Abstract
Peach fruit rapidly soften after harvest, a significant challenge for producers and marketers as it results in rotting fruit and significantly reduces shelf life. In this study, we identified two tandem genes, PpNAC1 and PpNAC5, within the sr (slow ripening) locus. Phylogenetic analysis showed that NAC1 and NAC5 are highly conserved in dicots and that PpNAC1 is the orthologous gene of Non-ripening (NOR) in tomato. PpNAC1 and PpNAC5 were highly expressed in peach fruit, with their transcript levels up-regulated at the onset of ripening. Yeast two-hybrid and bimolecular fluorescence complementation assays showed PpNAC1 interacting with PpNAC5 and this interaction occurs with the tomato and apple orthologues. Transient gene silencing experiments showed that PpNAC1 and PpNAC5 positively regulate peach fruit softening. Yeast one-hybrid and dual luciferase assays and LUC bioluminescence imaging proved that PpNAC1 and PpNAC5 directly bind to the PpPGF promoter and activate its transcription. Co-expression of PpNAC1 and PpNAC5 showed higher levels of PpPGF activation than expression of PpNAC1 or PpNAC5 alone. In summary, our findings demonstrate that the tandem transcription factors PpNAC1 and PpNAC5 synergistically activate the transcription of PpPGF to regulate fruit softening during peach fruit ripening.
Key message
PpNAC1 and PpNAC5 directly bind to the PpPGF promoters, synergistically activate its transcription to regulate peach fruit softening.
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References
Bonghi C, Trainotti L, Botton A, Tadiello A, Rasori A, Ziliotto F, Zaffalon V, Casadoro G, Ramina A (2011) A microarray approach to identify genes involved in seed-pericarp cross-talk and development in peach. BMC Plant Biol 11:107. https://doi.org/10.1186/1471-2229-11-107
Cao XM, Wei CY, Duan WY, Gao Y, Kuang JF, Liu MC, Chen KS, Klee H, Zhang B (2021) Transcriptional and epigenetic analysis reveals that NAC transcription factors regulate fruit flavor ester biosynthesis. Plant J 106:785–800. https://doi.org/10.1111/tpj.15200
Cao X, Li X, Su Y, Zhang C, Wei C, Chen K, Grierson D, Zhang B (2023) Transcription factor PpNAC1 and DNA demethylase PpDML1 synergistically regulate peach fruit ripening. Plant Physiol 22:kiad627. https://doi.org/10.1093/plphys/kiad627
Cheng X, Cui ZG, Jiang YB, Chen Y, Tan B, Cheng J, Zhang LL, Ye X, Wang XB, Zhang HP, Lian XD, Li JD, Li ZQ, Zheng XB, Feng JC (2023) PpeERF115 regulates peach fruit ripening by increasing polyamine turnover through up-regulation of genes involved in polyamine synthesis and catabolism. Postharvest Biol Technol 204:112432. https://doi.org/10.1016/j.postharvbio.2023.112432
Dai JY, Xu Z, Xu YT, Fang ZH, Shah K, Kang T, Wu HX, Zhang D, Xing LB, Ma JJ, Liu HK, Hu YN, Zhao CP (2023) A novel NAC transcription factor, PpNAP6, is involved in peach ripening by activating ethylene synthesis. Postharvest Biol Technol 201:112363. https://doi.org/10.1016/j.postharvbio.2023.112363
Dardick CD, Callahan AM, Chiozzotto R, Schaffer RJ, Piagnani MC, Scorza R (2010) Stone formation in peach fruit exhibits spatial coordination of the lignin and flavonoid pathways and similarity to Arabidopsis dehiscence. BMC Biol 8:13. https://doi.org/10.1186/1741-7007-8-13
Dettori MT, Quarta R, Verde I (2001) A peach linkage map integrating RFLPs, SSRs, RAPDs, and morphological markers. Genome 44:783–790. https://doi.org/10.1139/g01-065
Dirlewanger E, Cosson P, Boudehri K, Renaud C, Capdeville G, Tauzin Y, Laigret F, Moing A (2006) Development of a second-generation genetic linkage map for peach [Prunus persica (L.) Batsch] and characterization of morphological traits affecting flower and fruit. Tree Genet Genomes 3:1–13. https://doi.org/10.1007/s11295-006-0053-1
Eduardo I, Pacheco I, Chietera G, Bassi D, Pozzi C, Vecchietti A, Rossini L (2011) QTL analysis of fruit quality traits in two peach intraspecific populations and importance of maturity date pleiotropic effect. Tree Genet Genomes 7(2):323–335. https://doi.org/10.1007/s11295-010-0334-6
Eduardo I, Picañol R, Rojas E, Batlle I, Howad W, Aranzana MJ, Arús P (2015) Mapping of a major gene for the slow ripening character in peach: co-location with the maturity date gene and development of a candidate gene-based diagnostic marker for its selection. Euphytica 205:627–636. https://doi.org/10.1007/s10681-015-1445-9
Forlani S, Mizzotti C, Masiero S (2021) The NAC side of the fruit: tuning of fruit development and maturation. BMC Plant Biol 21:238. https://doi.org/10.1186/s12870-021-03029-y
Fu BL, Wang WQ, Liu XF, Duan XW, Allan AC, Grierson D, Yin XR (2021) An ethylene-hypersensitive methionine sulfoxide reductase regulated by NAC transcription factors increases methionine pool size and ethylene production during kiwifruit ripening. New Phytol 232(1):237–251. https://doi.org/10.1111/nph.17560
Gao Y, Wei W, Zhao XD, Tan XL, Fan ZQ, Zhang YP, Jing Y, Meng LH, Zhu BZ, Zhu HL, Chen JY, Jiang CZ, Grierson D, Luo YB, Fu DQ (2018) A NAC transcription factor, NOR-like1, is a new positive regulator of tomato fruit ripening. Hortic Res 5:75. https://doi.org/10.1038/s41438-018-0111-5
Gao Y, Wei W, Fan ZQ, Zhao XD, Zhang YP, Jing Y, Zhu BZ, Zhu HL, Shan W, Chen JY, Grierson D, Luo YB, Jemric T, Jiang CZ, Fu Q (2020) Re-evaluation of the nor mutation and the role of the NAC-NOR transcription factor in tomato fruit ripening. J Exp Bot 71:3560–3574. https://doi.org/10.1093/jxb/
Gu C, Guo ZH, Cheng HY, Zhou YH, Qi KJ, Wang GM, Zhang SL (2019) A HD-ZIP II HOMEBOX transcription factor, PpHB.G7, mediates ethylene biosynthesis during fruit ripening in peach. Plant Sci 278:12–19. https://doi.org/10.1016/j.plantsci.2018.10.008
Gu C, Wang L, Wang W, Zhou H, Ma BQ, Zheng HY, Fang T, Ogutu C, Vimolmangkang S, Han YP (2016) Copy number variation of a gene cluster encoding endopolygalacturonase mediates flesh texture and stone adhesion in peach. J Exp Bot 67(6):1993–2005. https://doi.org/10.1093/jxb/erw021
Guo J, Cao K, Deng C, Li Y, Zhu GR, Fang WC, Chen CW, Wang XW, Wu JL, Guan LP, Wu S, Guo WW, Yao JL, Fei ZJ, Wang LR (2020) An integrated peach genome structural variation map uncovers genes associated with fruit traits. Genome Biol 21:258. https://doi.org/10.1186/s13059-020-02169-y
Jin ZN, Wang JJ, Cao XM, Wei CY, Kuang JF, Chen KS, Zhang B (2022) Peach fruit PpNAC1 activates PpFAD3-1 transcription to provide ω-3 fatty acids for the synthesis of short-chain flavor volatiles. Hortic Res 9:85. https://doi.org/10.1093/hr/uhac085
Li XJ, Martín-Pizarro C, Zhou LL, Hou BZ, Wang YY, Shen YY, Li BB, Posé D, Qin GZ (2023) Deciphering the regulatory network of the NAC transcription factor FvRIF, a key regulator of strawberry (Fragaria vesca) fruit ripening. Plant Cell 35:4020–4045. https://doi.org/10.1093/plcell/koad210
Liu GS, Li HL, Grierson D, Fu DQ (2022) NAC Transcription factor family regulation of fruit ripening and quality: a review. Cells 11(3):525. https://doi.org/10.3390/cells11030525
Lü PT, Yu S, Zhu N, Chen YR, Zhou BY, Pan Y, Tzeng D, Fabi JP, Argyris J, Garcia-Mas J, Ye NH, Zhang JH, Grierson D, Xiang J, Fei ZJ, Giovannoni J, Zhong SL (2018) Genome encode analyses reveal the basis of convergent evolution of fleshy fruit ripening. Nat Plants 4(10):784–791. https://doi.org/10.1038/s41477-018-0249-z
Martin-Pizarro C, Vallarino JG, Osorio S, Meco V, Urrutia M, Pillet J, Casanal A, Merchante C, Amaya I, Willmitzer L, Fernie AR, Giovannoni JJ, Botella MA, Valpuesta V, Pose D (2021) The NAC transcription factor FaRIF controls fruit ripening in strawberry. Plant Cell 33:1574–1593. https://doi.org/10.1093/plcell/
Meneses C, Ulloa-Zepeda L, Cifuentes-Esquivel A, Infante R, Cantin CM, Batlle I, Arús P, Eduardo I (2016) A codominant diagnostic marker for the slow ripening trait in peach. Mol Breeding 36:77. https://doi.org/10.1007/s11032-016-0506-7
Migicovsky Z, Yeats TH, Watts S, Song J, Forney CF, Burgher-MacLellan K, Somers DJ, Gong YH, Zhang ZQ, Vrebalov J, Velzen RV, Giovannoni JG, Rose JKC, Myles S (2021) Apple ripening is controlled by a NAC transcription factor. Front Genet 12:671300. https://doi.org/10.3389/fgene.2021.671300
Monti LL, Bustamante CA, Osorio S, Gabilondo J, Borsani J, Lauxmann MA, Maulión E, Valentini G, Budde CO, Fernie AR, Lara MV, Drincovich MF (2016) Metabolic profiling of a range of peach fruit varieties reveals high metabolic diversity and commonalities and differences during ripening. Food Chem 190:879–888. https://doi.org/10.1016/j.foodchem.2015.06.043
Nuñez-Lillo G, Cifuentes-Esquivel A, Troggio M, Micheletti D, Infante R, Campos-Vargas R, Orellana A, Blanco-Herrera F, Meneses C (2015) Identification of candidate genes associated with mealiness and maturity date in peach [Prunus persica (L.) Batsch] using QTL analysis and deep sequencing. Tree Genet Genomes 11:86. https://doi.org/10.1007/s11295-015-0911-9
Ogundiwin EA, Peace CP, Gradziel TM, Parfitt DE, Bliss FA, Crisosto CH (2009) A fruit quality gene map of Prunus. BMC Genomics 10:587. https://doi.org/10.1186/1471-2164-10-587
Olsen AN, Ernst HA, Leggio LL, Skriver K (2005) NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci 10:79–87. https://doi.org/10.1016/j.tplants.2004.12.010
Peace C, Crisosto C, Gradziel T (2005) Endopolygalacturonase: a candidate gene for freestone and melting flesh in peach. Mol Breeding 16:21–31. https://doi.org/10.1007/s11032-005-0828-3
Pirona R, Eduardo I, Pacheco I, Linge CDS, Miculan M, Verde I, Tartarini S, Dondini L, Pea G, Bassi D, Rossini L (2013) Fine mapping and identification of a candidate gene for a major locus controlling maturity date in peach. BMC Plant Biol 13:166. https://doi.org/10.1186/1471-2229-13-166
Puranik S, Sahu PP, Srivastava PS, Prasad M (2012) NAC proteins: regulation and role in stress tolerance. Trends Plant Sci 17:369–381. https://doi.org/10.1016/j.tplants.2012.02.004
Qian M, Xu Z, Zhang ZH, Li Q, Yan XY, Liu HK, Han MY, Li FR, Zheng JC, Zhang D, Zhao CP (2021) The downregulation of PpPG21 and PpPG22 influences peach fruit texture and softening. Planta 254:22. https://doi.org/10.1007/s00425-021-03673-6
Schmidt GW, Delaney SK (2010) Stable internal reference genes for normalization of real-time RT-PCR in tobacco (Nicotiana tabacum) during development and abiotic stress. Mol Genet 283:233–241. https://doi.org/10.1007/s00438-010-0511-1
Tatsuki M, Haji T, Yamaguchi M (2007) The peach 1-aminocyclopropane-1-carboxylic acid synthase isogene, Pp-ACS1, is required for fruit softening. Springer, Dordrecht, pp 227–228. https://doi.org/10.1007/978-1-4020-6014-4_48
Tatsuki M, Nakajima N, Fujii H, Shimada T, Nakano M, Hayashi K, Hayama H, Yoshioka H, Nakamura Y (2013) Increased levels of IAA are required for system 2 ethylene synthesis causing fruit softening in peach (Prunus persica L. Batsch). J Exp Bot 64(4):1049–1059. https://doi.org/10.1093/jxb/ers381
Tong ZG, Gao ZH, Wang F, Zhou J, Zhang Z (2009) Selection of reliable reference genes for gene expression studies in peach using real-time PCR. BMC Mol Biol 10:71. https://doi.org/10.1186/1471-2199-10-71
Tonutti P, Casson P, Ramina A (1991) Ethylene biosynthesis during peach fruit development. J Am Soc Hortic Sci 116:274–279. https://doi.org/10.21273/jashs.116.2.274
Wang JF, Tian SW, Yu YT, Ren Y, Guo SG, Zhang J, Li MY, Zhang HY, Gong GY, Wang M, Xu Y (2022a) Natural variation in the NAC transcription factor NONRIPENING contributes to melon fruit ripening. J Integr Plant Biol 64(7):1448–1461. https://doi.org/10.1111/jipb.13278
Wang W, Liu SL, Cheng X, Cui ZG, Jiang YB, Zheng XB, Tan B, Cheng J, Ye X, Li JD, Li ZQ, Zhang LL, Wang XB, Zhang HP, Lian XD, Feng JC (2022b) Ethylene and polyamines form a negative feedback loop to regulate peach fruit ripening via the transcription factor PpeERF113 by regulating the expression of PpePAO1. Postharvest Biol Technol 190:111958. https://doi.org/10.1016/j.postharvbio.2022.111958
Wang XB, Pan L, Wang Y, Meng JR, Deng L, Niu L, Liu H, Ding YF, Yao JL, Nieuwenhuizen NJ, Ampomah-Dwamena C, Lu ZH, Cui GC, Wang ZQ, Zeng WF (2021) PpIAA1 and PpERF4 form a positive feedback loop to regulate peach fruit ripening by integrating auxin and ethylene signals. Plant Sci 313:111084. https://doi.org/10.1016/j.plantsci.2021.111084
Wei Y, Liu Z, Lv TX, Xu YX, Wei YJ, Liu WT, Liu L, Wang A, Li T (2023) Ethylene enhances MdMAPK3-mediated phosphorylation of MdNAC72 to promote apple fruit softening. Plant Cell 35(8):2887–2909. https://doi.org/10.1093/plcell/koad122
Zhang RX, Liu Y, Zhang X, Chen X, Sun J, Zhao Y, Zhang J, Yao JL, Liao L, Zhou H, Han Y (2023) Two adjacent NAC transcription factors regulate fruit maturity date and flavor in peach. New Phytol 241(2):632–649. https://doi.org/10.1111/nph.19372. (Epub. PMID: 37933224)
Zhang ST, Fu MM, Li ZQ, Li JW, Hai LF, Chen CY, Zheng XB, Tan B, Li JD, Cheng J, Wang W, Zhang LL, Ye X, Feng JC (2022) VvEIL2 and VvEIL4 regulate ethylene synthesis and carotenoid metabolism during senescence of grape rachis. Postharvest Biol Technol 187:111853. https://doi.org/10.1016/j.postharvbio.2022.111853
Zhou H, Kui LW, Wang H, Gu C, Dare AP, Espley RV, He H, Allan AC, Han Y (2015) Molecular genetics of blood-fleshed peach reveals activation of anthocyanin biosynthesis by NAC transcription factors. Plant J 82:105–121. https://doi.org/10.1111/tpj.12792
Funding
This work was supported by Innovative Research Group Project of the National Natural Science Foundation of China (32002014), the National Key Research and Development Program of China (2019YFD1000104), and the Special Fund for Henan Agriculture Research System (S2014-11-G02).
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Xiaofei Wang, Langlang Zhang and Kang Dong. The first draft of the manuscript was written by Langlang Zhang and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Zhang, L., Wang, X., Dong, K. et al. Tandem transcription factors PpNAC1 and PpNAC5 synergistically activate the transcription of the PpPGF to regulate peach softening during fruit ripening. Plant Mol Biol 114, 46 (2024). https://doi.org/10.1007/s11103-024-01429-w
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DOI: https://doi.org/10.1007/s11103-024-01429-w