Identification of the PMEI Gene Family of Pectin Methylesterase Inhibitor in Foxtail Millet and Analysis of Its Response to Abiotic Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-0018

Abstract

Abstract:

Objective PMEI (pectin methylesterase inhibitor) is a key component regulating the structure and properties of the cell wall, playing a significant role in plant stress response mechanisms. Therefore, studying the response mechanism of PMEI gene in foxtail millet under abiotic stress may provide a theoretical basis for the mechanism of resistance to stress in millet. Method Bioinformatics was used to identify PMEI gene family in foxtail millet, and quantitative real-time PCR was employed to analyze the expression patterns of family members under low temperature, drought, MeJA and ABA stress. Result A total of 68 SiPMEI gene family members were identified in the foxtail millet (Setaria italica) genome, unevenly distributed on nine chromosomes, and most of them were localized to the cell wall or chloroplasts. There were two main domains of SiPMEI family members, PMEI domain and pectinesteras + PMEI domain, as well as similar physicochemical properties, subcellular localization and gene structure of the family members containing the same domain. Promoter cis-acting element analysis suggested that the SiPMEI gene family members contained multiple abiotic stress and hormone responsive elements. In this study, eight SiPMEI family members containing more than two stress response elements were selected for quantitative real-time expression analysis. The results showed that SiPMEI was differentially expressed in the root, stem, leaf and ear of the foxtail millet. Under abiotic stress (low temperature, drought), hormone stress (MeJA, ABA), the expression of SiPMEI gene tended to increase within 0-24 h, and the highest expressions of member SiPMEI30, SiPMEI32, SiPMEI36 and SiPMEI63 containing PMEI domain and localized on the cell wall responded to stress at 8‒24 h. The highest expression distribution of member SiPMEI22, SiPMEI31, SiPMEI38, and SiPMEI47 with subcellular localization on chloroplasts was more dispersed and had more upregulation sites. Conclusion SiPMEI has a positive response under low temperature, ABA and MeJA stresses, and has a similar expression trend under drought and MeJA stresses. These differential expressions suggest that SiPMEI may respond to abiotic stress through different molecular mechanisms.

Key words: foxtail millet, PMEI gene family, drought stress, low-temperature stress, methyl jasmonate stress, abscisic acid stress, subcellular localization, domain

LI Xin-ni, LI Jun-yi, MA Xue-hua, HE Wei, LI Jia-li, YU Jia, CAO Xiao-ning, QIAO Zhi-jun, LIU Si-chen. Identification of the PMEI Gene Family of Pectin Methylesterase Inhibitor in Foxtail Millet and Analysis of Its Response to Abiotic Stress[J]. Biotechnology Bulletin, 2025, 41(7): 150-163.

Figures/Tables 14

References 35

[1]	刁现民. 育种创新造就谷子种业新发展 [J]. 中国种业, 2022(4): 4-7.
	Diao XM. Breeding innovation creates new development of millet seed industry [J]. China Seed Ind, 2022(4): 4-7.
[2]	周雪, 苏乐平, 李星星, 等. 25个谷子品种(系)主要农艺性状分析及评价 [J]. 中国种业, 2025(2): 95-100, 108.
	Zhou X, Su LP, Li XX, et al. Analysis and evaluation of main agronomic traits of 25 millet varieties(lines) [J]. China Seed Ind, 2025(2): 95-100, 108.
[3]	侯保俊, 何太, 杨红霞. 晋北高寒冷凉区谷子地膜覆盖栽培技术 [J]. 农业技术与装备, 2009(5): 34-35.
	Hou BJ, He T, Yang HX. Cultivation technology of millet in cold area of northern Shanxi [J]. Agric Technol Equip, 2009(5): 34-35.
[4]	张晓霞. 三种非生物胁迫下谷子qRT-PCR内参基因的筛选与鉴定 [D]. 太谷: 山西农业大学, 2023.
	Zhang XX. Screening and identification of qRT-PCR reference genes in foxtail millet under threeabiotic stresses [D]. Taigu: Shanxi Agricultural University, 2023.
[5]	汪明滔, 刘建伟, 赵春钊. 植物调控盐胁迫下细胞壁完整性的分子机制 [J]. 生物技术通报, 2023, 39(11): 18-27.
	Wang MT, Liu JW, Zhao CZ. Molecular mechanisms of cell wall integrity in plants under salt stress [J]. Biotechnol Bull, 2023, 39(11): 18-27.
[6]	关月, 费雅婷, 朱祝军, 茹磊. 果胶甲酯酶抑制子在植物生长发育和逆境胁迫中的作用 [J/OL]. 分子植物育种, 2024. .
	Guan Y, Fei YT, Zhu ZJ, Ru L. The role of pectin methylesterase inhibitors in plant growth, development, and stress response [J/OL]. Mol Plant Breed, 2024. .
[35]	Du XF, Wang ZL, Han KN, et al. Identification and analysis of RNA editing sites of chloroplast genes in foxtail millet [Setaria italica(L.) P.Beauv [J]. Acta Agron Sin, 2022, 48(4): 873-885.
[7]	Hothorn M, Wolf S, Aloy P, et al. Structural insights into the target specificity of plant invertase and pectin methylesterase inhibitory proteins [J]. Plant Cell, 2004, 16(12): 3437-3447.
[8]	熊媛. 毛果杨PMEI家族分子特征和功能性评估 [D]. 南京: 南京林业大学, 2023.
	Xiong Y. Molecular characteristics and functional evaluation of PMEI family of Populus tomentosa [D]. Nanjing: Nanjing Forestry University, 2023.
[9]	Balestrieri C, Castaldo D, Giovane A, et al. A glycoprotein inhibitor of pectin methylesterase in kiwi fruit (Actinidia chinensis) [J]. Eur J Biochem, 1990, 193(1): 183-187.
[10]	Wang JJ, Ling L, Cai H, et al. Gene-wide identification and expression analysis of the PMEI family genes in soybean (Glycine max) [J]. 3 Biotech, 2020, 10(8): 335.
[11]	Liu TT, Yu H, Xiong XP, et al. Genome-wide identification and characterization of pectin methylesterase inhibitor genes in Brassica oleracea [J]. Int J Mol Sci, 2018, 19(11): 3338.
[12]	王文玉, 施旭, 施琪, 等. 水稻PMEI基因家族的全基因组鉴定与生物信息学分析 [J]. 分子植物育种, 2023, 21(23): 7657-7666.
	Wang WY, Shi X, Shi Q, et al. Genome-wide identification and bioinformatics analysis of PMEI gene family in rice [J]. Mol Plant Breed, 2023, 21(23): 7657-7666.
[13]	Pinzón-Latorre D, Deyholos MK. Characterization and transcript profiling of the pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) gene families in flax (Linum usitatissimum) [J]. BMC Genomics, 2013, 14: 742.
[14]	Liu NN, Sun Y, Pei YK, et al. A pectin methylesterase inhibitor enhances resistance to Verticillium wilt [J]. Plant Physiol, 2018, 176(3): 2202-2220.
[15]	杨帅, 高尚珠, 卢晗, 等. 植物细胞壁形成及在非生物胁迫中的作用 [J]. 植物生理学报, 2023, 59(7): 1251-1264.
	Yang S, Gao SZ, Lu H, et al. Plant cell wall development and its function in abiotic stress [J]. Plant Physiol J, 2023, 59(7): 1251-1264.
[16]	仇婷婷. 木薯果胶甲基酯酶抑制因子MePMEI1功能分析及其互作蛋白的筛选 [D]. 海口: 海南大学, 2020.
	Qiu TT. Functional analysis of cassava pectin methyl esterase inhibitor MePMEI1 and screening of its interacting proteins [D]. Haikou: Hainan University, 2020.
[17]	王跃腾. PMEI抑制PME的机理及食品应用型PMEI的改造研究 [D]. 成都: 成都大学, 2023.
	Wang YT. Mechanism of PMEI inhibiting PME and modification of food application PMEI [D]. Chengdu: Chengdu University, 2023.
[18]	楚刘阳. 甘蓝型油菜PMEI和XTH基因家族的鉴定与转录谱分析 [D]. 武汉: 华中农业大学, 2021.
	Chu LY. Identification and transcription analysis of PMEI and XTH gene families in Brassica napus L [D]. Wuhan: Huazhong Agricultural University, 2021.
[19]	Bosch M, Hepler PK. Pectin methylesterases and pectin dynamics in pollen tubes [J]. Plant Cell, 2005, 17(12): 3219-3226.
[20]	盖芸芸, 何婵, 李娜, 等. 超声波预处理对贮藏过程中猕猴桃质构、果胶特性及果胶酶活性的影响 [J]. 食品科技, 2024, 49(5): 26-34.
	Gai YY, He C, Li N, et al. Effects of ultrasonic pretreatment on texture, pectin properties, and pectinase activity of kiwifruit during storage [J]. Food Sci Technol, 2024, 49(5): 26-34.
[21]	Wachsman G, Zhang JY, Moreno-Risueno MA, et al. Cell wall remodeling and vesicle trafficking mediate the root clock in Arabidopsis [J]. Science, 2020, 370(6518): 819-823.
[22]	Zhang GY, Feng J, Wu J, et al. BoPMEI1, a pollen-specific pectin methylesterase inhibitor, has an essential role in pollen tube growth [J]. Planta, 2010, 231(6): 1323-1334.
[23]	Coculo D, Lionetti V. The plant invertase/pectin methylesterase inhibitor superfamily [J]. Front Plant Sci, 2022, 13: 863892.
[24]	Nguyen HP, Jeong HY, Jeon SH, et al. Rice pectin methylesterase inhibitor28 (OsPMEI28) encodes a functional PMEI and its overexpression results in a dwarf phenotype through increased pectin methylesterification levels [J]. J Plant Physiol, 2017, 208: 17-25.
[25]	Müller K, Levesque-Tremblay G, Fernandes A, et al. Overexpression of a pectin methylesterase inhibitor in Arabidopsis thaliana leads to altered growth morphology of the stem and defective organ separation [J]. Plant Signal Behav, 2013, 8(12): e26464.
[26]	Peaucelle A, Louvet R, Johansen JN, et al. Arabidopsis phyllotaxis is controlled by the methyl-esterification status of cell-wall pectins [J]. Curr Biol, 2008, 18(24): 1943-1948.
[27]	An SH, Sohn KH, Choi HW, et al. Pepper pectin methylesterase inhibitor protein CaPMEI1 is required for antifungal activity, basal disease resistance and abiotic stress tolerance [J]. Planta, 2008, 228(1): 61-78.
[28]	Tan C, Liu ZY, Huang SN, et al. Pectin methylesterase inhibitor (PMEI) family can be related to male sterility in Chinese cabbage (Brassica rapa ssp. pekinensis) [J]. Mol Genet Genomics, 2018, 293(2): 343-357.
[29]	Vierstra RD. The ubiquitin-26S proteasome system at the nexus of plant biology [J]. Nat Rev Mol Cell Biol, 2009, 10(6): 385-397.
[30]	Zhang JZ. Evolution by gene duplication: an update [J]. Trends Ecol Evol, 2003, 18(6): 292-298.
[31]	Jeong HY, Nguyen HP, Eom SH, et al. Integrative analysis of pectin methylesterase (PME) and PME inhibitors in tomato (Solanum lycopersicum): Identification, tissue-specific expression, and biochemical characterization [J]. Plant Physiol Biochem, 2018, 132: 557-565.
[32]	廉雪菲. 柑橘果实囊衣发育形成规律研究 [D]. 长沙: 湖南农业大学, 2022.
	Lian XF. Study on the development and formation law of citrus fruit capsule [D]. Changsha: Hunan Agricultural University, 2022.
[33]	Hong MJ, Kim DY, Lee TG, et al. Functional characterization of pectin methylesterase inhibitor (PMEI) in wheat [J]. Genes Genet Syst, 2010, 85(2): 97-106.
[34]	陈逸飞, 王文博, 王赛赛, 祝建波. 甜瓜PMEI基因家族的鉴定和表达分析 [J/OL]. 分子植物育种, 2023. .
	Chen YF, Wang WB, Wang SS, Zhu JB. Identification and expression analysis of PMEI gene family in melon [J/OL]. Mol Plant Breed, 2023. .
[35]	杜晓芬, 王智兰, 韩康妮, 等. 谷子叶绿体基因RNA编辑位点的鉴定与分析 [J]. 作物学报, 2022, 48(4): 873-885.

基因名称 Gene name	基因序列 Gene sequence （5′‒3′）
25S-F	AGGCAACAGAAACTCCATACG
25S-R	ATGGCATAGCATTCATCACG
SiPMEI22-F	TGTTCTCGGGACAGGTTTCG
SiPMEI22-R	GTACAGCGTGTCCTGGTACG
SiPMEI30-F	GACCTCGGCTACGACTACTG
SiPMEI30-R	GGTTGTGCTCTTGGAGGCTA
SiPMEI31-F	TCTTCAGAGAGAGGGGGATCG
SiPMEI31-R	TAGCGCGACAAGCGAACAAT
SiPMEI32-F	TTTTCCGCGAACGTGTCCA
SiPMEI32-R	ACACCACCCCATATCGCTTG
SiPMEI36-F	CTCGGTACAGTCTTGCACCA
SiPMEI36-R	ATCCAGTGCAATGTCTCCGT
SiPMEI38-F	CCGAGTACATGAACCGTGGA
SiPMEI38-R	GTGGAGTTGAGCCAGAGGTC
SiPMEI47-F	CAGCCTTCGTGTTCCCTGTT
SiPMEI47-R	TCTTATTGATGACCGCCCTCG
SiPMEI63-F	AGCTTCGTTCGCTCATGCAA
SiPMEI63-R	AGTAATGAAGGCTGCAACACAC

基因名称 Gene name	基因序列 Gene sequence （5′‒3′）
25S-F	AGGCAACAGAAACTCCATACG
25S-R	ATGGCATAGCATTCATCACG
SiPMEI22-F	TGTTCTCGGGACAGGTTTCG
SiPMEI22-R	GTACAGCGTGTCCTGGTACG
SiPMEI30-F	GACCTCGGCTACGACTACTG
SiPMEI30-R	GGTTGTGCTCTTGGAGGCTA
SiPMEI31-F	TCTTCAGAGAGAGGGGGATCG
SiPMEI31-R	TAGCGCGACAAGCGAACAAT
SiPMEI32-F	TTTTCCGCGAACGTGTCCA
SiPMEI32-R	ACACCACCCCATATCGCTTG
SiPMEI36-F	CTCGGTACAGTCTTGCACCA
SiPMEI36-R	ATCCAGTGCAATGTCTCCGT
SiPMEI38-F	CCGAGTACATGAACCGTGGA
SiPMEI38-R	GTGGAGTTGAGCCAGAGGTC
SiPMEI47-F	CAGCCTTCGTGTTCCCTGTT
SiPMEI47-R	TCTTATTGATGACCGCCCTCG
SiPMEI63-F	AGCTTCGTTCGCTCATGCAA
SiPMEI63-R	AGTAATGAAGGCTGCAACACAC

重复基因1 Duplicated gene l	重复基因2 Duplicated gene 2	非同义替换率 Ka	同义替换率 Ks	比值 Ka/Ks
SiPMEI46	SiPMEI41	0.586 044 760	0.756 527 598	0.774 650 867
SiPMEI29	SiPMEI19	0.310 390 814	0.409 962 679	0.757 119 686
SiPMEI46	SiPMEI27	0.484 690 326	0.680 501 629	0.712 254 469
SiPMEI32	SiPMEI30	0.404 813 285	0.573 874 330	0.705 404 065
SiPMEI26	SiPMEI27	0.591 358 520	0.538 368 419	1.098 427 208
SiPMEI6	SiPMEI23	0.202 922 616	0.397 601 411	0.510 366 941
SiPMEI5	SiPMEI34	0.660 998 002	0.498 090 476	1.327 064 124
SiPMEI63	SiPMEI50	0.617 173 087	0.439 225 359	1.405 139 924

重复基因1 Duplicated gene l	重复基因2 Duplicated gene 2	非同义替换率 Ka	同义替换率 Ks	比值 Ka/Ks
SiPMEI46	SiPMEI41	0.586 044 760	0.756 527 598	0.774 650 867
SiPMEI29	SiPMEI19	0.310 390 814	0.409 962 679	0.757 119 686
SiPMEI46	SiPMEI27	0.484 690 326	0.680 501 629	0.712 254 469
SiPMEI32	SiPMEI30	0.404 813 285	0.573 874 330	0.705 404 065
SiPMEI26	SiPMEI27	0.591 358 520	0.538 368 419	1.098 427 208
SiPMEI6	SiPMEI23	0.202 922 616	0.397 601 411	0.510 366 941
SiPMEI5	SiPMEI34	0.660 998 002	0.498 090 476	1.327 064 124
SiPMEI63	SiPMEI50	0.617 173 087	0.439 225 359	1.405 139 924