Abstract
Main conclusion
The downregulation of PpPG21 and PpPG22 expression in melting-flesh peach delays fruit softening and hinders texture changes by influencing pectin solubilization and depolymerization.
Abstract
The polygalacturonase (PG)-catalyzed solubilization and depolymerization of pectin plays a central role in the softening and texture formation processes in peach fruit. In this study, the expression characteristics of 15 PpPG members in peach fruits belonging to the melting flesh (MF) and non-melting flesh (NMF) types were analyzed, and virus-induced gene silencing (VIGS) technology was used to identify the roles of PpPG21 (ppa006839m) and PpPG22 (ppa006857m) in peach fruit softening and texture changes. In both MF and NMF peaches, the expression of PpPG1, 10, 12, 23, and 25 was upregulated, whereas that of PpPG14, 24, 35, 38, and 39 was relatively stable or downregulated during shelf life. PpPG1 was highly expressed in NMF fruit, whereas PpPG21 and 22 were highly expressed in MF peaches. Suppressing the expression of PpPG21 and 22 by VIGS in MF peaches significantly reduced PG enzyme activity, maintained the firmness of the fruit during the late shelf life stage, and suppressed the occurrence of the “melting” stage compared with the control fruits. Moreover, the downregulation of PpPG21 and 22 expression also reduced the water-soluble pectin (WSP) content, increased the contents of ionic-soluble pectin (ISP) and covalent-soluble pectin (CSP) and affected the expression levels of ethylene synthesis- and pectin depolymerization-related genes in the late shelf life stage. These results indicate that PpPG21 and 22 play a major role in the development of the melting texture trait of peaches by depolymerizing cell wall pectin. Our results provide direct evidence showing that PG regulates peach fruit softening and texture changes.
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Data availability
The raw datasets generated during and/or analyzed during the current study are available from the first author on reasonable request. The data that support the finding of in this study are available in the supplementary material of this article.
Abbreviations
- CSP:
-
Covalent-soluble pectin
- DAH:
-
Days after harvest
- ISP:
-
Ionic-soluble pectin
- MF:
-
Melting flesh
- NMF:
-
Non-melting flesh
- PG:
-
Polygalacturonase
- VIGS:
-
Virus-induced gene silencing
- WSP:
-
Water-soluble pectin
References
Aday MS, Caner C, Rahvalı F (2011) Effect of oxygen and carbon dioxide absorbers on strawberry quality. Postharvest Biol Tec 62:179–187. https://doi.org/10.1016/j.postharvbio.2011.05.002
Atkinson RG, Sutherland PW, Johnston SL, Gunaseelan K, Hallett IC, Mitra D, Brummell DA, Schröder R, Johnston JW, Schaffer RJ (2012) Down-regulation of Polygalacturonase1 alters firmness, tensile strength and water loss in apple (Malus x domestica) fruit. BMC Plant Biol 12:129. https://doi.org/10.1186/1471-2229-12-129
Brummell DA (2006) Cell wall disassembly in ripening fruit. Funct Plant Biol 33:103–119. https://doi.org/10.1071/FP05234
Brummell DA, Cin VD, Crisosto CH, Labavitch JM (2004) Cell wall metabolism during maturation, ripening and senescence of peach fruit. J Exp Bot 55:2029–2039. https://doi.org/10.1093/jxb/erh227
Brummell DA, Labavitch JM (1997) Effect of antisense suppression of endopolygalacturonase activity on polyuronide molecular weight in ripening tomato fruit and in fruit homogenates. Plant Physiol 115:717–725. https://doi.org/10.1104/pp.115.2.717
Callahan AM, Scorza R, Bassett C, Nickerson M, Abeles FB (2004) Deletions in an endopolygalacturonase gene cluster correlate with non-melting flesh texture in peach. Funct Plant Biol 31:159–168. https://doi.org/10.1071/FP03131
Carrington CS, Greve LC, Labavitch JM (1993) Cell wall metabolism in ripening fruit. VI. Effect of the antisense polygalacturonase gene on cell wall changes accompanying ripening in transgenic tomatoes. Plant Physiol 103:429–434. https://doi.org/10.1104/pp.103.2.429
Chauhan OP, Raju PS, Dasgupta DK, Bawa AS (2006) Instrumental textural changes in banana (var. Pachbale) during ripening under active and passive modified atmosphere. Int J Food Prop 9:237–253. https://doi.org/10.1080/10942910600596282
Costa F, Peace CP, Stella S, Serra S, Musacchi S, Bazzani M, Sansavini S, de Weg WEV (2010) QTL dynamics for fruit firmness and softening around an ethylene-dependent polygalacturonase gene in apple (Malus x domestica Borkh.). J Exp Bot 61:3029–3039. https://doi.org/10.1093/jxb/erq130
Defilippi BG, Ejsmentewicz T, Covarrubias MP, Gudenschwager O, Campos-Vargas R (2018) Changes in cell wall pectins and their relation to postharvest mesocarp softening of “Hass” avocados (Persea americana Mill.). Plant Physiol Bioch 128:142–151. https://doi.org/10.1016/j.plaphy.2018.05.018
Fabi JP, Cordenunsi BR, Seymour GB, Lajolo FM, Do Nascimento JRO (2009) Molecular cloning and characterization of a ripening-induced polygalacturonase related to papaya fruit softening. Plant Physiol Bioch 47:1075–1081. https://doi.org/10.1016/j.plaphy.2009.08.002
Figueroa CR, Rosli HG, Civello PM, Martínez GA, Herrera R, Moya-León MA (2010) Changes in cell wall polysaccharides and cell wall degrading enzymes during ripening of Fragaria chiloensis and Fragaria× ananassa fruits. Sci Hortic-Amsterdam 124:454–462. https://doi.org/10.1016/j.scienta.2010.02.003
Fonseca S, Monteiro L, Barreiro MG, Pais MS (2005) Expression of genes encoding cell wall modifying enzymes is induced by cold storage and reflects changes in pear fruit texture. J Exp Bot 56:2029–2036. https://doi.org/10.1093/jxb/eri201
García-Gago JA, Posé S, Muñoz-Blanco J, Quesada MA, Mercado JA (2009) The polygalacturonase FaPG1 gene plays a key role in strawberry fruit softening. Plant Signal Behav 4:766–768. https://doi.org/10.4161/psb.4.8.9167
Ge T, Huang X, Pan X, Zhang J, Xie R (2019) Genome-wide identification and expression analysis of citrus fruitlet abscission-related polygalacturonase genes. 3 Biotech 9:250. https://doi.org/10.1007/s13205-019-1782-9
Ghiani A, Onelli E, Aina R, Cocucci M, Citterio S (2011) A comparative study of melting and non-melting flesh peach cultivars reveals that during fruit ripening endo-polygalacturonase (endo-PG) is mainly involved in pericarp textural changes, not in firmness reduction. J Exp Bot 62:4043–4054. https://doi.org/10.1093/jxb/err109
Giongo L, Poncetta P, Loretti P, Costa F (2013) Texture profiling of blueberries (Vaccinium spp.) during fruit development, ripening and storage. Postharvest Biol Tec 76:34–39. https://doi.org/10.1016/j.postharvbio.2012.09.004
Giovannoni JJ, DellaPenna D, Bennett AB, Fischer RL (1989) Expression of a chimeric polygalacturonase gene in transgenic rin (ripening inhibitor) tomato fruit results in polyuronide degradation but not fruit softening. Plant Cell 1:53–63. https://doi.org/10.2307/3869061
Glover H, Brady CJ (1995) Pectinesterases from mature, unripe peach fruit bind strongly to pectic polysaccharides. Aust J Plant Physiol 22:977–985
Gross KC (1982) A rapid and sensitive spectrophotometric method for assaying polygalacturonase using 2-cyanoacetamide. Hort Sci 17:933–934
Gu C, Wang L, Wang W, Zhou H, Ma B, Zheng H, Fang T, Ogutu C, Vimolmangkang S, Han Y (2016) Copy number variation of a gene cluster encoding endopolygalacturonase mediates flesh texture and stone adhesion in peach. J Exp Bot 67:1993–2005. https://doi.org/10.1093/jxb/erw021
Haji T, Yaegaki H, Yamaguchi M (2005) Inheritance and expression of fruit texture melting, non-melting and stony hard in peach. Sci Horti 105(2):241–248. https://doi.org/10.1016/j.scienta.2005.01.017
Haji T (2014) Inheritance of flesh texture in peach and effects of ethylene treatment on softening of the stony hard peach. Jpn Agric Res Q 48:57–61. https://doi.org/10.6090/jarq.48.57
Hayama H, Shimada T, Fujii H, Ito A, Kashimura Y (2006) Ethylene-regulation of fruit softening and softening-related genes in peach. J Exp Bot 57:4071–4077. https://doi.org/10.1093/jxb/erl178
Hiwasa K, Kinugasa Y, Amano S, Hashimoto A, Nakano R, Inaba A, Kubo Y (2003) Ethylene is required for both the initiation and progression of softening in pear (Pyrus communis L.) fruit. J Exp Bot 54:771–779. https://doi.org/10.1093/jxb/erg073
Huber DJ (1983) Polyuronide degradation and hemicellulose modifications in ripening tomato fruit. J Am Soc Hortic Sci (USA) 108:405–409
Jia H, Chai Y, Li C, Qin L, Shen Y (2011) Cloning and characterization of the H subunit of a magnesium chelatase gene (PpCHLH) in peach. J Plant Growth Regul 30:445–455. https://doi.org/10.1007/s00344-011-9207-y
Jiang F, Lopez A, Jeon S, Tonetto de Freitas S, Yu Q, Wu Z, Labavitch JM, Tian S, Powell ALT, Mitcham E (2019) Disassembly of the fruit cell wall by the ripening-associated polygalacturonase and expansin influences tomato cracking. Hort Res 6:17. https://doi.org/10.1038/s41438-018-0105-3
Kajuna ST, Bilanski WK, Mittal GS (1997) Textural changes of banana and plantain pulp during ripening. J Sci Food Agr 75:244–250. https://doi.org/10.1002/(SICI)1097-0010(199710)75:2<244::AID-JSFA861>3.0.CO;2-%23
Kan J, Liu J, Jin CH (2015) Changes in cell walls during fruit ripening in Chinese ‘honey’ peach. J Hort Sci B 88:37–46. https://doi.org/10.1080/14620316.2013.11512933
Kim J, Gross KC, Solomos T (1987) Characterization of the stimulation of ethylene production by galactose in tomato (Lycopersicon esculentum Mill.) fruit. Plant Physiol 85:804–807. https://doi.org/10.1104/pp.85.3.804
Kintner PK III, Van Buren JP (1982) Carbohydrate interference and its correction in pectin analysis using the m-hydroxydiphenyl method. J Food Sci 47:756–759. https://doi.org/10.1111/j.1365-2621.1982.tb12708.x
Layne DR, Bassi D (2008) The peach: botany, production and uses. CABI Publishing, Wallingford, p 615. https://doi.org/10.1079/9781845933869.0000
Lester DR, Speirs J, Orr G, Brady CJ (1994) Peach (Prunus persica) endopolygalacturonase cDNA isolation and mRNA analysis in melting and nonmelting peach cultivars. Plant Physiol 105:225–231. https://doi.org/10.1104/pp.105.1.225
Liguori G, Weksler A, Zutahi Y, Lurie S, Kosto I (2004) Effect of 1-methylcyclopropene on ripening of melting flesh peaches and nectarines. Postharvest Biol Tec 31:263–268. https://doi.org/10.1016/j.postharvbio.2003.09.007
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001
Longhi S, Hamblin MT, Trainotti L, Peace CP, Velasco R, Costa F (2013) A candidate gene based approach validates Md-PG1 as the main responsible for a QTL impacting fruit texture in apple (Malus × domestica Borkh). BMC Plant Biol 13:37. https://doi.org/10.1186/1471-2229-13-37
Longhi S, Moretto M, Viola R, Velasco R, Costa F (2012) Comprehensive QTL mapping survey dissects the complex fruit texture physiology in apple (Malus × domestica Borkh.). J Exp Bot 63:1107–1121. https://doi.org/10.1093/jxb/err326
Manganaris GA, Vasilakakis M, Diamantidis G, Mignani I (2006) Diverse metabolism of cell wall components of melting and non-melting peach genotypes during ripening after harvest or cold storage. J Sci Food Agr 86:243–250. https://doi.org/10.1002/jsfa.2310
Manganaris GA, Vasilakakis M, Diamantidis G, Mignani I (2010) Diverse metabolism of cell wall components of melting and non-melting peach genotypes during ripening after harvest or cold storage. J Sci Food Agric 86:243–250. https://doi.org/10.1002/jsfa.2310
Morgutti S, Negrini N, Nocito FF, Ghiani A, Bassi D, Cocucci M (2006) Changes in endopolygalacturonase levels and characterization of a putative endo-PG gene during fruit softening in peach genotypes with nonmelting and melting flesh fruit phenotypes. New Phytol 171:315–328. https://doi.org/10.1111/j.1469-8137.2006.01763.x
Murayama H, Arikawa M, Sasaki Y, Dal Cin V, Mitsuhashi W, Toyomasu T (2009) Effect of ethylene treatment on expression of polyuronide-modifying genes and solubilization of polyuronides during ripening in two peach cultivars having different softening characteristics. Postharvest Biol Tec 52:196–201. https://doi.org/10.1016/j.postharvbio.2008.11.003
Onelli E, Ghiani A, Gentili R, Serra S, Musacchi S, Citterio S (2015) Specific changes of exocarp and mesocarp occurring during softening differently affect firmness in melting (MF) and non melting flesh (NMF) fruits. PLoS ONE 10:e145341. https://doi.org/10.1371/journal.pone.0145341
Osorio S, Scossa F, Fernie A (2013) Molecular regulation of fruit ripening. Front Plant Sci 4:198. https://doi.org/10.3389/fpls.2013.00198
Paniagua C, Ric-Varas P, García-Gago JA, López-Casado G, Blanco-Portales R, Muñoz-Blanco J, Schückel J, Knox JP, Matas AJ, Quesada MA, Posé S, Mercado JA (2020) Elucidating the role of polygalacturonase genes in strawberry fruit softening. J Exp Bot 71:7103–7117. https://doi.org/10.1093/jxb/eraa398
Peng G, Wu J, Lu W, Li J (2013) A polygalacturonase gene clustered into clade E involved in lychee fruitlet abscission. Sci Hortic-Amsterdam 150:244–250. https://doi.org/10.1016/j.scienta.2012.10.029
Philosoph-Hadas S, Aharoni N (1987) Galactose inhibits the conversion of 1-aminocyclopropane-1-carboxylic acid to ethylene in aged tobacco leaf discs. Plant Physiol 83:8–11. https://doi.org/10.1104/pp.83.1.8
Posé S, Kirby AR, Paniagua C, Waldron KW, Morris VJ, Quesada MA, Mercado JA (2015) The nanostructural characterization of strawberry pectins in pectate lyase or polygalacturonase silenced fruits elucidates their role in softening. Carbohyd Polym 132:134–145. https://doi.org/10.1016/j.carbpol.2015.06.018
Posé S, Paniagua C, Cifuentes M, Blanco-Portales R, Quesada MA, Mercado JA (2013) 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 64:3803–3815. https://doi.org/10.1093/jxb/ert210
Pressey R, Hinton DM, Avants JK (1971) Development of polygalacturonase activity and solubilization of pectin in peaches during ripening. J Food Sci 36:1070–1073. https://doi.org/10.1111/j.1365-2621.1971.tb03348.x
Qian M, Zhang Y, Yan X, Han M, Li J, Li F, Li F, Zhang D, Zhao C (2016) Identification and expression analysis of polygalacturonase family members during peach fruit softening. Int J Mol Sci 17:1933. https://doi.org/10.3390/ijms17111933
Quesada MA, Blanco-Portales R, Posé S, García-Gago JA, Jiménez-Bermúdez S, Muñoz-Serrano A, Caballero JL, Pliego-Alfaro F, Mercado JA, Muñoz-Blanco J (2009) Antisense down-regulation of the FaPG1 gene reveals an unexpected central role for polygalacturonase in strawberry fruit softening. Plant Physiol 150:1022–1032. https://doi.org/10.1104/pp.109.138297
Rosli HG, Civello PM, Martinez GA (2004) Changes in cell wall composition of three Fragaria × ananassa cultivars with different softening rate during ripening. Plant Physiol Bioch 42:823–831. https://doi.org/10.1016/j.plaphy.2004.10.002
Santiago-Doménech N, Jiménez-Bemúdez S, Matas AJ, Rose JK, Muñoz-Blanco J, Mercado JA, Quesada MA (2008) Antisense inhibition of a pectate lyase gene supports a role for pectin depolymerization in strawberry fruit softening. J Exp Bot 59:2769–2779. https://doi.org/10.1093/jxb/ern142
Sekine D, Munemura I, Gao M, Mitsuhashi W, Toyomasu T, Murayama H (2006) Cloning of cDNAs encoding cell-wall hydrolases from pear (Pyrus communis) fruit and their involvement in fruit softening and development of melting texture. Physiol Plant 126:163–174. https://doi.org/10.1111/j.1399-3054.2006.00583.x
Siddiqui S, Brackmann A, Streif J, Bangerth F (1996) Controlled atmosphere storage of apples: cell wall composition and fruit softening. J Hort Sci 71:613–620. https://doi.org/10.1080/14620316.1996.11515441
Simpson SD, Ashford DA, Harvey DJ, Bowles DJ (1998) Short chain oligogalacturonides induce ethylene production and expression of the gene encoding aminocyclopropane 1-carboxylic acid oxidase in tomato plants. Glycobiology 8:579–583. https://doi.org/10.1093/glycob/8.6.579
Smith C, Watson CF, Ray J, Bird CR, Morris PC, Schuch W, Grierson D (1988) Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature 334:724. https://doi.org/10.1038/334724a0
Smith CJS, Watson CF, Morris PC, Bird CR, Seymour GB, Gray JE, Arnold C, Tucker GA, Schuch W, Harding S, Grierson D (1990) Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Mol Biol 14:369–379. https://doi.org/10.1007/BF00028773
Syamaladevi RM, Lupien SL, Bhunia K, Sablani SS, Dugan F, Rasco B, Killingerc K, Dhingrad A, Ross C (2014) UV-C light inactivation kinetics of Penicillium expansum on pear surfaces: influence on physicochemical and sensory quality during storage. Postharvest Biol Tec 87:27–32. https://doi.org/10.1016/j.postharvbio.2013.08.005
Tacken E, Ireland H, Gunaseelan K, Karunairetnam S, Wang D, Schultz K, Bowen J, Atkinson RG, Johnston JW, Putterill J, Hellens RP, Schaffer RJ (2010) The role of ethylene and cold temperature in the regulation of the apple Polygalacturonase1 gene and fruit softening. Plant Physiol 153:294–305. https://doi.org/10.1104/pp.109.151092
Tao S, Khanizadeh S, Zhang H, Zhang S (2009) Anatomy, ultrastructure and lignin distribution of stone cells in two Pyrus species. Plant Sci 176:413–419. https://doi.org/10.1016/j.plantsci.2008.12.011
Wang L, Zhu G, Fang W et al (2012) Peach genetic resource in China. China agriculture press, Beijing
Wei J, Qi X, Cheng Y, Guan J (2015) Difference in activity and gene expression of pectin-degrading enzymes during softening process in two cultivars of Chinese pear fruit. Sci Hortic-Amsterdam 197:434–440. https://doi.org/10.1016/j.scienta.2015.10.002
Wolf S, Hématy K, Höfte H (2012) Growth control and cell wall signaling in plants. Annu Rev Plant Biol 63:381–407. https://doi.org/10.1146/annurev-arplant-042811-105449
Yang Z, Zheng Y, Cao S, Tang S, Ma S, Li NA (2007) Effects of storage temperature on textural properties of Chinese bayberry fruit. J Texture Stud 38:166–177. https://doi.org/10.1111/j.1745-4603.2007.00092.x
Yoshioka H, Hayama H, Tatsuki M, Nakamura Y (2011) Cell wall modifications during softening in melting type peach “Akatsuki” and non-melting type peach “Mochizuki.” Postharvest Biol Tec 60:100–110. https://doi.org/10.1016/j.postharvbio.2010.12.013
Zaharah SS, Singh Z (2011) Mode of action of nitric oxide in inhibiting ethylene biosynthesis and fruit softening during ripening and cool storage of ‘Kensington Pride’mango. Postharvest Biol Tec 62:258–266. https://doi.org/10.1016/j.postharvbio.2011.06.007
Zerpa-Catanho D, Esquivel P, Mora-Newcomer E, Sáenz MV, Herrera R, Jiménez VM (2017) Transcription analysis of softening-related genes during postharvest of papaya fruit (Carica papaya L. ‘Pococí’hybrid). Postharvest Biol Tec 125:42–51. https://doi.org/10.1016/j.postharvbio.2016.11.002
Acknowledgements
We want to thank Minghui Lu for English language correction. This study was supported by grants from National Science Foundation of China (Grant number 31572079), the Key Research and Development Program of Shaanxi Province, China (Grant number 2018NY-048).
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425_2021_3673_MOESM2_ESM.docx
Supplementary file2 (DOCX 13 KB) Comparison of fruit firmness between MF group and NMF group. * and ** represent no significant (P > 0.05), significant (P ≤ 0.05) and highly significant differences (P ≤ 0.01), respectively. * of red and blue represent significant increase and decrease, respectively
425_2021_3673_MOESM3_ESM.docx
Supplementary file3 (DOCX 2293 KB) Sequencing analysis of PpPGs. a Coding DNA sequence (CDS) alignment between 45 PpPG genes and the consistent sequence of PpPG21/PpPG22. b CDS alignment between PpPG21 and PpPG22 and the consistent sequence of PpPG21/PpPG22. c Gene sequence alignment between PpPG21 and PpPG22
425_2021_3673_MOESM4_ESM.pdf
Supplementary file4 (PDF 359 KB) Control and RNAi fruit during shelf life. EP, IP and PES represent external fruit, internal fruit and paraffin-embedded sections, respectively. Bars = 200 μm. The circular patches on the fruit surface are freshly infected with Agrobacterium tumefaciens
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Supplementary file5 (PDF 1126 KB) Changes in the expression of PpACO1 and PpACS2 in the control and RNAi fruit during shelf life. The data are presented as the means ± standard errors (n = 3 biological replicates). High significant differences (P < 0.01) between means are indicated by “**”
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Qian, M., Xu, Z., Zhang, Z. et al. The downregulation of PpPG21 and PpPG22 influences peach fruit texture and softening. Planta 254, 22 (2021). https://doi.org/10.1007/s00425-021-03673-6
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DOI: https://doi.org/10.1007/s00425-021-03673-6