Analysis of Southern Blot and Genetic Character in Transgenic Cotton Expressing epsps-G6 Gene

doi:10.7668/hbnxb.2018.03.004

Abstract

Abstract: In order to identify the epsps-G6 gene successfully introduced into the cotton genome and analyze the copy number and genetic characteristics of the transgenic glyphosate-resistant cotton,eight transformation events(G6-1,G6-4,G6-5,G6-7,G6-11,G6-19,G6-20,G6-25)obtained by the pollen tube pathway method were used as materials. The results of southern Dot Blotting,double and single digestion enzyme Southern hybridization indicated that the exogenous epsps-G6 gene had been successfully integrated into the cotton genome and that four of the transformants contained a single copy of epsps-G6 gene. Three homozygous lines(G6-1,G6-5,G6-19)by self-pollination,with a single copy insertions and high glyphosate resistance,were selected to reciprocal cross with genetic background cultivar CCRI-49 and genetic standard line TM-1. The results of genetic analysis suggested that the three transformation events were inconsistent with the Mendelian separation rule of 3:1,which was not influenced by the cytoplasmic effect.

Key words: Cotton, epsps-G6 gene, Glyphosate, Herbicide, Pollen tube pathway

CLC Number:

S562.03

YAN Shufeng, XU Mengmeng, ZHU Shuijin, GUO Xinhai, TIE Shuanggui. Analysis of Southern Blot and Genetic Character in Transgenic Cotton Expressing epsps-G6 Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(3): 19-23. doi: 10.7668/hbnxb.2018.03.004.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2018.03.004

http://www.hbnxb.net/EN/Y2018/V33/I3/19

References

[1] Liang C,Sun B,Meng Z,et al. Co-expression of GR79 EPSPS and GAT yields herbicide-resistant cotton with low glyphosate residues[J]. Plant Biotechnology Journal,2017,15(12):1622-1629.
[2] 赵锦慧,郭婕,胡彦民. 应用诱变法筛选玉米抗草甘膦突变株系的研究[J]. 河南农业科学,2011,40(4):49-52.
[3] Wang Y Y,Zhou Z L,Wang X X,et al. Genome-wide association mapping of glyphosate-resistance in Gossypium hirsutum races[J]. Euphytica,2016,209(1):209-221.
[4] 李世民,沈海龙,刘迪,等. 盛蕾期转基因抗草甘膦棉花对草甘膦的抗性研究[J]. 河南农业科学,2010,39(10):46-48.
[5] 王旭静,张欣,刘培磊,等. 复合性状转基因植物的应用现状与安全评价[J]. 中国生物工程杂志,2016,36(4):18-23.
[6] 温广月. 抗草甘膦作物研究进展[J]. 世界农药,2010,32(1):10-17.
[7] 燕树锋,刘海芳,铁双贵,等. 通过草甘膦浸种鉴定抗草甘膦棉花[J]. 分子植物育种,2016,14(3):604-607.
[8] 王慧,闫晓红,徐杰,等. 我国抗草甘膦基因的发掘现状[J]. 农业生物技术学报,2014,22(1):109-118.
[9] Dill G M. Glyphosate-resistant crops:history,status and future[J]. Pest Management Science,2005,61(3):219-224.
[10] 燕树锋,刘海芳,铁双贵,等. EPSPS外源基因对陆地棉农艺性状和纤维品质的影响[J]. 棉花学报,2015,27(5):489-493.
[11] 欧婷,何秋伶,陈进红,等. 基于抗草甘膦基因的棉花茎尖农杆菌介导转化方法的研究[J]. 棉花学报,2013,25(5):410-416.
[12] 周长发,张锐,张晓,等. 地高辛随机引物法标记探针的Southern杂交技术优化[J]. 中国农业科技导报,2009,11(4):123-128.
[13] 张晓,张锐,于源华,等. 基于DIG-化学发光法的Southern blot方法优化[J]. 生物技术通报,2011(9):205-210.
[14] 刘禄,牛焱焱,雷昊,等. 基于地高辛标记对小麦进行Southern杂交分析主要影响因素的优化和验证[J]. 植物遗传资源学报,2012,13(2):182-188.
[15] 王小丽,杜建中,郝曜山,等. 转BADH基因玉米植株的获得及其耐盐性分析[J]. 作物学报,2014,40(11):1973-1979.
[16] 王秀红, 史向远, 白建荣, 等. 抗草甘膦基因的玉米转化及后代遗传分析[J]. 华北农学报, 2011, 26(1):117-121.
[17] 王守才,王国英,丁群星,等. 转基因在玉米中的遗传分离与整合特性的研究[J]. 遗传学报,1999,26(3):254-261.
[18] 马盾,周小云,黄乐平. 花粉管通道法转基因棉花后代的遗传特性[J]. 西北农业学报,2008,17(3):147-149.
[19] 刘昭军,李铁,刘丽艳,等. 花粉管通道介导的抗除草剂基因(bar)对大豆的遗传转化[J]. 大豆科学,2007,26(3):310-314.
[20] 燕树锋,祝水金,刘海芳,等. 转EPSPS基因抗草甘膦棉花的遗传分析[J]. 华北农学报,2015,30(3):54-57.

Please choose a citation manager

Content to export