‘壶瓶枣’果皮着色物质及其相关基因筛选

doi:10.16420/j.issn.0513-353x.2023-0527

摘要/Abstract

摘要：

以‘壶瓶枣’（Ziziphus jujuba‘Hupingzao’）白熟期、半红期、全红期果实为试材，测定果皮主要色素物质含量并进行转录组测序，从分子水平阐述果皮着色基因的变化。结果表明，白熟期后，果皮花色苷含量开始显著降低；3个时期果皮类黄酮含量均依次显著降低；叶绿素含量在白熟期到半红期变化不大，全红期显著下降；类胡萝卜素含量在转色阶段相对稳定。类黄酮、花色苷、类胡萝卜素和叶绿素是‘壶瓶枣’果皮的着色物质，且花色苷分为飞燕草素、矢车菊素、矮牵牛素、锦葵色素、芍药色素和天竺葵色素等6种。根据转录组测序中GO和KEGG富集分析发现，在类黄酮、花色苷、黄酮与黄酮醇、类胡萝卜素和卟啉代谢通路中差异基因个数分别为35、22、6、30和38。其中，白花青素还原酶基因（LOC107429483）、转录因子基因MYC2（LOC107430586）和二氢黄酮醇4-还原酶基因（LOC107420289）在果皮着色中发挥主要作用，β-胡萝卜素羟化酶基因（LOC107414761）是影响类胡萝卜素合成的关键基因；卟啉代谢中叶绿体A加氧酶基因（LOC107410684）是影响总叶绿素含量的关键基因。

关键词: 枣, 着色, 转录组, 类黄酮, 花色苷

Abstract:

The white ripening，semi-red and whole red fruits of Ziziphus jujuba‘Hupingzao’as materials，the contents of the main pigment substances in the peel were determined and transcriptome sequencing was carried out to elaborate the changes of coloring genes in the peel of jujube at the molecular level. The results showed that after the white ripening period，the anthocyanin content of jujube peel began to decrease significantly. In the three periods，the flavonoid content of jujube peel decreased significantly. The chlorophyll content did not change much from the white maturity stage to the semi-red stage，and decreased significantly at the full red stage. The content of carotenoids was relatively stable in the veraison stage. Flavonoids，anthocyanins，carotenoids and chlorophyll are the coloring substances of jujube peel，and the anthocyanins are divided into six types：delphinidin，cyanidin，petunidin，malvidin，peonidin and pelargonidin. According to the GO and KEGG enrichment analysis in transcriptome sequencing，the number of differential genes in flavonoids，anthocyanins，flavonoids and flavonols，carotenoids and porphyrin metabolic pathways were 35，22，6，30 and 38，respectively. Among them，the white anthocyanidin reductase gene（LOC107429483），transcription factor gene MYC2（LOC107430586）and dihydroflavonol 4-reductase gene（LOC107420289）played a major role in the coloring of jujube peel，and β-carotene hydroxylase gene（LOC107414761）was the key gene affecting carotenoid synthesis. The chloroplast A oxygenase gene（LOC107410684）in porphyrin metabolism is a key gene affecting total chlorophyll content.

Key words: Ziziphus jujuba, coloring, RNA-Seq, flavonoids, anthocyanins

李洁, 武超, 贾祥堑, 王娟. ‘壶瓶枣’果皮着色物质及其相关基因筛选[J]. 园艺学报, 2024, 51(8): 1728-1742.

LI Jie, WU Chao, JIA Xiangqian, WANG Juan. Screening of Ziziphus jujuba‘Hupingzao’Fruit Coloring Substances and Their Related Genes[J]. Acta Horticulturae Sinica, 2024, 51(8): 1728-1742.

图/表 13

图1 ‘壶瓶枣’白熟期（左）、半红期（中）和全红期（右）果实

Fig. 1 The white ripening（left），semi-red（middle）and whole red（right）fruits of Ziziphus jujuba‘Hupingzao’

图2 ‘壶瓶枣’3个时期果皮花色苷和类黄酮含量的变化不同小写字母表示不同时期同一指标差异显著（P < 0.05）。下同。

Fig. 2 Changes in the contents of anthocyanins and flavonoids in the peel of Ziziphus jujuba‘Hupingzao’in three periods Different lowercase letters indicated significant differences in the same index in different periods（P < 0.05）.The same below.

图3 ‘壶瓶枣’3个时期果皮叶绿素和类胡萝卜素含量的变化

Fig. 3 Changes in chlorophyll and carotenoid contents in the peel of Ziziphus jujuba‘Hupingzao’in three stages

表1 ‘壶瓶枣’果皮中黄酮类物质种类及其含量

Table 1 The content of various flavonoid substances in three kinds of Ziziphus jujuba‘Hupingzao’fruit

化合物种类 Compound type	含量/（ng · mL^-1） Content	化合物种类 Compound type	含量/（ng · mL^-1） Content
矢车菊素双葡萄糖苷Cyanidin 3,5-diglucoside	—	天竺葵色素Pelargonidin 锦葵色素Malvidin	6.566 15.392
矢车菊素半乳糖苷Cyanidin 3-galactoside	2.809	芍药色素Peonidin	7.794
飞燕草素Delphinidin	—	芦丁（芸香苷）Rutin	8.714
原花色素B4 Procyanidin B4	5.720	木犀草素Luteolin	136.234
矢车菊素Cyanidin	21.938	槲皮素Quercetin	44.376
双聚原矢车菊甘元Procyanidin B2	7.952	异鼠李素Isorhamnetin	32.522
矮牵牛色素Petunidin	23.033	山奈酚Kaempferol	31.923

表2 样品测序数据概述

Table 2 Overview of sample sequencing data

时期 Stage	样本 Sample	原始碱基数 Total bases	样本碱基总数/Gb Total giga bases	有效序列数 Total reads	Q20/%	Q30/%	比对率 Overall mapping rate
白熟期White ripening	1	6 469 880 608	6.47	44 889 082	97.9148	93.7792	0.946
	2	5 857 844 975	5.86	40 652 034	98.0241	94.0759	0.944
	3	6 166 062 009	6.17	42 803 420	97.8849	93.6980	0.944
半红期Semi-red	1	5 757 384 355	5.76	39 806 298	98.3328	94.8011	0.952
	2	6 114 634 623	6.11	42 321 102	98.0420	94.0131	0.951
	3	6 452 091 835	6.45	44 670 074	98.1427	94.3092	0.938
全红期Whole-red	1	5 731 716 448	5.73	39 656 256	98.1848	94.4039	0.953
	2	5 676 944 756	5.68	39 237 730	98.2798	94.6434	0.952
	3	6 040 803 819	6.04	41 780 350	98.2849	94.6781	0.954

图4 ‘壶瓶枣’3个时期果皮差异表达基因数 BSQ：白熟期；BHQ：半红期；QHQ：全红期。

Fig. 4 The number of differentially expressed genes in the Ziziphus jujuba‘Hupingzao’in the three stages BSQ：White ripening；BHQ：Semi-red；QHQ：Whole red.

图5 ‘壶瓶枣’3个时期果皮差异基因GO注释

Fig. 5 Three stages of GO annotation of Ziziphus jujuba‘Hupingzao’DEGs

图6 ‘壶瓶枣’白熟期与半红期果皮差异基因KEGG富集散点图

Fig. 6 Map of KEGG concentration and distribution point of different genes in pericarp of Ziziphus jujuba‘Hupingzao’at white ripening stage and semi-red stage

图7 ‘壶瓶枣’半红期与全红期果皮差异基因KEGG富集散点图

Fig. 7 Map of KEGG concentration and distribution point of pericarp differential gene in semi-red stage and whole red stage of Ziziphus jujuba‘Hupingzao’

图8 ‘壶瓶枣’白熟期与全红期果皮差异基因KEGG富集散点图

Fig. 8 Map of KEGG concentration and distribution point of different genes in pericarp of Ziziphus jujuba‘Hupingzao’ during white ripening stage and whole red stage

图9 KEGG富集到的不同时期‘壶瓶枣’果皮着色相关代谢通路中差异表达基因热图

Fig. 9 Heat map of differentially expressed genes in metabolic pathways related to Ziziphus jujuba‘Hupingzao’peel coloring at different stages enriched by KEGG

图10 KEGG富集到的‘壶瓶枣’不同时期果皮着色差异表达基因的聚类分析

Fig. 10 Cluster analysis of differentially expressed genes in Ziziphus jujuba‘Hupingzao’ peel coloring at different stages of KEGG enrichment

图11 花色苷合成通路图 3个色块从左到右表示白熟期、半红期和全红期的表达量。

Fig. 11 Diagram of anthocyanin synthesis pathway Three color blocks from left to right represent the expression of white ripe stage，semi-red stage and whole red stage.

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