Wu Zhaoxin, Wang Ping
College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot, 010018, Inner Mongolia, China.
Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Hohhot, 010031, Inner Mongolia, China.
Sci Rep. 2025 Jan 9;15(1):1501. doi: 10.1038/s41598-025-85615-w.
Pugionium cornutum (L.) Gaertn (P. cornutum) has strong tolerance to drought, salt and disease, but the tolerance mechanisms for such stresses in P. cornutum are largely unknown. In this study, we identified the PcNAC25 transcription factor gene in P. cornutum. Its open reading frame was revealed to comprise 891 bp, encoding a protein consisting of 297 amino acids, with an isoelectric point of 6.61. Phylogenetic analysis showed that PcNAC25 was most closely related to ANAC019. The expression of PcNAC25 was induced by dehydration, mannitol, heat, cold, salt stresses and abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (JA) treatments. A subcellular localization analysis confirmed that PcNAC25 was localized in the nucleus. The overexpressing PcNAC25 lines in Arabidopsis had longer roots than wild-type (WT) lines under drought and salt stress. The overexpression of PcNAC25 improved drought and salt tolerance in transgenic Arabidopsis. Under drought and salt stress, PcNAC25 transgenic lines exhibited higher the CAT, POD and SOD activities and scavenging ability of hydroxyl radical than WT, more proline accumulation than WT and less MDA and HO content and superoxide anion production rate than WT. PcNAC25 transgenic lines also exhibited greater reduced water loss rate of detached leaves than WT. Meanwhile, DAB and NBT staining showed that the accumulation of hydrogen peroxide and superoxide anion in PcNAC25 transgenic lines were also less than WT. In addition, overexpressing PcNAC25 enhanced the expression of drought response genes (DREB2A, SOD4, RD29A, NCED3, POD3, P5CS1, PYR1 and SAG13) and salt response genes NHX, SLAH1, SOS1 and NPF6.3. The mentioned above results indicated that PcNAC25 is a positive regulator that activates ROS-scavenging enzymes and enhances root formation in Arabidopsis, which provided a basis for further research on the molecular mechanism of PCNAC25-mediated regulation of drought and salt stress, and also provided gene resources of drought and salt tolerance.
沙芥(Pugionium cornutum (L.) Gaertn)对干旱、盐和病害具有较强的耐受性,但沙芥对这些胁迫的耐受机制在很大程度上尚不清楚。在本研究中,我们鉴定了沙芥中的PcNAC25转录因子基因。其开放阅读框显示由891个碱基对组成,编码一个由297个氨基酸组成的蛋白质,等电点为6.61。系统发育分析表明,PcNAC25与ANAC019关系最为密切。PcNAC25的表达受脱水、甘露醇、热、冷、盐胁迫以及脱落酸(ABA)、水杨酸(SA)和茉莉酸甲酯(JA)处理的诱导。亚细胞定位分析证实PcNAC25定位于细胞核。在干旱和盐胁迫下,拟南芥中过表达PcNAC25的株系比野生型(WT)株系的根更长。PcNAC25的过表达提高了转基因拟南芥的耐旱性和耐盐性。在干旱和盐胁迫下,PcNAC25转基因株系的CAT、POD和SOD活性以及羟自由基清除能力均高于WT,脯氨酸积累量高于WT,MDA和HO含量以及超氧阴离子产生率低于WT。PcNAC25转基因株系离体叶片的失水率也比WT降低得更多。同时,DAB和NBT染色表明,PcNAC25转基因株系中过氧化氢和超氧阴离子的积累也少于WT。此外,过表达PcNAC25增强了干旱响应基因(DREB2A、SOD4、RD29A、NCED3、POD3、P5CS1、PYR1和SAG13)和盐响应基因NHX、SLAH1、SOS1和NPF6.3的表达。上述结果表明,PcNAC25是一种正调控因子,可激活拟南芥中的ROS清除酶并促进根系形成,这为进一步研究PcNAC25介导的干旱和盐胁迫调控的分子机制提供了依据,也提供了耐旱和耐盐的基因资源。