陇绿棉3号胚性愈伤组织的诱导及植株再生

摘要

We investigated the influential factors in green cotton regeneration systems involving Longki 3, in order to establish a stable and efficient system, expanding renewable cotton genotypes. We observed and recorded the entire cultural process of Longlu 3 from explant to embryogenesis and seedling differentiation. We found that the optimal cultural conditions for the ex-plant were dark conditions for 4 d and light for 2 d. The optimal hormone combination for callus induction was MSB medium (MS inorganic salts + B5 organic ingredients) with 0.1 mg · L-1 2,4-D + 0.05-0.3 mg·L-1 KT or 0.1 mg· L-1 2,4-D + 0.1 mg · L-1 KT + 0.1 mg·L-1 IAA. The optimal conditions for callus differentiation were MSB with 1.9 g·L-1 KNO3 and 2.0 g·L-1 gelrite. In MSB with 0.1 mg · L-1 KT and 0.4 mg · L-1 IBA, both embryogenesis rate and cotyledon embryo generation rate reached peak values, and the cotyledon embryo was successfully differentiated into seedlings in 1/2 MSB medium.%以陇绿棉3号为试验材料,针对从外植体培养到胚状体发生及分化成苗整个组织培养过程的不同阶段,研究探索绿色棉—陇绿棉3号的再生条件,获得再生植株,为拓宽彩色棉再生基因型和转基因研究奠定了基础.研究结果表明:陇绿棉3号外植体的最适培养条件为暗4d/光2d处理；愈伤组织诱导最佳激素组合为0.1 mg·L-1 2,4-D+0.05～0.3 mg·L-1 KT或0.1 mg·L-1 2,4-D+0.1 mg·L-1 KT+0.1 mg·L-1 IAA的MSB培养基(MS培养基无机盐+B5培养基有机成分)；愈伤分化的最适条件为MSB中添加1.9 g·L-1 KNO3和2.0 g·L-1凝固剂gelrite;胚性愈伤增殖培养基为MSB(NH4NO3减半)中添加1.9 g·L-1KNO3并添加1.8 g·L-1凝固剂gclrite 、0.1g·L-1天冬酰胺和0.1 g·L-1谷氨酰胺.在去除NH4NO3,添加0.1 mg·L-1KT和0.4 mg·L-1 IBA的MSB培养基上,胚胎发生率和子叶胚萌发率均达到峰值,且子叶胚在1/2MSB培养基上可成功分化成苗.