【作者】 孙立军；

【导师】 毛鸿才； 郝秀英；

【作者基本信息】 新疆农业大学 ， 作物遗传育种， 2006， 硕士

【摘要】 棉花是植株再生最困难的作物之一,因此棉花组织培养和细胞培养落后于其它作物。虽然近些年,棉花的体细胞组织培养己经有了较大的发展,而且一些实验室己经建立了陆地棉再生体系,但是大多集中在少数品种上,尤其是一些模式品种,而且仍然存在着许多尚未解决的问题,如再生品种少、胚状体萌发率偏低、畸形苗率较高、再生周期长等问题,所以,不断发展新的再生体系和拓宽棉花再生植株的品种数量是目前棉花组织培养的新趋势。针对上述存在的诸多问题,本研究以新疆自育的中长绒陆地棉品种和特色长绒棉品种作为实验材料,着重从植物离体培养器官发生再生植株途径的角度考虑,进行子叶节和茎尖培养植株再生的研究,探讨棉花子叶节和茎尖再生,以及农杆菌介导遗传转化的主要影响因素。1.以中长绒陆地棉品种为实验材料,采用子叶节作外植体,分步添加6-BA来启动和诱导多芽的发生,经过伸长培养后,以1/2MS培养基添加10 mg/LIBA诱导生根,生根率可达72.2%。2.以新疆长绒棉品种为实验材料,采用子叶节作外植体,直接诱导多芽产生,在芽诱导培养基MSB+2.5mg/LKT+2.5mg/L6-BA上培养8周,再转入芽伸长培养基MSB+0.3mg/L6-BA培养4周,多芽诱导率达90%以上。3.以新陆中13的茎尖为材料,建立起茎尖培养体系:以3d苗龄,下胚轴长度为5-7mm的茎尖为外植体,接入附加0.1mg/LKT的MSB培养基中培养4周,成苗率能达到98.0%。4.棉花子叶节和茎尖分生组织培养植株成苗的时间短,培养方法简单,畸形苗少,并且不受基因型的限制,适合作为基因转化的受体系统,棉花茎尖分生组织更甚一筹。因此,本试验利用农杆菌介导法将绿色荧光蛋白(GFP)基因导入新陆中13茎尖分生组织,在含有Kan50mg/L的筛选培养基上进行直接筛选,最终得到抗性植株。更多还原

【Abstract】 Cotton is one of the most difficult regeneration in crops, and therefore lag the tissue and cell culture of other crops. Though a great deal of progress has been made in cotton tissue culture and the regenerative technique of upland cotton has been establish in some laboratorys for these years, these studies mostly focus on a few varieties, particularly some model varieties. And however, some problems have been not solved such as low germination rate of somatic embryos, high abnormal embryos’proportion, long cycle of regeneration and so on. So constantly developping newly regenerative system of cotton and broadening the number of regenerative varieties of cotton is presently a new trend to cotton tissue culture.The paper deal with the Gossypium hirsutum L. varieties with superior medium staple and the characteristic Gossypium barbadense L. varieties of Xinjiang cotton. Through organogenesis, carry on the research that regeneration from the cotyledon nodes and shoot apical meristems, inquiry into the cotton cotyledon nodes regeneration, the shoot apical meristems regeneration and the Agrobacterium -mediated transformation of main influence factor.1. The research deal with the Gossypium hirsutum L. varieties with superior medium staple of Xinjiang cotton that is one of three largest cotton production region in China. Cotyledon Nodes act as the explants In vitro. 6-BA of plant cytokinins is added into start-up medium and/or induction medium by two steps for preventing from browning effect on explants. Meanwhile, glucose serve as carbon resource in place of sucrose used usually. After elongating culture of shoots, rooting was induced in one half of MS medium added 10 mg/l IBA and its ratio can reach 72.2%.2.The research deal with the characteristic Gossypium barbadense L. varieties of Xinjiang cotton .Cotyledon Nodes act as the explants In vitro. The explants were cultured on the shoot inducement mediumMSB+2.5 mgs/LKT+2.5 mgs/L6-BAfor 8 weeks, then turn into the shoot elongation development the mediun for MSB+the 0.3 mgs/L6-BA develop for 4 weeks, having another the shoot inducement rate to reach to 90% above.更多还原