棉花胚性愈伤的快速诱导和繁殖（英文）

High Efficiently Inducing and Cloning Embryogenic Callus in Cotton

棉花（Gossypium hirsutum L.）是世界上最重要的商业纤维作物。与其他作物相比,棉花再生体系较难。通过生物技术对棉花进行遗传改良往往受制于缺乏高效的再生体系。高效、快速的再生体系的开发有助于提高棉花转化效率。在0.10mg/L的吲哚乙酸（IAA）、0.09mg/L细胞分裂素（KT）和0.09mg/L 2,4-二氯苯氧乙酸（2,4-D）的MSB5培养基上产生高效愈伤组织。胚性愈伤组织可在愈伤组织基础上与SM2培养基中诱导出来,也可在＂M1-SM1-SM2＂培养基交替使用过程中诱导出来。利用此方法可在2-3月内诱导出胚性愈伤,大大缩短再生周期。胚性愈伤在PM1或PM2培养基中可以维持良好的再生状态。可见建立的快速诱导和繁殖胚性愈伤的方法可以用于棉花基因工程遗传改良。

Cotton（Gossypium hirsutum L.）is one of the most important commercially fiber crops in the world.Compared with other crops,cotton represents a recalcitrant species for regeneration protocols.Genetic improvement in cotton via biotechnology is limited due to lack of an efficient regeneration system.The development of efficient and rapid regeneration protocol could help improve transformation efficiency in cotton.In this study,a novel protocol was developed for embryogenic callus inducing from hypocotyls of cotton cultivars.We also developed a novel method to keep and clone the embryogenic callus.Calli were efficiently produced on the MSB5 medium with 0.10mg/L indole-3-acetic acid（IAA）,0.09mg/L Kinetin（KT）and 0.09mg/L 2,4-Dichlorophenoxyacetic acid（2,4-D）.Embryogenic calli could be induced after calli were sub-cultured on SM2 medium.Embryogenic calli also could be induced by alternately cultured on＂M1-SM1-SM2＂media.Using this protocol,embryogenic calli could be induced in 2-3months.The important advantage of this presented method is shortening of regeneration time.Embryogenic calli could keep the good differentiation state when they were cultured on PM1 or PM2medium.The successful protocol of maintaining the differentiation state of embryogenic calli established in this study could be used to improve cotton cultivars by genetic engineering.