中国野生毛葡萄3个芪合酶基因表达分析  

Expression Analysis of Three Stilbene Synthase Genes from Chinese Wild Vitis quinquangularis

作　　者：吴凤颖 刘梦琦 王跃进[1] WU Feng-Ying;LIU Meng-Qi;WANG Yue-Jin(College of Horticulture/State Key Laboratory of Crop Stress Biology in Arid Areas/Key Laboratory of Horticultural Plant Germplasm Resource Utilization in Northwest China,Ministry ofAgriculture,NorthwestA&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学园艺学院/旱区作物逆境生物学国家重点实验室/农业部西北地区园艺作物生物与种质创制重点实验室,杨凌712100

出　　处：《农业生物技术学报》2020年第5期847-858,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31872055)。

摘　　要：白藜芦醇(resveratrol)是植物在生物和非生物胁迫条件下产生的一种植保素,是芪合酶(stilbene synthase, STS)表达的产物。中国野生毛葡萄’丹凤-2’(Chinese wild Vitis quinquangularis accession’Danfeng-2’)果实中白藜芦醇含量高,且植株抗病性强,研究其芪合酶基因的表达与功能,可以用其作为抗病种质资源进行抗病育种。本研究基于前期的研究成果,通过多种生物和非生物逆境处理探明芪合酶基因VqSTS12、VqSTS24和VqSTS25的表达模式。qRT-PCR结果表明在白粉菌(Uncinula necator)诱导12 h后VqSTS12、VqSTS24和VqSTS25基因就能被极显著地诱导表达,其次在96~120 h表达水平较高;同时,VqSTS12、VqSTS24和VqSTS25基因在茉莉酸甲酯(methyl jasmonate, MeJA)、水杨酸(salicylic acid, SA)、脱落酸(abscisic acid, ABA)、创伤、低温、高盐、干旱等胁迫处理下表达上调,表明这3个基因能广泛响应生物与非生物胁迫的诱导。瞬时转化烟草(Nicotiana benthamiana)表皮细胞进行亚细胞定位,分析结果表明这3个芪合酶定位在细胞质中。在过表达VqSTS12和VqSTS25的欧洲葡萄’无核白’(Vitis vinifera cv.’Thompson Seedless’)中,发现10%PEG6000模拟干旱胁迫处理下转基因植株中的芪合酶基因VqSTS12和VqSTS25表达显著上调,丙二醛(malondialdehyde, MDA)含量、过氧化物酶(peroxidase, POD)活性和过氧化氢酶(catalase, CAT)活性等生理指标均优于野生型植株,表明过表达VqSTS12和VqSTS25提高了无核白葡萄的抗旱性。综上所述,芪合酶基因VqSTS12、VqSTS24和VqSTS25可以在抗病育种中作为改良欧洲葡萄品种的抗性基因种质资源,研究结果为进一步改良提高欧洲葡萄的抗病、抗逆性提供了候选基因和参考依据。As a phytoalexin produced by plants under biotic and abiotic stresses,resveratrol is a metabolite of stilbene synthases(STSs).Chinese wild Vitis quinquangularis accession'Danfeng-2'contains a high concentration of resveratrol in its fruits,which also possesses strong plant resistance against diseases.The expressions and functions study of its STS genes can serve as the disease-resistant germplasm resources for resistance breeding.Based on the previous findings,this study explored the expression patterns of STS genes VqSTS12,VqSTS24 and VqSTS25 through a variety of biotic and abiotic stress treatments.As the qRT-PCR results revealed,the VqSTS12,VqSTS24 and VqSTS25 genes could be induced and expressed significantly just after 12 h of powdery mildew induction.Besides,their second-highest expressions were found during 96~120 h.Additionally,the 3 STS genes could be expressed under the following stress treatments:methyl jasmonate(MeJA),salicylic acid(SA),abscisic acid(ABA),trauma,low temperature,salt and drought.This indicated that the VqSTS12,VqSTS24 and VqSTS25 genes could be broadly induced in response to biotic and abiotic stresses.Subcellular localization in leaves of Nicotiana benthamiana observations showed that these 3 STSs were localized in the cytoplasm of leaf epidermal cells.In Vitis vinifera cv.'Thompson Seedless'overexpressing VqSTS12 and VqSTS25,significant up-regulations of STS genes(VqSTS12 and VqSTS25)were found in transgenic plants that were under drought stress simulated with 10%PEG6000.Moreover,the physiological indices like malondialdehyde(MDA)content,peroxidase(POD)activity and catalase(CAT)activity of the transgenic plants were all superior to the wild-type plants,which suggested that the over-expression of VqSTS12 and VqSTS25 genes improved the drought resistance of Thompson Seedless.In summary,the STS genes VqSTS12,VqSTS24 and VqSTS25 could be used as the disease-resistant germplasm resources of improved European grape varieties for resistance breeding.The results of the present study provide

关 键 词：中国野生毛葡萄 芪合酶基因 表达分析 生物胁迫 非生物胁迫 

分 类 号：S663.1[农业科学—果树学]