甜瓜CmPOD基因家族鉴定及叶组织响应低温胁迫的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0940

摘要/Abstract

摘要：

目的 Ⅲ类过氧化物酶（class Ⅲ peroxidases, POD）基因家族参与植物中多种生物与非生物胁迫。为探究甜瓜CmPOD家族基因对低温胁迫的响应，进行基因鉴定并对叶组织进行低温表达分析。方法以薄皮甜瓜‘通甜蜜4号’为材料，采用生物信息学方法，鉴定甜瓜POD基因家族并对其理化性质、进化树、基因结构、顺式作用元件进行系统分析。对部分CmPOD基因进行蛋白结构预测和低温胁迫下的表达分析。结果通过鉴定获得77个CmPOD基因家族成员，命名为CmPOD1-1-CmPOD12-7，氨基酸残基数范围为121‒416 aa，pI值范围为4.53‒9.50；亚细胞定位主要分布在叶绿体和细胞外基质。CmPOD基因非均匀分布在12条染色体上，在Chr01和Chr11分布较集中，分别有15个成员；进化树分析CmPOD基因家族分为13个亚族；启动子区富含MYB、MYC、ABRE和LTR等胁迫响应元件；RT-qPCR结果表明，低温4 ℃处理24、48 h后，CmPOD11-13、CmPOD12-6和CmPOD11-14等基因在叶片中上调表达，CmPOD7-3在叶片中下调表达。结论在甜瓜中鉴定得到77个POD基因家族成员，在低温胁迫下表达水平不同，推测CmPOD基因家族参与甜瓜低温胁迫响应。

关键词: 甜瓜, POD基因家族, 低温胁迫, 表达分析, 生物信息学分析

Abstract:

Objective The class Ⅲ peroxidase (POD) gene family is involved in various biotic and abiotic stresses in plants. This study aimed to identify the CmPOD family genes and analyze their expression in leaf tissues under low-temperature stress to explore their response to cold stress. Method The thin-skinned melon variety ‘Tongtianmi 4’ was used as the material. Bioinformatics methods were employed to identify the melon POD gene family, and a systematic analysis was conducted on their physicochemical properties, phylogenetic tree, gene structure, and cis-acting elements. Protein structure prediction and expression analysis under low-temperature stress were performed for selected CmPOD genes. Result A total of 77 members of the CmPOD gene family were identified and named CmPOD1-1 to CmPOD12-7. The amino acid residue number ranged from 121 to 416 aa, and the pI value ranged from 4.53 to 9.50. Subcellular localization was mainly predicted in chloroplasts and the extracellular matrix. The CmPOD genes were unevenly distributed across 12 chromosomes, with 15 members each on Chr01 and Chr11. Phylogenetic analysis divided the CmPOD gene family into 13 subfamilies. The promoter regions were rich in stress-responsive cis-elements such as MYB, MYC, ABRE, and LTR. RT-qPCR results showed that after treatment at 4 ℃ for 24 h and 48 h, genes including CmPOD11-13, CmPOD12-6, and CmPOD11-14 were upregulated in leaves, while CmPOD7-3 was downregulated. Conclusion Seventy-seven members of the POD gene family were identified in melon, and their expression levels varied under low-temperature stress. It is speculated that the CmPOD gene family is involved in the response of melon to low-temperature stress.

Key words: melon, POD gene family, low-temperature stress, expression analysis, bioinformatics analysis

梁智伦, 漆思庆, 李欣洋, 梁健浩, 刘骕骦. 甜瓜CmPOD基因家族鉴定及叶组织响应低温胁迫的表达分析[J]. 生物技术通报, 2026, 42(10): 188-197.

LIANG Zhi-lun, QI Si-qing, LI Xin-yang, LIANG Jian-hao, LIU Su-shuang. Identification of the CmPOD Gene Family in Melon and Expression Analysis of Leaf Tissues under Low-temperature Stress[J]. Biotechnology Bulletin, 2026, 42(10): 188-197.

图/表 10

表1 CmPOD家族顺式作用元件功能

Table 1 Cis-acting element functions in the CmPOD family

顺式作用元件 Cis-acting element	功能 Function
ABRE	脱落酸响应元件，参与ABA信号通路和胁迫响应
DRE core	脱水响应元件，响应低温与干旱
MYB	转录因子结合位点，参与发育、代谢和胁迫响应
MYC	转录因子结合位点，参与JA信号通路和胁迫响应
ARE	抗氧化响应元件，参与活性氧清除和氧化胁迫防御
LTR	低温响应元件，参与冷胁迫基因调控
ERE	乙烯响应元件，调控果实成熟和胁迫响应
GARE-motif	赤霉素响应元件，介导GA信号通路
AuxRR-core	生长素响应元件，调控生长素依赖基因表达
Box	光响应元件，调控光诱导基因表达
Box 4	光响应元件，参与光调控基因表达
Sp1	通用转录因子结合位点，增强基础转录
W box	WRKY转录因子结合位点，调控病原菌防御反应和SA信号通路
TC-rich repeats	防御和胁迫响应元件，参与病原体应答及机械损伤响应
MBS	MYB结合位点，参与干旱诱导基因表达
TATA-box	核心启动子元件，介导RNA聚合酶Ⅱ的转录起始定位
CAAT-box	增强子元件，提高基础转录效率
GC-motif	增强子元件，参与光诱导和胁迫响应

图1 CmPOD家族系统进化树构建

Fig. 1 Construction of the phylogenetic tree of the CmPOD family

图 2 CmPOD家族基因结构数量分析CDS：编码序列；exon：外显子；five prime UTR：5′非翻译区；mRNA：信使RNA；three prime UTR：3′非翻译区

Fig. 2 Quantitative analysis of gene structure in the CmPOD familyCDS: Coding sequence; exon: exon; five prime UTR: 5′ untranslated region; mRNA: messenger RNA; three prime UTR: 3′ untranslated region

图3 CmPOD家族基因结构及调控元件分析数字0、1、2表示内含子相位

Fig. 3 Analysis of the gene structure and regulatory elements in the CmPOD family0, 1, and 2 indicate intron phases

图4 CmPOD家族motif组成分析

Fig. 4 Motif composition of the CmPOD family

图5 CmPOD家族顺式作用元件统计

Fig. 5 Cis-acting element statistics in the CmPOD family

图6 CmPOD家族在染色体上的定位

Fig. 6 Chromosomal localization of the CmPOD family

图7 CmPOD家族物种内共线性分析

Fig. 7 Intraspecific collinearity analysis of the CmPOD family

图8 CmPOD家族叶组织表达热图

Fig. 8 Heatmap of CmPOD family leaf expression

图9 部分CmPOD家族成员叶组织表达差异* P<0.05， ** P<0.01， *** P<0.001. t-test， n=3

Fig. 9 Expression differences of selected CmPOD family members in leaves

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