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Gene Cloning and Expression Analysis of LoSorNINJAs in Oriental Lily (Lilium hybrid divisionⅦ) 'Sorbonne' |
ZHU Yun-Tao1, ZHAO Yi-Ran1, CHEN Ming-Hui2, HU Jun-Shu1, YANG Jie1, LIU Xiao-Lin1, SUN Ting-Ting1, KONG Xiang-Hong1, LUO Yuan-Fang1, NIE Yu-Wei1, GAO Xue1, HE Heng-Bin1,* |
1 School of Landscape Architecture/Beijing Key Laboratory of Flower Germplasm Innovation and Molecular Breeding/National Flower Engineering Technology Research Center/Urban and Rural Ecological Environment Beijing Laboratory/Ministry of Education Forest and Flower Breeding Laboratory, Beijing Forestry University, Beijing 100083, China; 2 China National Botanical Garden (North Garden),Beijing 100093, China |
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Abstract Jasmonic acid (JA) is an important plant endogenous hormone, which plays an important role in plant growth, development and defense response. NINJA (novel interactor of JAZ) protein is an inhibitor of the JA signal transduction pathway, involved in the regulation of plant growth, development, and stress response. In this study, 2 different NINJAs were identified from Oriental lily 'Sorbonne' (Lilium hybrid division Ⅶ cv. 'Sorbonne') based on transcriptome data, and named LoSorNINJA1 (GenBank No. OQ715340) and LoSorNINJA2 (GenBank No. OQ715341), respectively. Bioinformatics analysis showed that both LoSorNINJA1 and LoSorNINJA2 contained conserved EAR motif and Jas domain, which conformed to the characteristics of the NINJA gene family. Phylogenetic analysis indicated that LoSorNINJAs had the closest genetic relationship with the NINJA of Apostasia shenzhenica, Phalaenopsis equestris and Dendrobium catenatum. The results of subcellular localization in tobacco (Nicotiana tabacum) showed that both LoSorNINJA1 and LoSorNINJA2 were located in the nucleus. Expression analysis in different tissues of 'Sorbonne' showed that LoSorNINJA2 was highly expressed in the outer petals and anthers, and LoSorNINJA1 was highly expressed in the upper leaves. The shoot apical of 'Sorbonne' lily bulbs after low-temperature storage and greenhouse transplant were collected and analyzed by qRT-PCR. LoSorNINJA1 had 2 peaks of expression at 2 and 6 w cold storage, while LoSorNINJA2 had only 1 peak of expression at 6 w cold storage. Using GENIE3 to construct a potential gene regulatory network indicated that LoSorNINJA1 might be involved in tissue development and post-embryonic development, and LoSorNINJA2 might be involved in the response to the external environment. The above results suggest that LoSorNINJA1 and LoSorNINJA2 might have undergone functional differentiation, and their functional sites and developmental stages might also be different. The present study provides basic data for in-depth analysis of the function of NINJAs in lily dormancy release.
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Received: 07 March 2023
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Corresponding Authors:
* hengbinhe_1220@bjfu.edu.cn
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[1] 曹钦政. 2019. 百合早花群体的构建与FT基因功能研究[D]. 博士学位论文, 北京林业大学, 导师: 贾桂霞, pp. 43-50. (Cao Q Z.2019. Construction of early flowering populations and functional analysis of FLOWERING LOCUS T gene in lily[D]. Thesis for Ph.D., Beijing Forestry University, Supervisor: Jia G X, pp. 43-50.) [2] 贾星月. 2017. pCAMBIA1300-AFH16-GFP重组质粒的构建及纯合转基因拟南芥植株的筛选[D]. 硕士学位论文, 山西师范大学, 导师: 韩榕, pp. 21-22. (Jia X Y.2017. Construction of recombinant plasmid pCAMBIA1300-AFH16-GFP and screening of homozygous transgenic Arabidopsis plants[D]. Thesis for M.S., Shanxi Normal University, Supervisor: Han R, pp. 21-22.) [3] 李小方, 王洋. 2009. 植物休眠的分子调控[J]. 植物生理学通讯, 45(10): 1045-1049. (Li X F, Wang Y.2009. Molecular regulation of plant dormancy[J]. Plant Physiological Communications, 45(10): 1045-1049.) [4] 刘晓华. 2015. 东方百合低温诱导春化的分子调控机制[D]. 博士学位论文, 北京林业大学, 导师: 吕英民, pp. 15-16. (Liu X H.2015. The Molecular mechanism of vernalization induced by low temperature in oriental hybrid Lily 'Sorbonne'[D]. Thesis for Ph.D., Beijing Forestry University, Supervisor: Lv Y M, pp. 15-16.) [5] 廖永翠, 徐艳红, 张争, 等. 2014. 白木香AsNINJA1基因全长cDNA克隆及其在愈伤组织中的茉莉酸甲酯诱导表达[J]. 中草药, 45(20): 2968-2973. (Liao Y C, Xu Y H, Zhang Z, et al.Cloning of the full-length cDNA of AsNINJA1 gene of Baimuxiang and its methyl jasmonate-induced expression in callus[J]. Chinese Herbal Medicine, 45(20): 2968-2973.) [6] 吕嘉. 2018. CRISPR/Cas9编辑NtNINJA基因对低温胁迫下NtJAZ1的表达量的影响[D]. 硕士学位论文, 西南大学, 导师: 戴秀梅, pp. 14-15. (Lv J.2018. Effects of CRISPR/Cas9 editing of NtNINJA gene on the expression of NtJAZ1 under low temperature stress[D]. Thesis for M.S., Southwest University, Supervisor: Dai X M, pp. 14-15.) [7] 冉燕子. 2017. 苗期低温胁迫对烟草JA信号途径部分关键基因表达及JA含量的影响[D]. 硕士学位论文, 西南大学, 导师: 戴秀梅, pp. 43-51. (Ran Y Z.2017. Effects of low temperature stress at seedling stage on the expression and JA content of some key genes in tobacco JA signaling pathway[D]. Thesis for M.S., Southwest University, Supervisor: Dai X M, pp. 43-51.) [8] 杨琳, 张延龙. 2005. 低温对百合种球休眠的影响[J]. 陕西农业科学, (03): 49-51. (Yang L, Zhang Y L. 2005. Effects of low temperature on dormancy of lily bulbs[J]. Shaanxi Agricultural Science, (03): 49-51.) [9] Acosta I F, Gasperini D, Chételat A, et al.2013. Role of NINJA in root jasmonate signaling[J]. Proceedings of the National Academy of Sciences of the USA, 110(38): 15473-15478. [10] An C P, Deng L, Zhai H W, et al.2022. Regulation of jasmonate signaling by reversible acetylation of TOPLESS in Arabidopsis[J]. Molecular Plant, 15(8): 1329-1346. [11] Baekelandt A, Pauwels L, Wang Z B, et al.2018. Arabidopsis leaf flatness is regulated by PPD2 and NINJA through repression of CYCLIN D3 genes1[J]. Plant Physiology, 178: 16. [12] Browse J.2009. Jasmonate passes muster: A receptor and targets for the defense hormone[J]. Annual Review of Plant Biology, 60(1): 183-205. [13] Cantalapiedra C P, Hernández-Plaza A, Letunic I, Bork P, et al.2021. eggNOG-mapper v2: Functional annotation, orthology assignments, and domain prediction at the metagenomic scale[J]. Molecular Biology and Evolution, 38(12): 5825-5829. [14] Chen C J, Chen H, Zhang Y, et al.2020. TBtools: An integrative toolkit developed for interactive analyses of big biological data[J]. Molecular Plant, 13(8): 1194-1202. [15] Chico J M, Chini A, Fonseca S, et al.2008. JAZ repressors set the rhythm in jasmonate signaling[J]. Current Opinion in Plant Biology, 11(5): 486-494. [16] Chini A, Gimenez-Ibanez S, Goossens A, et al.2016. Redundancy and specificity in jasmonate signalling[J]. Current Opinion in Plant Biology, 33: 147-156. [17] Chouard P.1960. Vernalization and its relations to dormancy[J]. Annual Review of Plant Physiology, 11(1): 191-238. [18] Goossens J, Fernández-Calvo P, Schweizer F, et al.2016. Jasmonates: Signal transduction components and their roles in environmental stress responses[J]. Plant Molecular Biology, 91(6): 673-689. [19] Hadley W.2011. ggplot2: Elegant graphics for data analysis[J]. Journal of the Royal Statistical Society Series A: Statistics in Society, 174(1): 245-246. [20] Han X, Kui M Y, Xu T T, et al.2022. CO interacts with JAZ repressors and bHLH subgroup Ⅲd factors to negatively regulate jasmonate signaling in Arabidopsis seedlings[J]. The Plant Cell, 35(2): 852-873. [21] Howe G A, Major I T, Koo A J, 2018. Modularity in jasmonate signaling for multistress resilience[J]. Annual Review of Plant Biology, 69(1): 387-415. [22] Huynh-Thu V A, Irrthum A, Wehenkel L, et al.2010. Inferring regulatory networks from expression data using tree-based methods[J]. PLOS ONE, 5(9): e12776. [23] Kagale S, Rozwadowski K.2011. EAR motif-mediated transcriptional repression in plants: An underlying mechanism for epigenetic regulation of gene expression[J]. Epigenetics, 6(2): 141-146. [24] Li L L, Zhang H H, Chen C H, et al.2021. A class of independently evolved transcriptional repressors in plant RNA viruses facilitates viral infection and vector feeding[J]. Proceedings of the National Academy of Sciences of the USA, 118(11): e2016673118. [25] Li R, Wang M, Wang Y, et al.2017. Flower-specific jasmonate signaling regulates constitutive floral defenses in wild tobacco[J]. Proceedings of the National Academy of Sciences of the USA, 114(34): 7206-7214. [26] Love M, Huber W, Anders S, et al.2014. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2[J]. Genome Biology, 15(12): 550. [27] Pauwels L, Barbero G F, Geerinck J, et al.2010. NINJA connects the co-repressor TOPLESS to jasmonate signalling[J]. Nature, 464(7289): 788-791. [28] Seo J S, Joo J S, Kim M J, et al.2011. OsbHLH148, a basic helix-loop-helix protein, interacts with OsJAZ proteins in a jasmonate signaling pathway leading to drought tolerance in rice[J]. Plant Journal, 65(6): 907-921. [29] Shannon P, Markiel A, Ozier O, et al.2003. Cytoscape: A software environment for integrated models of biomolecular interaction networks[J]. Genome Research, 13(11): 2498-2504. [30] Shyu C, Figueroa P, DePew C L, et al.2012. JAZ8 lacks a canonical degron and has an EAR motif that mediates transcriptional repression of jasmonate responses in Arabidopsis[J]. The Plant Cell, 24(2): 536-550. [31] Thines B, Katsir L, Melotto M, et al.2007. JAZ repressor proteins are targets of the SCF-COI1 complex during jasmonate signalling[J]. Nature, 448(7154): 661-665. [32] Thireault C, Shyu C, Yoshida Y, et al.2015. Repression of jasmonate signaling by a non‐TIFY JAZ protein in Arabidopsis[J]. Plant Journal, 82(4): 669-679. [33] Turner J G, Ellis C, Devoto A.2002. The jasmonate signal pathway[J]. Plant Cell, 14(suppl 1): S153-S164. [34] Wan S, Xin X F.2022. Regulation and integration of plant jasmonate signaling: A comparative view of monocot and dicot[J]. Journal of Genetics and Genomics, 49(8): 704-714. [35] Wang L, Wu S M, Zhu Y, et al.2017. Functional characterization of a novel jasmonate ZIM-domain interactor (NINJA) from upland cotton (Gossypium hirsutum)[J]. Plant Physiology and Biochemistry, 112: 152-160. [36] Wang Y, Mostafa S, Zeng W, et al.2021. Function and mechanism of jasmonic acid in plant responses to abiotic and biotic stresses[J]. International Journal of Molecular Sciences, 22(16): 8568. [37] Wasternack C, Hause B.2013. Jasmonates: Biosynthesis, perception, signal transduction and action in plant stress response, growth and development[J]. Annals of Botany, 111(6): 1021-1058. [38] Wu H, Ye H Y, Yao R F.2015. OsJAZ9 acts as a transcriptional regulator in jasmonate signaling and modulates salt stress tolerance in rice[J]. Plant Science, 232: 1-12. [39] Xie D X, Feys B F, James S, et al.1998. COI1: An Arabidopsis gene required for jasmonate-regulated defense and fertility[J]. Science, 280(5366): 1091-1094. [40] Yamada S, Kano A, Tamaoki D, et al.2012. Involvement of OsJAZ8 in jasmonate-induced resistance to bacterial blight in rice[J]. Plant and Cell Physiology, 53(12): 2060-2072. [41] Yan J, Zhang C, Gu M, et al.2009. The Arabidopsis CORONATINE INSENSITIVE1 protein is a jasmonate receptor[J]. The Plant Cell, 21(8): 2220-2236. [42] Yang J, Duan G H, Li C Q, et al.2019. The crosstalks between jasmonic acid and other plant hormone signaling highlight the involvement of jasmonic acid as a core component in plant response to biotic and abiotic stresses[J]. Frontiers in Plant Science, 10: 1349. [43] Zhai Q Z, Zhang X, Wu F M, et al.2015. Transcriptional mechanism of jasmonate receptor COI1-mediated delay of flowering time in Arabidopsis[J]. The Plant Cell, 27(10): 2814-2828. [44] Zhou T S, Yu D, Dong H, et al.2020. Genome-wide identification and expression profile of NINJA and AFP genes in rice[J]. International Journal of Agriculture And Biology, 23(1): 171-182. |
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