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2025年8月3日 星期日
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转OsbHLH1和Bar基因水稻及相关特性分析
罗伯祥1,肖自友2,肖国樱2
1. 江苏徐淮地区淮阴农业科学研究所
2. 中国科学院亚热带农业生态研究所 亚热带农业生态过程重点实验室
Transgenic Rice with Bar and OsbHLH1 Genes and Its Agronomic Trait Analyses
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摘要 OsbHLH1基因编码bHLH(basic helix-loop-helix,碱性螺旋-环-螺旋)类转录因子,与水稻耐低温相关。草铵膦是一种高效低毒灭生性除草剂,Bar基因表达产物能解除草铵膦的毒性。本实验对通过农杆菌(Agrobacterium tumefaciens)浸染法获得的以粳型水稻(Oryza sativa L. subsp. japonica Kato) 恢复系淮C17为受体的转OsbHLH1和Bar基因水稻进行研究,鉴定其除草剂抗性和耐低温能力,考查其农艺性状。草铵膦筛选发现Bar基因正常表达;对草铵膦筛选获得的转基因植株进行PCR、Southern、RT-PCR和实时荧光定量PCR检测,发现OsbHLH1基因已经整合到水稻基因组中并且在水稻中表达。转基因T3代株系芽期(萌发芽长0.5 mm)在2℃条件下处理6 d,第6号转基因株系死苗率为17.8%,对照死苗率为61.1%。T3代苗期(一叶一心)在8~10℃下处理7d,第5、6、9、11号株系的根长显著长于对照,第3、5、11号株系的根数显著多于对照,第3号株系的平均鲜重显著重于对照。芽期、苗期耐冷性试验表明,OsbHLH1基因在水稻中过量表达显著提高了水稻芽期、苗期的耐低温能力。对转基因T2代的农艺性状进行考查,株高、千粒重、结实率、理论产量等农艺性状与对照相比较均出现了显著的变化,外源基因的导入对水稻的农艺性状有明显影响。研究结果将为选育抗除草剂、耐低温杂交粳稻新组合提供新种质。
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罗伯祥
肖自友
肖国樱
关键词 OsbHLH1基因Bar基因水稻耐低温抗除草剂    
Abstract:sbHLH1 gene encoded one of transcription factors in bHLH (basic helix-loop-helix) family, which is involved in cold tolerance. The expression of Bar gene can relieve the toxicity of glufosinate, a highly effective and low toxicity herbicide.The OsbHLH1 and Bar genes were transformed into rice cultivar Huai C17 (Oryza sativa L. subsp. japonica Kato) by Agrobacterium tumefaciens mediation. The herbicide resistance, cold tolerance and agronomic traits of transformants with Bar and OsbHLH1 genes were identified in this study. The existance of Bar gene and its expression in transgenic lines were identified by stress of glufosinate. The integration of OsbHLH1 gene and its expression were also detected and confirmed by PCR, Southern blot, RT-PCR and Real-time PCR. The T3 generation of the transgenic line No. 6 had been treated with 2℃ for 6 days at the germination stage. And the dead seedling of this transgenic line was 17.8% while that of control was 61.1%. Under stress of 8~10℃ for 7 days at the seedling stage, the root length of transgenic lines No. 5, 6, 9 and 11 of T3 generation were longer than that of control (P<0.05), and the root number of line No. 3, 5 and 11 were more than that of control (P<0.05) and the fresh weight of No. 3 line was higher than that of control (P<0.05). The increase of cold tolerance at germination and seedling stage of transgenic lines showed that the over-expression of OsbHLH1 gene could improve the cold tolerance of rice. Based on the investigation of the main agronomic traits, there were differences (P<0.05) in plant height, panicle length, 1000-grain weight, seed setting and theoretical yield between transgenic lines of T2 generation and control, which indicated that the over-expression of exogenous gene had obvious influence on rice agronomic traits. The new germplasm created in this study will be available for breeding of new japonica hybrid rice with cold and herbicide tolerance
Key wordsOsbHLH1 gene    Bar gene    rice    cold tolerance    herbicide-resistant
收稿日期: 2011-08-22     
通讯作者: 肖国樱   
引用本文:   
罗伯祥1,肖自友2,肖国樱2. 转OsbHLH1和Bar基因水稻及相关特性分析[J]. , 2012, 20(1): 30-37.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2012/V20/I1/30
 
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