转Cry1A基因海岛棉的获得和鉴定

doi:10.11924/j.issn.1000-6850.casb18030111

中国农学通报 ›› 2019, Vol. 35 ›› Issue (24): 122-127.doi: 10.11924/j.issn.1000-6850.casb18030111

所属专题：生物技术；棉花

转Cry1A基因海岛棉的获得和鉴定

邓莉

新疆农业大学农学院

收稿日期:2018-03-22 修回日期:2018-05-16 接受日期:2018-05-24 出版日期:2019-08-26 发布日期:2019-08-26
通讯作者: 邓莉
基金资助:
自治区重点研发项目“棉花优质、抗逆功能基因研究与育种新材料创制”（2016B01001-1）；国家自然科学基金“新疆海岛棉新海16体细胞胚发生体系的优化”（31660078）

The Acquisition of Transgenic Gossypium barbadense with Cry1A Gene and Its Identification

Received:2018-03-22 Revised:2018-05-16 Accepted:2018-05-24 Online:2019-08-26 Published:2019-08-26

摘要/Abstract

摘要： 进行农杆菌介导转抗虫基因试验，以期选育抗虫性优良的海岛棉新品系(品种)；本实验在海岛棉体细胞胚再生体系的基础上，将携带Cry1A基因的表达载体pBI121通过农杆菌介导法转化到‘新海33号’的胚性愈伤组织中。经培养基筛选、PCR检测、RT-PCR检测、抗虫性鉴定等筛选抗虫后代。经培养基筛选共获得12棵抗性植株。PCR检测结果显示，这12棵抗性植株均能扩增出了大小相符的片段，检测结果呈阳性；RT-PCR结果显示，12棵转基因植株目的基因已经整合入基因组，且能反转录出mRNA；Bt-Cry1Ab/Ac转基因试纸条检测结果表明这12棵转基因植株含有目的蛋白；抗虫性鉴定结果证实了这些转基因植株对棉铃虫幼虫具有一定的抗性。本研究成功将抗虫基因导入至海岛棉的体内，获得了具有抗虫性的植株，其结果对转基因抗虫海岛棉新品种的培育具有一定的参考价值。

关键词: 扭黄茅, 扭黄茅, 土壤水分胁迫, 光合作用光响应, 光响应模型

Abstract: Agrobacterium mediated transgenic insect-resistant gene test is conducted in order to select new island cotton lines (varieties) with excellent insect resistance. On the basis of embryonic regeneration system of island cotton somatic cells, the expression vector pBI121 with Cry1A gene was transferred to the embryogenic callus of‘Xinhai 33’by Agrobacterium-mediated method. The insect-resistant offspring were screened by the culture medium screening, PCR detection, RT-PCR detection and insect-resistance identification. 12 resistant plants were obtained. The PCR result revealed that the 12 resistant plants could amplify the expected-size DNA fragments, and the test results were positive. RT- PCR result showed that the target gene had been integrated into 12 transgenic plant genomes, and could reverse transcription of mRNA. The test result of Bt- Cry1Ab/Ac transgenic paper indicated that the 12 transgenic plants contained the target protein. Transgenic plants had certain resistance to cotton bollworm larvae by insect- resistant identification. This study successfully transforms insect-resistant genes into island cotton, obtains the insect-resistant plants, and the results have some reference value for the breeding of new transgenic insect-resistant island cotton varieties.

邓莉. 转Cry1A基因海岛棉的获得和鉴定[J]. 中国农学通报, 2019, 35(24): 122-127.

参考文献

[1]姚贺盛,张亚黎,易小平等.海岛棉和陆地棉叶片光合特性、冠层结构及物质生产的差异[J].中国农业科学,2015,48(2):251-261.
[2]孔庆平.我国海岛棉生产概况及比较优势分析[J].中国棉花,2002,29(12):19-22.
[3]宁新民,孔庆平,阿里甫,等.新疆海岛棉杭虫转基因棉花的筛选及杭虫鉴定[C].中国棉花学会2009年年会论文汇编.
[4]束春娥,刘贤金,柏立新,等.Bt 转基因棉花抗棉铃虫毒性机理研究[J].棉花学报,1996,8(4):219-222.
[5]崔洪志,郭三堆.Bt毒蛋白抗虫植物基因工程研究(综述) [J].农业生物技术学报,1998,6(2):166-172.
[6]Schuler T H, Popygm, Kerryb Y, et al. Insect-resistant transgenic plants[J].Tibtech,1998(16):168- 175.
[7]Schnepf H E, Whiteley H R. Cloning and Expression of the Bacillus Thuringiensis. Crystalprotein gene in Escherichiacoli [J]. Proc. Natl. Acad. Sci. USA 1981,78(5):2893-2897.
[8]Perlak, F J, Deaton R W, Armstrong T A, et al. Insect Resistant Cotton Plants [J]. Bio/Technology 1990,8:939-942.
[9]王继磊,刘迪秋,丁元明,等.BT转基因抗虫植物研究进展[J].生物学杂志,2010,27(4):75-78.
[10]崔洪志,郭三堆.我国抗虫转基因棉花研究取得重大进展[J].中国农业科学,1996,29( 1):93-95.
[11]贺雅婷,曲延英,孔庆平,等.海岛棉愈伤组织诱导及分化的影响因素初探[J].分子植物育种,2008,6(3):597-602.
[12]李琼.棉花体细胞胚胎发生机制研究及p53基因的遗传转化[D].新疆农业大学硕士论文,2012.
[13]周丽容.农杆菌介导转Bt基因海岛棉技术体系的建立及抗虫性鉴定[D].新疆农业大学,2011.
[14] 蓝海燕,刘桂珍,王长海.棉叶总DNA提取的改良方法[J].棉花学报,2000,12(1):53-56.
[15]何素平,赵静,界国香.BT毒蛋白免疫金标试纸条的制备与应用[C].中国棉花学会2006年年会暨论文汇编,2006,254-256.
[16] NY/T 2162-2012,棉花抗棉铃虫性鉴定方法[S].2016.
[17]苏豫梅.新疆海岛棉转基因育种的研究进展[J].南方农业,2015,9(24):252-256.
[18]Kumria R, Sunnichan V G, Das D K, et al. High-frequency somatic embryo production and maturation into normal plants in cotton(Gossypium hirsutum)through metabolic stress. Plant Cell Rep, 2003, 21:635-639.
[19]谢德意,金双侠,郭小平,等.棉花胚性愈伤组织的转化及转基因胚状体的有效萌发与成苗技术研究[J].作物学报,2007,33(5):751-756.
[20]李长福,吴振廷,沈基长,等.转BT基因棉的检测和抗虫性鉴定研究进展[J].安徽农业科学,2002,30(3):346-349.
[21]Mathiasc, Martine, Annevc. A semi-quantitative RT-PCR method to readily compare expression levels with in Botrytis cinerea multigenic families in vitro and in planta. Curr Genet, 2003(1):303-309.
[22]聂新辉,尤春源,陈惠瑜,等.金标Bt-CryIAb/Ac试纸条定性检测转Bt基因棉花的方法研究[J].中国棉花,2013,40(1):15-17.
[23]Halcomb J H, Benedict J H, Cook B, et al. Surviral and growth of bollworm and tobacco budworm on nontransgenic and transgenic cotton expressing a Cry IA insecticidal protein[J]. Enviro Entom, 1996,25(2):250- 255.
[24]Wilson, F D, et al. Resistance of cotton lines containing a Bacillus thuringiensis toxin to pink bollworm and other insects[J]. Econ Entomol, 1992, 85(4):1516-1521.

转Cry1A基因海岛棉的获得和鉴定

The Acquisition of Transgenic Gossypium barbadense with Cry1A Gene and Its Identification

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