College of Horticulture, Gansu Agricultural University, Lanzhou, 730070, China.
Plant Cell Rep. 2024 Oct 17;43(11):265. doi: 10.1007/s00299-024-03353-1.
Twenty-five VvIAA genes and eighteen VaIAA genes were identified from Pinot Noir and Shanputao, respectively. The overexpression of VaIAA3 in transgenic Arabidopsis increased cold tolerance by regulating auxin, ABA and ethylene signaling. Aux/IAA genes are key genes involved in regulating auxin signal transduction in plants. Although IAA genes have been characterized in various plant species, the role of IAA genes in grape cold resistance is unclear. To further explore the members of the Aux/IAA gene family in grape and their functions, in this study, using genomic data for Pinot Noir (Vitis vinifera cv. 'Pinot Noir') and Shanputao (Vitis amurensis), 25 VvIAA genes and 18 VaIAA genes were identified. The VaIAA genes presented different expression patterns at five different temperatures (28 ± 1 °C, 5 ± 1 °C, 0 ± 1 °C, -5 ± 1 °C, and -10 ± 1 °C) according to qRT‑PCR results. VaIAA3 was selected as a candidate gene for further functional analysis because of its high expression level under low-temperature stress. Subcellular localization experiments revealed that VaIAA3 was localized in the nucleus. Additionally, under 4 °C treatment for 24 h, relative expression level of VaIAA3, antioxidant enzyme activity, survival rate, and cold-responsive gene expression in three transgenic lines (OE-1, OE-2, OE-3) were greater, whereas relative electrolytic conductivity (REC), malondialdehyde (MDA) content and hydrogen peroxide (HO) content were lower than those of the wild type (WT). Transcriptome sequencing analysis revealed that VaIAA3 regulated cold stress resistance in Arabidopsis thaliana (Arabidopsis) through pathways involving auxin, ABA, JA, or ethylene. Importantly, heterologous overexpression of VaIAA3 increased the resistance of Arabidopsis to cold stress, which provides a theoretical basis for the further use of VaIAA3 to improve cold resistance in grape.
从黑比诺和山葡萄中分别鉴定出 25 个 VvIAA 基因和 18 个 VaIAA 基因。过表达转基因拟南芥中的 VaIAA3 通过调节生长素、ABA 和乙烯信号来提高耐冷性。Aux/IAA 基因是植物中参与调节生长素信号转导的关键基因。尽管已在各种植物物种中鉴定出 IAA 基因，但 IAA 基因在葡萄耐寒性中的作用尚不清楚。为了进一步探讨葡萄 Aux/IAA 基因家族的成员及其功能，本研究利用黑比诺（Vitis vinifera cv. 'Pinot Noir'）和山葡萄（Vitis amurensis）的基因组数据，鉴定出 25 个 VvIAA 基因和 18 个 VaIAA 基因。根据 qRT-PCR 结果，VaIAA 基因在五个不同温度（28±1°C、5±1°C、0±1°C、-5±1°C 和-10±1°C）下表现出不同的表达模式。由于 VaIAA3 在低温胁迫下表达水平较高，因此选择其作为进一步功能分析的候选基因。亚细胞定位实验表明，VaIAA3 定位于细胞核内。此外，在 4°C 处理 24 h 后，三个转基因株系（OE-1、OE-2、OE-3）的 VaIAA3 相对表达水平、抗氧化酶活性、存活率和冷响应基因表达均较高，而相对电导率（REC）、丙二醛（MDA）含量和过氧化氢（HO）含量均低于野生型（WT）。转录组测序分析表明，VaIAA3 通过涉及生长素、ABA、JA 或乙烯的途径调节拟南芥的冷胁迫抗性。重要的是，VaIAA3 的异源过表达提高了拟南芥对冷胁迫的抗性，这为进一步利用 VaIAA3 提高葡萄的耐寒性提供了理论依据。
