VvWRKY13 from Vitis vinifera Regulate Fruit Development by Controlling Ethylene Biosynthesis

doi:10.7668/hbnxb.20194673

Abstract

Abstract:

Fruit development is the key stage which determines the yield and quality of grape.WRKY family transcription factors play important roles in regulating plant development and environment adaptation.Ethylene is the important plant hormone which participates in regulation of fruit development,and ACC synthase is the key enzyme that limits ethylene synthesis.Using grape variety Zuoyouhong, VvWRKY13 overexpressing grape callus,as well as VvWRKY13 heterologous over-expressed tomato lines as materials,the role of VvWRKY13 from grape in fruit development and its relationship with ethylene were studied though plant physiological and biochemical methods as well as molecular biological techniques.The results displayed that the expressions of VvWRKY13 as well as ACC synthase genes VvACS2 and VvACS7 significantly upregulated at early stage of fruit development,and in VvWRKY13 over-expression grape callus,the expression of VvACS7C was significantly higher than control,but the expression of VvACS2 displayed no significant difference compared with control.Yeast one hybrid experiment showed that VvWRKY13 could directly bind to the VvACS7 promoter,VvWRKY13 had no direct interaction with VvACS2.We also found that the ethylene content and the expression of ACS family members,such as SlACS1b,SlACS4 and SlACS6,the ethylene synthesis genes,were significantly induced in VvWRKY13 heterologous over-expressed tomato,and the time from flowering to breaking of tomato was 3—6 days shorter than wild type;the above results indicated that,VvWRKY13 could regulate ethylene synthesis by promoting the expression of ACC synthase gene to participate in regulation of fruit development.

Key words: Vitis vinifera, VvWRKY13, VvACS7, Ethylene, Fruit development

CHE Yongmei, CHEN Huiting, ZHANG Suifang, HUI Mengling, YE Qing, HOU Lixia, LIU Xin. VvWRKY13 from Vitis vinifera Regulate Fruit Development by Controlling Ethylene Biosynthesis[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(2): 99-105. doi: 10.7668/hbnxb.20194673.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20194673

http://www.hbnxb.net/EN/Y2024/V39/I2/99

Figures/Tables 8

Tab.1 Quantitative PCR primer sequence

引物名称 Primer name	引物序列(5'—3') Primer sequences(5'—3')
VvWRKY13-FP	GGTTGCCAACAATCCCT
VvWRKY13-RP	GTCATCTCCACCGATACTTC
VvActin-FP	ATAGAAGCAGCAAGGGA
VvActin-RP	TGAGGCTCTTACTAATG
VvACS2-FP	GAGGTGGAGAACAGGAGTGC
VvACS2-RP	GAGCAAGCCTTTCACTTTGG
VvACS7-FP	GATGAAGTTTACGCTGGAACTGTG
VvACS7-RP	CCAGAGTCGGAAAGAAGGCAC
SlACS2-FP	GGAGGTTCGTAGGTGTTGAG
SlACS2-RP	ATGGTGAGGGAGGAATAGG
SlACS4-FP	GGGTTATTCAGATGGGTCTC
SlACS4-RP	TCCACCAGCCATTACTACAC
SlACS6-FP	GCCACAAATGATGGACATGG
SlACS6-RP	GAGGCTTTTGGGTTGTTGACT
SlACS1a-FP	CATTGCTCAGAGCCAGG
SlACS1a-RP	AAGACATAGGTGAATGAGGAG
SlACS1b-FP	TGTTCTGAACCTGGTTGGT
SlACS1b-RP	TAAAATCATCCAATCTTCGA
SlActin-FP	GGAATGGGACAGAAGGAT
SlActin-RP	CAGTCAGGAGAACAGGGT

Fig.1 The relative expression level of VvWRKY13 in grape fruit at different development stage Different lowercase letters indicate significant differences at 0.05 level.The same as Fig.2—3,5.

Fig.2 The transcription profiels of ethylene biosynthesis genes during the development of grape fruit

Fig.3 Expression levels of ethylene biosynthesis genes in VvWRKY13 over-expressing grape callus A.VvWRKY13 over-expression grape callus(OEWRKY13);B.The relative expression of VvWRKY13 in OEWRKY13;C.Relative expression of ethylene biosynthesis genes in OEWRKY13.

Fig.4 The binding activity of VvWRKY13 to promoters of VvACS2 (A)and VvACS7 (B)in a yeast one-hybrid system

Fig.5 Ethylene content and expression levels of ethylene biosynthesis genes in fruit of VvWRKY13 ectopic over-expressing tomato plants

Fig.6 The fruit of VvWRKY13 ectopic over-expressing tomato dpa.Days post anthesis;dpk.Days post breaking.

Tab.2 The times from flowering to breaking and breaking to maturity of VvWRKY13 ectopic over-expressing tomato

株系 Lines	开花—转色时间/d Time from flowering to breaking	转色—成熟时间/d Time from breaking to maturity
WT	42.67±1.15a	10±1.15a
L3	36.33±1.52b	10±1.52a
L9	39.33±4.0ab	10±4.04a
L14	36.33±1.52b	10±1.52a

References 23

[1]	Conde C, Silva P, Fontes N, Dias A, Tavares R, Sousa M, Agasse A, Delrot S, Geros H. Biochemical changes throughout grape berry development and fruit and wine quality[J]. Food Biophysics, 2007,1:1-22.
[2]	Seymour G B, Østergaard L, Chapman N H, Knapp S, Martin C. Fruit development and ripening[J]. Annual Review of Plant Biology, 2013,64:219-241.doi:10.1146/annurev-arplant-050312-120057.
[3]	He L, Meng N, Castellarin S D, Wang Y, Sun Q, Li X Y, Dong Z G, Tang X P, Duan C Q, Pan Q H. Combined metabolite and transcriptome profiling reveals the norisoprenoid responses in grape berries to abscisic acid and synthetic auxin[J]. International Journal of Molecular Sciences, 2021, 22(3):1420.doi:10.3390/ijms22031420. URL
[4]	Cara B, Giovannoni J J. Molecular biology of ethylene during tomato fruit development and maturation[J]. Plant Science, 2008, 175(1/2):106-113.doi:10.1016/j.plantsci.2008.03.021. URL
[5]	Osorio S, Alba R, Damasceno C M B, Lopez-Casado G, Lohse M, Zanor M I, Tohge T, Usadel B, Rose J K C, Fei Z J, Giovannoni J J, Fernie A R. Systems biology of tomato fruit development:combined transcript,protein,and metabolite analysis of tomato transcription factor(nor,rin)and ethylene receptor(Nr)mutants reveals novel regulatory interactions[J]. Plant Physiology, 2011, 157(1):405-425.doi:10.1104/pp.111.175463. pmid: 21795583
[6]	Cao H H, Chen J, Yue M, Xu C, Jian W, Liu Y D, Song B Q, Gao Y Q, Cheng Y L, Li Z G. Tomato transcriptional repressor MYB70 directly regulates ethylene-dependent fruit ripening[J]. The Plant Journal, 2020, 104(6):1568-1581.doi:10.1111/tpj.15021. pmid: 33048422
[7]	Wang P P, Ge M Q, Yu A S, Song W, Fang J G, Leng X P. Effects of ethylene on berry ripening and anthocyanin accumulation of Fujiminori grape in protected cultivation[J]. Journal of the Science of Food and Agriculture, 2022, 102(3):1124-1136.doi:10.1002/jsfa.11449. URL
[8]	Xin T X, Zhang Z, Li S, Zhang S, Li Q, Zhang Z H, Huang S W, Yang X Y. Genetic regulation of ethylene dosage for cucumber fruit elongation[J]. The Plant Cell, 2019, 31(5):1063-1076.doi:10.1105/tpc.18.00957. pmid: 30914499
[9]	Kashyap K, Banu S. Characterizing ethylene pathway genes during the development,ripening,and postharvest response in Citrus reticulata Blanco fruit pulp[J]. Turkish Journal of Botany, 2019, 43(2):173-184.doi:10.3906/bot-1711-45. URL
[10]	Chervin C, El-Kereamy A, Roustan J P, Latch A, Lamon J, Bouzayen M. Ethylene seems required for the berry development and ripening in grape,a non-climacteric fruit[J]. Plant Science, 2004, 167(6):1301-1305.doi:10.1016/j.plantsci.2004.06.026. URL
[11]	Deluc L G, Grimplet J, Wheatley M D, Tillett R L, Quilici D R, Osborne C, Schooley D A, Schlauch K A, Cushman J C, Cramer G R. Transcriptomic and metabolite analyses of cabernet sauvignon grape berry development[J]. BMC Genomics, 2007, 8(1):429.doi:10.1186/1471-2164-8-429.
[12]	任永娟, 王东姣, 苏亚春, 王玲, 张旭苏, 炜华阙, 友雄. 2021. 植物WRKY转录因子:结构、分类、进化和功能[J]. 农业生物技术学报,2021, 29(1):105-124.doi:10.3969/j.issn.1674-7968.2021.01.011.
	Ren Y J, Wang D J, Su Y C, Wang L, Zhang X S, Wei H Q, You X. Structure,classification,evolution and function of plant WRKY transcription factors[J]. Journal of Agricultural Biotechnology, 2021, 29(1):105-124.
[13]	Tiika R J, Wei J, Ma R, Yang H S, Cui G X, Duan H R, Ma Y J. Identification and expression analysis of the WRKY gene family during different developmental stages in Lycium ruthenicum Murr.fruit[J]. PeerJ, 2020,8:e10207.doi:10.7717/peerj.10207.
[14]	Wang M, Vannozzi A, Wang G, Liang Y H, Tornielli G B, Zenoni S, Cavallini E, Pezzotti M, Cheng Z M M. Genome and transcriptome analysis of the grapevine(Vitis vinifera L.) WRKY gene family[J]. Horticulture Research, 2014,1:14016.doi:10.1038/hortres.2014.16.
[15]	Wei Y, Meng N, Wang Y C, Cheng J, Duan C Q, Pan Q H. Transcription factor VvWRKY70 inhibits both norisoprenoid and flavonol biosynthesis in grape[J]. Plant Physiology, 2023, 193(3):2055-2070.doi:10.1093/plphys/kiad423. pmid: 37471439
[16]	Liu F, Dou T X, Hu C H, Zhong Q F, Sheng O, Yang Q S, Deng G M, He W D, Gao H J, Li C Y, Dong T, Liu S W, Yi G J, Bi F C. WRKY transcription factor MaWRKY49 positively regulates pectate lyase genes during fruit ripening of Musa acuminata[J]. Plant Physiology and Biochemistry, 2023, 194:643-650.doi:10.1016/j.plaphy.2022.12.015. URL
[17]	Singh D, Debnath P, Roohi, Sane A P, Sane V A. Expression of the tomato WRKY gene,SlWRKY23,alters root sensitivity to ethylene,auxin and JA and affects aerial architecture in transgenic Arabidopsis[J]. Physiology and Molecular Biology of Plants, 2020, 26(6):1187-1199.doi:10.1007/s12298-020-00820-3.
[18]	Sun X M, Zhang L L, Wong D C J, Wang Y, Zhu Z F, Xu G Z, Wang Q F, Li S H, Liang Z C, Xin H P. The ethylene response factor VaERF092 from Amur grape regulates the transcription factor VaWRKY33,improving cold tolerance[J]. The Plant Journal, 2019, 99(5):988-1002.doi:10.1111/tpj.14378. URL
[19]	Ma Q, Zhang G K, Hou L X, Wang W J, Hao J, Liu X. Vitis vinifera VvWRKY13 is an ethylene biosynthesis-related transcription factor[J]. Plant Cell Reports, 2015, 34(9):1593-1603.doi:10.1007/s00299-015-1811-z. URL
[20]	Li M, Chen R, Gu H, Cheng D W, Guo X Z, Shi C Y, Li L, Xu G Y, Gu S C, Wu Z Y, Chen J Y. Grape small auxin upregulated RNA(SAUR)041 is a candidate regulator of berry size in grape[J]. International Journal of Molecular Sciences, 2021, 22(21):11818.doi:10.3390/ijms222111818. URL
[21]	Fenn M A, Giovannoni J J. Phytohormones in fruit development and maturation[J]. The Plant Journal, 2021, 105(2):446-458.doi:10.1111/tpj.15112. pmid: 33274492
[22]	Nitsch L, Kohlen W, Oplaat C, Charnikhova T, Cristescu S, Michieli P, Wolters-Arts M, Bouwmeester H, Mariani C, Vriezen W H, Rieu I. ABA-deficiency results in reduced plant and fruit size in tomato[J]. Journal of Plant Physiology, 2012, 169(9):878-883.doi:10.1016/j.jplph.2012.02.004. pmid: 22424572
[23]	Hao J, Ma Q, Hou L X, Zhao F G, Xin L. VvWRKY13 enhances ABA biosynthesis in Vitis vinifera[J]. Acta Societatis Botanicorum Poloniae, 2017, 86(2):1-14.doi:10.5586/asbp.3546.

Please choose a citation manager

Content to export