Selection and Agrobaeterium-mediated lines with high frequency of somatic (Gossypium hirsutum L.) in Xinjiang transformation of regenerated embryogenesis of upland cotton | 新疆陆地棉高效体细胞胚胎发生系的培育及其遗传转化

Chino. 为了筛选高频体细胞胚胎再生能力的棉花品种（系），提高农杆菌介导转化效率，本研究以新疆陆地棉03298品系经组织培养产生的三个不同世代的再生系03298R1、03298R2和03298R3为材料，分别进行农杆菌介导的遗传转化。在相同的培养体系下，03298再生系下胚轴和子叶外植体的分化率由5％（一代再生系03298R1）提高到10％-15％（二代再生系03298R2）再到23％～33％（三代再生系03298R3），表明连续组织培养定向筛选提高了03298再生系体细胞胚胎再生能力。PCR和southern杂交检测显示抗卡那霉素的再生植株成功整合了NPTII基因。本研究经组织培养筛选获得了高频体细胞再生能力的03298再生系，并建立了相应的高效遗传转化体系，为新疆陆地棉遗传改良提供了新受体材料及其配套的遗传转化体系。

Inglés. In an effort to improve the frequency of Agrobacterium-mediated transformation in upland cotton, three different generations of regenerated lines (03298R1, 03298R2 and 03298R3) generated through somatic embryogenesis from upland cotton cultivar 03298 were subjected to Agrobacterium-mediated transformation with hypocotyl and cotyledonary explants, respectively. The differentiation rate in term of the ratio of independent kanamycin-resistant events to total explants increased from 5% in the first generation of regenerated line 03298R1 to 10% - 15% in the second generation of regenerated line 03298R2 and to 23% - 33% in the third generation of regenerated line 03298R3, suggesting that the frequency of somatic embryogenesis of 03298 regenerated lines was improved under the selection of tissue culture through somatic embryogenesis. PCR and southern blot further confirmed the integration of NPTII gene into the cotton genome of the kanamycin-resistant plants. The regenerated line 03298R3 with high frequency of somatic embryogenesis as well as its corresponding protocol of Agrobacterium mediated transformation presented herein would be conductive to the genetic improvement of upland cotton in Xinjiang.