中文摘要：

【目的】研究中国野生毛葡萄‘丹凤-2＇ （Vitis quinquangularis ‘Danfeng-2＇ ）芪合成酶（stilbene synthase）因双向启动子的启动活性与差异。【方法】通过染色体步移技术克隆中国野生毛葡萄‘丹凤-2’芪合成酶基因双向启动子，并与U／dA基因（β-葡萄糖苷酶基因，GUS）融合，利用真空渗透法在葡萄叶片中瞬时表达，组织化学染色和GUS蛋白定量法分别检测白粉菌接种、温度处理、茉莉酸甲酯、脱落酸和水杨酸处理对双向启动子活性的影响。以毛葡萄‘丹凤-2’和欧洲葡萄‘赤霞珠’不同成熟时期果实的cDNA为模板，通过荧光实时定量PCR的方法分析VqSTSl2／VvSTS31和VqSTS33／VvSTS32的表达情况。[结果]PvqDSTSl2受到白粉菌、茉莉酸甲酯、高温和低温的诱导后，启动活性增强，对水杨酸和脱落酸处理不敏感；PvqDSTS33在茉莉酸甲酯处理下启动活性增强，在高温和低温的处理下启动活性降低，对水杨酸、脱落酸和白粉病处理不敏感。荧光实时定量PCR结果显示，在果实发育的6个时期中，‘丹凤-2’VqSTSl2和VqSTS33的表达量均高于‘赤霞珠’VvSTS31和VvSTS32。4个基因的基本表达趋势为：浆果转色期之前持续增长，转色后期下降，浆果半熟期达到最高峰，浆果成熟期表达降低。其中毛葡萄‘丹凤-2’VqSTSl2的表达量在转色后期并未降低且增加到上一时期的2．3倍；欧洲葡萄VvSTS32基因在成熟期表达量升高为前一时期的1．42倍。【结论】双向启动子活性的研究为利用植物天然双向启动子提供依据。

英文摘要：

[Objective] To learn of the activities and differences of bidirectional stilbene synthase promot- ers from the Chinese wild V. quirutuangularis ＇ Danfeng-2＇. As the position of the 48 genes of the stilbene synthase family from V. vinifera has been reported, we analyzed and found that the stilbene synthase genes VvSTS31 and VvSTS32 space 1 737 bp, and started expressing in the opposite direction. We noted that the sequence between these two genes is a bidirectional promoter. As China is one of the original ar- eas for the grape, it is filled with extensive resistance germplasm resources. Among them, V. quinquangu- laris ＇ Danfeng-2＇ not only has good resistance, but also a high content of resveratrol. Since the genomic DNA sequences of Chinese wild V. quinquangularis have a high degree of similarity with V. vlnifera, pro- moters, cloning of bidirectional stilbene synthase from Chinese wild V. quinquangularis ＇ Danfeng-2＇ can provide a novel resistance gene resource for the grapes. [Methods]Bidirectional stilbene synthase promot- ers from wild V. quinquangularis ＇ Danfeng-2＇ were cloned by genome walking. The sequences of bidirec- tional stilbene synthase promoters from ＇Danfeng-2＇ were submitted to the promoter prediction websitePlantCARE （http：//bioinformatics.psb.ugent.be/webtools/plantcare/html/）, and the corresponding cis-act- ing elements were obtained and marked on the sequences. In order to analyze the promoter activity of bidi- rectional stilbene synthase promoters, they were fused with a UidA gene （[3-glucuronidase, GUS）. Recom- binant plasmid PvqDSTS31：：GUS and PvqDSTS32：：GUS were transient transformed into grape leaves us- ing Agrobacterium-mediated vacuum infiltration. The expression patterns of GUS in grape leaves under different stress treatments, including powdery mildew, high temperature, low temperature, MeJA, ABA and SA, were analyzed using histochemical staining and GUS quantification. Treatment of the biotic and abiotic stresses were conducted as follows：