四个抗草甘膦基因的抗性比较

doi:10.11686/cyxb20150519

草业学报 ›› 2015, Vol. 24 ›› Issue (5): 159-166.DOI: 10.11686/cyxb20150519

四个抗草甘膦基因的抗性比较

刘健^{1, 2**, *}, 肖雅文^{2**, *}, 芦佳^{1, 2**, *}, 吴忠义², 张中保², 徐杰^{1*, *}, 姚磊^{2*, *}

1.内蒙古师范大学生命科学与技术学院,内蒙古呼和浩特 010022;
2.北京市农林科学院农业生物技术研究中心,北京100097

收稿日期:2014-05-16 出版日期:2015-05-20 发布日期:2015-05-20
作者简介:刘健(1988-),男,山西吕梁人,在读硕士。E-mail: 906047892@qq.com。肖雅文(1988-),女,四川成都人,硕士。E-mail: xyw_1020@126.com。芦佳(1987-),女,山西大同人,在读硕士。E-mail: 905696441@qq.com。
基金资助:
北京市农林科学院科技创新能力建设专项(No.KJCX201102003)资助

Comparison of four glyphosate tolerance genes

LIU Jian^{1, 2, **}, XIAO Ya-Wen^{2, **}, LU Jia^{1, 2, **}, WU Zhong-Yi², ZHANG Zhong-Bao², XU Jie^{1, *}, YAO Lei^{2, *}

1.College of Life Science and Technology, Inner Mongolia Normal University, Hohhot 010022, China;
2.Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry, Beijing 100097, China

Received:2014-05-16 Online:2015-05-20 Published:2015-05-20

摘要/Abstract

摘要： 草甘膦抗性是当代农业重要的农艺性状。为了选择最佳的抗性基因用于植物筛选标记,通过选取cp4-epsps、GR79-epsps、gat4621和GAT四个草甘膦抗性基因进行比较,在拟南芥中验证各基因对草甘膦的抗性。4个基因的转化苗经4个浓度的草甘膦喷施检测,结果显示,GAT类基因的抗性明显高于epsps类基因,基因间的差异达到了极显著水平。gat4621基因对草甘膦的抗性最好,在各浓度下存活率均表现为最高值,且抗性稳定。国内发掘的GAT基因对草甘膦也表现出良好的抗性。GAT类基因的拟南芥转化苗在喷施高浓度草甘膦条件下出现严重抽苔抑制;但在停施草甘膦后抑制解除,且可以开花结实。转epsps类基因可以获得高抗草甘膦植株,但在花期喷施草甘膦将造成败育。从基因大小及抗性强弱考虑,GAT类基因作为筛选标记基因应该更为适合。

Abstract: Glyphosate tolerance is an important agronomic trait in modern agriculture. In order to determine the best glyphosate tolerance gene for plant selectable markers we compared four glyphosate tolerance genes, cp4-epsps, GR79-epsps, gat4621 and GAT, in Arabidopsis. Four transgenic plants harboring each gene were treated with four different rates of glyphosate. The results showed that the resistance of plants with GAT gene was significantly higher than plants with the epsps gene. Plants with the gat4621 gene had the highest survival rate at all herbicide rates. The plants with the domestic GAT gene also had strong resistance. Reproductive development of Arabidopsis plants with the GAT gene was suppressed at higher glyphosate application rates.It was concluded that the GAT gene was more suitable as a selectable marker gene.

刘健, 肖雅文, 芦佳, 吴忠义, 张中保, 徐杰, 姚磊. 四个抗草甘膦基因的抗性比较[J]. 草业学报, 2015, 24(5): 159-166.

LIU Jian, XIAO Ya-Wen, LU Jia, WU Zhong-Yi, ZHANG Zhong-Bao, XU Jie, YAO Lei. Comparison of four glyphosate tolerance genes[J]. Acta Prataculturae Sinica, 2015, 24(5): 159-166.

参考文献

[1] Amrhein N, Deus B, Gehrke P, et al . The site of the inhibition of the shikimate pathway by glyphosate: II. Interference of glyphosate with chorismate formation in vivo and in vitro . Plant Physiology, 1980, 66(5): 830-834.
[2] Gruys K J, Walker M C, Sikorski J A. Substrate synergism and the steady-state kinetic reaction mechanism for EPSP synthase from Escherichia coli . Biochemistry, 1992, 31(24): 5534-5544.
[3] Wang H, Yan X H, Xu J, et al . Current status of excavation glyphosate-resistant gene in China. Journal of Agricultural Biotechnology, 2014, 22(1): 109-118.
[4] Padgette S R, Kolacz K H, Delannay X, et al . Development, identification, and characterization of a glyphosate tolerant soybean line. Crop Science, 1995, 35: 1451-1461.
[5] Forlani G, Mangiagalli A, Nielsen E, et al . Degradation of the phosphonate herbicide glyphosate in soil: evidence for a possible involvement of unculturable microorganisms. Soil Biology and Biochemistry, 1999, 31(7): 991-997.
[6] Green J M, Hazel C B, Forney D R, et al . New multiple-herbicide crop resistance and formulation technology to augment the utility of glyphosate. Pest Management Science, 2008, 64(4): 332-339.
[7] Lin M, Liang A M, Lu W, et al . High Glyphosate Tolerant EPSP Synthase and its Coding Sequence[P]. China Patent, ZL200710177090.1, 2010.
[8] Cao G, Liu Y, Zhang S, et al . A novel 5-enolpyruvylshikimate-3-phosphate synthase shows high glyphosate tolerance in Escherichia coli and tobacco plants. PloS one, 2012, 7(6): e38718.
[9] Lin M, Dun B Q, Lu W, et al . Glyphosate Acetyltransferase Gene and its Application[P]. China Patent, ZL200510086626.X, 2007.
[10] Lin M, Wang X J, Dun B Q. A Bivalent Expression Vector for Generating Glyphosate-resistant Plant[P]. China Patent, ZL200710118968.4, 2010.
[11] Komori T, Imayama T, Kato N, et al . Current status of binary vectors and superbinary vectors. Plant Physiology, 2007, 145(4): 1155-1160.
[12] Zhou H, Arrowsmith J W, Fromm M E, et al . Glyphosate-tolerant CP4 and GOX genes as a selectable marker in wheat transformation. Plant Cell Reports, 1995, 15(3-4): 159-163.
[13] Yao L, Yan X, Wang H, et al . pYBA: A series ease-of-use binary vectors for plant transformation. Plant Genomics in China XIII[C]. 2012: 212.
[14] Preuss S B, Meister R, Xu Q, et al . Expression of the Arabidopsis thaliana BBX32 gene in soybean increases grain yield. PloS one, 2012, 7(2): 30717.
[15] Yan X H, Wang H, Ye Y Y, et al . YBA100: An ease-of-use binary vector with LoxP-FRT recombinase site for plant transformation. Molecular Plant Breeding, 2012, 10(3): 371-379.
[16] Clough S J, Bent A F. Floral dip: a simplified method for Agrobacterium -mediated transformation of Arabidopsis thaliana . The Plant Journal, 1998, 16(6): 735-743.
[17] Zeng G, Ye Y Y, Yan X H, et al . Quick detection of turnip mosaic virus by one-step multiple RTPCR. Acta Agriculture Boreali-Sinica, 2012, 27(3): 102-107.
[18] Joshi C P, Zhou H, Huang X, et al . Context sequences of translation initiation codon in plants. Plant Molecular Biology, 1997, 35(6): 993-1001.
[19] Pline-Srnic W. Technical performance of some commercial glyphosate-resistant crops. Pest Management Science, 2005, 61(3): 225-234.
[20] Gaines T A, Zhang W, Wang D, et al . Gene amplification confers glyphosate resistance in Amaranthus palmeri . Proceedings of the National Academy of Sciences, 2010, 107(3): 1029-1034.
[3] 王慧, 闫晓红, 徐杰, 等. 我国抗草甘膦基因的发掘现状. 农业生物技术学报, 2014, 22(1): 109-118.
[7] 林敏, 梁爱敏, 陆伟, 等. 高耐受草甘膦的EPSP合酶及其编码序列[P]. 中国专利,ZL200710177090.1, 2010.
[9] 林敏, 顿宝庆, 陆伟, 等. 草甘膦乙酰转移酶基因及其应用[P]. 中国专利,ZL200510086626.X., 2007.
[10] 林敏, 王旭静, 顿宝庆. 培育抗草甘膦植物的双价表达载体[P]. 中国专利,ZL200710118968.4. 2010.
[15] 闫晓红, 王慧, 叶艳英, 等. 一个含LoxP-FRT重组酶位点及易于操作的植物表达双元载体pYBA100. 分子植物育种, 2012, 10(3): 371-379.
[17] 曾钢, 叶艳英, 闫晓红, 等. 简化一步多重RT-PCR法快速检测芜菁花叶病毒. 华北农学报, 2012, 27(3): 102-107.

四个抗草甘膦基因的抗性比较

Comparison of four glyphosate tolerance genes

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics