陆地棉抗病品种中521植株再生的研究

Study on the Establishment of Plant Regeneration Method in Upland Cotton CCRI 521

以农艺性状优良且抗枯黄萎病的陆地棉品种中521的下胚轴为外植体,采用固液结合的体细胞培养方法建立细胞悬浮系并进行植株再生的研究。结果表明,愈伤组织的诱导以MS+1.0 mg.L-1IBA+0.05 mg.L-1KT+0.1 mg.L-12,4-D+30g.L-1葡萄糖的固体培养基为最佳;体细胞胚的形成以MS+0.01 mg.L-1IBA+0.01 mg.L-1KT+0.005 mg.L-12,4-D+1.0g.L-1谷氨酰胺的液体培养基为最佳,悬浮30 d后可以发现体细胞胚的形成。之后转接入四种不同的MSB固体培养基上诱导再生植株形成,发现MSB+2 g.L-1谷氨酰胺+0.5 g.L-1天冬酰胺+30g.L-1蔗糖+6.0 g.L-1琼脂为最佳培养基,且发现此方法比全程固体培养缩短了体细胞再生的周期,此品种在本研究建立的再生体系下植株再生周期为6个月左右。

At present, many regeneration plant systems have been established. But only a few model varieties were fit to those regenerative systems and the genetic types were the main restrictive factor in. the process of cotton somatic culture and plant regeneration, and the upland cotton variety CCRI521 with better agronomic properties and tolerance to Verticilliurn wilt had not been studied. This study obtained cell suspending lines and regenerative plants from upland cotton CCRI521 by the approach of solid and liquid culturing. The results showed that the solid medium MS＋ 1.0 mg·L^-1 IBA＋0.05 mg·L^-1 KT＋0. 1 mg·L^-1 2,4-D＋30g·L^-1 glucose was the best for callus induction from hypocotyls in our study. While the liquid medium MS＋0. 01 mg·L^-1 IBA ＋0. 01 mg·L^-1 KT ＋0. 005 mg·L^-1 2,4-D ＋1.0 g·L^-1 glutamine was fit for somatic embryogenesis, somatic embryo could be seen after suspending for 30 days. Then these somatic embryo were transferred into four different MSB media for regenerative plants induction and found the medium MSB＋ 2 g·L^-1 glutamine＋0.5 g·L^-1 asparagines＋30 g·L^-1 sucrose＋6. 0 g·L^-1 agar was the best for regenerative plants induction in these four different MSB media. And the cycle of plant regeneration was about 6 months in this system.