绿色棉新彩棉7号体细胞胚胎发生及其植株再生

以绿色棉新彩棉7号的子叶、下胚轴为外植体,MSB(MS培养基附加B5维生素)基本培养基附加不同激素组合,诱导愈伤组织及调控分化,通过体细胞胚胎发生方式获得再生植株.结果表明:0.1 mg· L-1 KT(Kinetin,激动素)+ 0.1 mg·L-12,4-D(2,4-dichlorophenoxyacetic,2,4-二氯苯氧乙酸)为诱导愈伤组织的最适植物激素组合,不同外植体处理出愈率均达到100％,但下胚轴纵切面背向培养基放置培养更有利于诱导愈伤组织形成;分化调控阶段的最佳植物激素组合为0.15 mg·L-1 KT+ 0.3 mg·L-1 IBA(Indole-3-butyric acid,吲哚丁酸),胚性愈伤分化率可达23.33％; MSB中去除NH4NO3同时KNO3加倍,附加0.5 g·L-1Asn(Asparagine,天冬酰胺)和lg· L-1 Gln(Glutamine,谷氨酰胺),胚性愈伤可进一步分化获得体细胞胚,将成熟的子叶胚接种于1/2MS获得完整的再生植株. 本研究通过体细胞胚发生途径获得了新彩棉7号的再生植株,为天然彩色棉基因工程研究奠定了一定基础.

Somatic Embryogenesis and Regeneration of Green Cotton(Gossypium hirsutum L.) Xincaimian No.7

Using hypocotyls and cotyledon of Xincaimian No.7(Green fiber cotton cultivar) as explants, MSB(MS salts and B5 vitamins) medium supplemented with different phytohormone combinations as callus induction media, and by embryogenic callus differentiation, regeneration experiment was carried out. The results indicated that the calli were easily induced in medium supplemented with 0.1 mg·L^-1 2，4-D and 0.1 mg·L^-1 KT, the frequency of callus induction was 100%. Split hypocotyl segments reversely cultured on medium was the best mode for callus induction. 0.3 mg·L^-1 IBA and 0.15 mg·L^-1 KT was the best phytohormone combination for embryogenic callus differentiation, and the highest differentiation frequency was 23.33%. Somatic embryos were acquired in MSB medium with NH₄NO₃ free and doubled KNO₃, supplemented with 0.5 g·L^-1 Asn and 1 g·L^-1 Gln. Complete cotyledon embryos were isolated and cultured on hormone-free 1/2MS medium for germination and development. The regeneration protocol of Xincaimian No.7 via somatic embryogenesis was established, laying a solid foundation for gene engineering in naturally colored cotton.