College of Forestry, Shanxi Agricultural University, Jinzhong 030801, China.
Int J Mol Sci. 2023 Oct 22;24(20):15437. doi: 10.3390/ijms242015437.
Salt stress is one of the major abiotic stresses that limits plant growth and development. The MYB transcription factor family plays essential roles in plant growth and development, as well as stress tolerance processes. In this study, the cDNA of the 84K poplar ( × ) was used as a template to clone the full length of the gene fragment, and transgenic poplar lines with overexpression (OX) or inhibited expression (RNAi, RNA interference) were cultivated. The role of PagMYB205 in poplar growth and development and salt tolerance was detected using morphological and physiological methods. The full-length CDS sequence of was 906 bp, encoding 301 amino acids, and the upstream promoter sequence contained abiotic stress-related cis-acting elements. The results of subcellular localization and transactivation assays showed that the protein had no self-activating activity and was localized in the nucleus. Under salt stress, the rooting rate and root vitality of RNAi were higher than OX and wild type (WT). However, the malondialdehyde (MDA) content of the RNAi lines was significantly lower than that of the wild-type (WT) and OX lines, but the reactive oxygen species (ROS) scavenging ability, such as the peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) enzyme activities, was dramatically more powerful. Most significantly of all, the RNAi3 line with the lowest expression level of had the lowest MDA content, the best enzyme activity and root vitality, and the best salt stress tolerance compared to the other lines. The above results suggest that the transcription factor PagMYB205 could negatively regulate salt stress tolerance by regulating antioxidant enzyme activity and root vitality.
盐胁迫是限制植物生长和发育的主要非生物胁迫之一。MYB 转录因子家族在植物生长发育以及胁迫耐受过程中发挥着重要作用。本研究以 84K 杨( × )cDNA 为模板,克隆得到全长基因片段,培育过表达(OX)或抑制表达(RNAi,RNA 干扰)的转基因杨树株系。采用形态和生理方法检测 PagMYB205 在杨树生长发育和耐盐性中的作用。全长 CDS 序列为 906bp,编码 301 个氨基酸,上游启动子序列含有非生物胁迫相关顺式作用元件。亚细胞定位和转录激活实验结果表明,该蛋白无自激活活性,定位于细胞核。在盐胁迫下,RNAi 的生根率和根活力均高于 OX 和野生型(WT)。然而,RNAi 系的丙二醛(MDA)含量明显低于 WT 和 OX 系,但活性氧(ROS)清除能力,如过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)酶活性,显著增强。最为重要的是,与其他株系相比,表达水平最低的 RNAi3 系 MDA 含量最低,酶活性和根活力最好,耐盐性最强。上述结果表明,转录因子 PagMYB205 可以通过调节抗氧化酶活性和根活力来负调控盐胁迫耐受性。