辣椒子叶外植体高效离体再生及抗性愈伤组织诱导  

High-frequency regeneration from cotyledon explants and optimized resistant callus formation of chili pepper

作　　者：杨博智 宋钰 童辉[2] 肖佳林 黄湖荣 周书栋[2] 邹学校[1] YANG Bozhi;SONG Yu;TONG Hui;XIAO Jialin;HUANG Hurong;ZHOU Shudong;ZOU Xuexiao(College of horticulture,Hunan Agricultural University,Changsha 410128,China;Hunan Vegetable Research Institute,Changsha 410125,China)

机构地区：[1]湖南农业大学园艺学院,长沙410128 [2]湖南省蔬菜研究所,长沙410125

出　　处：《植物生理学报》2022年第4期788-796,共9页Plant Physiology Journal

基　　金：国家自然科学基金项目(31601757和U19A2028);湖南省蔬菜产业技术体系项目。

摘　　要：为提高辣椒E29子叶外植体遗传转化效率,本研究建立了从E29子叶外植体愈伤组织中获得植株的高效离体再生体系,并优化了子叶外植体抗性愈伤组织获得的条件。结果表明,在添加2.0 mg·L^(-1)6-BA和0.5 mg·L^(-1)IAA的MS培养基中,大于98%的子叶外植体切口处可诱导愈伤组织。添加0.1 mg·L^(-1)2,4-D的MS培养基能高效促进愈伤组织增殖。添加9.0 mg·L^(-1)6-BA的MS培养基中,愈伤组织诱导芽点效果最好,20 d后芽点诱导率达11.27%。添加3.0 mg·L^(-1)6-BA、0.1 mg·L^(-1)IAA及0.3 mg·L^(-1)GA_(3)的MS培养基中,不定芽的伸长效果最佳,平均每块带芽点愈伤组织上可以产生2.01个伸长的不定芽。不添加任何激素的1/2MS培养基适宜生根,生根率可达100%。生根后的小植株炼苗后成活率可达98%。在此基础上优化E29子叶外植体抗性愈伤组织获得的条件,发现潮霉素B与特美汀(Timentin)最适筛选浓度分别为30mg·L^(-1)、300 mg·L^(-1),共培养最适时间为3 d,此条件下抗性愈伤组织诱导率可达到36.24%。该研究可为今后辣椒遗传转化体系的建立奠定基础。In order to improve genetic transformation efficiency of cotyledon explants of pepper E29,a high-frequency regeneration from cotyledon-induced callus was established and the resistant callus formation protocol was optimized.The result showed that more than 98%response in calli induction from the cuts of cotyledons explants on MS medium containing 2.0 mg·L^(-1)6-benzylaminopurine(6-BA)and 0.5mg·L^(-1)indolyl-3-acetic acid(IAA);The calli were successfully proliferated on MS medium containing 0.1mg·L^(-1)2,4-dichlorophenoxyacetic acid(2,4-D);The callus had the best bud induction effect on MS medium supplemented with 9.0 mg·L^(-1)6-BA for 20 d.Under this condition,the adventitious buds induction rate could reach 11.27%;Adventitious buds had the best elongation effect on MS medium supplemented with 3.0 mg·L^(-1)6-BA,0.3 mg·L^(-1)gibberellin A3(GA_(3)),and 0.1 mg·L^(-1)IAA.An average of 2.01 shoots were obtained on each adventitious bud from cotyledons explants.All shoots could take roots on half-strength MS medium and the seedling survival rate was 98%.For resistant callus formation,the optimal concentration of hygromycin B and Timentin was 30 mg·L^(-1)and 300 mg·L^(-1),respectively.The co-cultivation of the hypocotyl explants with Agrobacterial for the duration of 3 d was optimum for obtaining high transformation efficiency,the induction rate of resistant callus reached 36.24%under this culture condition.We expect this method will be beneficial to the establishment of a genetic transformation system for chili peppers.

关 键 词：辣椒 子叶外植体 再生培养 农杆菌介导 抗性愈伤组织 

分 类 号：S641.3[农业科学—蔬菜学]