Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (11): 2202-2213.doi: 10.3864/j.issn.0578-1752.2022.11.010

• HORTICULTURE • Previous Articles     Next Articles

Peptidome Analysis of Mesocarp in Melting Flesh and Stony Hard Peach During Fruit Ripening

LI Ang(),MIAO YuLe,MENG JunRen,NIU Liang,PAN Lei,LU ZhenHua,CUI GuoChao,WANG ZhiQiang(),ZENG WenFang()   

  1. Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences/National Peach and Grape Improvement Center/Key Laboratory of Fruit Breeding Technology of Ministry of Agriculture and Rural Affairs, Zhengzhou 450009
  • Received:2021-09-29 Accepted:2022-01-12 Online:2022-06-01 Published:2022-06-16
  • Contact: ZhiQiang WANG,WenFang ZENG E-mail:1125351664la@gmail.com;wangzhiqiang@caas.cn;zengwenfang@caas.cn

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Abstract:

【Objective】 The aim of this study was to explore the differences between melting flesh and stony hard peaches at the peptide level and precursor protein level during fruit ripening, so as to provide a theoretical basis for mining the key peptides of determining or regulating the ripening process.【Method】 The characteristics of peptides and precursor protein functions in CN13 (melting flesh, MF) and CN16 (stony hard, SH) peaches were analyzed by peptidome, the relative contents of precursor proteins and peptides during MF and SH peach fruit ripening were compared, and the precursor proteins of different peptide segments were analyzed by function enrichment.【Result】 In this study, the peptides of CN13 and CN16 (S3 and S4III) were extracted for mass spectrometry. A total of 473 precursor proteins were identified, including 2 580 specific peptide sequences. The molecular weight, isoelectric point and cleavage sites of the peptide were summarized. In addition, the high-abundance precursor proteins corresponding to endogenous peptides were explored by COG function annotations and pathway enrichment analysis, and the results showed that the precursor proteins were mainly involved in the processes of general function prediction, post-translational modification, protein turnover, energy production and conversion, carbohydrate transport and metabolism. The enrichment analysis showed that the differential peptide precursor proteins of CN13 were related to biological processes, such as oxidation reduction, oxygen and oxygen and electron transport chain, which were mainly involved in glycolysis/gluconeogenesis, pentose phosphate pathway and RNA transport; the differential peptide precursor proteins in CN16 were related to biological processes, such as response to metal ion, response to inorganic substance, and response to cadmium ion, which were mainly involved in microbial metabolism in diverse environments, spliceosome and RNA transport; the differential peptide precursor proteins at the same stage in CN13 and CN16 at S4III were related to gene expression, translation and cellular macromolecular biological processes, which were mainly involved in RNA degradation, RNA transport and splicing.【Conclusion】 There were significant differences in peptides between CN13 and CN16 during fruit ripening. The precursor proteins of differential peptide were involved in starch/sucrose metabolism, glycolysis and ribosome synthesis, and it was suggested that these metabolic pathways were closely related to peach fruit ripening, which provided a theoretical reference for further exploring the key peptides of regulating peach fruit ripening and senescence.

Key words: peach, melting flesh, stony hard, fruit ripening, peptidomics

Table 1

Overview of the identification results of CN13 and CN16"

样本名称 Sample name 图谱数目 Spectrum number 多肽数目 Peptide number 蛋白数目 Protein number
CN13_S3 1855 1265 288
CN13_S4III 2121 1450 298
CN16_S3 2051 1359 313
CN16_S4III 1409 988 269
归并 Combined 7436 2580 473

Fig. 1

Reliability analysis of appraisal results"

Fig. 2

CN13 and CN16 polypeptide characteristics"

Fig. 3

Statistical distribution map of polypeptide cleavage sites"

Fig. 4

The COG function annotation analysis of the identified proteins in CN13 and CN16"

Table 2

The differential expressed proteins of CN13 and CN16 in different comparison groups"

CN13_S4III/CN13_S3 CN16_S4III/CN13_S4III CN16_S4III/CN16_S3
定量的蛋白数量Quantitative protein number 196 189 191
表达量显著差异蛋白数目Significant difference number 74 54 67
表达量显著上调蛋白数目Up-regulated number 42 23 42
表达量显著下调蛋白数目Down-regulated number 32 31 25

Table 3

The differential expressed peptide of CN13 and CN16 in different comparison groups"

CN13_S4III/CN13_S3 CN16_S4III/CN13_S4III CN16_S4III/CN16_S3
定量的多肽数量 Quantitative peptide number 642 576 555
表达量显著差异多肽数目 Significant difference number 88 72 65
表达量显著上调多肽数目 Up-regulated number 117 40 60
表达量显著下调多肽数目 Down-regulated number 205 112 125

Fig. 5

The KEGG enrichment analysis of the differential expressed proteins in CN13 and CN16 peach A：CN13_S4III vs CN13_S3;B：CN16_S4III vs CN16_S3;C：CN16_S4III vs CN13_S4III"

Table 4

KEGG pathyway of differential protein in CN13_S4III vs CN13_S3"

通路
Map name		 Uniprot号
Uniprot ID		 蛋白描述
Protein description		 差异倍数
Fold change
淀粉和蔗糖代谢
Starch and sucrose metabolism		 A0A251R9S2 葡萄糖-6-磷酸异构酶Glucose-6-phosphate isomerase 2.76
M4QFX1 UDP-葡萄糖6-脱氢酶UDP-glucose 6-dehydrogenase 2.14
M5XDI1 磷酸葡萄糖变位酶Phosphoglucomutase 0.21
糖酵解/糖异生
Glycolysis/Gluconeogenesis		 A0A251MRE3 含有ADH_zinc_N结构域的蛋白ADH_zinc_N domain-containing protein 2.19
M5VPR4 磷酸丙酮酸水合酶Phosphopyruvate hydratase 3.12
M5XFH2 果糖二磷酸醛缩酶Fructose-bisphosphate aldolase 2.97
核糖体
Ribosome		 A0A251NA79 未表征蛋白Uncharacterized protein 0.47
M5WAA2 含有RRM结构域的蛋白RRM domain-containing protein 0.20
M5WJM0 40S核糖体蛋白S25 40S ribosomal protein S25 2.38

Table 5

KEGG Pathyway of differential protein in CN16_S4III vs CN16_S3"

通路
Map name		 Uniprot号
Uniprot ID		 蛋白描述
Protein description		 差异倍数
Fold change
淀粉和蔗糖代谢Starch and sucrose metabolism M5XH82 葡萄糖-6-磷酸异构酶Glucose-6-phosphate isomerase 0.39
糖酵解/糖异生Glycolysis/Gluconeogenesis A0A251PJZ3 磷酸甘油酸激酶Phosphoglycerate kinase 0.44
M5XQ59 甘油醛-3-磷酸脱氢酶Glyceraldehyde-3-phosphate dehydrogenase 0.19
核糖体Ribosome A0A251NA79 为表征蛋白Uncharacterized protein 0.26
M5WAA2 含有RRM结构域的蛋白RRM domain-containing protein 0.14
A0A251MVP0 含有RRM结构域的蛋白RRM domain-containing protein 2.51

Table 6

KEGG pathyway of differential protein in CN16_S4III vs CN13_S4III"

通路
Map name		 Uniprot号
Uniprot ID		 蛋白描述
Protein description		 差异倍数
Fold change
淀粉和蔗糖代谢Starch and sucrose metabolism M5XH82 葡萄糖-6-磷酸异构酶Glucose-6-phosphate isomerase 0.35
糖酵解/糖异生Glycolysis/Gluconeogenesis A0A251PJZ3 磷酸甘油酸激酶Phosphoglycerate kinase 0.44
M5VPR4 磷酸丙酮酸水合酶Phosphopyruvate hydratase 0.44
核糖体Ribosome A0A251NA79 未表征蛋白Uncharacterized protein 0.34
M5WAA2 含有RRM结构域的蛋白RRM domain-containing protein 0.36
M5WJM0 40S核糖体蛋白S2540S ribosomal protein S25 0.34
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