State Key Laboratory of Cotton Biology, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, China.
Int J Mol Sci. 2023 Mar 6;24(5):5057. doi: 10.3390/ijms24055057.
The CCCH zinc-finger protein contains a typical C3H-type motif widely existing in plants, and it plays an important role in plant growth, development, and stress responses. In this study, a CCCH zinc-finger gene, , was isolated and thoroughly characterized to regulate salt stress in cotton and . The expression of was up-regulated under salt, drought, and ABA treatments. GUS activity was detected in the root, stem, leaves, and flowers of Pro::GUS transgenic . Compared with the control, the GUS activity of Pro::GUS transgenic seedlings under NaCl treatment was stronger. Through the genetic transformation of , three transgenic lines of 35S- were obtained. Under NaCl and mannitol treatments, the roots of the transgenic lines were significantly longer than those of the wild-type (WT) . The leaves of the WT turned yellow and wilted under high-concentration salt treatment at the seedling stage, while the leaves of the transgenic lines did not. Further investigation showed that compared with the WT, the content of catalase (CAT) in the leaves of the transgenic lines was significantly higher. Therefore, compared with the WT, overexpression of enhanced the salt stress tolerance of transgenic . A virus-induced gene silencing (VIGS) experiment showed that compared with the control, the leaves of pYL156- plants were wilted and dehydrated. The content of chlorophyll in pYL156- leaves was significantly lower than those of the control. Therefore, silencing of reduced salt stress tolerance in cotton. Two interacting proteins (GhPP2CA and GhHAB1) of GhC3H20 have been identified through a yeast two-hybrid assay. The expression levels of and in transgenic were higher than those in the WT, and pYL156- had expression levels lower than those in the control. and are the key genes involved in the ABA signaling pathway. Taken together, our findings demonstrate that GhC3H20 may interact with GhPP2CA and GhHAB1 to participate in the ABA signaling pathway to enhance salt stress tolerance in cotton.
CCCH 锌指蛋白含有一个广泛存在于植物中的典型 C3H 基序，它在植物的生长、发育和应激反应中起着重要作用。本研究分离并深入研究了一个 CCCH 锌指基因 ，以调节棉花的盐胁迫。在盐、干旱和 ABA 处理下， 的表达上调。Pro::GUS 转基因 的根、茎、叶和花中检测到 GUS 活性。与对照相比，NaCl 处理下 Pro::GUS 转基因 幼苗的 GUS 活性更强。通过 的遗传转化，获得了三个 35S- 的转基因株系。在 NaCl 和甘露醇处理下，转基因株系的根明显长于野生型（WT）。在幼苗期高浓度盐处理下，WT 的叶片变黄萎蔫，而转基因 株系的叶片没有。进一步研究表明，与 WT 相比，转基因株系叶片中的过氧化氢酶（CAT）含量显著升高。因此，与 WT 相比，过表达 增强了转基因 的耐盐性。病毒诱导的基因沉默（VIGS）实验表明，与对照相比，pYL156- 植株的叶片萎蔫脱水。pYL156-叶片中的叶绿素含量明显低于对照。因此，沉默 降低了棉花的耐盐性。通过酵母双杂交试验鉴定了 GhC3H20 的两个互作蛋白（GhPP2CA 和 GhHAB1）。转基因 中 和 的表达水平高于 WT，而 pYL156- 的表达水平低于对照。 和 是 ABA 信号通路的关键基因。综上所述，我们的研究结果表明，GhC3H20 可能与 GhPP2CA 和 GhHAB1 相互作用，参与 ABA 信号通路，增强棉花的耐盐性。
