Genome-wide Identification of MaMKK Family in Banana (Musa spp.)and Their Expression Analysis in Response to Low Temperature Stress
LIN Wei1, WEI Mi2, LI Yue-Sen1, WU Shui-Jin1*
1 Subtropical Agriculture Research Institute, Fujian Academy of Agricultural Sciences, Zhangzhou 363005, China; 2 Zhangzhou Institute of Agricultural Sciences, Zhangzhou 363005, China
Abstract：The mitogen-activated protein kinase (MAPK) cascade is crucial to plant growth, development, and stress response, with MAPK kinase (MKK) serving a critical intermediary role in the cascade. Low temperature is one of the main reasons affecting banana (Musa spp.) yield. To explore the key genes for cold resistance in bananas, systematic evolutionary, gene structure assessment, chromosome localization andcollinearity analysis were conducted on the MaMKK gene family, clarifying the expression of the MaMKKgene family induced by low temperature in the 'Cavendish' (Musa AAA Cavendish) and 'Fenjiao' (MusaABB Pisang Awak). Subsequent analyses included sequence alignment and promoter element identification of the candidate genes. The result showed that A total of 13 members of the MaMKK gene family were identified and divided into 4 subclasses. Collinearity analysis showed that the members of the MaMKK gene family homologous had a strong connection, which had been purified and selected. After exposure to 7 ℃ , theexpression levels of MaMKK2a, MaMKK2b, and MaMKK5a significantly increased in both varieties, with agreater upregulation in 'Fenjiao' than in 'Cavendish'. However, the expression levels of MaMKK1, MaMKK2c, MaMKK3, and MaMKK5d showed a negative or unrelated relationship in the 2 varieties. Further analysis of these 7 candidate genes showed that MaMKK2a, MaMKK2b, and MaMKK5a responded more rapidly to low temperatures in 'Cavendish' than in 'Fenjiao'. The expression trends of MaMKK1, MaMKK2c, MaMKK3 and MaMKK5b showed significant differences within 48 h of low temperature treatment in 'Cavendish'/'Fenjiao'.The tissue expression results showed that there were significant differences in the tissue distribution of the 7 candidate genes in 'Cavendish'/'Fenjiao'. Analysis of the functional domains of 7 candidate genes revealed that they all contained phosphorylation targets for MEKK and MAPK, as well as multiple stress-related promotercis-acting elements. Among them, MaMKK2a and MaMKK5a contained low-temperature promoter cis-actingelements. The results indicated that the MaMKK gene in banana responded to low temperature stress, and the7 candidate genes screened may be related to the differences in cold resistance between 'Cavendish' and'Fenjiao'. This study provides a theoretical basis for developing cold-resistant banana varieties.
林蔚, 韦幂, 李跃森, 吴水金. 香蕉MaMKK家族全基因组鉴定及其低温胁迫响应表达分析[J]. 农业生物技术学报, 2025, 33(6): 1220-1233.
LIN Wei, WEI Mi, LI Yue-Sen, WU Shui-Jin. Genome-wide Identification of MaMKK Family in Banana (Musa spp.)and Their Expression Analysis in Response to Low Temperature Stress. 农业生物技术学报, 2025, 33(6): 1220-1233.
[1] 高洁.2017. 大蕉与香牙蕉抗寒性差异的分子机理解析[D]. 博士学位论文, 华南农业大学, 导师: 易干军, pp. 47-61. (Gao J.2017. Analysis of molecular mechanism of the different cold tolerance between Dajiao and Caven-dish banana[D]. Thesis for Ph. D., South China Agricul-tural University, Supervisor: Yi G J, pp. 47-61.) [2] 谷思, 刘璐, 李安然, 等. 2021. 草莓果实酵母双杂交文库的构建及 FvM4K1 互作蛋白的筛选[J]. 园艺学报, 48(6): 1067-1078. (Gu S, Liu L, Li A R, et al.2021. Construc-tion of yeast two hybrid library and screening of FvM4K1 interacting protein in the fruit of Fragraria ves-ca[J]. Acta Horticulturae Sinica, 48(6): 1067-1078.) [3] 郝丽叶, 路琦, 阿力木•阿木提, 等. 2024. 低温胁迫对扁桃优株生理特性的影响[J]. 山西农业科学 , 52(1): 68-78. (Hao L Y, Lu Q, Amti-Alimu, et al.2024. Effect of low temperature on physiological characteristics of almond strains[J]. Journal of Shanxi Agricultural Sciences, 52(1): 68-78.) [4] 李盼盼, 江松霖, 薛宁涵, 等. 2024. 自然低温下红茴香和双蕊野扇花的耐寒性研究[J]. 江西农业学报, 36(09): 52-59. (Li P P, Jiang S L, Xue L H, et al.2024. Cold toler-ance of illicium henryi diels and Sarcococca hookeriana var. digyna Franch at natural low temperature[J]. Acta Agriculturae Jiangxi, 36(9): 52-59.) [5] 李倩, 沈春生, 林启昉, 等. 2021. 采后香蕉果实冷害发生与控制技术研究进展[J]. 果树学报, 38(5): 817-827. (Li Q, Shen C S, Lin Q F, et al.2021. Advances in research on thechilling injury occurrence and control technolo-gies of postharvest banana fruit[J]. Journal of Fruit Sci-ence, 38(5): 817-827.) [6] 龙琴, 谢宇, 雷天刚, 等. 2020. 柑橘 MAPK 基因的生物信息及其响应溃疡病菌侵染的表达分析[J]. 园艺学报, 47(11): 2095-2106. (Long Q, Xie Y, Lei T G, et al.2020. Bioinformatics analysis of MAPK genes in Citrus and their expression in response to canker disease[J]. Acta Horticulturae Sinica, 47(11): 2095-2106.) [7] 王安邦, 金志强, 刘菊华, 等. 2014. 香蕉寒害研究现状及展望[J]. 生物技术通报,(8): 28-33. (Wang A B, Jin Z Q, Liu J H, et al. 2014. The current situation and prospects of banana chilling stress[J]. Biotechnology Bulletin,(8): 28-33.) [8] 徐慧妮, 王秀峰, 孙旭东, 等. 2008. 黄瓜促分裂原活化蛋白激酶基因的生物信息学分析及原核表达[J]. 园艺学报 , 35(07): 1017-1022. (Xu H N, Wang X F, Sun X D, et al.2008. Bioinformatics analysis and prokaryotic ex-pression of MAPK in cucumber[J]. Acta Horcitulturae Sinica, 35: 1017-1022.) [9] 徐子怡, 罗晨宇, 占坤, 等. 2024. 猕猴桃 OSCA 基因家族鉴定及其在非生物胁迫下的表达分析[J]. 果树学报, 41(3): 436-447. (Xu Z Y, Luo C Y, Zhan K, et al.2024. Ge-nome-wide identification of OSCA gene family mem-bers and their expression under different abiotic stresses in kiwifruit[J]. Journal of Fruit Science, 41(3): 436-447.) [10] Barker M S, Baute G J, Liu S L.2012. Duplications and turn-over in plant genomes[J]. Plant Genome Diversity, 1:155-169. [11] Chen C J, Chen H, Zhang Y, et al.2020. TBtools: An integra-tive toolkit developed for interactive analyses of big bio-logical data[J]. Molecular Plant, 13(8): 1194-1202. [12] Chen L, Sun H, Wang F J, et al.2020. Genome-wide identifi-cation of MAPK cascade genes reveals the Gh-MAP3K14-GhMKK11-GhMPK31 pathway is involved in the drought response in cotton[J]. Plant Molecular Bi-ology, 103(9): 211-223. [13] Colcombet J, Hirt H.2008. Arabidopsis MAPKs: A complex signaling network involved in multiple biological pro-cesses[J]. Biochemical Journal, 413: 217-226. [14] Dhindsa R S, Plumb P D, Thorpe T A.1981. Leaf Senescence: Correlated with increased levels of membrane permea-bility and lipid peroxidation, and decreased levels of su-peroxide dismutase and catalase[J]. Journal of Experi-mental Botany, 32(1): 93-101. [15] Dutilleul C, Benhassaine K G, Demandre Z A, et al.2012.Phytosphingosine-phosphate is a signal for AtMPK6 ac-tivation and Arabidopsis response to chilling[J]. New Phytologist, 194: 181-191. [16] Gao J, Dou T X, He W D, et al.2021. MaMAPK3-MaICE1-MaPOD P7 pathway, a positive regulator of cold toler-ance in banana[J]. BMC Plant Biology, 21(1): 97. [17] He W D, Gao J, Dou T X, et al.2018. Early cold-induced per-oxidases and aquaporins are associated with high cold tolerance in Dajiao(Musa spp. 'Dajiao')[J]. Frontiers in Plant Science,9: 282. [18] Li H, Ding Y, Shi Y, et al.2017. MPK3-and MPK6-mediated ICE1 phosphorylation negatively regulates ICE1 stabili-ty and freezing tolerance in Arabidopsis[J]. Developmen-tal Cell, 43(5): 630-642. [19] Liu Z N, Lv Y X, Zhang M, et al.2013. Identification, expres-sion, and comparative genomic analysis of the IPT and CKX gene families in Chinese cabbage(Brassica rapa ssp. pekinensis)[J]. BMC Genomics, 14(1): 594. [20] Luo J, Wang X, Feng L, et al.2017. The mitogen-activated protein kinase kinase 9(MKK9)modulates nitrogen ac-quisition and anthocyanin accumulation under nitrogen-limiting condition in Arabidopsis[J]. Biochemical and Biophysical Research Communications, 487: 539-544. [21] Luo Z B, Calfapietra C, Scarascia-Mugnozza G, et al.2008.Carbon-based secondary metabolites and internal nitro-gen pools in Populus nigra under free air CO2 enrich-ment(FACE)and nitrogen fertilisation[J]. Plant Soil,304(1): 45-57. [22] Hamel L P, Nicole M C, Sritubtim S, et al.2006. Ancient sig-nals: Comparative genomics of plant MAPK and MAP-KK gene families[J]. Trends Plant Science, 11: 192-198. [23] Ichimura K, Shinozaki K, Tena G, et al.2002. Mitogen-acti-vated protein kinase cascades in plants: A new nomen-clature[J]. Trends in Plant Science, 7: 301-308. [24] Jackson S, Chen Z J.2010. Genomic and expression plasticity of polyploidy[J]. Current Opinion in Plant Biology, 13(2): 153-159. [25] Jonak C, Okresz L, Bogre L, et al.2002. Complexity, cross talk and integration of plant MAP kinase signalling[J]. Current Opinion in Plant Biology, 5: 415-424. [26] Kong X, Pan J, Zhang M, et al.2011. ZmMKK4, a novel group C mitogen-activated protein kinase kinase in maize(Zea mays), confers salt and cold tolerance in transgenic Arabidopsis[J]. Plant Cell and Environment,34: 1291-1303. [27] Kong X P, Lv W, Zhang D, et al.2013a. Genome-wide identi-fication and analysis of expression profiles of maize mi-togen-activated protein kinase kinase kinase[J]. PLOS ONE, 8: e57714. [28] Kong X P, Pan J W, Zhang D, et al.2013b. Identification of mitogen-activated protein kinase kinase gene family and MKK-MAPK interaction network in maize[J]. Biochem-ical and Biophysical Research Communications, 441:964-969. [29] Rao K P, Richa T, Kumar K, et al.2010. In silico analysis re-veals 75 members of mitogen-activated protein kinase kinase kinase gene family in rice[J]. DNA Research, 17:139-153. [30] Reyna N S, Yang Y.2006. Molecular analysis of the rice MAP kinase gene family in relation on Magnaporthe grisea in-fection[J]. Molecular Plant-Microbe Interactions 19:530-540. [31] Rodriguez M C S, Petersen M, Mundy J.2010. Mitogen-acti-vated protein kinase signaling in plants[J]. Annual Re-view of Plant Biology, 61: 621-649. [32] Schaeffer H J, Weber M J.1999. Mitogen-activated protein ki-nases: Specific messages from ubiquitous messengers[J]. Molecular and Cellular Biology, 19: 2435-2444. [33] Schefe J H, Lehmann K E, Buschmann I R, et al.2006. Quan-titative real-time RT-PCR data analysis: Current con-cepts and the novel "gene expression's CT difference" formula[J]. Journal of Molecular Medicine, 84(11): 901-910. [34] Singh R, Lee M O, Lee J E, et al.2012. Rice mitogen-activat-ed protein kinase interactome analysis using the yeast two-hybrid system[J]. Plant Physiology, 160: 477-487. [35] Teige M, Scheikl E, Eulgem T, et al.2004. The MKK2 path-way mediates cold and salt stress signaling in Arabidopsis[J]. Molecular Cell, 15: 141-152. [36] Tena G, Asai T, Chiu WL, et al.2001. Plant mitogen-activated protein kinase signaling cascades[J]. Current Opinion in Plant Biology, 4: 392-400. [37] Wang H B, Gong M, Guo J Y, et al.2018. Genome-wide iden-tification of Jatropha curcas MAPK, MAPKK, and MAPKKK gene families and their expression profile un-der cold stress[J]. Scientific Reports, 8(1): 16163. [38] Wang J, Pan C T, Wang Y, et al.2015. Genome-wide identifi-cation of MAPK, MAPKK, and MAPKKK gene fami-lies and transcriptional profiling analysis during devel-opment and stress response in cucumber[J]. BMC Ge-nomics, 16: 386. [39] Widmann C, Gibson S, Jarpe M B, et al.1999. Mitogen-acti-vated protein kinase: Conservation of a three-kinase module from yeast to human[J]. Physiological Reviews,79: 143-180. [40] Xiao X M, Li L L, Kuang J F, et al.2023. Cold pretreatment promotes chlorophyll degradation of green ripening ba-nana peel by activating MaCBF1 to MaCBR and MaS-GR1[J]. Food Chemistry, 413: 135575. [41] Yang C, Wang R, Gou L, et al.2017. Overexpression of Popu-lus trichocarpa mitogen-activated protein kinase kinase4 enhances salt tolerance in tobacco[J]. International Jour-nal of Molecular Sciences, 18: 2090. [42] Yang Q S, Gao J, He W D, et al.2015. Comparative transcrip-tomics analysis reveals difference of key gene expres-sion between banana and plantain in response to cold stress[J]. BMC Genomics, 16: 446. [43] Zhao C Z, Wang P C, Si T, et al.2017. MAP kinase cascades regulate the cold response by modulating ICE1 protein stability[J]. Developmental Cell, 43(5): 618-629. [44] Zhao L M, Wang J N, Shan W, et al.2013. Induction of jasmo-nate signalling regulators MaMYC2s and their physical interactions with MaICE1 in methyl jasmonate-induced chilling tolerance in banana fruit[J]. Plant Cell and Envi-ronment, 36: 30-51.
