XIAO Yu,ZHANG Jian-xia,ZHOU Qi,et al.Research on the STS Gene Promoter of Chinese Wild Vitis?pseudoreticulata Transformed Tomato[J].Northern Horticulture,2013,37(01):89-94.
中国野生华东葡萄芪合成酶基因启动子转化番茄的研究
- Title:
- Research on the STS Gene Promoter of Chinese Wild Vitis?pseudoreticulata Transformed Tomato
- 文章编号:
- 1001-0009(2013)01-0089-06
- 分类号:
- S 663.1
- 文献标志码:
- A
- 摘要:
- 中国野生华东葡萄“白河-35-1”中的芪合成酶基因启动子被证明具有病原菌和胁迫诱导表达活性。现将芪合成酶基因置于增添不同增强子的该启动子下,构建4个植物表达载体,通过农杆菌介导法进行了转化番茄的研究,并对转基因株系抗番茄灰霉病表现及其白藜芦醇含量变化进行了测定。结果表明:经PCR-Southern鉴定,获得转基因株系4个共54株;不同载体转化的番茄株系对灰霉病抗性均高于野生型,但抗性存在差异;添加增强元件enhancer启动子启动的芪合成酶基因表达的番茄株系,可使白藜芦醇含量提高4.4倍。
- Abstract:
- Stilbene synthase gene (STS) and its specific promoter which was related to disease resistance and stress were previously isolated from Chinese wild Vitis pseudoreticulata accession ‘Baihe-35-1’.In this paper,the promoter was added different enhancement components for construction of three enhanced expression vectors.The results showed that four lines including 54 transgenic tomato plants which were tested by PCR-Southern were obtained by Agrobacterium-mediated genetic transformation.The disease resistance index of transgenic tomato plants was measured after inoculation of Botrytis cinerea,and the resistance of all transgenic plants was increased to some degree.Resveratrol content analysis showed that addition of enhancer element could increase the amount of resveratrol expression to 4.4 times.
参考文献/References:
[1]许志刚.普通植物病理学[M].4版.北京:高等教育出版社,2009. [2]李知行.葡萄病虫害防治[M].北京:金盾出版社,2009. [3]Serazetdinova L,Oldach K,Lorz H.Expression of transgenic stilbene synthases in wheat causes the accumulation of unknown stilbene derivatives with antifungal activity[J].J Plant Physiol,2005,162:985-1002. [4]Jeandet P,Bessis R,Gautheron B.The Production of resveratrol (3’5’4’-trihydroxystilbene ) by grape berries in different developmental stages[J].Am J Enol Vitie,1991,42(1):41-46. [5]Sarig P,Zutkhi Y,Monjauze A,et al.Phytoalexin elicitation in grape berries and their susceptibility to Rhizopus stolonifer[J].Physiol Mol Plant P,1997,50:337-347. [6]Delmas D,Lancon A,Colin D,et al.Resveratrol as a chemopreventive agent:Apromising molecule for fighting cancer[J].Curr Drug Targets,2006,7:423-442. [7]Hain R,Reif H J,Krause E,et al.Disease resistance results from foreign phytoalexin expression in a novel plant[J].Nature,1993,361:153-156. [8]Liang H,Zheng J,Duan X Y,et al.A transgenic wheat with a stilbene synthase gene resistant to powdery mildew obtained by biolistic method [J].Chin Sci Bull,2000,45:634-638. [9]Leckband G,Lorz H.Transformation and expression of a stilbene synthase gene of Vitis vinifera L.in barley and wheat for increased fungal resistance[J].Theor Appl Genet,1998,96:1004-1012. [10]Richter A,de Kathen A,de Lorenzo G,et al.Transgenic peas (Pisum sativum) expressing polygalacturonase inhibiting protein from raspberry (Rubus idaeus) and stilbene synthase from grape (Vitis vinifera) [J].Plant Cell Rep,2006,25:1166-1173. [11]Zhu Y J,Agbayani R,Jazckson M.C,et al.Expression of the grapevine stilbenesynthase gene VST1 in papaya provides increased resistance against diseases caused by?Phytophthora?palmivora [J].Planta,2004,12:807-812. [12]王跃进,贺普超.中国葡萄属野生种叶片抗白粉病遗传研究[J].中国农业科学,1997,30(1):19-25. [13]Xu W,Yu Y H,Ding J H,et al.Characterization of a novel stilbene synthase promoter involved in pathogen-and stress-inducible expression from Chinese wild Vitis pseudoreticulata[J].Planta,2010,231:475-487. [14]徐伟荣.中国野葡萄抗白粉病芪合成酶基因启动子克隆与功能分析[D].杨凌:西北农林科技大学,2010. [15]Kasajima I,Ide Y,Ohkama-Ohtsu N,et al.A protocol for rapid DNA extraction from?Arabidopsis thaliana?for PCR analysis[J].Plant Mol Biol Rep,2004,22:49-52. [16]王琦,姜道宏.植物病理学研究进展.中国植物病理学会第八届青年学术研讨会论文选编[C].北京:中国农业科学技术出版社,2007:415-419. [17]陈双臣,刘爱荣,邹志荣.转葡聚糖酶和防御素基因的番茄植株及其对番茄灰霉病的抗性[J].植物保护学报,2006,33(4):357-362. [18]罗小英,曾雪嘉,肖月华,等.抗菌肽和几丁质酶基因提高烟草对黑痉病菌和赤星病菌的抗性研究[J].植物病理学报,2005,35(3):249-255. [19]方中达.植病研究方法[M].3版.北京:中国农业出版社,1998. [20]路静,赵华燕,何奕昆,等.高等植物启动子及其应用研究进展[J].自然科学进展,2004,14(8):856-962. [21]Xu W R,Yu Y H,Zhou Q,et al.Expression pattern,genomic structure and promoter analysis of the gene encoding stilbene synthase from Chinese wild Vitis pseudoreticulata[J].Journal of Experimental Botany,2011,62:2745-2761. [22]Gallie D R,Sleat D E,Watts J W,et al.The 5’ leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcript in vivo?and in vitro[J].Nucleic Acids Res,1987,15(8):3257-3273. [23]侯丙凯,夏光敏,陈正华.植物基因工程表达载体的改进和优化策略[J].遗传,2001,23(5):492-497. [24]郭殿京,傅荣昭,李文彬,等.小麦中外源基因瞬间表达调控研究及兔防御素(NP-1)基因的转化[J].遗传学报,1999,26(2):168-173. [25]刘根齐,李秀丽,杨春玲,等.不同调控序列控制下的GUS基因在水稻和毛白杨愈伤组织中的瞬时表达[J].植物生理与分子生物学学报,2005,31(3):327-330.
备注/Memo
第一作者简介:肖宇(1988-),女,硕士,研究方向为果树种质资源与生物技术育种。