共转化6-SFT和bar表达框小麦的获得及检测

doi:10.14088/j.cnki.issn0439-8114.2022.24.038

湖北农业科学 ›› 2022, Vol. 61 ›› Issue (24): 185-188.doi: 10.14088/j.cnki.issn0439-8114.2022.24.038

共转化6-SFT和bar表达框小麦的获得及检测

贺晓岚^1a, 王建伟^1b, 陈新宏², 李文旭³, 秦绍钊^1a, 汤宏^1b, 吴显芝^1a

1.凯里学院,a.大健康学院,b.理学院,贵州凯里 556011;
2.西北农林科技大学农学院/陕西省植物遗传工程育种重点实验室, 陕西杨凌 712100;
3.河南省农业科学院小麦研究所,郑州 450002

收稿日期:2022-02-07 出版日期:2022-12-25 发布日期:2023-01-18
通讯作者: 王建伟（1982-）,男,甘肃平凉人,副教授,博士,主要从事植物营养学研究,（电话）18212333515（电子信箱）agan1982@126.com。
作者简介:贺晓岚（1980-）,女,内蒙古乌兰察布人,博士,主要从事植物基因克隆及遗传转化研究,（电话）18285549260（电子信箱）helingzhi123@126.com。
基金资助:
贵州省教育厅科技拔尖人才支持计划项目（黔教合KY字[2017]094）; 贵州省科技支撑计划项目（黔科合支撑[2017]2522 ）; 贵州省基础研究计划项目（黔科合基础[2017]1167）; 贵州省教育厅黔教合人才团队项目（[2015]70）

Acquisition and detection of co-transformed 6-SFT and bar expression frames in wheat

HE Xiao-lan^1a, WANG Jian-wei^1b, CHEN Xin-hong², LI Wen-xu³, QIN Shao-zhao^1a, TANG Hong^1b, WU Xian-zhi^1a

1a. School of Life and Health Science, 1b. School of Science, Kaili University, Kaili 556011, Guizhou, China;
2. College of Agronomy/Shaanxi Key Laboratory of Genetic Engineering for Plant Breeding, Northwest A&F University, Yangling 712100, Shaanxi, China;
3. Institute for Wheat Research, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China

Received:2022-02-07 Online:2022-12-25 Published:2023-01-18

摘要/Abstract

摘要： 为了探究源于华山新麦草和簇毛麦的果聚糖合成酶基因6-SFT改良小麦（Triticum aestivum L.）抗非生物胁迫的可行性,以bar为筛选标记基因,通过基因枪介导法将Ph-6-SFT或Dv-6-SFT基因导入普通小麦（科农199）幼胚愈伤组织中,用除草剂草丁膦作为筛选剂筛选得到抗性植株,并对其进行PCR及GUS组织化学染色鉴定。结果显示,通过PPT筛选体系分别得到146株和187株抗性植株,其中PCR阳性植株分别为4株和6株,GUS组织化学染色阳性植株均为1株,转化率分别为0.24%和0.19%。结果表明,Ph-6-SFT和Dv-6-SFT基因已分别整合到小麦基因组中,获得了无抗生素标记基因的转基因小麦。

关键词: 小麦（Triticum aestivum L.）, 共转化, Ph-6-SFT基因, Dv-6-SFT基因, bar基因, 表达盒

Abstract: To investigate the feasibility of fructan synthase gene 6-SFT derived from Psathyrostachys huashanica and Haynaldia villosa to improve the resistance of wheat （Triticum aestivum L.） to abiotic stress, the Ph-6-SFT or Dv-6-SFT gene was introduced into the young embryonic healing tissue of the common wheat family Kenong 199 by gene gun-mediated method using bar as a screening marker gene, and resistant plants were obtained by using the herbicide glufosinate as a screening agent and identified by PCR and GUS histochemical staining. The results showed that 146 and 187 resistant plants were obtained by the PPT screening system, of which 4 and 6 were PCR positive and 1 was GUS histochemically stained, respectively, and the transformation rates were 0.24% and 0.19%, respectively. The results showed that Ph-6-SFT and Dv-6-SFT genes had been integrated into the wheat genome, respectively, and transgenic wheat without antibiotic marker genes was obtained.

Key words: Triticum aestivum L., co-transformation, Ph-6-SFT gene, Dv-6-SFT gene, bar gene, expression cassettes

中图分类号:

S188

贺晓岚, 王建伟, 陈新宏, 李文旭, 秦绍钊, 汤宏, 吴显芝. 共转化6-SFT和bar表达框小麦的获得及检测[J]. 湖北农业科学, 2022, 61(24): 185-188.

HE Xiao-lan, WANG Jian-wei, CHEN Xin-hong, LI Wen-xu, QIN Shao-zhao, TANG Hong, WU Xian-zhi. Acquisition and detection of co-transformed 6-SFT and bar expression frames in wheat[J]. HUBEI AGRICULTURAL SCIENCES, 2022, 61(24): 185-188.

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共转化6-SFT和bar表达框小麦的获得及检测

Acquisition and detection of co-transformed 6-SFT and bar expression frames in wheat

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