彩色棉再生体系影响因子及抗病基因NP-1转化的研究  

Study on Factors Influencing Regeneration System and NP-1 Gene Transformation in Colored Cotton

作　　者：张红艳[1] 李冠[1] 李仁敬[2] 

机构地区：[1]新疆大学生命科学与技术学院,新疆乌鲁木齐830046 [2]新疆天彩科技股份有限公司,新疆乌鲁木齐830016

出　　处：《新疆大学学报（自然科学版）》2004年第1期94-98,共5页Journal of Xinjiang University(Natural Science Edition)

摘　　要：选用彩色棉优良品种(棕色):新彩1号、新彩2号.以下胚轴和子叶作为外植体,确立了5～6日苗龄的彩色棉下胚轴是较好的愈伤组织诱导外植体.不同激素组合及浓度对愈伤组织的诱导有重要作用,2,4-D是愈伤组织诱导中重要的激素,在附加有0.1mol/L2,4-D+0.1mol/LKT的MSB培养基上,诱导形成的愈伤组织质地疏松、生长旺盛,有利于分化为胚性愈伤.比较了不同糖源在愈伤组织诱导中的效应,在蔗糖和葡萄糖为糖源的培养基中,愈伤组织诱导率均为100%,但葡萄糖更有利于愈伤组织的形成和生长,蔗糖的诱导效果次之;麦芽糖和乳糖对愈伤组织的诱导效应不及蔗糖和葡萄糖;甘露醇作为糖源则抑制了愈伤组织的诱导和生长.在胚性愈伤组织的诱导培养中,KNO3加倍和NH4NO3减半的组合利于胚性愈伤组织的发生.在根癌农杆菌介导的抗病基因兔防御素NP-1转化过程中,外植体经农杆菌浸染10min,共培养48h较为合适,在添加头孢霉素500mg/L、卡那霉素50mg/L的培养基中筛选抗性愈伤组织.Explants derived from hypocotyls and cotyledons segments of 5~6 day seedlings , and hypocotyls are superior to cotyledons in the induction of callus. Different hormones and concentrations play an important role in the callus induction and growth. 2,4-D is an important hormone of induced callus. State of the callus growth is better on basic medium MSB medium containing 0.1 mol/L2,4-D and 0.1 mol/L KT. The effects of various sugar sources is different on the formation and growth of cotton callus. The results indicat that glucose is benifit to the induction and proliferation of cotton callus, sucrose comes second. Maltose and lactose are not good to the induction and growth of callus. The addition of mannitol restrain the induction of callus. The medium MSB containing twice of KNO3 and NH4NO3 reduced by half can induce a number of embryogenic callus. Furthermore, we discuss Agrobacterium-mediated carrying NP-1 transformation and some factors affecting the transformation frequency. It is appropriate for Agrobacterium infecting explants 10min and coculturing 48h, then transferred into the medium supplemented with 500 mg/L Cefotaxime and 50 mg/L kanamycin to select resistant callus.

关 键 词：彩色棉 影响因子 愈伤组织 兔防御素NP-1 外植体确定 根癌农杆菌 遗传转化 糖源诱导效应 

分 类 号：S562.037[农业科学—作物学]