Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (23): 4728-4742.doi: 10.3864/j.issn.0578-1752.2022.23.013

• Food Science and engineering • Previous Articles     Next Articles

Physiological Metabolic Rol e of Nanocomposite Packaged Agaricus bisporus During Postharvest Cold Storage Analyzed by TMT-Based Quantitative Proteomics

WANG Chao1(),FANG DongLu2,ZHANG PanRong1,JIANG Wen1,PEI Fei1,HU QiuHui1,MA Ning1,*()   

  1. 1College of Food Science and Engineering, Nanjing University of Finance and Economics/Jiangsu Collaborative Innovation Center for Modern Grain Circulation and Safety/Key Laboratory of Grains and Oils Quality Control and Processing, Nanjing 210023
    2College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037
  • Received:2022-03-09 Accepted:2022-05-25 Online:2022-12-01 Published:2022-12-06
  • Contact: Ning MA;


【Background】 Agaricus bisporus (A. bisporus) is prone to quality deterioration, such as umbrella opening, water loss, and browning after harvest, which seriously affects the storage quality and commercial value. Our previous research has confirmed that the nanocomposite packaging material (Nano-PM) could effectively delay the postharvest quality deterioration of A. bisporus, but the preservation mechanism is still unclear. 【Objective】 In this study, the differentially expressed proteins of A. bisporus in Nano-PM and polyethylene packaging material (Normal-PM) during storage were analyzed by Tandem Mass Tag (TMT) quantitative proteomics technology. The preservation mechanism of Nano-PM on A. bisporus was further explored. 【Method】 A. bisporus was taken as the research object. The Nano-PM was used for the preservation of A. bisporus, and the Normal-PM was used as the control. The protein extraction and trypsin hydrolysis were performed on A. bisporus during storage. The differentially expressed proteins were screened by TMT labeling and liquid chromatography-tandem mass spectrometry detection. Combined with bioinformatics analysis, the main metabolic pathways involved in differential proteins were studied. Quantitative real-time polymerase chain reaction (qPCR) technology was used to determine the gene expression levels of differential proteins..【Result】 The Nano-PM effectively maintained the appearance quality of A. bisporus and delayed the increase of cell membrane permeability. The number of differential proteins in two groups increased during storage. In the middle (6 d) and late (10 d) stages of storage, the numbers of differential proteins were 62 and 148, respectively. Among them, 22 differential proteins were common. Combined with bioinformatics analysis, these differential proteins were mainly related to pathways, such as energy metabolism and lipid metabolism. The lipid metabolism pathway was mainly analyzed, and the results showed that the Nano-PM had a regulatory effect on the membrane lipid metabolism of A. bisporus. Compared with the Normal-PM, the protein expression of fatty acid synthase, phosphorylcholine cytidylyltransferase, and phosphatidic acid phosphatase under Nano-PM were up-regulated, while the protein expression of key enzymes in membrane lipid degradation, such as phospholipase D and lipase, were down-regulated. At the gene level, the expression of genes encoding these proteins were consistent with the proteomics results..【Conclusion】 The differential proteins of different packaged A. bisporus during storage could be screened and analyzed by TMT-based quantitative proteomics technology. Nano-PM regulated the membrane lipid metabolism of A. bisporus, and inhibited the expression of membrane lipid degradation-related enzymes, which effectively delayed the increase in cell membrane permeability of A. bisporus, maintained the structure and function of the cell membrane, and delayed the quality deterioration of A. bisporus during storage.

Key words: nanocomposite packaging, Agaricus bisporus, preservation, proteomics

Table 1

Primer sequences used for qPCR"

引物名称Primer name 引物序列Primes sequence (5′-3′)

Table 2

qPCR condition settings"

步骤 Step 循环次数 Cycle 温度 Temperature (℃) 时间 Time (s)
预变性 Initial denaturation 1 95 30
循环反应 Cycling stage 40 95 10
60 30
融解曲线 Melt curve 1 95 15
60 60
95 15

Fig. 1

Flow chart of TMT quantitative proteomics analysis of metabolic law of A. bisporus with different packages"

Fig. 2

Changes in appearance quality (A) and cell membrane permeability (B) of A. bisporus during storage in different packages *indicate sinificant difference (P<0.05). The same as below"

Fig. 3

Quantities of differentially expressed proteins during storage of A. bisporus in different packages A: The differentially expressed proteins between the Nano-PM group and the Normal-PM group at 6 d; B: The differentially expressed proteins between the Nano-PM group and the Normal-PM group at 10 d"

Fig. 4

Differentially expressed proteins of A. bisporus during storage in different packaging groups A: The volcano plot of differentially expressed proteins between the Nano-PM group and the Normal-PM group at 6 d; B: The volcano plot of the differentially expressed proteins between the Nano-PM group and the Normal-PM group at 10 d; C: Venn diagram of differentially expressed proteins between Nano-PM group and Normal-PM group during storage. N: Nano-PM group; P: Normal-PM group; CK: Fresh group"

Fig. 5

GO (A) and KEGG (B) analysis of differentially expressed proteins during storage of A. bisporus in different packaging groups"

Table 3

Main differentially expressed proteins of A. bisporus during storage in different packaging groups"

Protein accession
Protein description
Gene name
差异倍数Fold change
6N/6P 10N/10P
XP_006456433.1 吡喃糖脱氢酶受体蛋白Pyranose dehydrogenase receptor protein AGABI1DRAFT_116682 0.58 /
XP_006457863.1 鸟嘌呤核苷酸结合蛋白Guanine nucleotide-binding protein AGABI1DRAFT_82206 1.95 1.82
XP_006458170.1 核糖体生物合成蛋白Ribosome biogenesis protein AGABI1DRAFT_33715 / 0.55
XP_006459487.1 未表征蛋白质Uncharacterized protein AGABI1DRAFT_114418 0.58 /
XP_006460807.1 PRA家族蛋白PRA family protein AGABI1DRAFT_112435 0.62 0.55
XP_006463403.1 未表征蛋白质Uncharacterized protein AGABI1DRAFT_108190 / 1.68
XP_006454671.1 膜组分蛋白Membrane component protein AGABI1DRAFT_103710 1.76 2.09
XP_006453769.1 未表征蛋白质Uncharacterized protein AGABI1DRAFT_124747 0.53 0.64
XP_006453957.1 几丁质合成酶Chitin synthase AGABI1DRAFT_52110 / 0.59
XP_006460122.1 未表征蛋白质Uncharacterized protein AGABI1DRAFT_127941 0.55 0.66
XP_006462684.1 磷脂酶D Phospholipase D AGABI1DRAFT_58967 0.68 0.51
XP_006454339.1 ATP激酶ATP kinase AGABI1DRAFT_110490 1.65 1.51
XP_006454403.1 膜组分蛋白Membrane component protein AGABI1DRAFT_66797 1.54 1.65
XP_006456787.1 磷脂酸磷酸酯酶Phosphatidic acid phosphatase AGABI1DRAFT_127573 1.52 1.67
XP_006455355.1 超氧化物歧化酶Superoxide dismutase AGABI1DRAFT_114765 1.82 1.56
XP_006454789.1 线粒体膜转运蛋白Mitochondrial membrane transporter AGABI1DRAFT_65989 1.77 1.59
XP_006455653.1 未表征蛋白质Uncharacterized protein AGABI1DRAFT_115714 / 1.56
XP_006459063.1 脂肪酸合酶Fatty acid synthase AGABI1DRAFT_61049 1.61 1.87
XP_006454643.1 膜组分蛋白Membrane component protein AGABI1DRAFT_50813 1.92 1.73
XP_006453790.1 脂肪酶蛋白Lipase protein AGABI1DRAFT_67808 0.65 0.57
XP_006455684.1 ATP结合蛋白ATP-binding protein AGABI1DRAFT_102264 1.64 1.55
XP_006456057.1 乌头酸水合酶Aconitate hydratase AGABI1DRAFT_101681 / 1.82
XP_006456371.1 膜组分蛋白Membrane component protein AGABI1DRAFT_46804 / 2.12
XP_006458132.1 DNA解旋酶活性蛋白DNA helicase active protein AGABI1DRAFT_118129 0.36 0.59
XP_006457745.1 膜水解酶活性蛋白Membrane hydrolase active protein AGABI1DRAFT_124786 0.41 0.61
XP_006459487.1 膜转运蛋白Membrane transporter AGABI1DRAFT_114418 0.62 0.48
XP_006453826.1 CTP活性蛋白CTP active protein AGABI2DRAFT_181860 1.56 1.68
XP_006459152.1 柠檬酸合酶蛋白Citrate synthase protein AGABI1DRAFT_129944 0.64 0.56
XP_006454232.1 RNA结合蛋白RNA binding protein AGABI1DRAFT_96937 1.59 1.52
XP_006457601.1 氧化还原酶活性Oxidoreductase active protein AGABI1DRAFT_116005 0.36 0.52
XP_006453831.1 组蛋白H3 Histone H3 AGABI1DRAFT_52286 / 1.87

Fig. 6

Membrane lipid metabolism pathway and differential protein expression of A. bisporus during storage in different packaging groups Red means up-regulation, and green means down-regulation"

Fig. 7

qPCR analysis of differential proteins in membrane lipid metabolism pathway during storage of A. bisporus in different packaging groups"

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