过量表达转录因子OsbZIP60对水稻抗热和抗旱能力的研究

摘要

[目的]利用转基因技术研究转录因子OsbZIP60的生物学功能,为培育抗逆水稻、减轻高温干旱对水稻的伤害奠定基础.[方法]构建水稻OsbZIP60的过表达载体pHB-0sbZIP60,通过根癌农杆菌介导的遗传转化法将其导入水稻品种日本晴,通过观察转基因植株和野生型植株在胁迫条件下的表型差异,分析其生物学功能.采用半定量PCR和荧光定量PCR检测OsbZIP60的表达模式以及其在转基因植株中的表达水平.[结果]在转基因植株中,OsbZIP60表达量明显高于野生型日本晴植株.OsbZIP60的表达受热信号诱导,在热激条件下,转基因植株的OsbZIP60表达量进一步升高.OsbZIP60在水稻各组织中均有表达,且在成熟植株叶片中表达量最高.转基因植株与野生型相比具有更强的抗胁迫能力,并且离体部分失水率更低.[结论]水稻转录因子OsbZIP60在水稻热胁迫和干旱胁迫应答中具有重要的作用,过量表达转录因子OsbZIP60能够增强水稻的抗热和抗旱能力.%[Objective] Dissect biological function of transcription factor OsbZIP60 by transgenic technology, making preparation for developing stress tolerance rice to reduce the hazard caused by heat and drought stresses. [Method] The overexpression vector containing the full-length coding region of OsbZIP60 gene was constructed and introduced into rice cv. Nipponbare by Agmbacterium-medizted genetic transformation. The biological function of OsbZIP60 was analyzed by observing the phenotypes of transgenic and wild type plants under stress conditions. The expression pattern of OsbZIP60 and its expression level in transgenic line was analyzed by semi-quantitative RT-PCR and real-time quantitative PCR. [Result] The expression level of OsbZIP60 in overexpression transgenic lines is distinctly increased compared to that of WT plants. OsbZIP60 can be induced by heat, and it's further up-regulated in transgenic lines under stress condition. OsbZIP60 is constitutively expressed in rice and has the highest abundance in leaves. Transgenic plants showed significant enhanced stress tolerance. Detached aerial parts of transgenic plants also showed a slower water loss rate compared to that of WT plants. [Conclusion] Transcription factor OsbZIP60 plays an important role in response to heat and drought. Overexpression of OsbZIP60 might be utilized in genetic improvement of heat and drought tolerance in rice.