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利用花粉管通道转基因技术,将DE35S启动子驱动的野生荠菜凝集素(WSA)基因导入10个高品质棉花品种(系)。所使用的转基因表达载体还含有选择标记基因NPTⅡ(卡那霉素抗性基因)和Ω序列,以利于转基因植株的筛选以及WSA基因的高效表达。对转化当代3197棵棉苗的检测结果表明,4.88%植株具有卡那霉素抗性(Kan+)。根据野生荠菜凝集素基因序列设计一对特异性引物,对156个Kan+植株进行PCR检测,筛选出45个转WSA基因棉花工程植株。45个株系纤维品质测定结果表明,获得了9个高品质转WSA基因棉花株系。并对植物基因工程研究中以NPTⅡ作为标记基因的局限性以及一些转WSA基因棉花株系纤维强度变劣的原因进行了探讨。
Using the pollen tube pathway gene transfer technique, 10 high quality cotton cultivars (lines) were induced by DE35S promoter-driven wild Plutella xylostella lectin (WSA) gene. The transgene expression vectors used also contain the selectable marker gene NPTII (kanamycin resistance) and the omega sequence to facilitate screening of transgenic plants and efficient expression of the WSA gene. The test results of 3197 cotton seedlings transplanted in modern times showed that 4.88% of the plants had kanamycin resistance (Kan +). A pair of specific primers was designed according to the sequence of the wild Capsella lectin gene and PCR was carried out on 156 Kan + plants. 45 WSA transgenic cotton plants were screened out. The results of fiber quality measurement of 45 lines showed that 9 high-quality WSA transgenic cotton lines were obtained. In addition, the limitations of NPT Ⅱ as a marker gene in plant genetic engineering and the reasons for the deterioration of fiber strength of some WSA transgenic cotton lines were also discussed.