Vannozzi Alessandro, Dry Ian B, Fasoli Marianna, Zenoni Sara, Lucchin Margherita
Department of Agronomy, Food, Natural resources, Animals and Environment, University of Padova, Agripolis, viale dell'Università 16, 35020, Legnaro, Padova, Italy.
BMC Plant Biol. 2012 Aug 3;12:130. doi: 10.1186/1471-2229-12-130.
Plant stilbenes are a small group of phenylpropanoids, which have been detected in at least 72 unrelated plant species and accumulate in response to biotic and abiotic stresses such as infection, wounding, UV-C exposure and treatment with chemicals. Stilbenes are formed via the phenylalanine/polymalonate-route, the last step of which is catalyzed by the enzyme stilbene synthase (STS), a type III polyketide synthase (PKS). Stilbene synthases are closely related to chalcone synthases (CHS), the key enzymes of the flavonoid pathway, as illustrated by the fact that both enzymes share the same substrates. To date, STSs have been cloned from peanut, pine, sorghum and grapevine, the only stilbene-producing fruiting-plant for which the entire genome has been sequenced. Apart from sorghum, STS genes appear to exist as a family of closely related genes in these other plant species.
In this study a complete characterization of the STS multigenic family in grapevine has been performed, commencing with the identification, annotation and phylogenetic analysis of all members and integration of this information with a comprehensive set of gene expression analyses including healthy tissues at differential developmental stages and in leaves exposed to both biotic (downy mildew infection) and abiotic (wounding and UV-C exposure) stresses. At least thirty-three full length sequences encoding VvSTS genes were identified, which, based on predicted amino acid sequences, cluster in 3 principal groups designated A, B and C. The majority of VvSTS genes cluster in groups B and C and are located on chr16 whereas the few gene family members in group A are found on chr10. Microarray and mRNA-seq expression analyses revealed different patterns of transcript accumulation between the different groups of VvSTS family members and between VvSTSs and VvCHSs. Indeed, under certain conditions the transcriptional response of VvSTS and VvCHS genes appears to be diametrically opposed suggesting that flow of carbon between these two competing metabolic pathways is tightly regulated at the transcriptional level.
This study represents an overview of the expression pattern of each member of the STS gene family in grapevine under both constitutive and stress-induced conditions. The results strongly indicate the existence of a transcriptional subfunctionalization amongst VvSTSs and provide the foundation for further functional investigations about the role and evolution of this large gene family. Moreover, it represents the first study to clearly show the differential regulation of VvCHS and VvSTS genes, suggesting the involvement of transcription factors (TFs) in both the activation and repression of these genes.
植物芪类化合物是一小类苯丙烷类化合物,已在至少72种不相关的植物物种中被检测到,并在应对生物和非生物胁迫(如感染、创伤、紫外线-C照射和化学处理)时积累。芪类化合物通过苯丙氨酸/聚丙二酸途径形成,该途径的最后一步由芪合酶(STS)催化,STS是一种III型聚酮合酶(PKS)。芪合酶与类黄酮途径的关键酶查尔酮合酶(CHS)密切相关,这两种酶共享相同底物这一事实就说明了这一点。迄今为止,已从花生、松树、高粱和葡萄中克隆出STS,葡萄是唯一一种已对其全基因组进行测序的产芪结果实植物。除高粱外,STS基因在这些其他植物物种中似乎以一个密切相关的基因家族形式存在。
在本研究中,对葡萄中STS多基因家族进行了全面表征,首先对所有成员进行了鉴定、注释和系统发育分析,并将这些信息与一组全面的基因表达分析相结合,包括不同发育阶段的健康组织以及受到生物(霜霉病感染)和非生物(创伤和紫外线-C照射)胁迫的叶片。鉴定出至少33个编码VvSTS基因的全长序列,根据预测的氨基酸序列,这些序列聚集成3个主要组,分别命名为A、B和C组。大多数VvSTS基因聚集在B组和C组,位于第16号染色体上,而A组中少数基因家族成员位于第10号染色体上。微阵列和mRNA测序表达分析揭示了VvSTS家族不同组之间以及VvSTS与VvCHS之间转录本积累的不同模式。事实上,在某些条件下,VvSTS和VvCHS基因的转录反应似乎截然相反,这表明这两个相互竞争的代谢途径之间的碳流在转录水平上受到严格调控。
本研究概述了葡萄中STS基因家族每个成员在组成型和胁迫诱导条件下的表达模式。结果有力地表明VvSTS之间存在转录亚功能化,并为进一步研究这个大基因家族的作用和进化提供了基础。此外,这是第一项清楚显示VvCHS和VvSTS基因差异调控的研究,表明转录因子(TFs)参与了这些基因的激活和抑制。