甜瓜CmPIPs基因家族鉴定和表达模式分析

doi:10.7668/hbnxb.20195142

摘要/Abstract

摘要：

对甜瓜CmPIPs基因家族鉴定和表达模式分析,为进一步探究甜瓜CmPIPs基因家族功能及甜瓜遗传改良提供理论依据和支持。利用TBtools、MEME、MEGA X、Plant-CARE工具对CmPIPs进行生物信息学分析,利用GraphPad Prism 10软件对CmPIP2;7在甜瓜授粉后不同时期中果皮中的表达量以及CmPIPs各成员在不同组织和不同浓度的植物激素处理幼叶中的表达量进行可视化解析。结果显示,CmPIP2;7与CsPIP2;8亲缘关系最近;12个CmPIPs家族成员主要分布在1,3,4,5,9,10,11号染色体上;除CmPIP2;8有3个外显子区,其余成员均有4个外显子区;CmPIPs各成员的启动子区域有多个顺式作用元件,如生长素、赤霉素、脱落酸等激素应答元件;CmPIP2;7在甜瓜的快速生长期以及成熟期的表达量显著上调;CmPIPs各家族成员在甜瓜的不同组织中均有表达;生长素浓度为40.0 μmol/L时,CmPIP2;4表达量显著上调,CmPIP1;1、CmPIP2;1、CmPIP2;2和CmPIP2;3的表达量极显著下调;脱落酸浓度为0.4,4.0,40.0 μmol/L时,CmPIP1;1、CmPIP2;1、CmPIP2;3、CmPIP2;9表达量均显著下调;茉莉酸甲酯浓度为44.640 μmol/L时,CmPIP2;1和CmPIP2;5表达量显著下调,CmPIP2;2、CmPIP2;3、CmPIP2;7、CmPIP2;9表达量显著上调;乙烯利浓度为4.0 mmol/L时,CmPIPs各成员表达量均显著上调。明确了CmPIPs基因家族成员的基因结构、序列特征、进化关系以及共线性关系,对其表达模式进行了解析。

关键词: 甜瓜, 基因家族, 质膜内在蛋白, 植物激素, 表达特性

Abstract:

The identification and expression pattern analysis of CmPIPs gene family in melon could provide theoretical basis and support for further exploring the function of CmPIPs gene family and the genetic improvement of melon.TBtools,MEME,MEGA X and Plant-CARE tools were used to analyze the bioinformatics of CmPIPs,and the expression level of CmPIP2;7 in the pericarp of melon at different stages after pollination,and the expression level of each member of CmPIPs in different tissues and different concentrations of plant hormone treated young leaves were visualized in the software GraphPad Prism 10.The results showed that CmPIP2;7 and CsPIP2;8 had nearest kinship;the 12 members of the CmPIPs family were mainly distributed on chromosomes 1,3,4,5,9,10 and 11;except that CmPIP2;8 had 3 CDS regions,the other members had 4 CDS regions.The promoter regions of each member of CmPIPs had multiple cis-acting elements,hormone responsive elements,such as auxin,gibberellin,and abscisic acid.The expression level of CmPIP2;7 was significantly up-regulated during rapid development and maturity of melon fruit.Members of various families of CmPIPs were expressed in different tissues of melon.After treatment with 40.0 μmol/L auxin,the expression level of CmPIP2;4 was significantly up-regulated,while the expression level of CmPIP1;1,CmPIP 2;1,CmPIP2;2 and CmPIP2;3 were extremely significantly down-regulated,and when the concentrations of abscisic acid were 0.4,4.0 and 40.0 μmol/L,the expression level of CmPIP1;1,CmPIP2;1,CmPIP2;3,CmPIP2;9 were significantly down-regulated.After treatment with 44.640 μmol/L methyl jasmonate,the expression level of CmPIP2;1 and CmPIP2;5 were significantly down-regulated,while the expression level of CmPIP2;2,CmPIP2;3,CmPIP2;7 and CmPIP2;9 were significantly up-regulated.When the concentration of ethylene glycol was 4.0 mmol/L,the expression level of each member of CmPIPs was significantly up-regulated.The gene structure,sequence characteristics,evolutionary relationship and collinearity of CmPIPs gene family members were clarified,and their expression pattern were analyzed.

Key words: Melon, Gene family, Integral membrane protein, Plant hormone, Expression characteristics

贺姜姜, 金兰, 金苏日古嘎, 李永, 红雨, 高峰. 甜瓜CmPIPs基因家族鉴定和表达模式分析[J]. 华北农学报, 2025, 40(1): 53-62. doi: 10.7668/hbnxb.20195142.

HE Jiangjiang, JIN Lan, JIN Suriguga, LI Yong, HONG Yu, GAO Feng. Identification and Expression Pattern Analysis of CmPIPs Gene Family in Melon[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(1): 53-62. doi: 10.7668/hbnxb.20195142.

http://www.hbnxb.net/CN/Y2025/V40/I1/53

图/表 12

表1 RT-qPCR引物序列

Tab.1 Sequences of primers used in RT-qPCR

基因名 Gene name	上游引物序列(5'—3') Forward primer sequence (5'—3')	下游引物序列(5'—3') Reverse primers sequence(5'—3')	大小/bp Size
CmGAPDH	ATCATTCCTAGCAGCACTGG	TTGGCATCAAATATGCTTGACCTG	278
CmPIP1;1	GGTGACTTTCGGGCTGTTTCT	CTTTGGTGTATCCCTCGCTCA	176
CmPIP1;2	TCTCCGCCACCGATGCTAA	AATGGTCATCCCAAGCGTGTT	189
CmPIP2;1	CGGTGTTCATGGTTCATTTGG	GCTTTTGCTGCTCCTGCTCTTA	199
CmPIP2;2	AGTAGCCAGTCCGACATCAAACA	TTCCACCGAAAGCCCAAGC	82
CmPIP2;3	ATTCGCCGTCAAGGATTACCA	TGCCACCAAATGCCCAAGC	233
CmPIP2;4	CCAAAGCAAAGACTACCAAGACC	CCCAAGCGATACCGAGAATG	212
CmPIP2;5	TAATAGACGGCGTGGAGATAAGG	ACATACAGGAAAAGCAGAGTCGC	85
CmPIP2;6	TGATGGAAGAAGCAATGTCAAGG	GAGAAGCGTGGCAACGAACT	124
CmPIP2;7	AAAGAAGTGACGGAAGAGGGACA	GCGATGAACTCAGCGATAAGG	131
CmPIP2;8	GGAGGGCACATAAATCCAGCAG	CCCACAATCTCAACAGCAAGG	224
CmPIP2;9	GATCATCCTCCGGCTCCCTT	CGGCAGAATCCTGACTTTGG	148
CmPIP2;10	ACAACAGCCAGACCGATACCA	GCACTAACGACACTTTACGAGCC	189

图1 甜瓜PIPs家族成员的基因结构和保守基序分析 A.保守基序;B.Motif 序列微标(每个字母的高度表示该位置氨基酸的出现频率);C.基因结构。

Fig.1 Gene structure and conservative motif analysis of PIPs family members in melon A.Conserved motif;B.Motif sequence micro label(The height of each letter indicates the frequency of occurrence of amino acids at that location);C.Gene structure.

图2 CmPIPs基因家族成员染色体定位

Fig.2 Chromosomal mapping of CmPIPs gene family members

图3 CmPIPs基因家族成员共线性分析

Fig.3 Synteny analysis of CmPIPs gene family members

图4 甜瓜、黄瓜、西瓜PIPs基因家族的系统发育树

Fig.4 Phylogenetic tree of PIPs family in melon,cucumber and watermelon

图 5 CmPIPs基因家族成员的启动子区域顺式作用元件

Fig.5 Promoter region cis-acting elements of various members of the CmPIPs gene family

图6 CmPIP2;7在甜瓜授粉后不同天数中果皮的表达特性不同小写字母表示在P<0.05水平上差异显著。试验结果为1个生物、3个技术重复。图7—11同。

Fig.6 Expression characteristics of CmPIP2;7 in the pericarp of melon at different days after pollination Different lowercase letters indicate significant differences at P<0.05 level. The results were 1 biological and 3 technical replicates. The same as Fig.7—11.

图7 不同组织中CmPIPs的表达特性

Fig.7 Expression characteristics of CmPIPs in different tissues

图8 生长素处理时CmPIPs的表达特性

Fig.8 Expression characteristics of CmPIPs under auxin treatment

图9 脱落酸处理时CmPIPs的表达特性

Fig.9 Expression characteristics of CmPIPs under abscisic acid treatment

图10 茉莉酸甲酯处理时CmPIPs的表达特性

Fig.10 Expression characteristics of CmPIPs under methyl jasmonate treatment

图11 乙烯利处理时CmPIPs的表达特性

Fig.11 Expression characteristics of CmPIPs under ethylene treatment

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