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利用农杆菌介导法将蚕丝心蛋白基因(fibroin)导入陆地棉品系WC,共获得了9块愈伤组织系的30株再生植株.PCR检测、卡那霉素抗性筛选和GUS组织化学分析均为阳性的有17株.经连续4代的卡那霉素抗性筛选和PCR检测,在T3代获得了6个转化事件的转基因纯系.Southern blot和Northern blot结果表明,fibroin已整合到6个纯合株系基因组,且能稳定遗传和表达.扫描电子显微镜和纤维品质检测结果显示,所有转基因纯合株系棉纤维的转曲数和纤维伸长率比对照明显增加,部分株系比强度提高,说明蚕丝心蛋白基因在棉纤维中特异表达影响了棉纤维的结构和品质,可以获得纤维品质改良的株系,有望在生产上得到应用.
A total of 30 regenerated plants from 9 callus lines were obtained by introducing fibroin into G. hirsutum line WC using Agrobacterium tumefaciens-mediated method.PCR, kanamycin resistance screening and GUS histochemical analysis 17 were positive.After 4 generations of kanamycin resistance screening and PCR detection, transgenic lines of 6 transformation events were obtained in T3 generation.The results of Southern blot and Northern blot showed that fibroin has been integrated into 6 homozygous lines were obtained and their genomes could be stably inherited and expressed.The results of scanning electron microscopy and fiber quality test showed that the number of rotations and fiber elongation of all the transgenic homozygous cotton lines were significantly increased compared with the control, The results showed that the specific expression of silk fibroin gene in cotton fiber affected the structure and quality of cotton fiber and improved the fiber quality. It is expected to be applied in production.