Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (22): 4869-4879.doi: 10.3864/j.issn.0578-1752.2021.22.013

• HORTICULTURE • Previous Articles     Next Articles

Analysis of Changes in Phenolic Acids of Luffa cylindrica Pulp During Browning Based on Metabolomics

WANG YaHui1,2(),LIU XiaoHong1,YONG MingLi1,XIONG AiSheng2,SU XiaoJun1()   

  1. 1Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Nanjing 210014
    2College of Horticulture, Nanjing Agricultural University/State Key Laboratory of Crop Genetics and Germplasm Enhancement/Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing 210095
  • Received:2021-01-28 Accepted:2021-03-24 Online:2021-11-16 Published:2021-11-19
  • Contact: XiaoJun SU E-mail:2019204036@njau.edu.cn;xiaojunsu@jaas.ac.cn

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Abstract:

【Objective】Browning is the main element affecting the commercial value of Luffa cylindrica. The principal consideration of browning is linked to the metabolism of phenolic acids. Studying the changes of phenolic acid during the browning process of Luffa is helpful to understand the physiological mechanism of browning, so as to provide the basis for the storage and processing, breed selection and utilization of germplasm resources of Luffa. 【Method】Browning-resistant Luffa cultivar 2D-2 and browning-prone cultivar 35D-7 were used as experiment materials. Their browning degree, total phenol content and polyphenol oxidase activity after fresh cutting for different time were measured. Ultra high-performance liquid chromatography-mass spectrometry (UPLC-MS) was utilized to analyze the metabolomic of Luffa pulp before and after browning in boiling water bath. 【Result】 After fresh-cut browning treatment, the content of polyphenols and the polyphenol oxidase activity in the flesh of both two Luffa cultivars were increased. Luffa 35D-7 had severer browning degree. The total phenol content after 24 h storage was much more than 3 times of that of 2D-2, and the polyphenol oxidase activity achieved the top at 12 h, reaching 174.23 U·g-1·min-1. Based on metabolomics, the substrate of enzymatic browning reaction and the difference of metabolites in browning-prone Luffa 35D-7 before and after browning were analyzed. A total of 420 metabolites were detected, 229 of which were differential metabolites. After browning, the contents of 140 kinds of differential metabolites were substantially decreased, while 89 kinds were increased. KEGG pathway enrichment analysis demonstrated that metabolites related to phenylpropanoid biosynthesis, proline metabolism and flavonoid biosynthesis displayed significant changes. The results of qualitative and quantitative analysis of metabolomics suggested the content of p-coumaric acid was the highest in the phenolic acids identified in the flesh of the tested Luffa, followed by coniferin and gentisic acid. After browning treatment, 35 different metabolites were classified as phenolic acid metabolites. Among them, the content of coniferaldehyde, 3-aminosalicylic acid and phenethyl caffeate decreased most obviously, while the content of syringin, 2,5-dihydroxy benzoic acid O-hexside and isochlorogenic acid A increased significantly, and the log2 fold change was more than 10. 【Conclusion】The main phenolic substances in the flesh of 35D-7 were p-coumaric, and the main phenolic acid metabolites involved in the browning process of 35D-7 were coniferaldehyde, syringin and isochlorogenic acid A etc. The results could help further reveal the physiological mechanism of Luffa browning and provide basis for the improvement of Luffa variety.

Key words: Luffa, pulp, enzymatic browning, metabolomics, phenolic acids

Fig. 1

Browning of the Luffa pulp at different placement time after fresh cutting The white lines in the lower right corner represent 2 cm"

Fig. 2

Phenols content in Luffa pulp at different placement time after fresh cutting Different lowercase letters indicate significant differences at the P<0.05 level. The same as below"

Fig. 3

PPO enzyme activity in Luffa pulp at different placement time after fresh cutting"

Fig. 4

Overlap total ion chromatogram of all samples"

Fig. 5

Cluster heatmap of metabolite content changes during Luffa browning treatment"

Fig. 6

Enrichment of differential metabolite pathway during Luffa browning treatment"

Table 1

Partial differential metabolites during Luffa browning treatment"

KEGG通路
KEGG pathway		 代谢物
Metabolite		 结构式
Formula		 变化倍数
（褐变处理/对照）
Fold change (BR/CK)		 Log2变化倍数
Log2 Fold change		 变化趋势
Change trends
苯丙素的生物合成
Phenylpropanoid biosynthesis		 松柏醇 Coniferyl alcohol C10H12O3 2.06E-01 -2.28 下降 Down
咖啡酸 Caffeic acid C9H8O4 7.17E-02 -3.80 下降 Down
对香豆酸 p-coumaric acid C9H8O3 1.40E-01 -2.84 下降 Down
紫丁香苷 Syringin C17H24O9 1.93E+05 17.56 升高 Up
松柏醛 Coniferaldehyde C10H10O3 3.46E-05 -14.82 下降 Down
芥子酸 Sinapic acid C11H12O5 1.49E-01 -2.75 下降 Down
阿魏酸 Ferulic acid C10H10O4 9.54E-02 -3.39 下降 Down
脯氨酸代谢
Purine metabolism		 腺苷-3′,5′-环单磷酸水合物 Cyclic AMP C10H12N5O6P 4.00E+02 8.65 升高 Up
腺苷-3′-磷酸 3′-Aenylic acid C10H14N5O7P 8.32E+00 3.06 升高 Up
鸟苷3′,5′-环单磷酸 Guanosine 3′,5′-cyclic monophosphate C10H12N5O7P 4.95E+00 2.31 升高 Up
2′-脱氧鸟苷 Deoxyguanosine C10H13N5O4 5.65E+00 2.50 升高 Up
2′-脱氧腺苷 Deoxyadenosine C10H13N5O3 2.01E+02 7.65 升高 Up
鸟嘌呤 Guanine C5H5N5O 3.35E-01 -1.58 下降 Down
2′-脱氧腺苷-5′-单磷酸 2′-Deoxyadenosine-5′-monophosphate C10H14N5O6P 5.65E+00 2.50 升高 Up
腺苷 Adenosine C10H13N5O4 2.15E+02 7.75 升高 Up
黄嘌呤 Xanthine C5H4N4O2 2.93E-01 -1.77 下降 down
类黄酮生物合成
Flavonoid biosynthesis		 木犀草素 Luteolin C15H10O6 4.84E-05 -14.34 下降 Down
表儿茶素 L-epicatechin C15H14O6 4.68E-01 -1.10 下降 Down
短叶松素 Pinobanksin C15H12O5 1.16E-02 -6.42 下降 Down
柚皮素查尔酮 Naringenin chalcone C15H12O5 2.80E-02 -5.16 下降 Down
芹菜素 Apigenin C15H10O5 1.12E-05 -16.45 下降 Down

Fig. 7

Relative content of phenolic acid metabolites in Luffa pulp (Top 10)"

Fig. 8

Top 10 fold change differential metabolites of phenolic acids during Luffa browning treatment"

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