华北农学报 ›› 2017, Vol. 32 ›› Issue (3): 13-20. doi: 10.7668/hbnxb.2017.03.003

所属专题: 水稻 生物技术

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基于比较转录组的水稻苗期耐冷相关基因BGIOSGA032296 TALEN基因组编辑技术构建

沈春修   

  1. 宜春学院 生命科学与资源环境学院, 江西 宜春 336000
  • 收稿日期:2016-04-20 出版日期:2017-06-28
  • 作者简介:沈春修(1979-),男,湖南溆浦人,讲师,博士,主要从事水稻抗逆性研究。
  • 基金资助:
    国家自然科学基金项目(31660379);江西省教育厅科学技术研究项目(GJJ151039)

Construction of TALEN Genome-editing Technique for BGIOSGA032296 Related to Cold Tolerance in Rice Seedling Based on Comparative Transcriptome

SHEN Chunxiu   

  1. College of Life Sciences, Resources and Environment Sciences, Yichun University, Yichun 336000, China
  • Received:2016-04-20 Published:2017-06-28

摘要: 为了挖掘水稻苗期耐冷基因,基于比较转录组分析,参考公共数据库中的基因序列信息设计靶位点,对冷胁迫条件下东乡野生稻苗期下调表达耐冷相关基因BGIOSGA032296进行TALEN基因组编辑技术构建,克隆酶切鉴定和序列比对结果表明,BGIOSGA032296 TALEN敲除植物表达载体1301TALEN-032296构建成功,利用农杆菌介导法将TALEN敲除植物表达载体1301TALEN-032296转入水稻受体品种TP309,获得了17株转基因水稻植株,经潮霉素抗性标记基因和Fok I基因特异引物PCR检测,表明获得的转基因植株皆为携带1301TALEN-032296的阳性植株。该研究为水稻BGIOSGA032296基因后续耐冷表型鉴定奠定了前期基础。

关键词: 比较转录组, 东乡野生稻, TALEN基因组编辑技术

Abstract: To dig cold tolerance genes at seedling stage of rice,based on the analysis of comparative transcriptome,TALEN genome-editing technique was constructed for BGIOSGA032296 which was down-regulated in Dongxiang wild rice seedling under cold stress.The analysis of enzyme identification and sequence alignment showed that TALEN knockout plant expression vector 1301TALEN-032296 was constructed successfully.By the agrobacterium-mediated method,the recombination clone 1301TALEN-032296 was transformed into the receptor rice variety TP309.17 transgenic rice plants were obtained.After PCR detection of hygromycin resistance gene and Fok I gene,they were found to be positive transgenic plants carrying 1301TALEN-032296.This research laid some basis for subsequent cold resistant phenotype identification.

Key words: Comparative transcriptome, Dongxiang wild rice, TALEN genome-editing technique

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引用本文

沈春修. 基于比较转录组的水稻苗期耐冷相关基因BGIOSGA032296 TALEN基因组编辑技术构建[J]. 华北农学报, 2017, 32(3): 13-20. doi: 10.7668/hbnxb.2017.03.003.

SHEN Chunxiu. Construction of TALEN Genome-editing Technique for BGIOSGA032296 Related to Cold Tolerance in Rice Seedling Based on Comparative Transcriptome[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(3): 13-20. doi: 10.7668/hbnxb.2017.03.003.

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