Full
Outline
To investigate the dynamic changes of metabolites in Betula pendula Roth juice during the harvesting period, and analyze their intrinsic associations with physicochemical indices including soluble solid content,pH and electrical conductivity.
Betula pendula Roth juice collected on 7 consecutive days from the Yakeshi production region in Inner Mongolia, China, was pretreated by methanol-water (7:3, V:V) ultrasonic extraction in an ice-water bath, centrifugation and 0.22 μm membrane filtration, and was then analyzed by ultra performance liquid chromatography-quadrupole-orbitrap high resolution mass spectrometry (UPLC-Q-Exactive-MS) based untargeted metabolomics. Principal component analysis (PCA) was used to examine the overall variation of the metabolome,differential metabolites were screened with variable importance in projection (VIP)>1 and P<0.05, key metabolic pathways were resolved by UpSet plots, volcano plots and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis, and the soluble solid content, pH and electrical conductivity were determined to construct a metabolite-physicochemical index correlation network by Spearman rank correlation analysis.
The results showed that the metabolome of Betula pendula Roth juice could be divided into 3 stages along the harvesting period:D1–D3, stress-responsive metabolites such as alkaloids and organic acids were up-regulated, with log2FC of +0.88 for pilocarpine and +2.68 for alpha-ketoglutaric acid, and as many as 119 kinds of differential metabolites (41 kinds of them unique to this comparison) were detected between D1 and D2; D4–D6, carbohydrates, lipids and amino acids predominated, the pentose phosphate pathway, starch and sucrose metabolism and phenylpropanoid biosynthesis were significantly enriched and alpha-linolenic acid (log2FC=+4.21) and methyl jasmonate (log2FC=+2.95) peaked at this stage; D7, most functional metabolites were down-regulated whereas aroma-associated benzenoids increased selectively,and only 4 kinds of differential metabolites were detected between D6 and D7, indicating that the metabolome tended to be stable. During the harvesting period, the pH, electrical conductivity and soluble solid content of Betula pendula Roth juice ranged from 4.11 to 6.99, from 211.16 to 608.40 μS/cm and from 0.80 to 2.66 °Brix, respectively; the correlation network showed that the soluble solid content was strongly and positively correlated with various carbohydrates and organic acids, and DL-malic acid and acetic acid acted as metabolic hub compounds simultaneously linking multiple physicochemical indices.
The metabolite composition of Betula pendula Roth juice undergoes a clearly stage-dependent evolution within a harvesting period of only 7 d, D4–D6 represent the stage richest in functional metabolites, and DL-malic acid and acetic acid can serve as candidate markers for quality evaluation; these findings provide a theoretical basis for scientifically defining the harvesting window and for the quality evaluation and high-value utilization of Betula pendula Roth juice.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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