【作者】 迟吉娜；

【导师】 马峙英；

【作者基本信息】 河北农业大学 ， 作物遗传育种， 2004， 硕士

【摘要】 棉花是我国主要的经济作物之一，棉花生产的兴衰对于国民经济的发展具有重要意义。随着基因工程和分子生物学的发展，以转基因技术和分子选择技术为核心的现代育种，能够实现作物设计育种，将外源有益基因导入受体植物细胞，并使其定向而稳定地遗传。小麦、玉米、水稻、大豆、番茄等作物的基因工程都取得了很大的进展。目前转基因的方法很多，在棉花的遗传转化中应用的主要有基因枪法、农杆菌介导法和花粉管通道法，其中前两种方法都需要建立基因转移的受体系统。建立高效而稳定的植物遗传转化体系，是保证植物转基因获得大量可供筛选群体的前提条件。因此，只有不断地优化棉花体细胞胚胎发生和植株再生体系，才能有效的发挥基因遗传转化在棉花现代育种中的作用。 棉花一直被公认为是植株再生最困难的植物之一，棉花组织和细胞培养落后于其它植物。虽然棉花体细胞组织培养经过20多年的发展，在一些实验室已基本建立了陆地棉植株再生体系，但仍然有许多尚未解决的问题，如再生品种少、胚状体的萌发率偏低、畸形胚率高等。本试验对陆地棉体细胞胚胎发生和植株再生体系进行了优化，并对再生植株的变异进行了初步研究： 1 以13个陆地棉(Gossypium hirsutum L.)品种为材料，对体细胞胚胎发生和植株再生体系进行了研究，本试验已得到珂字312、冀无2031和冀合321共3个品种的再生植株和珂字201的胚性愈伤组织。 2 对多种激素及其配比对愈伤组织的诱导效应进行了研究，并与前人的研究进行比较，建立了一套比较完善的培养程序。 3 研究了珂字201幼花提取物在愈伤组织诱导及其分化中的作用，珂字201幼花提取物促进愈伤组织诱导和分化的最适浓度为40 mg／L。 4 对聚乙二醇(PEG)在胚性愈伤组织悬浮培养和固体培养中的作用进行了研究，液体悬浮培养基中添加50g／L的PEG6000对胚性愈伤组织的增殖和胚状体的正常萌发有促进作用；固体培养基中添加80～160mg／LPEG6000并在黑暗中培养1～2周对悬浮培养后的胚性愈伤组织起到保持胚性的作用，恢复正常光照培养后还能促进胚状体的正常萌发。 5 在珂字201继代过程中观察到了非胚性愈伤组织向胚性愈伤组织的转化现象；发现了前人未曾报道过的体胚苗变异类型。 6 对棉花体细胞胚胎发生和植株再生基因型范围的拓宽进行了研究，有望得到丰产抗病棉花品种农大94-7的胚性愈伤组织。更多还原

【Abstract】 Cotton is one of the most important cash crops in China, playing an important role in the national economy. With the development of genetic engineering and molecular biotechnology, molecular breeding becomes a new kind of method to realize crop designing. Some exotic genes coming from different kinds of plant, animal and microbecterium can be introduced to target plant cell and the exotic genes can be inherited truly. Genetic engineering provided the new and effective approach to facilitate plant breeding. Crop biotechnology has been successfully made in recent years, and considerable amounts of genetically engineered plants have been produced, such as wheat, rice, maize, soybean, etc. There were more than ten transformation systems used in plant genetic transformation. Agrobacterium-mediated gene transfer, particle bombardment and pollen-tube pathway system were often used in cotton transformation. The former two genetic transformation systems both need efficient and repeatable somatic embryogenesis and plant regeneration system.Cotton was one of the most difficult regenerated plants. Compared to other crops, cotton tissue culture was lagging behind. In vitro cultured cotton, cells had been induced to produce somatic embryogenesis using varied strategies in some laboratories. Over the past 20 years, a great deal of progress was made. However, there were still some problems to be solved. The rate of germination on embryoids was on the low side, and the abnormal embryos proportion was high. There was a need to broaden the number of regenerable cotton species. One of the objective of this work was to optimize the somatic embryogenesis and plant regeneration system in upland cotton, and somaclonal variation was studied elementarily.1 Thirteen varieties of upland cotton were studied on somatic embryogenesis and plant regeneration. Coker312, Jiwu2031, Jihe321 and Coker201 produced different amount of embryogenic callus, and the former three varieties induced regenerated plant.2 Effects of several phytohormones and their combinations were studied on callus induction, and the process were established.3 The impact of different method extracting solution of Coker 201 young inflorescence on callus induction were studied, more and better callus were produced in the media with extracting solution in water than without it. The proper concentration was 40mg/L.4 The function of various concentration of PEG6000 on germination in liquid culture and solid culture were studied. PEG6000 markedly raised embryoids germination, which got the highest rate of embryoids germination in the treatments when 50g/L in liquid culture. It promoted embryogenic callus germination in normal light period in two weeks after one or two weeks in darkness with 80~160mg/L PEG6000 in solid culture.5 Wide variation was observed in the regenerated plant. Some new kinds of variations were found in the regenerated plants. And the new transform from non-embryogenic callus to embryogenic callus was observed in Coker 201 callus induction.6 To broad the number of regenerable cotton species, a new high yield and disease resistant variety, Nongda94-7 was studied, and Embryogenic callus of it could be induced for the future.更多还原