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2025年8月14日 星期四
  2018, Vol. 26 Issue (5): 756-763    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
转基因水稻B2A68中抗虫蛋白Cry2Aa的表达特征和抗性分析
翁绿水1,王作平2,肖国樱1
1. 中国科学院亚热带农业生态研究所
2.
Expression Profile of Insecticidal Protein Cry2Aa and Lepidopteran Resistance in Transgenic Rice (Oryza sativa) B2A68
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摘要 摘 要 苏云金芽胞杆菌(Bacillus thuringiensis, Bt)基因的高效表达和稳定遗传对抗虫转基因水稻(Oryza sativa)的培育和商业化生产至关重要。转基因水稻B2A68是通过农杆菌介导法把双丙氨酰磷抗性基因(biplaphos resistance gene, Bar)和Bt杀虫晶体蛋白基因(insecticidal crystal proteins gene)Cry2Aa同时转入到恢复系D68中,培育出的抗除草剂、抗螟虫的两系早稻恢复系。本研究运用酶联免疫吸附测定(enzyme linked immunosorbent assay, ELISA)检测法研究了B2A68纯杂合基因型、不同组织及不同发育时期叶片中Cry2Aa蛋白的时空表达特征,并结合室内生测实验,揭示抗虫蛋白的表达水平、时期、部位与抗虫性的相关性。研究表明,转基因抗虫水稻B2A68纯合植株叶片中Cry2Aa蛋白表达量极显著高于杂合植株。乳熟期各器官中Cry2Aa蛋白含量为:叶片>颖壳和脱壳籽粒>茎秆和叶鞘(P<0.05);腊熟期各器官中Cry2Aa蛋白含量为:叶片>脱壳籽粒>茎秆和颖壳(P<0.05),但脱壳籽粒和叶鞘之间、叶鞘和茎秆之间、茎秆和颖壳之间的Cry2Aa蛋白含量差异不显著。从秧苗期到腊熟期的叶片中Cry2Aa蛋白含量表现为先升后降再升高的波动,且T3、T4两代波动趋势一致。二化螟幼虫(Chilo suppressalis)死亡率与乳熟期各器官中的Cry2Aa蛋白含量高度相关(r=0.837),稻纵卷叶螟幼虫(Cnaphalocrocis medinali)死亡率与各发育时期叶片中的Cry2Aa蛋白含量高度相关(r=0.988)。Cry2Aa蛋白含量较低的杂合植株叶片对稻纵卷叶螟幼虫的全部致死时间稍长,但不超过5 d。本研究结果对田间害虫的防治以及转基因水稻的育种都具有参考价值。
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翁绿水
王作平
肖国樱
关键词 水稻Cry2Aa蛋白抗螟虫基因型时空表达    
Abstract:Abstract The high-efficiency expression and stable inheritance of Bacillus thuringiensis gene (Bt) are crucial to transgenic rice breeding and commercial production. A transgenic line B2A68 (Oryza sativa) was obtained by introducing biplaphos resistance gene (Bar) and insecticidal crystal proteins gene (Cry2Aa) into the early season rice restorer line D68 via Agrobacterium mediated method. In this study, the temporal and spatial expression profile of Cry2Aa protein between homozygous and heterozygous genotype, different tissues and various developmental stages were studied by enzyme linked immunosorbent assay (ELISA), and the relationship between expression level, stage and tissue of insecticidal protein and insect resistance was revealed by referring bioassay results, in order to provide technical parameters for developing excellent lepidopteran resistant rice. The results showed that the expression level of Cry2Aa protein in homozygous plant leaf was significantly higher than that in heterozygous plant leaf (P<0.01). At milk stage, the contents of Cry2Aa protein in each organ were: leaf > glume and hulled grain > stem and sheath (P<0.05). At hard dough stage, the contents of Cry2Aa protein in each organ were: leaf > hulled grain > stem and glume (P<0.05), but there were no significant differences in the contents of Cry2Aa protein between hulled grain and sheath, sheath and stem, and stem and glume, respectively. From seedling to ripening phase, the Cry2Aa protein content in leaf increased first, then decreased, and later increased again, fluctuating in a same pattern in T3 and T4 generation. The mortality of striped rice borer larvae was highly correlated with the Cry2Aa protein content in different organs at milk stage (r=0.837), and the mortality of rice leaf roller larvae was highly correlated with the Cry2Aa protein content in leaves at different stages (r=0.988). For the heterozygous plant leaf with low Cry2Aa protein content, the lethal time of the rice leaf roller feeding with heterozygous plant leaf was slightly longer than that of feeding with homozygous plant leaf, but shorter than 5 days. This study will contribute to controlling Lepidopteran pests in field and developing transgenic rice in the future.
Key wordsRice    Cry2Aa protein    Lepidopteran resistance    Genotype    Temporal and spatial expression
收稿日期: 2017-10-30      出版日期: 2018-05-02
基金资助:国家转基因生物新品种培育科技重大专项
通讯作者: 肖国樱     E-mail: xiaoguoying@isa.ac.cn
引用本文:   
翁绿水 王作平 肖国樱. 转基因水稻B2A68中抗虫蛋白Cry2Aa的表达特征和抗性分析[J]. , 2018, 26(5): 756-763.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2018/V26/I5/756
 
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