Journal of Integrative Agriculture ›› 2014, Vol. 13 ›› Issue (10): 2081-2090.DOI: 10.1016/S2095-3119(13)60538-9

• 论文 • 上一篇    下一篇

Development of Insect-Resistant Hybrid Rice by Introgressing the Bt Gene from Bt Rice Huahui 1 into II-32A/B, a Widely Used Cytogenic Male Sterile System

 LAI  Yun-song, HUANG  Hai-qing, XU  Meng-yun, WANG  Liang-chao, ZHANG  Xiao-bo, ZHANG  Ji-wen , TU  Ju-min   

  1. 1、Institute of Crop Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, P.R.China
    2、Institute of Pomology & Olericulture, Sichuan Agricultural University, Chengdu 611130, P.R.China
    3、Rice Research Institute of Enshi Academy of Agriculture Sciences, Enshi 445000, P.R.China
    4、Chinese National Rice Research Institute, Hangzhou 310006, P.R.China
  • 收稿日期:2013-05-20 出版日期:2014-10-01 发布日期:2014-10-10
  • 通讯作者: TU Ju-min, Tel/Fax: +86-571-88982835, E-mail: jtu@zju.edu.cn
  • 作者简介:LAI Yun-song, Tel/Fax: +86-28-86291136, E-mail: ronson521@hotmail.com
  • 基金资助:

    Acknowledgements This work was supported by the National High-Tech R&D Program of China (2006AA10Z159).

Development of Insect-Resistant Hybrid Rice by Introgressing the Bt Gene from Bt Rice Huahui 1 into II-32A/B, a Widely Used Cytogenic Male Sterile System

 LAI  Yun-song, HUANG  Hai-qing, XU  Meng-yun, WANG  Liang-chao, ZHANG  Xiao-bo, ZHANG  Ji-wen , TU  Ju-min   

  1. 1、Institute of Crop Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, P.R.China
    2、Institute of Pomology & Olericulture, Sichuan Agricultural University, Chengdu 611130, P.R.China
    3、Rice Research Institute of Enshi Academy of Agriculture Sciences, Enshi 445000, P.R.China
    4、Chinese National Rice Research Institute, Hangzhou 310006, P.R.China
  • Received:2013-05-20 Online:2014-10-01 Published:2014-10-10
  • Contact: TU Ju-min, Tel/Fax: +86-571-88982835, E-mail: jtu@zju.edu.cn
  • About author:LAI Yun-song, Tel/Fax: +86-28-86291136, E-mail: ronson521@hotmail.com
  • Supported by:

    Acknowledgements This work was supported by the National High-Tech R&D Program of China (2006AA10Z159).

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摘要： Huahui 1 is an elite transgenic male sterile restorer line of wild rice abortive-type that expresses a Bacillus thuringiensis (Bt) δ-endotoxin and provides effective and economic control of lepidopteran insects. To exploit Huahui 1 to develop a new Bt rice, the insertion site of the Bt gene was determined by thermal asymmetric interlaced PCR (TAIL-PCR). Bt was located in the promoter region of LOC.Os10g10360, approximately 5.35 Mb from the telomere of the short arm of chromosome 10. For the first time, a Bt cytoplasmic male sterile (CMS) system was developed by introgressing Bt from Huahui 1. The recipient CMS system used consisted of Indonesia paddy rice-type II-32B (maintainer line) and II-32A (male sterile line). Marker-assisted selection was used to increase selection efficiency in the backcrossing program. In BC5F1, the Bt plant 85015-8 was selected for further analyses, as it had the highest SSR marker homozygosity. In addition, the linkage drag of the foreign Bt gene in 85015-8 was minimized to 8.01-11.46 Mb. The foreign Bt gene was then delivered from 85015-8 into II-32A. The resultant Bt II-32A and Bt II-32B lines were both resistant to lepidopteran in field trials, and agronomic traits were not disturbed. The maintainability of II-32B, and the male sterility and general combining ability of II-32A, were not affected by the Bt introgression. This study demonstrates a simple and fast approach to develop Bt hybrid rice.

关键词: Bt rice , male sterile line , genome walker , marker-assisted backcross (MAB) , marker-assisted selection (MAS) , SSR

Abstract: Huahui 1 is an elite transgenic male sterile restorer line of wild rice abortive-type that expresses a Bacillus thuringiensis (Bt) δ-endotoxin and provides effective and economic control of lepidopteran insects. To exploit Huahui 1 to develop a new Bt rice, the insertion site of the Bt gene was determined by thermal asymmetric interlaced PCR (TAIL-PCR). Bt was located in the promoter region of LOC.Os10g10360, approximately 5.35 Mb from the telomere of the short arm of chromosome 10. For the first time, a Bt cytoplasmic male sterile (CMS) system was developed by introgressing Bt from Huahui 1. The recipient CMS system used consisted of Indonesia paddy rice-type II-32B (maintainer line) and II-32A (male sterile line). Marker-assisted selection was used to increase selection efficiency in the backcrossing program. In BC5F1, the Bt plant 85015-8 was selected for further analyses, as it had the highest SSR marker homozygosity. In addition, the linkage drag of the foreign Bt gene in 85015-8 was minimized to 8.01-11.46 Mb. The foreign Bt gene was then delivered from 85015-8 into II-32A. The resultant Bt II-32A and Bt II-32B lines were both resistant to lepidopteran in field trials, and agronomic traits were not disturbed. The maintainability of II-32B, and the male sterility and general combining ability of II-32A, were not affected by the Bt introgression. This study demonstrates a simple and fast approach to develop Bt hybrid rice.

Key words: Bt rice , male sterile line , genome walker , marker-assisted backcross (MAB) , marker-assisted selection (MAS) , SSR