Importance of FaWRKY71 in Strawberry (Fragaria × ananassa) Fruit Ripening
Abstract
:1. Introduction
2. Results
2.1. Sequence Analysis and Subcellular Localization of FaWRKY71
2.2. Analysis of FaWRKY71 Promoter
2.3. Expression Patterns of FaWRKY71 in Different Tissues under Multiple Abiotic Stresses
2.4. Accumulation of Anthocyanin, Lignin and Fruit Ripening
2.5. Antioxidant Ability of Strawberry Fruit
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Treatments
4.2. Amplification of FaWRKY71 and the Promoter of FaWRKY71
4.3. Bioinformatics Analysis
4.4. qRT-PCR
4.5. Subcellular Localization of FaWRKY71 in Nicotiana Benthamiana
4.6. Transient Overexpression in Strawberry Fruits
4.7. Determination of ROS Scavenging-Related Enzyme Activity, MDA Content, and Anthocyanin Content
4.8. Section Staining and Microstructure Observation
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Eulgem, T.; Rushton, P.J.; Robatzek, S.; Somssich, I.E. The WRKY superfamily of plant transcription factors. Trends Plant Sci. 2000, 5, 199–206. [Google Scholar] [CrossRef]
- Rushton, P.J.; Somssich, I.E.; Ringler, P.; Shen, Q.J. WRKY transcription factors. Trends Plant Sci. 2010, 15, 247–258. [Google Scholar] [CrossRef]
- Viana, V.E.; Busanello, C.; da Maia, L.C.; Pegoraro, C.; Costa de Oliveira, A. Activation of rice WRKY transcription factors: An army of stress fighting soldiers? Curr. Opin. Plant Biol. 2018, 45, 268–275. [Google Scholar] [CrossRef] [PubMed]
- Ishiguro, S.; Nakamura, K. Characterization of a cDNA encoding a novel DNA.binding protein, SPF1, that recognizes SP8 sequences in the 5′ upstream regions of genes coding for sporamin and p.amylase from sweet potato. Mol. Gen. Genet. 1994, 244, 563–571. [Google Scholar] [CrossRef] [PubMed]
- Wang, M.; Vannozzi, A.; Wang, G.; Liang, Y.H.; Tornielli, G.B.; Zenoni, S.; Cavallini, E.; Pezzotti, M.; Cheng, Z.M. Genome and transcriptome analysis of the grapevine (Vitis vinifera L.) WRKY gene family. Hortic. Res. 2014, 1, 14016. [Google Scholar] [CrossRef] [Green Version]
- Meng, D.; Li, Y.Y.; Bai, Y.; Li, M.J.; Cheng, L.L. Genome-wide identification and characterization of WRKY transcriptional factor family in apple and analysis of their responses to waterlogging and drought stress. Plant Physiol. Biochem. 2016, 103, 71–83. [Google Scholar] [CrossRef] [PubMed]
- Jiang, J.J.; Ma, S.H.; Ye, N.H.; Jiang, M.; Cao, J.S.; Zhang, J.H. WRKY transcription factors in plant responses to stresses. J. Integr. Plant Biol. 2017, 59, 86–101. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wei, W.; Liang, D.W.; Bian, X.H.; Shen, M.; Xiao, J.H.; Zhang, W.K.; Ma, B.; Lin, Q.; Lv, J.; Chen, X.; et al. GmWRKY54 improves drought tolerance through activating genes in abscisic acid and Ca2+ signaling pathways in transgenic soybean. Plant J. 2019, 100, 384–398. [Google Scholar] [CrossRef]
- Zhu, H.; Jiang, Y.N.; Guo, Y.; Huang, J.B.; Zhou, M.H.; Tang, Y.Y.; Sui, J.M.; Wang, J.S.; Qiao, L.X. A novel salt inducible WRKY transcription factor gene, AhWRKY75, confers salt tolerance in transgenic peanut. Plant Physiol. Biochem. 2021, 160, 175–183. [Google Scholar] [CrossRef] [PubMed]
- Luo, D.L.; Ba, L.J.; Shan, W.; Kuang, J.F.; Lu, W.J.; Chen, J.Y. Involvement of WRKY Transcription Factors in ABA-Induced Cold Tolerance of Banana Fruit. J. Agric. Food Chem. 2017, 65, 3627–3635. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.H.; Zhao, F.X.; Zhang, G.; Jia, S.Z.; Yan, Z.M. FaWRKY11 transcription factor positively regulates resistance to Botrytis cinerea in strawberry fruit. Sci. Hortic. 2021, 279, 109893. [Google Scholar] [CrossRef]
- Lei, Y.Y.; Sun, Y.P.; Wang, B.T.; Yu, S.; Dai, H.Y.; Li, H.; Zhang, Z.H.; Zhang, J.X. Woodland strawberry WRKY71 acts as a promoter of flowering via a transcriptional regulatory cascade. Hortic Res. 2020, 7, 137. [Google Scholar] [CrossRef] [PubMed]
- An, J.P.; Zhang, X.W.; You, C.X.; Bi, S.Q.; Wang, X.F.; Hao, Y.J. MdWRKY40 promotes wounding-induced anthocyanin biosynthesis in association with MdMYB1 and undergoes MdBT2-mediated degradation. New Phytol. 2019, 224, 380–395. [Google Scholar] [CrossRef] [PubMed]
- Li, C.; Wu, J.; Hu, K.D.; Wei, S.W.; Sun, H.Y.; Hu, L.Y.; Han, Z.; Yao, G.F.; Zhang, H. PyWRKY26 and PybHLH3 cotargeted the PyMYB114 promoter to regulate anthocyanin biosynthesis and transport in red-skinned pears. Hortic. Res. 2020, 7, 37. [Google Scholar] [CrossRef] [Green Version]
- Lin-Wang, K.; McGhie, T.K.; Wang, M.; Liu, Y.; Warren, B.; Storey, R.; Espley, R.V.; Allan, A.C. Engineering the anthocyanin regulatory complex of strawberry (Fragaria vesca). Front. Plant Sci. 2014, 5, 651. [Google Scholar] [CrossRef] [Green Version]
- Zhang, J.J.; Wang, X.; Yu, O.; Tang, J.J.; Gu, X.G.; Wan, X.C.; Fang, C.B. Metabolic profiling of strawberry (Fragaria × ananassa Duch.) during fruit development and maturation. J. Exp. Bot. 2011, 62, 1103–1118. [Google Scholar] [CrossRef]
- Medina-Puche, L.; Blanco-Portales, R.; Molina-Hidalgo, F.J.; Cumplido-Laso, G.; García-Caparrós, N.; Moyano-Cañete, E.; Caballero-Repullo, J.L.; Muñoz-Blanco, J.; Rodríguez-Franco, A. Extensive transcriptomic studies on the roles played by abscisic acid and auxins in the development and ripening of strawberry fruits. Funct. Integr. Genomics 2016, 16, 671–692. [Google Scholar] [CrossRef]
- Symons, G.M.; Chua, Y.J.; Ross, J.J.; Quittenden, L.J.; Davies, N.W.; Reid, J.B. Hormonal changes during non-climacteric ripening in strawberry. J. Exp. Bot. 2012, 63, 4741–4750. [Google Scholar] [CrossRef] [Green Version]
- Li, T.Y.; Dai, Z.R.; Zeng, B.Z.; Li, J.; Ouyang, J.Y.; Kang, L.; Wang, W.; Jia, W.S. Autocatalytic biosynthesis of abscisic acid and its synergistic action with auxin to regulate strawberry fruit ripening. Hortic. Res. 2022, 9, uhab076. [Google Scholar] [CrossRef]
- Jiang, Y.M.; Joyce, D.C. ABA effects on ethylene production, PAL activity, anthocyanin and phenolic contents of strawberry fruit. Plant Growth Regul. 2003, 39, 171–174. [Google Scholar] [CrossRef]
- Moya-León, M.A.; Mattus-Araya, E.; Herrera, R. Molecular Events Occurring During Softening of Strawberry Fruit. Front. Plant Sci. 2019, 10, 615. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Paniagua, C.; Ric-Varas, P.; García-Gago, J.A.; López-Casado, G.; Blanco-Portales, R.; Muñoz-Blanco, J.; Schückel, J.; Knox, J.P.; Matas, A.J.; Quesada, M.A.; et al. Elucidating the role of polygalacturonase genes in strawberry fruit softening. J. Exp. Bot. 2020, 71, 7103–7117. [Google Scholar] [CrossRef] [PubMed]
- Posé, S.; Paniagua, C.; Cifuentes, M.; Blanco-Portales, R.; Quesada, M.A.; Mercado, J.A. Insights into the effects of polygalacturonase FaPG1 gene silencing on pectin matrix disassembly, enhanced tissue integrity, and firmness in ripe strawberry fruits. J. Exp. Bot. 2013, 64, 3803–3815. [Google Scholar] [CrossRef] [PubMed]
- Wei, W.; Hu, Y.; Han, Y.T.; Zhang, K.; Zhao, F.L.; Feng, J.Y. The WRKY transcription factors in the diploid woodland strawberry Fragaria vesca: Identification and expression analysis under biotic and abiotic stresses. Plant Physiol. Biochem. 2016, 105, 129–144. [Google Scholar] [CrossRef]
- Chen, P.; Liu, Q.Z. Genome-wide characterization of the WRKY gene family in cultivated strawberry (Fragaria × ananassa Duch.) and the importance of several group III members in continuous cropping. Sci. Rep. 2019, 9, 8423. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zou, X.H.; Dong, C.; Liu, H.T.; Gao, Q.H. Genome-wide characterization and expression analysis of WRKY family genes during development and resistance to Colletotrichum fructicola in cultivated strawberry (Fragaria × ananassa Duch.). J. Integr. Agr. 2022, 21, 1658–1672. [Google Scholar] [CrossRef]
- Zhang, Y.T.; Jiang, L.Y.; Li, Y.L.; Chen, Q.; Ye, Y.T.; Zhang, Y.; Luo, Y.; Sun, B.; Wang, X.R.; Tang, H.R. Effect of Red and Blue Light on Anthocyanin Accumulation and Differential Gene Expression in Strawberry (Fragaria × ananassa). Molecules 2018, 23, 820. [Google Scholar] [CrossRef] [Green Version]
- Wang, L.; Luo, Z.S.; Yang, M.Y.; Liang, Z.; Qi, M.; Dong, Y.Y.; Xu, Y.Q.; Lin, X.Y.; Li, L. The action of RED light: Specific elevation of pelargonidin-based anthocyanin through ABA-related pathway in strawberry. Postharvest Biol. Technol. 2022, 186, 111835. [Google Scholar] [CrossRef]
- Malekzadeh Shamsabad, M.R.; Esmaeilizadeh, M.; Roosta, H.R.; Dąbrowski, P.; Telesiński, A.; Kalaji, H.M. Supplemental light application can improve the growth and development of strawberry plants under salinity and alkalinity stress conditions. Sci. Rep. 2022, 12, 9272. [Google Scholar] [CrossRef]
- Zhang, Y.T.; Hu, W.J.; Peng, X.R.; Sun, B.; Wang, X.R.; Tang, H.R. Characterization of anthocyanin and proanthocyanidin biosynthesis in two strawberry genotypes during fruit development in response to different light qualities. J. Photochem. Photobiol. B 2018, 186, 225–231. [Google Scholar] [CrossRef]
- Fei, J.; Wang, Y.S.; Cheng, H.; Su, Y.B.; Zhong, Y.J.; Zheng, L. The Kandelia obovata transcription factor KoWRKY40 enhances cold tolerance in transgenic Arabidopsis. BMC Plant Biol. 2022, 22, 274. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Y.; Yu, H.J.; Yang, X.R.; Li, Q.; Ling, J.; Wang, H.; Gu, X.F.; Huang, S.W.; Jiang, W.J. CsWRKY46, a WRKY transcription factor from cucumber, confers cold resistance in transgenic-plant by regulating a set of cold-stress responsive genes in an ABA-dependent manner. Plant Physiol. Biochem. 2016, 108, 478–487. [Google Scholar] [CrossRef] [PubMed]
- Misson, J.; Raghothama, K.G.; Jain, A.; Jouhet, J.; Block, M.A.; Bligny, R.; Ortet, P.; Creff, A.; Somerville, S.; Rolland, N.; et al. A genome-wide transcriptional analysis using Arabidopsis thaliana Affymetrix gene chips determined plant responses to phosphate deprivation. PNAS 2005, 102, 11934–11939. [Google Scholar] [CrossRef] [Green Version]
- Devaiah, B.N.; Karthikeyan, A.S.; Raghothama, K.G. WRKY75 transcription factor is a modulator of phosphate acquisition and root development in Arabidopsis. Plant Physiol. 2007, 143, 1789–1801. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jaakola, L. New insights into the regulation of anthocyanin biosynthesis in fruits. Trends Plant Sci. 2013, 18, 477–483. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhang, J.K.; Wang, Y.C.; Mao, Z.L.; Liu, W.N.; Ding, L.C.; Zhang, X.N.; Yang, Y.W.; Wu, S.Q.; Chen, X.S.; Wang, Y.L. Transcription factor McWRKY71 induced by ozone stress regulates anthocyanin and proanthocyanidin biosynthesis in Malus crabapple. Ecotoxicol. Environ. Saf. 2022, 232, 113274. [Google Scholar] [CrossRef] [PubMed]
- Schaart, J.G.; Dubos, C.; Romero De La Fuente, I.; van Houwelingen, A.M.M.L.; de Vos, R.C.H.; Jonker, H.H.; Xu, W.; Routaboul, J.M.; Lepiniec, L.; Bovy, A.G. Identification and characterization of MYB-bHLH-WD40 regulatory complexes controlling proanthocyanidin biosynthesis in strawberry (Fragaria × ananassa) fruits. New Phytol. 2013, 197, 454–467. [Google Scholar] [CrossRef] [PubMed]
- Wang, H.; Zhang, H.; Yang, Y.; Li, M.; Zhang, Y.T.; Liu, J.S.; Dong, J.; Li, J.; Butelli, E.; Xue, Z.; et al. The control of red colour by a family of MYB transcription factors in octoploid strawberry (Fragaria × ananassa) fruits. Plant Biotechnol. J. 2019, 18, 1169–1184. [Google Scholar] [CrossRef] [Green Version]
- An, X.H.; Tian, Y.; Chen, K.Q.; Liu, X.J.; Liu, D.D.; Xie, X.B.; Cheng, C.G.; Cong, P.H.; Hao, Y.J. MdMYB9 and MdMYB11 are involved in the regulation of the JA-induced biosynthesis of anthocyanin and proanthocyanidin in apples. Plant Cell Physiol. 2015, 56, 650–662. [Google Scholar] [CrossRef] [Green Version]
- Carlos, R.F.; Hernán, G.R.; Pedro, M.C.; Gustavo, A.M.; Raúl, H.; María, A.M. Changes in cell wall polysaccharides and cell wall degrading enzymes during ripening of Fragaria chiloensis and Fragaria × ananassa fruits. Sci. Hortic. 2010, 124, 454–462. [Google Scholar] [CrossRef]
- Zhang, W.W.; Zhao, S.Q.; Zhang, L.C.; Xing, Y.; Jia, W.S. Changes in the cell wall during fruit development and ripening in Fragaria vesca. Plant Physiol. Biochem. 2020, 154, 54–65. [Google Scholar] [CrossRef] [PubMed]
- Castro, R.I.; Muñoz-Vera, M.; Parra-Palma, C.; Valenzuela-Riffo, F.; Figueroa, C.R.; Morales-Quintana, L. Characterization of cell wall modification through thermogravimetric analysis during ripening of Chilean strawberry (Fragaria chiloensis) fruit. Cellulose 2021, 28, 4611–4623. [Google Scholar] [CrossRef]
- Given, N.K.; Venis, M.A.; Gierson, D. Hormonal regulation of ripening in the strawberry, a non-climacteric fruit. Planta 1988, 174, 402–406. [Google Scholar] [CrossRef] [PubMed]
- Li, B.J.; Grierson, D.; Shi, Y.N.; Chen, K.S. Roles of abscisic acid in regulating ripening and quality of strawberry, a model non-climacteric fruit. Hortic. Res. 2022, 9, uhac089. [Google Scholar] [CrossRef]
- Xu, X.B.; Ma, X.Y.; Le, H.H.; Yin, L.L.; Shi, X.Q.; Song, H.M. MicroRNAs play an important role in the regulation of strawberry fruit senescence in low temperature. Postharvest Biol. Technol. 2015, 108, 39–47. [Google Scholar] [CrossRef]
- Nagpal, P.; Ellis, C.M.; Weber, H.; Ploense, S.E.; Barkawi, L.S.; Guilfoyle, T.J.; Hagen, G.; Alonso, J.M.; Cohen, J.D.; Farmer, E.E.; et al. Auxin response factors ARF6 and ARF8 promote jasmonic acid production and flower maturation. Development 2005, 132, 4107–4118. [Google Scholar] [CrossRef] [Green Version]
- Apel, K.; Hirt, H. Reactive oxygen species: Metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biol. 2004, 55, 373–399. [Google Scholar] [CrossRef] [Green Version]
- Shen, N.; Wang, T.F.; Gan, Q.; Liu, S.; Wang, L.; Jin, B. Plant flavonoids: Classification, distribution, biosynthesis, and antioxidant activity. Food Chem. 2022, 383, 132531. [Google Scholar] [CrossRef] [PubMed]
- Chen, Q.; Yu, H.W.; Wang, X.R.; Xie, X.L.; Yue, X.Y.; Tang, H.R. An alternative cetyltrimethylammonium bromide-based protocol for RNA isolation from blackberry (Rubus L.). Genet. Mol. Res. 2012, 11, 1773–1782. [Google Scholar] [CrossRef] [PubMed]
- Jiang, L.Y.; Yue, M.L.; Liu, Y.Q.; Ye, Y.Y.; Zhang, Y.T.; Lin, Y.X.; Wang, X.R.; Chen, Q.; Tang, H.R. Alterations of Phenylpropanoid Biosynthesis Lead to the Natural Formation of Pinkish-Skinned and White-Fleshed Strawberry (Fragaria × ananassa). Int. J. Mol. Sci. 2022, 23, 7375. [Google Scholar] [CrossRef]
- Raab, T.; López-Ráez, J.A.; Klein, D.; Caballero, J.L.; Moyano, E.; Schwab, W.; Muñoz-Blanco, J. FaQR, required for the biosynthesis of the strawberry flavor compound 4-hydroxy-2,5-dimethyl-3(2H)-furanone, encodes an enone oxidoreductase. Plant Cell. 2006, 18, 1023–1037. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT Method. Methods 2001, 25, 402–408. [Google Scholar] [CrossRef] [PubMed]
- Lin, Y.X.; Jiang, L.Y.; Chen, Q.; Li, Y.L.; Zhang, Y.T.; Luo, Y.; Zhang, Y.; Sun, B.; Wang, X.R.; Tang, H.R. Comparative Transcriptome Profiling Analysis of Red- and White-Fleshed Strawberry (Fragaria × ananassa) Provides New Insight into the Regulation of the Anthocyanin Pathway. Plant Cell Physiol. 2018, 59, 1844–1859. [Google Scholar] [CrossRef]
- Stewart, R.R.; Bewley, J.D. Lipid Peroxidation Associated with Accelerated Aging of Soybean Axes. Plant Physiol. 1980, 65, 245–248. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, Z.F.; Zheng, Y.H.; Cao, S.F. Effect of High Oxygen Atmosphere Storage on Quality, Antioxidant Enzymes, and DPPH-Radical Scavenging Activity of Chinese Bayberry Fruit. J. Agric. Food Chem. 2009, 57, 176–181. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.S.; Tian, S.P.; Xu, Y. Effects of high oxygen concentration on pro- and anti-oxidant enzymes in peach fruits during postharvest periods. Food Chem. 2005, 99, 99–104. [Google Scholar] [CrossRef]
- Giusti, M.M.; Wrolstad, R.E. Characterization and Measurement of Anthocyanins by UV-Visible Spectroscopy. In Current Protocols in Food Analytical Chemistry; Thermo Fisher Scientific: Waltham, MA, USA, 2001; pp. F1.2.1–F1.2.13. [Google Scholar] [CrossRef]
Cis-Acting Element | Function of Site | Sequence | Number |
---|---|---|---|
ABRE | cis-acting element involved in the abscisic acid responsiveness | CACGTG | 1 |
AE-box | part of a module for light response | AGAAACAA | 1 |
ARE | cis-acting regulatory element essential for the anaerobic induction | AAACCA | 5 |
Box 4 | part of a conserved DNA module involved in light responsiveness | ATTAAT | 1 |
CGTCA-motif | cis-acting regulatory element involved in the MeJA-responsiveness | CGTCA | 3 |
G-Box | cis-acting regulatory element involved in light responsiveness | CACGTG | 1 |
MBS | MYB binding site involved in drought-inducibility | CAACTG | 1 |
MRE | MYB binding site involved in light responsiveness | AACCTAA | 1 |
MYB | MYB binding site | CAACCA/ TAACTG | 1/1 |
MYC | MYB binding site | CATTTG/CATGTG | 2/1 |
Sp1 | light responsive element | GGGCGG | 1 |
TGA-box | part of an auxin-responsive element | TGACGTAA | 1 |
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Yue, M.; Jiang, L.; Zhang, N.; Zhang, L.; Liu, Y.; Wang, Y.; Li, M.; Lin, Y.; Zhang, Y.; Zhang, Y.; et al. Importance of FaWRKY71 in Strawberry (Fragaria × ananassa) Fruit Ripening. Int. J. Mol. Sci. 2022, 23, 12483. https://doi.org/10.3390/ijms232012483
Yue M, Jiang L, Zhang N, Zhang L, Liu Y, Wang Y, Li M, Lin Y, Zhang Y, Zhang Y, et al. Importance of FaWRKY71 in Strawberry (Fragaria × ananassa) Fruit Ripening. International Journal of Molecular Sciences. 2022; 23(20):12483. https://doi.org/10.3390/ijms232012483
Chicago/Turabian StyleYue, Maolan, Leiyu Jiang, Nating Zhang, Lianxi Zhang, Yongqiang Liu, Yan Wang, Mengyao Li, Yuanxiu Lin, Yunting Zhang, Yong Zhang, and et al. 2022. "Importance of FaWRKY71 in Strawberry (Fragaria × ananassa) Fruit Ripening" International Journal of Molecular Sciences 23, no. 20: 12483. https://doi.org/10.3390/ijms232012483