新疆大叶苜蓿PYL-PP2C-SnRK2基因家族鉴定及对干旱胁迫的响应分析

doi:10.11686/cyxb2025161

摘要/Abstract

摘要：

全球气候变暖导致干旱加剧，严重影响着牧草产业的可持续发展。PYL-PP2C-SnRK2是ABA信号转导通路的核心模块，在植物适应干旱胁迫的过程中发挥重要作用。本研究利用生物信息学和分子生物学的研究方法对新疆大叶苜蓿PYL-PP2C-SnRK2基因家族成员进行全基因组鉴定和干旱胁迫下的调控模式分析。结果表明：新疆大叶苜蓿基因组共包含40个MsPYLs、216个MsPP2Cs和36个MsSnRK2s基因，3个基因家族成员分别分布在22、21和24条染色体上。系统进化分析将MsPYL-PP2C-SnRK2分为3、13和3个亚家族。MsPYLs均包含PYR-PYL-RCAR-like结构域；大多数MsPP2Cs都包含PP2Cc结构域；MsSnRK2s均包含STKc结构域。在启动子区域发现了多个与胁迫反应和植物生长相关的顺式作用元件。在不同程度干旱胁迫下，MsPYL-PP2C-SnRK2s基因家族成员的表达模式存在显著差异。加权基因共表达网络分析将干旱胁迫下新疆大叶苜蓿内源ABA含量与基因表达量相关联，进一步筛选出核心调控基因。分子对接结果验证表明，MsSnRK2-14.1/14.2/1.2和MsPP2C88.4可能在干旱逆境中起关键的正向调控作用。本研究为深入探究MsPYL-PP2C-SnRK2基因在干旱胁迫响应中的功能机制提供了重要理论依据，为优良牧草抗旱性的遗传改良提供了优异的基因资源。

关键词: 干旱胁迫, PYL-PP2C-SnRK2, 脱落酸, 加权基因共表达网络分析, 新疆大叶苜蓿

Abstract:

Global climate change has led to the intensification of drought， which has had a serious impact on the sustainability of the forage industry. Among the various physiological and molecular mechanisms that plants employ to adapt to drought， the PYL-PP2C-SnRK2 module stands out because it constitutes the core components of the abscisic acid （ABA） signaling pathway， which plays an essential role in mediating plant adaptation to drought. In this study， genome-wide identification of PYL-PP2C-SnRK2 gene family members in Medicago sativa ‘Xinjiangdaye’ and analysis of their regulatory patterns under drought stress were conducted using bioinformatics and molecular biological methods. Searches of the M. sativa ‘Xinjiangdaye’ genome revealed 40 MsPYL， 216 MsPP2C， and 36 MsSnRK2 genes. The members of these three gene families were distributed across different chromosomes： MsPYLs across 22 chromosomes； MsPP2Cs across 21 chromosomes； and MsSnRK2s across 24 chromosomes. Domain architecture analysis of the putative proteins encoded by these genes confirmed the presence of the PYR-PYL-RCAR-like structural domain inall of the MsPYLs； the PP2Cc structural domain in most of the MsPP2Cs； and the STKc structural domainin all the MsSnRK2s. Analyses of gene promoter regions identified multiple stress-responsive and growth-related cis-regulatory elements. Under progressive drought stress， the MsPYL-PP2C-SnRK2 genes exhibited significantly divergent transcript profiles. A weighted gene co-expression network analysis established robust correlations between endogenous ABA accumulation and the transcript levels of specific genes， which allowed us to identify candidate core regulatory factors involved in the drought adaptation process. Molecular docking analyses demonstrated that MsSnRK2-14.1/14.2/1.2 and MsPP2C88.4 had functional predominance as key positive regulators during the drought adaptation of M. sativa ‘Xinjiangdaye’. The results of this study provide a foundational theoretical framework for further studies on the functional mechanisms of the MsPYL-PP2C-SnRK2 module and its components in the response to drought stress. Furthermore， it offers an excellent set of genetic resources for breeding new varieties of drought-resistant pasture grasses.

Key words: drought stress, PYL-PP2C-SnRK2, abscisic acid, weighted gene co-expression network analysis, Medicago sativa ‘Xinjiangdaye’

周晓彤, 罗永忠, 齐建伟, 邱应德, 马超. 新疆大叶苜蓿PYL-PP2C-SnRK2基因家族鉴定及对干旱胁迫的响应分析[J]. 草业学报, 2026, 35(4): 169-196.

Xiao-tong ZHOU, Yong-zhong LUO, Jian-wei QI, Ying-de QIU, Chao MA. Genome-wide identification of PYL-PP2C-SnRK2 gene family members in Medicago sativa ‘Xinjiangdaye’ and their transcript profiles under drought stress[J]. Acta Prataculturae Sinica, 2026, 35(4): 169-196.

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基因 Gene	正向引物 Forward primer （5′-3′）	反向引物 Reverse primer （5′-3′）
MsPP2C10.1	GATGGCTTGTGGGGTGTAATGT	GCAGCTTCAGCAGCACGACTAC
MsPP2C25	TGAAACGCCTATCCTCAAAGCT	TGATTGCTCAAGTGCTCCCATA
MsPP2C66.3	ATGTCTGATACTTGCGAGTGATGG	TCGCCAATGCCAACTTCGT
MsPP2C66.4	TTACAACTGGGCTGCAACGAG	TCACAAATGGCGTCCAGAATAG
MsActin	GCCAATGATGCTTAGCTGGTCTT	CCCTTCTTCACTCACGCGATAT

基因 Gene	正向引物 Forward primer （5′-3′）	反向引物 Reverse primer （5′-3′）
MsPP2C10.1	GATGGCTTGTGGGGTGTAATGT	GCAGCTTCAGCAGCACGACTAC
MsPP2C25	TGAAACGCCTATCCTCAAAGCT	TGATTGCTCAAGTGCTCCCATA
MsPP2C66.3	ATGTCTGATACTTGCGAGTGATGG	TCGCCAATGCCAACTTCGT
MsPP2C66.4	TTACAACTGGGCTGCAACGAG	TCACAAATGGCGTCCAGAATAG
MsActin	GCCAATGATGCTTAGCTGGTCTT	CCCTTCTTCACTCACGCGATAT