XJIPC OpenIR  > 省部共建新疆特有药用资源利用重点实验室
Alternative TitleEstablishment of Genetic Transformation System on Cotton (Gossypium hirsutum L.) Cultivar ‘Xinluzao 33’ and Transformation of AtPGIP1 Gene
肖向文; 刘海峰; 李雪源; 曹艳艳; 王俊铎; 黄芳; 郑巨云; 谢旗; 李晓波
Source Publication西北植物学报

利用生物技术方法对棉花进行遗传改良主要限于有效的遗传转化系统.以新疆主栽优良陆地棉品种‘新陆早33号’为材料,利用下胚轴作为外植体对影响农杆菌介导的棉花遗传转化及体细胞胚胎发生的因素进行研究,成功建立了除草剂Basta筛选的棉花遗传转化技术体系.同时将植物抗病相关基因多聚半乳糖醛酸酶抑制蛋白基因AtPGIP1导入棉花,经过对再生转化植株的PCR鉴定,初步证明外源基因已经整合到棉花基因组.研究发现:Basta是棉花遗传转化中很有效的筛选剂,低浓度Basta(2.5 mg/L)就能够获得很好的筛选效果;较低的共培养温度(20℃)及合适的农杆菌浓度(OD600=0.5)有助于提高转化效率.该研究结果表明,‘新陆早33号’具备作为棉花优良遗传转化受体的基本特征,研究中获得的15株AtPGIP1转基因植株经PCR分子检测均为阳性植株.该研究为新疆棉区棉花分子生物学研究及转基因育种研究奠定了重要基础.

Other Abstract

Genetic improvement in cotton via biotechnology is limited due to lack of an efficient transformation and regeneration system.A reliable and high-efficiency system of genetic transformation mediated by Agrobacterium tumefaciens was developed in cotton(Gossypium hirsutum L.)cultivarXinluzao 33(an elite cultivar in Xinjiang area),using hypocotyl as explants.Various aspects of transformation were examined in efforts to improve the efficiency of producing transgenic plants.Meanwhile,a foreign gene Polyga-lacturonase-inhibiting protein(AtPGIP1),which is related to plant disease resistance,was induced to this cultivar.The integration of the interest gene and BAR transgenes into the genome of putative transgenic plants was confirmed by polymerase chain reaction(PCR).Our results showed Basta is an effective selection agent in cotton genetic transformation,which can be applied at a low concentration of 2.5mg/L. Agrobacteriumat a relatively low concentration(OD_(600)=0.5)helped to improve the efficiency of transformation. Relatively low co-cultivation temperature(20℃)was optimal for obtaining a higher efficient transformation. This result suggested thatXinluzao 33can be used as a good receptor variety in cotton genetic transformation mediated by A.tumefaciens,increasing the range of cotton genotypes that can be transformed. Using this protocol,more than 15normal plants were obtained through transformation of AtPGIP1, of which all regenerated plants were transgene-positive confirmed by PCR analysis.The protocol established in this study can be used in cotton functional genomics and in improving cotton cultivars by genetic engineering.

Keyword棉花 组织培养 体细胞胚胎发生 遗传转化 Cotton Tissue Culture Somatic Embryogenesis Genetic Transformation
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Document Type期刊论文
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GB/T 7714
肖向文,刘海峰,李雪源,等. 新陆早33号’棉花转基因体系建立及AtPGIP1基因的导入[J]. 西北植物学报,2014,34(4):658-664.
APA 肖向文.,刘海峰.,李雪源.,曹艳艳.,王俊铎.,...&李晓波.(2014).新陆早33号’棉花转基因体系建立及AtPGIP1基因的导入.西北植物学报,34(4),658-664.
MLA 肖向文,et al."新陆早33号’棉花转基因体系建立及AtPGIP1基因的导入".西北植物学报 34.4(2014):658-664.
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