Functional Analysis of Fusarium wilt Resistance Gene GhERF14 in Cotton

doi:10.7668/hbnxb.20195360

Abstract

Abstract:

This study explored the relationship between cotton GhERF14 gene and Fusarium oxysporum pathogenicity,analyzed the molecular mechanism of F.oxysporum pathogenicity,and tentatively explored the response of cotton GhERF14 gene to Fusarium wilt disease and its regulatory effect on related resistance genes,to provides some theoretical basis for breeding new cotton cultivars resistant to wilt.Gene cloning and virus-induced gene silencing(VIGS)were used to construct the non-conserved domain interference vector pTRV2-GhERF14.Using Real-time fluorescence quantification(qRT-PCR)technology and VIGS technology,the expression characteristics of GhERF14 and downstream genes related to lignin,ethylene(ET),jasmonic acid(JA),salicylic acid(SA),antioxidant enzymes and disease progression-related protein(PR)were analyzed after F.oxysporum stress and hormone treatment,and the role of GhERF14 in the process of cotton disease resistance was analyzed.The results indicated that inhibition of GhERF14 gene expression could significantly reduce the synthesis of jasmonic acid(JA),salicylic acid(SA)and ethylene(ET)and the expression of genes related to the signaling pathway.After GhERF14 gene silencing by VIGS technology,cotton plants were more susceptible to Fusarium wilt.These results suggested that GhERF14 may play an important role in the pathogenesis and host-pathogen interaction of F.oxysporum.

Key words: Cotton, GhERF14, Blight, VIGS, Fusarium oxysporum

CLC Number:

ZHAO Longfei, ZHANG Jianhua, ZHAO Zengqiang, CAI Shudong, ZHANG Wei. Functional Analysis of Fusarium wilt Resistance Gene GhERF14 in Cotton[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(3): 133-140. doi: 10.7668/hbnxb.20195360.

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References 34

[1]	吴征彬, 杨业华, 刘小丰, 王强. 枯萎病对棉花产量和纤维品质的影响[J]. 棉花学报, 2004, 16(4):236-239.doi:10.3969/j.issn.1002-7807.2004.04.009.
	Wu Z B, Yang Y H, Liu X F, Wang Q. Effect of Fusarium wilt on the cotton yield and fiber quality[J]. Acta Gossypii Sinica, 2004, 16(4):236-239.
[2]	Li F G, Fan G Y, Lu C R, et al. Genome sequence of cultivated Upland cotton(Gossypium hirsutum TM-1)provides insights into genome evolution[J]. Nature Biotechnology, 2015, 33(5):524-530.doi:10.1038/nbt.3208.
[3]	Zhang T Z, Hu Y, Jiang W K, et al. Sequencing of allotetraploid cotton(Gossypium hirsutum L.acc.TM-1)provides a resource for fiber improvement[J]. Nature Biotechnology, 2015, 33(5):531-537.doi:10.1038/nbt.3207.
[4]	鞠林芳, 段尚尧, 张妍, 哈斯阿古拉, 牛一丁. 甜瓜乙烯应答因子基因CmERF Ⅰ-14的生物信息学分析[J]. 基因组学与应用生物学, 2016, 35(9):2467-2474.doi:10.13417/j.gab.035.002467.
	Ju L F, Duan S Y, Zhang Y, Hasi A, Niu Y D. Bioinformatics analysis of melon ethylene response factor gene CmERF Ⅰ-14[J]. Genomics and Applied Biology, 2016, 35(9):2467-2474.
[5]	Ohme-Takagi M, Shinshi H. Ethylene-inducible DNA binding proteins that interact with an ethylene-responsive element[J]. The Plant Cell, 1995, 7(2):173-182.doi:10.1105/tpc.7.2.173.
[6]	董娜, 张增艳, 辛志勇. 病原诱导的小麦ERF转录因子TaERF1b的原核表达及纯化[J]. 植物遗传资源学报, 2008, 9(3):283-287.doi:10.13430/j.cnki.jpgr.2008.03.003.
	Dong N, Zhang Z Y, Xin Z Y. Expression and purification of pathogen-induced wheat ERF transcription factor TaERF1b in Escherichia coli[J]. Journal of Plant Genetic Resources, 2008, 9(3):283-287.
[7]	庄静, 彭日荷, 高峰, 付晓燕, 朱波, 金晓芬, 熊爱生, 姚泉洪. 沪油15中两个AP2/ERF-B1亚族转录因子的克隆与分析[J]. 核农学报, 2009, 23(3):435-441,453. doi: 10.11869/hnxb.2009.03.0435
	Zhuang J, Peng R H, Gao F, Fu X Y, Zhu B, Jin X F, Xiong A S, Yao Q H. Cloning and analysis of Ap2/Erf-B1 subfamily transcription factors from Brassica napus L.Huyou 15[J]. Journal of Nuclear Agricultural Sciences, 2009, 23(3):435-441,453.
[8]	郭文婷, 张鹏飞, 李潇玲, 张析, 张薇. 棉花ERF-B3亚组转录因子基因GhERF5-1的克隆及表达[J]. 江苏农业科学, 2018, 46(19):20-24.doi:10.15889/j.issn.1002-1302.2018.19.005.
	Guo W T, Zhang P F, Li X L, Zhang X, Zhang W. Cloning and expression of cotton ERF-B3 subgroup transcription factor gene GhERF5-1[J]. Jiangsu Agricultural Sciences, 2018, 46(19):20-24.
[9]	Huang P Y, Catinot J, Zimmerli L. Ethylene response factors in Arabidopsis immunity[J]. Journal of Experimental Botany, 2016, 67(5):1231-1241.doi:10.1093/jxb/erv518.
[10]	Wang F J, Wang C L, Liu P Q, Lei C L, Hao W, Gao Y, Liu Y G, Zhao K J. Enhanced rice blast resistance by CRISPR/Cas9-targeted mutagenesis of the ERF transcription factor gene OsERF922[J]. PLoS One, 2016, 11(4):e0154027.doi:10.1371/journal.pone.0154027.
[11]	Fujimoto S Y, Ohta M, Usui A, Shinshi H, Ohme-Takagi M. Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box-mediated gene expression[J]. The Plant Cell, 2000, 12(3):393-404.doi:10.1105/tpc.12.3.393.
[12]	金莉. 棉花GbERF1基因的克隆与功能鉴定[D]. 武汉: 华中农业大学, 2012.doi:10.7666/d.Y2162848.
	Jin L. Isolation and characterization identification of GbERF1 in cotton[D]. Wuhan: Huazhong Agricultural University, 2012.
[13]	Anderson J P, Lichtenzveig J, Gleason C, Oliver R P, Singh K B. The B-3 ethylene response factor MtERF1-1 mediates resistance to a subset of root pathogens in Medicago truncatula without adversely affecting symbiosis with rhizobia[J]. Plant Physiology, 2010, 154(2):861-873.doi:10.1104/pp.110.163949.
[14]	Oñate-Sánchez L, Anderson J P, Young J, Singh K B. AtERF14,a member of the ERF family of transcription factors,plays a nonredundant role in plant defense[J]. Plant Physiology, 2007, 143(1):400-409.doi:10.1104/pp.106.086637. pmid: 17114278
[15]	Champion A, Hebrard E, Parra B, Bournaud C, Marmey P, Tranchant C, Nicole M. Molecular diversity and gene expression of cotton ERF transcription factors reveal that group IXa members are responsive to jasmonate, ethylene and Xanthomonas[J]. Molecular Plant Pathology, 2009, 10(4):471-485.doi:10.1111/j.1364-3703.2009.00549.x.
[16]	Qin J, Zuo K J, Zhao J Y, Ling H, Cao Y F, Qiu C X, Li F P, Sun X F, Tang K X. Overexpression of GbERF confers alteration of ethylene-responsive gene expression and enhanced resistance to Pseudomonas syringae in transgenic tobacco[J]. Journal of Biosciences, 2006, 31(2):255-263.doi:10.1007/BF02703918.
[17]	Zuo K J, Qin J, Zhao J Y, Ling H, Zhang L D, Cao Y F, Tang K X. Over-expression GbERF2 transcription factor in tobacco enhances brown spots disease resistance by activating expression of downstream genes[J]. Gene, 2007, 391(1/2):80-90.doi:10.1016/j.gene.2006.12.019.
[18]	Jia J H, Liang Y F, Gou T Y, Hu Y H, Zhu Y X, Huo H Q, Guo J, Gong H J. The expression response of plasma membrane aquaporins to salt stress in tomato plants[J]. Environmental and Experimental Botany, 2020, 178:104190.doi:10.1016/j.envexpbot.2020.104190.
[19]	Liu D F, Chen X J, Liu J Q, Ye J C, Guo Z J. The rice ERF transcription factor OsERF922 negatively regulates resistance to Magnaporthe oryzae and salt tolerance[J]. Journal of Experimental Botany, 2012, 63(10):3899-3911.doi:10.1093/jxb/ers079.
[20]	Yang C L, Liang S, Wang H Y, Han L B, Wang F X, Cheng H Q, Wu X M, Qu Z L, Wu J H, Xia G X. Cotton major latex protein 28 functions as a positive regulator of the ethylene responsive factor 6 in defense against Verticillium dahliae[J]. Molecular Plant, 2015, 8(3):399-411.doi:10.1016/j.molp.2014.11.023.
[21]	李继东, 倪静, 叶霞, 陈鹏, 谭彬, 张梦洋, 郑先波, 张玉, 冯建灿. 枣AP2/ERF转录因子鉴定及其响应枣疯病植原体表达分析[J]. 园艺学报, 2020, 47(8):1463-1474.doi:10.16420/j.issn.0513-353x.2019-0965.
	Li J D, Ni J, Ye X, Chen P, Tan B, Zhang M Y, Zheng X B, Zhang Y, Feng J C. Genomic identification of jujube AP2/ERF transcription factors and their expression pattern during jujube witches' broom pathogenesis process[J]. Acta Horticulturae Sinica, 2020, 47(8):1463-1474.
[22]	张莉. 苹果抗白粉病转录因子MdERF100克隆与功能研究[D]. 杨凌: 西北农林科技大学, 2020.doi:10.27409/d.cnki.gxbnu.2020.000980.
	Zhang L. Cloning and function analysis of MdERF100 transcription factor in apple[D]. Yangling: Northwest A&F University, 2020..
[23]	张曦, 钱玉源, 王广恩, 刘祎, 权月伟, 崔淑芳, 米换房, 李俊兰. 棉花色素腺体发育相关基因GhERF105-like的克隆与表达分析[J]. 华北农学报, 2022, 37(6):34-41.doi:10.7668/hbnxb.20193130.
	Zhang X, Qian Y Y, Wang G E, Liu Y, Quan Y W, Cui S F, Mi H F, Li J L. Cloning and expression analysis of a cotton pigment gland formation related gene GhERF105-like[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(6):34-41.
[24]	李文正, 张海文, 王俊英, 黄荣峰. ERF转录因子及其在烟草抗逆性改良中的应用[J]. 生物技术通报, 2006, 22(4):30-34.doi:10.3969/j.issn.1002-5464.2006.04.007.
	Li W Z, Zhang H W, Wang J Y, Huang R F. Ethylene responsive factors and their application in improving tobacco tolerance to biotic and abiotic stresses[J]. Biotechnology Bulletin, 2006, 22(4):30-34.
[25]	Glazebrook J. Genes controlling expression of defense responses in Arabidopsis[J]. Current Opinion in Plant Biology, 1999, 2(4):280-286.doi:10.1016/s1369-5266(99)80050-8. pmid: 10458996
[26]	赵龙飞, 明聪, 张中荣, 袁玥, 刘戈辉, 张薇. 棉花AP2/ERF-B3亚组转录因子基因GhERF14的克隆与表达分析[J]. 分子植物育种, 2022, 20(9):2804-2811.doi:10.13271/j.mpb.020.002804.
	Zhao L F, Ming C, Zhang Z R, Yuan Y, Liu G H, Zhang W. Cloning and expression analysis of a transcription factor gene GhERF14 from cotton AP2/ERF-B3 subgroup[J]. Molecular Plant Breeding, 2022, 20(9):2804-2811.
[27]	Yang H H, Sun Y G, Wang H X, Zhao T T, Xu X Y, Jiang J B, Li J F. Genome-wide identification and functional analysis of the ERF2 gene family in response to disease resistance against Stemphylium lycopersici in tomato[J]. BMC Plant Biology, 2021, 21(1):72.doi:10.1186/s12870-021-02848-3.
[28]	Wang G N, Wang X F, Zhang Y, Yang J, Li Z K, Wu L Z, Wu J H, Wu N, Liu L X, Liu Z W, Zhang M, Wu L Q, Zhang G Y, Ma Z Y. Dynamic characteristics and functional analysis provide new insights into long non-coding RNA responsive to Verticillium dahliae infection in Gossypium hirsutum[J]. BMC Plant Biology, 2021, 21(1):68.doi:10.1186/s12870-021-02835-8.
[29]	Wang Y Q, Umer M J, Cai X Y, Yang M Y, Hou Y Q, Xu Y C, Batool R, Mehari T G, Zheng J, Wang Y H, Wang H, Li Z K, Zhou Z L, Liu F. Dynamic characteristics and functional analysis provide new insights into the role of GauERF105 for resistance against Verticillium dahliae in cotton[J]. BMC Plant Biology, 2023, 23(1):501.doi:10.1186/s12870-023-04455-w.
[30]	Wang P P, Zhao Y P, Wu N, Azhar M T, Hou Y X, Shang H H. GhERF.B4-15D:a member of ERF subfamily B4 group positively regulates the resistance against Verticillium dahliae in upland cotton[J]. Biomolecules, 2023, 13(9):1348.doi:10.3390/biom13091348.
[31]	张启燕. 千年桐(Vernicia montana)对枯萎病应激反应及抗病基因功能鉴定[D]. 北京: 中国林业科学研究院, 2019.doi:10.27625/d.cnki.gzlky.2019.000028.
	Zhang Q Y. Response rules and novel potential resistance factors to Fusarium wilt disease in Vernicia montana[D]. Beijing: Chinese Academy of Forestry, 2019.
[32]	郭伟锋. 海岛棉乙烯响应因子GbERF1-like在抗黄萎病反应中的功能解析[D]. 武汉: 华中农业大学, 2016.doi:10.7666/d.Y3053005.
	Guo W F. Functional analysis of ethylene response factor GbERF1-like in island cotton against Verticillium wilt[D]. Wuhan: Huazhong Agricultural University, 2016.
[33]	杨德鑫. 水稻ERF蛋白OsERF109的特性及功能分析[D]. 北京: 中国农业科学院, 2012.
	Yang D X. The characterization and function analysis of rice ERF protein OsERF109[D]. Beijing: Chinese Academy of Agricultural Sciences, 2012.
[34]	Zhu Z G, Shi J L, Xu W R, Li H E, He M Y, Xu Y, Xu T F, Yang Y Z, Cao J L, Wang Y J. Three ERF transcription factors from Chinese wild grapevine Vitis pseudoreticulata participate in different biotic and abiotic stress-responsive pathways[J]. Journal of Plant Physiology, 2013, 170(10):923-933.doi:10.1016/j.jplph.2013.01.017.

基因名称 Gene name	正向引物序列(5'—3') Forward primer sequence(5'—3')	反向引物序列(5'—3') Reverse primer sequence(5'—3')
GhPR1	TTTAGTAAGGTTTTTGACCGACGAA	GACTCTGTCCATCTTGGTTGTGCTA
GhPR2	TTCAAGGTTTGCACTCGGAAGA	CCACCAGCAGCAGAAGTTATCG
GhPR3	GATGACTCCACAATCACCGAAGC	GCGGTCTTCTACCTGGGCATT)
GhPR5	TTGGCTCTTACTTCCGACCATCT	GCCGTGATTCATACAGTTATCCTCA
GhICS1	AAGAATGCCAGAGGTAAGAGGAGGA	ATGGATGAATGGGTGCGAAGG
GhEDS1	GCAGCAACAGCTCCTCTACCTCAA	GGCAGACCAAGACGCTACAGATACA
GhNPR1	CACGTGGTGCTGTTGTTGTTACTG	GCGAATCGTTGCTTTCTTCTTCA
GhPAD4	GGATGGAAGAATGGAAAGAAATGAA	GAACTAGGAAAGCAGACTAAGGAACCA
GhWRKY70	TGGCAACATACGAAGGACAACACA	CTTCGCTCGTCCCGGAAAATATCGC
GhPDF1.2	AAAATCGACTGAAATGGCTA	TGTCGGTTACAAAGTAAATTGC
GhJAZ1	CTTTGGTTCCACTGCTGCTGATT	GACCAAATCTTGTGGCATCTACCTC
GhAOS	CGATGTCCCAACTTCAATCTCAAAC	ATCCGACGGTGGAGAATAAACAGT
GhAOC4	CAAAAATGCCAGACCCACCAGTA	CAAAAATGCCAGACCCACCAGTA
GhEIN2	CCCGAGCTGTGGGGAAAATACACT	GGGATTTGAAGGCAGAAGCAGGAT
GhEIN3	CCACCAATCTTCGTTCCCGGTA	AAACGGAAACCCATGTAGCAG
GhACO	TGACCTCAAGTTCCAAGCCAA	AGAATTTATCCATCAGCCGACT
GhPAL	TGGTGGCTGAGTTTAGGAAA	TGAGTGAGGCAATGTGTGA
GhC4H1	CCGAACCCGACACCCATAAGC	GCAGGGATGTCATACCCACCAAG
GhERF1	CTTTAGATCACAACTCGCTTC	TTTTCTTTCGTCGGGCTT
GhF5H2	GGCAGATGCGAAAGATTTGTGTA	ACATCCATCATTTCTTGCTGCTC
Gh4CL	ATTCAAAAGGGAGATGCC	GAGAAGGGCAAAGCAACA
GhCCR1	GCACTGTAAGGAACCCAGACGAT	CATTTGTTCAGGATCATCAGTCACA
GhCCoAOMT	TACGACAACACGCTGTGGAATGGG	CATCGCCGACAGGAAACATACAGA
GhSOD	CATCTCACGCACTCTGTC	CCTTAGCCATTTCTGTCTGTG
GhPOD	ATATCCTTGTTCTGTCTGCTA	CTCCTTCTACCGTCTCTTC
GhCAT	TGATAAGTTGCTCCAGACTCG	CTTCGTGGTGATTGTTGTGA
GhUBQ7	GAAGACCTACACCAAGCCCAA	CGGACTCTACTCAATCCCCACC
qPCRERF14	GGAGTATCCAATGGGTAGCGGC	CCATCGACGACGACGAAGAAGA

基因名称 Gene name	正向引物序列(5'—3') Forward primer sequence(5'—3')	反向引物序列(5'—3') Reverse primer sequence(5'—3')
GhPR1	TTTAGTAAGGTTTTTGACCGACGAA	GACTCTGTCCATCTTGGTTGTGCTA
GhPR2	TTCAAGGTTTGCACTCGGAAGA	CCACCAGCAGCAGAAGTTATCG
GhPR3	GATGACTCCACAATCACCGAAGC	GCGGTCTTCTACCTGGGCATT)
GhPR5	TTGGCTCTTACTTCCGACCATCT	GCCGTGATTCATACAGTTATCCTCA
GhICS1	AAGAATGCCAGAGGTAAGAGGAGGA	ATGGATGAATGGGTGCGAAGG
GhEDS1	GCAGCAACAGCTCCTCTACCTCAA	GGCAGACCAAGACGCTACAGATACA
GhNPR1	CACGTGGTGCTGTTGTTGTTACTG	GCGAATCGTTGCTTTCTTCTTCA
GhPAD4	GGATGGAAGAATGGAAAGAAATGAA	GAACTAGGAAAGCAGACTAAGGAACCA
GhWRKY70	TGGCAACATACGAAGGACAACACA	CTTCGCTCGTCCCGGAAAATATCGC
GhPDF1.2	AAAATCGACTGAAATGGCTA	TGTCGGTTACAAAGTAAATTGC
GhJAZ1	CTTTGGTTCCACTGCTGCTGATT	GACCAAATCTTGTGGCATCTACCTC
GhAOS	CGATGTCCCAACTTCAATCTCAAAC	ATCCGACGGTGGAGAATAAACAGT
GhAOC4	CAAAAATGCCAGACCCACCAGTA	CAAAAATGCCAGACCCACCAGTA
GhEIN2	CCCGAGCTGTGGGGAAAATACACT	GGGATTTGAAGGCAGAAGCAGGAT
GhEIN3	CCACCAATCTTCGTTCCCGGTA	AAACGGAAACCCATGTAGCAG
GhACO	TGACCTCAAGTTCCAAGCCAA	AGAATTTATCCATCAGCCGACT
GhPAL	TGGTGGCTGAGTTTAGGAAA	TGAGTGAGGCAATGTGTGA
GhC4H1	CCGAACCCGACACCCATAAGC	GCAGGGATGTCATACCCACCAAG
GhERF1	CTTTAGATCACAACTCGCTTC	TTTTCTTTCGTCGGGCTT
GhF5H2	GGCAGATGCGAAAGATTTGTGTA	ACATCCATCATTTCTTGCTGCTC
Gh4CL	ATTCAAAAGGGAGATGCC	GAGAAGGGCAAAGCAACA
GhCCR1	GCACTGTAAGGAACCCAGACGAT	CATTTGTTCAGGATCATCAGTCACA
GhCCoAOMT	TACGACAACACGCTGTGGAATGGG	CATCGCCGACAGGAAACATACAGA
GhSOD	CATCTCACGCACTCTGTC	CCTTAGCCATTTCTGTCTGTG
GhPOD	ATATCCTTGTTCTGTCTGCTA	CTCCTTCTACCGTCTCTTC
GhCAT	TGATAAGTTGCTCCAGACTCG	CTTCGTGGTGATTGTTGTGA
GhUBQ7	GAAGACCTACACCAAGCCCAA	CGGACTCTACTCAATCCCCACC
qPCRERF14	GGAGTATCCAATGGGTAGCGGC	CCATCGACGACGACGAAGAAGA

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