橄榄氨基酸组分差异对鲜食品质的潜在影响

doi:10.13560/j.cnki.biotech.bull.1985.2026-0148

摘要/Abstract

摘要：

目的揭示橄榄（Canarium album Raeusch.）种质间鲜食品质差异形成的影响因素，为优异鲜食种质筛选与品种改良提供理论依据。方法通过整合代谢组和转录组分析，研究24份不同类型橄榄种质资源的代谢物谱和基因表达谱差异，同时采用RT-qPCR对候选调控基因进行表达模式验证。结果从鲜食与非鲜食橄榄种质资源群体间共鉴定出152种差异富集代谢物（differentially accumulated metabolites, DAMs），主要包括氨基酸、酚酸、萜类、类黄酮、有机酸和糖类等。基于DAMs，共富集到16条氨基酸生物合成或代谢相关的KEGG代谢途径。进一步分析发现，鲜味氨基酸（天冬氨酸和谷氨酸）及芳香族氨基酸（苯丙氨酸）在鲜食与非鲜食橄榄种质资源群体间差异性富集，其中天冬氨酸在‘孔江3号’‘思光种’‘福榄2号’等鲜食种质中显著积累，谷氨酸在‘马坑3号’鲜食种质中特异性富集，苯丙氨酸在‘福榄2号’‘清榄3号’‘棱尖’等鲜食种质中高水平富集。利用转录组分析共鉴定到106个氨基酸相关的差异表达基因（differentially expressed genes, DEGs），其中L-天冬酰胺酶基因（L-asparaginase, L-ASN）、谷氨酸脱羧酶基因（glutamatedecarboxylase, GAD）、莽草酸脱水酶基因（arogenatedehydratase, ADT）和苯丙氨酸解氨酶基因（phenylalanineammonia lyase, PAL）是调控氨基酸生物合成的重要候选基因。基于DEGs，共富集到19条氨基酸相关的KEGG代谢途径。RT-qPCR试验结果表明，L-ASN、GAD、ADT和PAL的表达趋势与转录组测序结果一致。结论鲜味或芳香族氨基酸的差异性积累是影响橄榄鲜食品质的潜在因子。

关键词: 橄榄, 种质资源, 鲜食品质, 鲜味氨基酸, 芳香族氨基酸, 代谢组学, 转录组学

Abstract:

Objective To reveal factors influencing fresh-eating quality differences among germplasms of Chinese olive (Canarium album Raeusch.) and to provide a theoretical basis for germplasm selection and cultivar improvement. Method Integrated metabolomic and transcriptomic analyses were performed to assess differences in metabolite profiles and gene expression profiles across 24 Chinese olive germplasms with varying types. RT-qPCR was also used to validate the expression patterns of candidate regulatory genes. Result A total of 152 differentially accumulated metabolites (DAMs) were identified between fresh-edible and non-fresh-edible germplasm groups, primarily including amino acids, phenolic acids, terpenoids, flavonoids, organic acids, and sugars. These DAMs were enriched in 16 KEGG pathways related to amino acid biosynthesis or metabolism. Further analysis revealed that umami amino acids (aspartate and glutamate) and the aromatic amino acid (phenylalanine) were differentially enriched between the two groups. Specifically, aspartic acid significantly accumulated in fresh-edible germplasms such as ‘Kongjiang 3’, ‘Siguangzhong’, and ‘Fulan 2’; glutamic acid was specifically enriched in ‘Makeng 3’; and phenylalanine was highly enriched in ‘Fulan 2’, ‘Qinglan 3’, and ‘Lengjian’. Transcriptomic analysis detected 106 differentially expressed genes (DEGs) involved in amino acid biosynthesis and metabolism, among which L-asparaginase (L-ASN), glutamate decarboxylase (GAD), arogenate dehydratase (ADT), and phenylalanine ammonia-lyase (PAL) were identified as important candidate genes regulating amino acid biosynthesis. These DEGs were enriched in 19 KEGG pathways associated with amino acid metabolism. RT-qPCR results showed that the expression trends of L-ASN, GAD, ADT, and PAL were consistent with the transcriptome sequencing data. Conclusion Differential accumulation of umami or aromatic amino acids is a potential factor affecting the fresh-eating quality of Chinese olive fruits.

Key words: Canarium album Raeusch., germplasm resources, fresh-eating quality, umami amino acids, aromatic amino acids, metabolomics, transcriptomics

沈朝贵, 肖维强, 赖瑞联, 韦晓霞. 橄榄氨基酸组分差异对鲜食品质的潜在影响[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0148.

SHEN Chao-gui, XIAO Wei-qiang, LAI Rui-lian, WEI Xiao-xia. Potential Effect of Differential Amino Acid Composition on Fresh-eating Quality in Chinese Olive （Canarium album）[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0148.

图/表 8

表1 试验所用橄榄种质资源样本

Table 1 Germplasm resource samples of Chinese olive used in the experiment

序号 No.	名称 Name	代号 Code	来源 Resource	类型 Types
1	福榄2号	FL2	福建闽侯	鲜食
2	国道种	GDZ	福建闽侯
3	孔江3号	KJ3	福建闽清
4	棱尖	LJ	广东饶平
5	灵峰橄榄	LF	福建闽侯
6	马坑3号	MK3	福建闽清
7	闽清2号	MQ2	福建闽清
8	清榄1号	QL1	福建闽清
9	清榄3号	QL3	福建闽侯
10	三棱榄	SLL	广东潮阳
11	思光种	SGZ	福建闽侯
12	小美人	XMR	福建闽侯
13	橄榄王	GLW	福建闽清	非鲜食
14	惠圆	HY	福建闽侯
15	两头尖	LTJ	福建闽侯
16	七粒尺	QLC	福建闽侯
17	青皮长营	QPCY	福建闽侯
18	瑞安3号	RA3	浙江瑞安
19	瑞安4号	RA4	浙江瑞安
20	檀头	TT	福建闽清
21	羊矢	YS	福建闽清
22	漳浦11号	ZP11	福建漳浦
23	猪榄3号	ZL3	广西浦北
24	自来圆	ZLY	福建闽侯

图1 鲜食与非鲜食橄榄间DAMs鉴定A：正离子模式下主成分分析；B：正离子模式下偏最小二乘法‒判别分析；C：正离子模式下正交偏最小二乘法‒判别分析；D：差异积累代谢物分类；FE和NFE分别表示鲜食橄榄和非鲜食橄榄，下同

Fig. 1 DAM identification between fresh edible and non-fresh edible Chinese olivesA: Principal component analysis in positive ion mode. B: Partial least squares-discriminant analysis in positive ion mode. C: Orthogonal partial least squares-discriminant analysis in positive ion mode. D: DAMs classification. FE and NFE indicate fresh edible and non-fresh edible Chinese olives. The same below

图2 基于差异积累代谢物的聚类分析颜色越接近图例上端表示积累水平越高，而颜色越接近图例下端表示积累水平越低

Fig. 2 Cluster analysis based on DAMsColors closer to the upper end of the legend indicate higher accumulation levels, while colors closer to the lower end of the legend represent lower accumulation levels

图3 基于NFE vs. FE比较组中DAMs的KEGG前20代谢途径富集分析由红到蓝的颜色梯度表示P值由低到高，点的大小反映了代谢物的数量

Fig. 3 Enrichment of top 20 KEGG metabolic pathways based on DAMs in comparison group of NFE vs. FEColor gradient from red to blue indicates P‑values ranging from low to high, and dot size reflects the number of metabolites

图4 鲜食与非鲜食橄榄转录组分析及DEGs鉴定A：PCA主成分分析；B：相关性分析；C：聚类分析；D：KEGG代谢途径富集分析；颜色越接近图例上端表示相关性或表达水平越高，颜色越接近图例下端表示相关性或表达水平越低

Fig. 4 Transcriptomic analysis and DEGs identification of fresh edible and non-fresh edible types of Chinese oliveA: PCA analysis. B: Correlation analysis. C: Cluster analysis. D: KEGG metabolic pathway enrichment analysis. Colors closer to the upper end of the legend indicate higher correlation or expression levels, while colors closer to the lower end of the legend represent lower correlation or expression levels

表2 基于DAMs与DEGs的氨基酸相关KEGG代谢途径变化

Table 2 Changes in amino acid-related KEGG metabolic pathways based on DAMs and DEGs

代谢通路ID Pathway ID	代谢通路 Pathway	代谢组学 Metabolome			转录组学 Transcriptome
代谢通路ID Pathway ID	代谢通路 Pathway	DAMs	P-value	Impact	DEGs	P-value	FDR
ko00350	Tyrosine metabolism	5	0.200	0.090	16	0.005	0.072
ko00270	Cysteine and methionine metabolism	5	0.128	0.136	29	0.023	0.166
ko00330	Arginine and proline metabolism	3	0.526	0.124	19	0.035	0.187
ko00380	Tryptophan metabolism	3	0.653	0.168	12	0.040	0.201
ko00360	Phenylalanine metabolism	4	0.131	0.200	11	0.056	0.251
ko00400	Phenylalanine， tyrosine and tryptophan biosynthesis	3	0.163	0.089	12	0.081	0.323
ko00290	Valine， leucine and isoleucine biosynthesis	—	—	—	5	0.108	0.388
ko00280	Valine， leucine and isoleucine degradation	—	—	—	15	0.253	0.656
ko00250	Alanine， aspartate and glutamate metabolism	4	0.024	0.350	11	0.444	0.897
ko00260	Glycine， serine and threonine metabolism	4	0.124	0.067	11	0.589	0.942
ko00310	Lysine degradation	3	0.390	0.044	7	0.607	0.942
ko00220	Arginine biosynthesis	4	0.012	0.104	5	0.698	0.959
ko00340	Histidine metabolism	2	0.567	0.008	3	0.714	0.959
ko00300	Lysine biosynthesis	4	0.049	0.174	2	0.800	0.996
ko00430	Taurine and hypotaurine metabolism	1	0.626	0.031	5	0.032	0.183
ko00410	β-Alanine metabolism	2	0.368	0.073	14	0.163	0.525
ko00460	Cyanoamino acid metabolism	5	0.032	0.048	7	0.320	0.749
ko00480	Glutathione metabolism	1	0.790	0.005	12	0.657	0.955
ko00450	Selenocompound metabolism	—	—	—	2	0.856	1.000

图5 鲜食与非鲜食橄榄间氨基酸相关代谢途径中DAMs与DEGs的变化红色和蓝色分别表示代谢物的上调与下调积累。颜色越接近图例右端表示基因表达水平越高，颜色越接近图例左端则表示基因表达水平越低

Fig. 5 Changes in DAMs and DEGs in amino acid‑related metabolic pathways between fresh edible and non‑fresh edible types of Chinese oliveRed and blue colors indicate up‑accumulation and down‑accumulation of metabolites, respectively. Colors closer to the right end of the legend indicate higher gene expression levels, while colors closer to the left end of the legend represent lower gene expression levels

图7 氨基酸影响橄榄鲜食品质的潜在作用网络红色箭头代表上调，蓝色箭头表示下调。实线表示基于本研究中揭示的互作关系，虚线表示通过多组学整合分析推测的潜在关联

Fig. 7 Potential network of amino acids affecting fresh edible quality of Chinese oliveRed- and blue-arrows indicate up- and down-regulation. Solid lines represent interactions revealed in this study, while dashed lines suggest potential interactions

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