthesis. With further growth of the embryos, the GUS staining became stronger and spread to throughout the vascular tissues in the cotyledon. Especially GUS positive expression is detected in the pollens and special fiber cells. It showed CaMV 35S promoter was the same active in these specialized epidermal cells. Thus, our evidences suggest that CaMV 35S promoter was developmentally regulated during embryogenesis in cotton. The promoter could express in most cell and tissue types in cotton at various developmental stages.

关 键 词：GUS基因 花椰菜花叶病毒(CaMV)35S启动子 转基因棉花 胚胎发育 表皮细胞 

分 类 号：S562[农业科学—作物学]