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作者	单位
董陈文华，周洪斌，郎雨萌，罗亮扎，李仕金，姜瑞梅，毛孝强，赵海云，唐进，李勇，王玫郦，陈升位	（云南农业大学农学与生技术学院，云南昆明 650201）

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中文摘要:

为了解大麦LRR型类受体蛋白激酶基因HvLRR-RLK510启动子及其功能，以特异性PCR引物扩增大麦材料Morex和保大麦8号的HvLRR-RLK510基因启动子并构建pEASY^-510重组质粒，基于大肠杆菌Trans1-T₁菌株的阳性克隆测定获得2 368 bp的启动子序列。结果表明，通过在线软件Plant CARE预测，该启动子携带茉莉酸甲酯、脱落酸、低温等非生物因素的响应元件。经拟南芥转基因阳性植株和本氏烟草瞬时表达植株的根、茎、叶和花的GUS染色分析，所克隆启动子及其截短片段（-1 458、-868和-460 bp）可等效驱动GUS基因在花药中表达。qRT-PCR检测发现，茉莉酸甲酯(JAs)、脱落酸(ABA)、低温、聚乙二醇(PEG-2000)和光照能有效改变大麦叶片和根中HvLRR-RLK510基因的表达特性。HvLRR-RLK510基因启动子属诱导型启动子，携带茉莉酸、脱落酸、低温、干旱和光的响应元件，其-460 bp的截断片段可替代全长启动子；该启动子及其截断片段可用于HvLRR-RLK510基因转录调控机制解析。

英文摘要:

To elucidate the promoter of the barley LRR-type receptor-like protein kinase gene HvLRR-RLK510 and its function, specific PCR primers were used to amplify the promoter of the HvLRR-RLK510 gene from Morex and Baodamai 8. The pEASY^-510 recombinant plasmid was constructed, and the promoter sequence of 2 368 bp was sequenced based on the positive colonies of the Escherichia coli Trans1-T₁ strain. The prediction results from the online software PlantCARE indicated that the promoter contains response elements to non-biotic factors such as methyl jasmonate, abscisic acid, and low temperature. GUS staining results in roots, stems, leaves, and flowers of transgenic Arabidopsis thaliana and transiently expressed Nicotiana benthamiana plants demonstrated that the cloned promoter and its truncated fragments(-1 458, -868, and -460 bp) can equivalently drive the expression of the GUS gene in anthers. Quantitative real-time PCR(qRT-PCR) results showed that methyl jasmonate(MeJA), abscisic acid(ABA), low temperature, polyethylene glycol(PEG-2000), and light can effectively alter the expression characteristics of the HvLRR-RLK510 gene in barley leaves and roots. The promoter of the HvLRR-RLK510 gene cloned in this study is an inducible promoter containing response elements to jasmonic acid, abscisic acid, low temperature, drought, and light. Its -460 bp truncated fragment can replace the full-length promoter. The promoter and its truncated fragments can be used to elucidate the transcriptional regulatory mechanisms of the HvLRR-RLK510 gene.

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