Table_3_Translational profile of coding and non-coding RNAs revealed by genome wide profiling of ribosome footprints in grapevine.xls

Translation is a crucial process during plant growth and morphogenesis. In grapevine (Vitis vinifera L.), many transcripts can be detected by RNA sequencing; however, their translational regulation is still largely unknown, and a great number of translation products have not yet been identified. Here, ribosome footprint sequencing was carried out to reveal the translational profile of RNAs in grapevine. A total of 8291 detected transcripts were divided into four parts, including the coding, untranslated regions (UTR), intron, and intergenic regions, and the 26 nt ribosome-protected fragments (RPFs) showed a 3 nt periodic distribution. Furthermore, the predicted proteins were identified and classified by GO analysis. More importantly, 7 heat shock-binding proteins were found to be involved in molecular chaperone DNA J families participating in abiotic stress responses. These 7 proteins have different expression patterns in grape tissues; one of them was significantly upregulated by heat stress according to bioinformatics research and was identified as DNA JA6. The subcellular localization results showed that VvDNA JA6 and VvHSP70 were both localized on the cell membrane. Therefore, we speculate that DNA JA6 may interact with HSP70. In addition, overexpression of VvDNA JA6 and VvHSP70, reduced the malondialdehyde (MDA) content, improved the antioxidant enzyme activity of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), increased the content of proline, an osmolyte substance, and affected the expression of the high-temperature marker genes VvHsfB1, VvHsfB2A, VvHsfC and VvHSP100. In summary, our study proved that VvDNA JA6 and the heat shock protein VvHSP70 play a positive role in the response to heat stress. This study lays a foundation for further exploring the balance between gene expression and protein translation in grapevine under heat stress.

翻译是植物生长与形态建成过程中的关键环节。在葡萄（Vitis vinifera L.）中，通过RNA测序（RNA sequencing）可检测到大量转录本，但目前其翻译调控机制仍未明确，且大量翻译产物尚未被鉴定。本研究采用核糖体印迹测序（ribosome footprint sequencing）技术，解析了葡萄体内RNA的翻译谱特征。本研究共检测到8291条转录本，将其划分为编码区、非翻译区（untranslated regions, UTR）、内含子区与基因间区四类；同时发现26 nt的核糖体保护片段（ribosome-protected fragments, RPFs）呈现3 nt的周期性分布特征。此外，通过基因本体（Gene Ontology, GO）分析，本研究对预测得到的蛋白质进行了鉴定与分类。尤为重要的是，研究发现7种热休克结合蛋白隶属于分子伴侣DNA J家族，参与非生物胁迫应答过程。这7种蛋白在葡萄组织中呈现不同的表达模式；经生物信息学分析，其中1种可被热胁迫显著上调表达，经鉴定为DNA JA6。亚细胞定位结果显示，VvDNA JA6与VvHSP70均定位于细胞膜上，据此推测DNA JA6可能与HSP70存在相互作用。此外，过表达VvDNA JA6与VvHSP70可降低丙二醛（malondialdehyde, MDA）含量，提升超氧化物歧化酶（superoxide dismutase, SOD）、过氧化氢酶（catalase, CAT）与过氧化物酶（peroxidase, POD）的抗氧化酶活性，增加渗透调节物质脯氨酸的含量，并影响高温标记基因VvHsfB1、VvHsfB2A、VvHsfC与VvHSP100的表达水平。综上，本研究证实VvDNA JA6与热休克蛋白VvHSP70在植物应对热胁迫的过程中发挥正向调控作用。本研究为进一步解析热胁迫下葡萄体内基因表达与蛋白质翻译间的平衡关系奠定了基础。