Scientia Agricultura Sinica ›› 2024, Vol. 57 ›› Issue (13): 2635-2650.doi: 10.3864/j.issn.0578-1752.2024.13.011

• HORTICULTURE • Previous Articles     Next Articles

Differential Analysis of Aroma Substance Content and Gene Expression in the Berry Skins of Different Grape Germplasms

XU MengYu1(), WANG JiaYang1, WANG JiangBo2, TANG Wen1, CHEN YiHeng1, SHANGGUAN LingFei1, FANG JingGui1, LU SuWen1()   

  1. 1 College of Horticulture, Nanjing Agricultural University, Nanjing 210095
    2 College of Plant Science, Tarim University, Alar 843300, Xinjiang
  • Received:2023-11-22 Accepted:2024-02-02 Online:2024-07-09 Published:2024-07-09
  • Contact: LU SuWen

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Abstract:

【Objective】 This study aimed to explore the differences in the types, content, and related gene expression levels of volatile aroma compounds in the mature grape skins of different grape germplasms, so as to provide a reference basis for the analysis of the metabolic regulation mechanism of grape volatile aroma substances and the full utilization of aroma resources. 【Method】 In this study, GC-MS was used to identify the types and content of volatile aroma substances in the pericarp of 13 grape germplasms. The transcriptional level of the entire genome was also analyzed by using Illumina high-throughput sequencing platform.【Result】 A total of 154 volatile aroma substances were detected in the 13 grape germplasms, mainly including aldehydes, esters, terpenoids and other compounds. The germplasm with the highest content of volatile aroma substances in the skin was Kyoho, while the lowest was Cabernet Sauvignon. Esters were the most abundant volatile aroma substances in the skins of Kyoho and Petit Verdot, while aldehydes were the most abundant volatile aroma substances in the other germplasms skins. Twelve characteristic aroma substances such as hexanal, ethyl caprylate and linalool contributed more to the aroma of grape skins. The results of principal component analysis showed that there were large differences in volatile aroma substances in the skins of Kyoho, Thompson Seedless, Red Globe and other germplasm. The results of transcriptome analysis showed that the number of differentially expressed genes (DEGs) varied considerably among different germplasms. DEGs were significantly enriched in the pathways of linoleic acid metabolism, α-linolenic acid metabolism, phenylalanine metabolism, tyrosine metabolism, and monoterpene biosynthesis. The expressions of lipoxygenase (LOX), alcohol acyltransferase (AMAT), deoxyd-xylose-5-phosphate reductase (DXS), alcohol dehydrogenase (ADH) and other genes were highly correlated with the content of aroma compounds. Further analysis revealed that the transcription factors WRKY7, WRKY28, ARF4, and ARF22 were potential regulators of aroma synthesis in grapes.【Conclusion】 The content of aroma substances and the expression of related genes in the pericarp of different grape germplasm varied greatly, and the results of the joint multi-omics analysis provided a reference for further research on the synthesis mechanism of volatile aroma substances in grapes.

Key words: grape (Vitis vinifera), skin, aroma, volatile aroma substance, metabolome, transcriptome

Table 1

Detailed information of 13 grape germplasms"

序号
Number		 葡萄品种名称
Name of grape variety		 属性
Attribute		 果皮颜色
Skin color		 类型
Type
1 无核白Thompson Seedless 欧亚Eurasian 白White 鲜食 Table grape
2 阳光玫瑰Shine Muscat 欧美Europe and America 金黄Golden yellow 鲜食 Table grape
3 红地球Red Globe 欧亚Eurasian 紫红Purplish red 鲜食Table grape
4 玫瑰香Muscat Hamburg 欧亚Eurasian 紫红Purplish red 鲜食Table grape
5 巨峰Kyoho 欧美Europe and America 紫黑Atropurpureus 鲜食Table grape
6 贵人香Italian Riesling 欧亚Eurasian 白White 酿酒Wine grape
7 小芒森Petit Manseng 欧亚Eurasian 白White 酿酒Wine grape
8 威代尔Vidal 欧美Europe and America 白White 酿酒Wine grape
9 赤霞珠Cabernet Sauvignon 欧亚Eurasian 紫黑Atropurpureus 酿酒Wine grape
10 蛇龙珠Cabernet Gernischet 欧亚Eurasian 紫黑Atropurpureus 酿酒Wine grape
11 梅鹿辄Merlot 欧亚Eurasian 紫黑Atropurpureus 酿酒 Wine grape
12 小味儿多Petit Verdot 欧亚Eurasian 紫黑Atropurpureus 酿酒 Wine grape
13 马瑟兰Marselan 欧亚Eurasian 深蓝Dark blue 酿酒 Wine grape

Fig. 1

Berries of different grape cultivars at the maturation TS: Thompson Seedless; SM: Shine Muscat; RG: Red Globe; MH: Muscat Hamburg; KY: Kyoho; IR: Italian Riesling; PM: Petit Manseng; VD: Vidal; CS: Cabernet Sauvignon; CG: Cabernet Gernischet; ML: Merlot; PV: Petit Verdot; MS: Marselan. The same as below"

Table 2

Primer sequence"

基因名称
Gene name		 基因号
Gene ID		 正向引物
Forward primer (5′-3′)		 反向引物
Reverse primer (5′-3′)
LOXA VIT_06s0004g01510 TTTTCTTGGTTTAGGGATGA CTTTGATTTGCCTTACTTCG
LOXC VIT_14s0128g00780 TGAGCCTACCACATCCTAAT TAGCCAGAGTCATTCACAGC
HPL VIT_12s0059g01060 CCGTCCCACTCCAATACGCT TTCGGGTCTGTCATCGCCAC
ADH1 VIT_18s0001g15410 AGCCATTAGTGATAGAAGAAGTG GACATAGGGAGGTGTAGAGGA
LOXO VIT_09s0002g01080 CTCAACGACCCACAACGCTAC CGCACCTGTTTCTTCGGCTA
DXS VIT_05s0020g02130 TTTGGTTGCGTCTTCTTTGC GGACTCCACTTTGTTGTGCC

Table 3

Aroma thresholds for characteristic aroma substances"

化学成分
Compound		 阈值[15,20]
Threshold (μg∙L-1)		 香气特征[15,20-21]
Aromatic characteristic
正己醛 Hexanal 4.500 绿色蔬菜 Green vegetables
2-己烯醛 2-hexenal 17.000 青草，草本 Grass, herbaceous
反式-2-壬烯醛 Trans-2-Nonenal 0.080 潮湿的，土壤味 Moist, soil odor
(E,Z)-2,6-壬二烯醛 (E,Z)-2,6-nonadienal 0.020 紫罗兰和黄瓜香味 Violet and cucumber aromas
壬醛 Nonanal 1.000 脂肪，柑橘，青菜，果味 Fat, citrus, vegetables, fruity taste
异丁酸乙酯 Ethyl isobutyrate 0.100 果香 Fruity aroma
正己酸乙酯 Ethyl caproate 1.000 水果，青苹果，香蕉，葡萄酒，白兰地 Fruits, green apples, bananas, wine, brandy
辛酸乙酯 Ethyl octanoate 0.100 甜味，花香，水果味，香蕉，梨，白兰地 Sweet, floral, fruity, banana, pear, brandy
芳樟醇 Linalool 6.000 柑橘，花香，甜味 Citrus, floral aroma, sweet taste
香叶醇 Geraniol 10.000 柠檬味，花香，橙子味，玫瑰味 Lemon flavor, floral aroma, orange flavor, rose flavor
(E)-氧化玫瑰 (E)-Oxidized roses 0.100 玫瑰香，天竺兰香 Rose fragrance, geranium fragrance
β-紫罗兰酮 β-ionone 0.007 脂膏香，玫瑰香味，紫罗兰香 Cream fragrance, rose fragrance, violet fragrance

Fig. 2

Characters of volatile substances in grape berry skins among different cultivars A: Types of volatile aroma substances; B: Content of volatile aroma substances; C: Content of volatile aroma substances in each metabolic pathway; D: OAV of volatile aroma substances"

Fig. 3

Principal component analysis of volatile aroma substances in different grape cultivars"

Table 4

Quality assessment of grape skin transcriptome sequencing"

样品 Sample 原始数据 Raw reads 高质量数据 Clean reads Q20 (%) Q30 (%) GC (%)
TS 7509114100 7377606868 97.63 93.23 46.38
SM 7984925800 7885274082 97.64 93.11 45.46
RG 6797327000 6629690349 97.57 93.21 46.49
MH 6863553200 6699407763 97.57 93.00 46.20
KY 6529118900 6378759691 97.83 93.82 46.20
IR 8716216500 8624849243 97.62 93.02 45.81
PM 6548941700 6424235773 97.72 93.61 46.40
VD 7417100700 7300033789 97.55 93.14 46.24
CS 7019247100 6845200698 97.39 92.48 46.67
CG 6581499100 6444723077 97.69 93.63 46.12
ML 7391406100 7271554342 97.56 93.24 46.43
PV 8206753300 8086178375 97.63 93.15 46.36
MS 7488201600 7299543929 97.36 92.31 46.20

Fig. 4

Statistics of DEGs among different grape cultivars A: DEGs; B: Down-regulated DEGs; C: Up-regulated DEGs; D: Venn diagram of DEGs"

Fig. 5

KEGG enrichment pathways of DEGs in five comparison groups"

Fig. 6

Metabolic pathways and related genes’ expression of aroma in grape A: Fatty acid metabolic pathway; B: Amino acid metabolic pathway; C: Terpenoid synthetic pathway; D: Heat map of expression of aroma-related genes"

Fig. 7

Relative expression levels of aroma related genes in grape"

Fig. 8

WGCNA analysis A: Hierarchical cluster tree illustrating the modules identified by WGCNA; B: KEGG enrichment analysis of genes in the darkorange module; C: Diagram of the gene regulatory network"

Fig. 9

Conjoint analysis of transcriptome and metabolome A: Association between characteristic aroma substances and DEGs; B: The O2PLS loading plots for volatile aroma substances; C: The O2PLS loading plots for DEGs. The top ten main aromatic substances and DEGs that had greater effect on the other omics were indicated by red dots"

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