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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。
GUS as a reporter gene was used to detect the expression of GUS gene in cotton transformed callus, somatic embryos, R0 generation cotton roots, stems, leaves, floral organs and developing embryos by GUS histochemical localization method. The expression of CaMV 3 5S Promoter expression profile in cotton cells. The results showed that the GUS gene was stably expressed and passed on to progeny cells during the mitosis and proliferation of callus cells. GUS expression activity was detected in root, stem and leaf cells. During the embryonic development, the earliest GUS expression was detected in the torpedo and endosperm blooming for 11 days. As the embryo developed, the GUS expression activity gradually increased and extended to the cotyledonary microtubules, indicating that the CaMV 3 5S promoter is gradually regulated as the embryonic development progresses. In pollen and special fibroblasts, GUS expression activity was also detected. Data show that the promoter is expressed in most tissues and cells at different developmental stages of cotton.