壶瓶枣低温贮藏期间红变规律及转录组学分析

    Reddening and transcriptome analysis of Huping jujube during storage at low temperature

    • 摘要: 为探究枣果采后贮藏过程红变原因,该研究以半红期壶瓶枣为试材,测定枣果冷藏过程中色素物质、酚类物质以及褐变相关酶的变化并进行转录组学分析,从分子水平研究低温贮藏过程中壶瓶枣色泽变化机制。结果表明:低温贮藏有效延缓了枣果叶绿素、总酚、总黄酮、原花青素、可溶性糖的降解,减缓了多酚氧化酶(polyphenol oxidase,PPO)活性上升,有效抑制了枣果转色。贮藏过程中枣果红变与果实着色物质以及褐变反应相关,通过GO(gene ontology)和KEGG (kyoto encyclopedia of genes and genomes)富集分析发现低温贮藏枣果差异基因主要集中在细胞成分(cellular component,CC)和分子功能(molecular function,MF)部分,其中与枣果转色相关的差异基因主要集中在生物过程(biological process,BP);在卟啉和叶绿素代谢以及黄酮类化合物生物合成代谢途径中, 反式肉桂酸4-单加氧酶基因(LOC107411586、LOC107417662)、黄酮醇合酶基因(LOC107415994)、类黄酮3’,5’-羟化酶1-like基因(LOC107403962)参与氧化还原过程,调控黄酮类化合物生物合成,查尔酮异构酶样蛋白2基因(LOC107413997)调控多酚类化合物生物合成,原叶绿素还原酶基因(LOC107432449)参与新陈代谢过程,与卟啉、叶绿素a合成相关,这些基因在枣果贮藏红变过程中发挥重要作用。该研究可为壶瓶枣采后贮藏色泽品质调控提供理论依据。

       

      Abstract: This study aims to explore the effect of low-temperature storage on the color change of Huping jujube. The natural half-red Huping jujube was taken as the raw material and then stored at 0°C (0±1°C) low temperature, while setting 18 °C (18±2°C) room temperature storage as control. A series of experiments were carried out to explore the color mechanism of Huping jujube. Therefore, the physiological and biochemical indexes were monitored to determine the combination of pigment and browning, including the color difference (L, a, b, and △E), chlorophyll, total phenols, total flavonoids, proanthocyanidins, soluble sugar content, and polyphenol oxidase (PPO) activity. The samples were selected for transcriptome sequencing. The results showed that the state of low-temperature storage Huping jujube was significantly better than that of normal-temperature storage. The color of the jujube fruit peel gradually deepened with the extension of storage time, and the chlorophyll was degraded continuously. The contents of total phenols, total flavonoids, and soluble sugar decreased gradually, indicating that the transformation of fruit color was related to its tissue oxidation. The gradual increase of PPO activity accelerated the consumption of total polyphenols in tissues. The transcriptome analysis showed that the differential genes annotated by GO were concentrated mainly in the cellular component (CC) and molecular function (MF). The differentially expressed genes related to color were concentrated in the biological process (BP); The pathways annotated by KEGG were closely related to Porphyrin and chlorophyll metabolism, Flavonoid biosynthesis, and metabolism. A total of 18 differentially expressed genes with high expression levels were further screened. Among them, the expression levels of trans-cinnamate 4-monooxygenase gene (LOC107411586, LOC107417662), flavonol synthase gene (LOC107415994), flavonoid 3',5'-hydroxylase 1-like gene (LOC107403962) were up-regulated, whereas, the expression level of chalcone isomerase-like protein 2 gene (LOC107413997) was down-regulated. The expression levels of the porphobilinogen deaminase gene (LOC107405622), uroporphyrinogen-III synthase gene (LOC107420915), uroporphyrinogen decarboxylase gene (LOC107413960), oxygen-dependent coproporphyrinogen-III oxidase gene (LOC107416652), magnesium-chelatase subunit Chll gene (LOC107411201), magnesium-chelatase subunit ChlH gene (LOC107431688), magnesium protoporphyrin IX methyltransferase gene (LOC107407353), magnesium-protoporphyrin IX monomethyl ester oxidative cyclase gene ( LOC107423584 ), protochlorophyllide reductase gene (LOC107432449), and glutamate-tRNA ligase gene (LOC107411689) were all down-regulated. Trans-cinnamate 4-monooxygenase gene (LOC107411586, LOC107417662), flavonol synthase gene (LOC107415994), and flavonoid 3',5'-hydroxylase 1-like gene (LOC107403962) were involved in the oxidation-reduction as the key genes for the biosynthesis of flavonoids. The flavonoid 3',5'-hydroxylase 1-like gene (LOC107403962) was a key gene for anthocyanin synthesis. The up-regulation of its expression led to the up-regulation of the expression of dihydroquercetin, the precursor of anthocyanin synthesis, leading to the content of anthocyanin. The chalcone isomerase-like protein 2 gene (LOC107413997) was a key gene for the biosynthesis of polyphenolic compounds. The down-regulation of its expression led to the down-regulation of the precursor of polyphenolic compounds, which ultimately dominated the content of polyphenolic compounds. The protochlorophyllide reductase gene (LOC107432449) was involved in the metabolic process as the key gene for chlorophyll synthesis. The down-regulation of its expression directly dominated the biosynthesis of chlorophyll a. At the same time, the expression of differential genes in the chlorophyll synthesis metabolic pathway was down-regulated, and the expression of most differential genes related to anthocyanin synthesis showed an upward trend, indicating that the content of chlorophyll and polyphenols decreased in the later stage of storage. The up-regulation of anthocyanin synthesis led to the color change of jujube fruit in the later stage of storage. This finding can provide a theoretical basis for the variation in the color quality of Huping jujube.

       

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