Cloning and function analysis of DlWRKY9 gene in longan (Dimocarpus longan)

Dengwei JUE; Yuxin XIA; Qigao GUO; Huan LIU; Xuelian SANG

doi:10.15835/nbha50312916

Authors

Dengwei JUE Southwest University, College of Horticulture and Landscape, Chongqing, 400715; Chongqing University of Arts and Sciences, College of Landscape Architecture and Life Science, Yongchuan 402160, Chongqing (CN)
Yuxin XIA Chongqing University of Arts and Sciences, College of Landscape Architecture and Life Science, Yongchuan 402160, Chongqing (CN)
Qigao GUO Southwest University, College of Horticulture and Landscape, Chongqing, 400715 (CN)
Huan LIU Chongqing University of Arts and Sciences, College of Landscape Architecture and Life Science, Yongchuan 402160, Chongqing (CN)
Xuelian SANG Chongqing University of Arts and Sciences, College of Landscape Architecture and Life Science, Yongchuan 402160, Chongqing (CN) https://orcid.org/0000-0001-5975-063X

DOI:

https://doi.org/10.15835/nbha50312916

Keywords:

DlWRKY9, flowering, longan, transgenic Arabidopsis

Abstract

WRKY is one of the largest plant transcription factors (TFs) which is widely involved in plant growth, development, and responses to stresses. In the present study, a WRKY TF DlWRKY9 was cloned from longan (Dimocarpus longan). The coding sequence (CDS) of DlWRKY9 is 762 bp in length and encodes 253 amino acids. It has a typical WRKY domain and zinc finger structure which belongs to type IIa WRKY protein. The molecular weight of DlWRKY9 protein was 30.27kda and the theoretical isoelectric point (PI) was 5.24. It is an unstable hydrophilic protein. The secondary structure of DlWRKY9 protein consists of helical structure (17.39%), extended chain (8.70%) and other structures (turn and random coil) (73.91%). The amino acid sequence of DlWRKY9 protein had the highest similarity with DlWRKY9 (xp_006450293.1) of citrus Clementina. DlWRKY9 gene promoter elements contain light, abscisic acid, gibberellin, jasmonic acid and other response elements. The results of qRT-PCR showed that the relative expression level of DlWRKY9 gene in pericarp was higher, followed by young fruits and floral organs. Meanwhile, DlWRKY9 gene specifically down-regulated in the early stage of flower induction in ‘Sijimi’ (SJ) longan. The results of transient expression of Arabidopsis protoplasts showed that the fluorescence signal was mainly concentrated in the nucleus. Moreover, overexpression of DlWRKY9 in Arabidopsis promoted early flowering. These results provide useful information for revealing the biological roles of DlWRKY9 in longan and increase our understanding of the WRKY family in fruit trees.

Metrics

Metrics Loading ...

References

Baranwal V, Negi N, Khurana P (2016). Genome-wide identification and structural, functional and evolutionary analysis of WRKY components of mulberry. Scientific Reports 6(1):1-13. https://doi.org/10.1038/srep30794

Cai X, Ballif J, Endo S, Davis E, Liang M, Chen D, … Wu Y (2007). A putative CCAAT-binding transcription factor is a regulator of flowering timing in Arabidopsis. Plant Physiology 145(1):98-105. https://doi.org/10.1104/pp.107.102079

Cai Y, Chen X, Xie K, Xing Q, Wu Y, Li J, Du C, Sun Z, Guo Z (2014). Dlf1, a WRKY transcription factor, is involved in the control of flowering time and plant height in rice. PloS One 9(7):e102529. https://doi.org/10.1371/journal.pone.0102529

Chen F, Hu Y, Vannozzi A, Wu K, Cai H, Qin Y, Mullis A, Lin Z, Zhang L (2017). The WRKY transcription factor family in model plants and crops. Critical Reviews in Plant Sciences 36(5-6):311-335. https://doi.org/10.1080/07352689.2018.1441103

Clough S (2005). Floral dip: agrobacterium-mediated germ line transformation. In: Transgenic Plants: Methods and Protocols. Springer, pp 91-101. https://doi.org/10.1385/1-59259-827-7:091

Gocal G, Sheldon C, Gubler F, Moritz T, Bagnall D, MacMillan C, … King R (2001). GAMYB-like genes, flowering, and gibberellin signaling in Arabidopsis. Plant Physiology 127(4):1682-1693. https://doi.org/10.1104/pp.010442

Guo C, Guo R, Xu X, Gao M, Li X, Song J, Zheng Y, Wang X (2014). Evolution and expression analysis of the grape (Vitis vinifera L.) WRKY gene family. Journal of Experimental Botany 65(6):1513-1528. https://doi.org/10.1093/jxb/eru007

Huang R, Liu D, Huang M, Ma J, Li Z, Li M, Sui S (2019). CpWRKY71, a WRKY transcription factor gene of Wintersweet (Chimonanthus praecox), promotes flowering and leaf senescence in Arabidopsis. International Journal of Molecular Sciences 20(21):5325. https://doi.org/10.3390/ijms20215325

Hwang K, Susila H, Nasim Z, Jung J-Y, Ahn J-H (2019). Arabidopsis ABF3 and ABF4 transcription factors act with the NF-YC complex to regulate SOC1 expression and mediate drought-accelerated flowering. Molecular Plant 12(4):489-505. https://doi.org/10.1016/j.molp.2019.01.002

Jue D, Sang X, Liu L, Shu B, Wang Y, Liu C, Wang Y, Xie J, Shi S (2019). Comprehensive analysis of the longan transcriptome reveals distinct regulatory programs during the floral transition. BMC Genomics 20(1):1-18. https://doi.org/10.1186/s12864-019-5461-3

Jue, D., Sang, X., Liu, L., Shu, B., Wang, Y., Liu, C., et al. (2018). Identification of WRKY gene family from Dimocarpus longan and its expression analysis during flower induction and abiotic stress responses. International Journal of Molecular Sciences 19(8):2169. https://doi.org/10.3390/ijms19082169

Kim S-G, Kim S-Y, Park C-M (2007). A membrane-associated NAC transcription factor regulates salt-responsive flowering via FLOWERING LOCUS T in Arabidopsis. Planta 226(3):647-654. https://doi.org/10.1007/s00425-007-0513-3

Lee JH, Ryu H-S, Chung KS, Posé D, Kim S, Schmid M, Schmid M, Ahn J-H (2013). Regulation of temperature-responsive flowering by MADS-box transcription factor repressors. Science 342(6158):628-632. https://doi.org/10.1126/science.124109

Li W, Wang H, Yu D (2016). Arabidopsis WRKY transcription factors WRKY12 and WRKY13 oppositely regulate flowering under short-day conditions. Molecular Plant 9(11):1492-1503. https://doi.org/10.1016/j.molp.2016.08.003

Lin Y, Min J, Lai R, Wu Z, Chen Y, Yu L, … Lai Z (2017). Genome-wide sequencing of longan (Dimocarpus longan Lour.) provides insights into molecular basis of its polyphenol-rich characteristics. Gigascience 6(5):gix023. https://doi.org/10.1093/gigascience/gix023

Ling J, Jiang W, Zhang Y, Yu H, Mao Z, Gu X, Huang S, Xie B (2011). Genome-wide analysis of WRKY gene family in Cucumis sativus. BMC Genomics 12(1):1-20. https://doi.org/10.1186/1471-2164-12-471

Ma N, An Y, Li J, Wang L (2020a). Cloning and characterization of a homologue of the FLORICAULA/LEAFY gene in Ficus carica L., FcLFY, and its role in flower bud differentiation. Scientia Horticulturae 261:109014. https://doi.org/10.1016/j.scienta.2019.109014

Ma Z, Li W, Wang H, Yu D (2020b). WRKY transcription factors WRKY12 and WRKY13 interact with SPL10 to modulate age‐mediated flowering. Journal of Integrative Plant Biology 62(11):1659-1673. https://doi.org/10.1111/jipb.12946

Moon J, Lee H, Kim M, Lee I (2005). Analysis of flowering pathway integrators in Arabidopsis. Plant and Cell Physiology 46(2):292-299. https://doi.org/10.1093/pcp/pci024

Phukan U-J, Jeena G-S, Shukla R-K (2016). WRKY transcription factors: molecular regulation and stress responses in plants. Frontiers in Plant Science 7:760. https://doi.org/10.3389/fpls.2016.00760

Rinerson C-I, Rabara R-C, Tripathi P, Shen Q, Rushton P-J (2015). The evolution of WRKY transcription factors. BMC Plant Biology 15(1):1-18. https://doi.org/10.1186/s12870-015-0456-y

Rishmawi L, Pesch M, Juengst C, Schauss AC, Schrader A, Hülskamp M (2014). Non-cell-autonomous regulation of root hair patterning genes by WRKY75 in Arabidopsis. Plant Physiology 165(1):186-195. https://doi.org/10.1104/pp.113.233775

Ross C-A, Liu Y, Shen Q-J. (2007). The WRKY gene family in rice (Oryza sativa). Journal of Integrative Plant Biology 49(6):827-842. https://doi.org/10.1111/j.1744-7909.2007.00504.x

Rushton P-J, Somssich I-E, Ringler P, Shen Q-J (2010). WRKY transcription factors. Trends in Plant Science 15(5):247-258. https://doi.org/10.1016/j.tplants.2010.02.006

Shabala S, Bose J, Hedrich R (2014). Salt bladders: do they matter? Trends in Plant Science 19(11):687-691. https://doi.org/10.1016/j.tplants.2014.09.001

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013). MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30(12):2725-2729. https://doi.org/10.1093/molbev/mst197

Turck F, Zhou A, Somssich I-E (2004). Stimulus-dependent, promoter-specific binding of transcription factor WRKY1 to its native promoter and the defense-related gene PcPR1-1 in parsley. The Plant Cell 16(10):2573-2585. https://doi.org/10.1105/tpc.104.024810

Wani S-H, Anand S, Singh B, Bohra A, Joshi R (2021). WRKY transcription factors and plant defense responses: latest discoveries and future prospects. Plant Cell Reports 40(7):1071-1085. https://doi.org/10.1007/s00299-021-02691-8

Wu Y, Zhang S, Huang X, Lyu L, Li W, Wu W (2022). Genome-wide identification of WRKY gene family members in black raspberry and their response to abiotic stresses. Scientia Horticulturae 304:111338. https://doi.org/10.1016/j.scienta.2022.111338

Yoo S-D, Cho Y-H, Sheen J (2007). Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nature Protocols 2(7):1565-1572. https://doi.org/10.1038/nprot.2007.199

You X, Wang L, Liang W, Gai Y, Wang X, Chen W (2012). Floral reversion mechanism in longan (Dimocarpus longan Lour.) revealed by proteomic and anatomic analyses. Journal of Proteomics 75(4):1099-1118. https://doi.org/10.1016/j.jprot.2011.10.023

Yu Y, Liu Z, Wang L, Kim S-G, Seo P-J, Qiao M, Wang N, Li S, Cao X, Park C-M, Xiang F (2016). WRKY 71 accelerates flowering via the direct activation of FLOWERING LOCUS T and LEAFY in Arabidopsis thaliana. The Plant Journal 85(1):96-106. https://doi.org/10.1111/tpj.13092

Zhang H, Shi S, Li W, Shu B, Liu L, Xie J, Wei Y (2016). Transcriptome analysis of ‘Sijihua’longan (Dimocarpus longan L.) based on next-generation sequencing technology. The Journal of Horticultural Science and Biotechnology 91(2):180-188. https://doi.org/10.1080/14620316.2015.1133539

Zhang Y, Zhang B, Yang T, Zhang J, Liu B, Zhan X, Liang Y (2020). The GAMYB-like gene SlMYB33 mediates flowering and pollen development in tomato. Horticulture Research 7:133. https://doi.org/10.1038/s41438-020-00366-1