Cloning of Cuticular Wax Synthesis-Related Protein VvWSD1 Gene in Grape (Vitis vinifera) and Its Function Analysis of Low Temperature Response
YE Qing1,2, ZONG Quan-Ying1,2, ZHU Xiao-Min1,2, TIAN Ye1,2, HOU Li-Xia1,2, LIU Xin1,2*
1 College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China; 2 Key Laboratory of Plant Biotechnology in Universities of Shandong Province, Qingdao 266109, China
Abstract：Low temperature is the major stress factor limiting the high-quality development of the grape (Vitis vinifera) industry. The cuticular wax on leaves is closely related to the plant's stress resistance, but the biological functions and synthesis pathways of cuticular wax during cold stress response is still unclear. The wax ester synthase 1 (WSD1) catalyzes the production of wax esters, an important component of cuticular wax, which play a crucial role in the synthesis of cuticular wax. The present study initially examined the correlation between leaf cuticular wax and cold resistance of grape by utilizing 24 diverse grape germplasm with varying levels of cold resistance. The results showed that the total wax content of leaf epidermis of cold resistant grape varieties was significantly higher than that of sensitive grape varieties, low temperature could promote the wax accumulation in grape leaves, and the wax content in the epidermis of cold resistant grape varieties increased significantly, with the most obvious increase in esters and alkanes. Meanwhile, the wax ester synthase gene VvWSD1 (GenBank No. XM_002263373.5) induced by low temperature stress was cloned from the cold-resistant grape variety 'Zuoyouhong', bioinformatics analysis and expression characteristic analysis of the VvWSD1 were conducted. Using transgenic Arabidopsis plants, the function of VvWSD1 in cold tolerance was studied. The results showed that VvWSD1 cDNA was 1 524 bp in length, encoding 514 amino acids. VvWSD1 was expressed in the stems, leaves, buds and berries of grapes, with the highest expression level observed in the mature leaves. VvWSD1 was induced by cold stress, and the expression levels in resistant grape varieties were significantly higher than those in sensitive ones. Under cold stress, the content of cuticular wax increased significantly after VvWSD1 transiently transformed, the growth of the VvWSD1 overexpressing lines was better than wild-type. Compared with the wild type, the VvWSD1 transgenic lines accumulated wax in the leaf epidermis at low temperatures, decreased leaf water loss rate and chlorophyll leaching, decreased malondialdehyde (MDA) and reactive oxygen species (ROS) content, and decreased relative permeability of cell membranes, indicating that VvWSD1 mediated wax accumulation and participated in grape response to low temperature stress. This study establishes a theoretical framework for elucidating the molecular mechanisms through which leaf cuticular waxes and their associated synthesis enzyme genes contribute to plant responses to cold stress, thereby holding significant implications for the advancement of cold-resistant grape cultivation and germplasm selection.
叶青, 宗全颖, 朱晓敏, 田也, 侯丽霞, 刘新. 葡萄表皮蜡质合成相关基因VvWSD1克隆及其低温胁迫响应功能分析[J]. 农业生物技术学报, 2024, 32(12): 2740-2754.
YE Qing, ZONG Quan-Ying, ZHU Xiao-Min, TIAN Ye, HOU Li-Xia, LIU Xin. Cloning of Cuticular Wax Synthesis-Related Protein VvWSD1 Gene in Grape (Vitis vinifera) and Its Function Analysis of Low Temperature Response. 农业生物技术学报, 2024, 32(12): 2740-2754.
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