浙江农业学报 ›› 2017, Vol. 29 ›› Issue (2): 177-185.DOI: 10.3969/j.issn.1004-1524.2017.02.01

• 作物科学 • 上一篇    下一篇

水稻LOC_Os10g05490位点冷胁迫条件下表达分析及CRISPR/Cas9定向编辑

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:2016-09-09 出版日期:2017-02-15 发布日期:2017-03-06
  • 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。E-mail:shenchunxiu@126.com
  • 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)

CRISPR/Cas9 editing and expression analysis of LOC_Os10g05490 in rice under cold stress

SHEN Chunxiu   

  1. College of Life Sciences, Resources and Environment, Yichun University, Yichun 336000, China
  • Received:2016-09-09 Online:2017-02-15 Published:2017-03-06

摘要: 为探明LOC_Os10g05490位点冷胁迫条件下的表达特性和耐冷性功能,以耐冷东乡野生稻和冷敏感水稻93-11为试验材料进行了半定量分析,结果表明,LOC_Os10g05490在冷敏感水稻93-11冷处理前后表达丰度相当,而在耐冷东乡野生稻中冷处理后表达丰度下调,这一结果与前期转录组测序(RNA-seq)结果相符,进一步验证了该位点可能与水稻耐冷相关,随后利用CRISPR/Cas9基因组编辑技术对LOC_Os10g05490编码区于同样具有较强耐冷性的粳稻品种台北309中实施定向基因敲除,并成功获得了26株转基因阳性植株。

关键词: 冷胁迫, 转录组测序, 东乡野生稻, CRISPR/Cas9基因组编辑技术

Abstract: To explore the expression and cold tolerance function of LOC_Os10g05490 in rice under cold stress, semi-quantitative analysis was performed for Dongxiang wild rice and 93-11. The results showed that expression level of LOC_Os10g05490 in 93-11 was the same before and after cold stress, but it was down regulated in Dongxiang wild rice after cold stress. This was consistent with RNA-seq data and further study indicated that this gene might be associated with cold tolerance in Dongxiang wild rice. Plant expression vectors with CRISPR/Cas9 genome editing technique were subsequently constructed to target the LOC_Os10g05490 locus, and were transformed into the japonica rice variety TP309. A total of 26 transgenic positive plants were obtained. This study may lay the foundation for subsequent screening of mutant plants and identification of genes associated with cold tolerance in rice.

Key words: cold stress, RNA-seq, Dongxiang wild rice, CRISPR/Cas9 genome editing technique

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