薄皮甜瓜果实香味差异的关键酯类物质鉴定及相关酶基因家族筛选

doi:10.7668/hbnxb.20195590

摘要/Abstract

摘要： 为了明确导致薄皮甜瓜果实香味差异的酯类物质及其关键酶活性和基因的相关性,以薄皮甜瓜DX108和DX3-5果实为试验材料,研究了果实酯类代谢物的差异及羧酸酯酶(CXE)和醇酰基转移酶(AAT)活性和基因表达特性。结果表明:薄皮甜瓜果实中共检测到644种代谢物,其中酯类物质114种;成熟果实中共检测到差异代谢物108种,差异的酯类物质23种;与DX3-5相比,DX108成熟果实中酯类物质种类与含量增加明显,表明酯类物质是薄皮甜瓜成熟果实香气形成差异的关键物质;K-均值聚类分析法发现乙酸异戊酯、乙酸2-甲基丁酯、β-苯乙酸乙酯、乙酸对甲酚酯、乙酸己酯、苯乙酸甲酯、邻氨基苯甲酸甲酯和己酸乙酯是造成2个薄皮甜瓜品种成熟果实香味差异的关键酯类物质。此外,果实AAT和CXE活性变化趋势相反,暗示了AAT和CXE协调影响了酯类物质的代谢;随着果实的发育,AAT和CXE基因家族成员的表达特性存在差异,且相关性分析暗示了CmCXE5、CmCXE6和CmAAT5参与了薄皮甜瓜成熟果实中8种差异酯类物质的代谢。由上可知,8种关键酯类物质合成与积累差异是导致2种薄皮甜瓜成熟果实香味差异的重要原因,且CmCXE5、CmCXE6和CmAAT5可能发挥了主要调节作用。

关键词: 薄皮甜瓜, 酯类物质, 羧酸酯酶, 醇酰基转移酶, 基因表达

Abstract:

To determine the relationship between the esters compounds and key enzyme activities and genes that involved in the differences in aromas of oriental melon fruits,the disparities of ester metabolites and the enzyme activities and gene expression characteristics of CXE and AAT were investigated,with the fruits of oriental melon,DX108 and DX3-5,used as experimental materials.The results showed that a total of 644 metabolites were detected in oriental melon fruits,including 114 esters.A total of 108 metabolite variations and 23 different esters were detected in mature fruits.Compared with DX3-5,the types and contents of esters in ripe fruits of DX108 increased significantly,indicating that esters were the key substances in the formation of aroma difference in ripe fruits of oriental melons.K-means clustering analysis showed that isoamyl acetate,ethyl 2-methybuyrate,β-ethyl phenylacetate,p-cresol acetate,hexyl acetate ester,methyl phenylacetate,methyl anthranilate and ethyl caproate were the key esters,which caused the difference of ripe fruit aroma between two oriental melons.In addition,the activities of AAT and CXE in fruits showed opposite trend,suggesting that the coordination of AAT and CXE affected the metabolism of esters.The expression characteristics of AAT and CXE gene family members were different with fruit development,and correlation analysis suggested that CmCXE5,CmCXE6 and CmAAT5 were involved in the metabolism of eight different esters in oriental melon ripe fruits.From those findings,the variations in synthesis and accumulation of eight key esters are important reasons that direct the difference in aroma of two kinds of oriental melon,and CmCXE5,CmCXE6 and CmAAT5 may play a major regulatory role.

Key words: Oriental melon, Esters, Carboxylesterase, Alcohol acyltransferase, Gene expression

中图分类号:

S652.2

欧阳世贤, 刘胜博, 孙超, 陈凯, 何淑平, 李春霞, 靳亚忠. 薄皮甜瓜果实香味差异的关键酯类物质鉴定及相关酶基因家族筛选[J]. 华北农学报, 2025, 40(5): 62-73. doi: 10.7668/hbnxb.20195590.

OUYANG Shixian, LIU Shengbo, SUN Chao, CHEN Kai, HE Shuping, LI Chunxia, JIN Yazhong. Identification of Key Ester Substances and Screening of Enzyme Gene Family Members for Aroma Differences in Fruits of Oriental Melons[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(5): 62-73. doi: 10.7668/hbnxb.20195590.

http://www.hbnxb.net/CN/Y2025/V40/I5/62

图/表 12

表1 实时荧光定量PCR分析基因表达的引物序列

Tab.1 Sequence of primers used for gene expression analysis by real-time quantitative PCR

基因 Gene	引物 Primer	序列(5'—3') Sequence(5'—3')
CmCXE1	CmCXE1-F	TCACCATCAACCGCTGTCTC
	CmCXE1-R	AGGTCGTGGGAAGAGGAAGA
CmCXE2	CmCXE2-F	TGTGGCGGAGAAGGACAATC
	CmCXE2-R	GTTCCACCCCATCCCGATTT
CmCXE3	CmCXE3-F	TGGTGGTGATTGGGGGTTTG
	CmCXE3-R	GTTGGCGAATTGTTTGGGGG
CmCXE4	CmCXE4-F	TTTATCTCCGTCGAGCACCG
	CmCXE4-R	TTTCGTTACCGAGAGCCACC
CmCXE5	CmCXE5-F	CCGAATCCACGTCCCAATCA
	CmCXE5-R	GAGGAGTTTGGAATCGGGGG
CmCXE6	CmCXE6-F	CGAGGTGGTGGAGACGAAAG
	CmCXE6-R	CAATCTCGGGATCCACCGTT
CmAAT1	CmAAT1-F	TCAAATCTCCACCCTTCGCC
	CmAAT1-R	AAACCGCACTTCCTCCTCTG
CmAAT2	CmAAT2-F	GGTCCTCGCTGCCTATGTTT
	CmAAT2-R	AAACCGCACTTCCTCCTCTG
CmAAT3	CmAAT3-F	CTGTTGGTGAGATGGCTCGT
	CmAAT3-R	TCAAGCGGGATGATAGTGCC
CmAAT5	CmAAT5-F	CTTCTTTGGGCTGACCGAGA
	CmAAT5-R	AAGGCAAAGCAAACGCACTT
18S rRNA	18S rRNA-F	AAACGGCTACCACATCCA
	18S rRNA-R	CACCAGACTTGCCCTCCA

图1 薄皮甜瓜果实中挥发性代谢物质的种类占比

Fig.1 The proportion of volatile metabolites in oriental melon fruits

图2 薄皮甜瓜品种不同发育时期差异代谢物火山图 A.开花后10 d;B.开花后20 d;C.开花后30 d。

Fig.2 Volcanic maps of different metabolites of oriental melon at different developmental stages A.10 days after flowering;B.20 days after flowering;C.30 days after flowering

表2 DX108与DX3-5成熟果实中显著上调物质的种类及数量

Tab.2 Types and quantities of significantly up-regulated substances in ripe fruits of DX108 and DX3-5

种类 Species	数量 Quantity	占比/% Proportion
酯Ester	18	25.71
萜类Terpenoids	17	24.29
杂环化合物Heterocyclic compound	7	10.00
酮Ketone	6	8.57
醇Alcohol	5	7.14
芳烃Aromatics	4	5.71
烃类Hydrocarbons	4	5.71
醛Aldehyde	2	2.86
胺Amine	2	2.86
酚Phenol	2	2.86
含氮化合物Nitrogen compounds	1	1.43
含硫化合物Sulfur compounds	1	1.43
酸Acid	1	1.43

图3 薄皮甜瓜果实发育过程中各种挥发性代谢物的K均值聚类分析

Fig.3 K-means clustering analysis of various volatile metabolites during the development of oriental melon

图4 薄皮甜瓜果实成熟过程中AAT和CXE活性的变化

Fig.4 Changes of AAT and CXE activities during fruit ripening of oriental melon

图5 薄皮甜瓜果实成熟过程中AAT基因相对表达量变化不同小写字母表示处理之间差异显著(P<0.05)。图6同。

Fig.5 Changes of relative AAT gene expression level during fruit ripening of oriental melons Different Lowercase letters indicate the significance of the difference between treatments(P<0.05).The same as Fig.6.

图6 薄皮甜瓜果实成熟过程中CXE基因相对表达量变化

Fig.6 Changes of relative CXE gene expression level during fruit ripening of oriental melons

图7 DX108中关键差异酯类物质含量和差异基因相对表达量相关性分析 *.P<0.05。图8同。

Fig.7 Correlation analysis between the content of key differential ester compounds and the relative expressions of differential genes in DX108 *.P<0.05.The same as Fig.8.

图8 DX3-5中关键差异酯类物质含量和差异基因相对表达量相关性分析

Fig.8 Correlation analysis between the content of key differential ester compounds and the relative expressions of differential genes in DX3-5

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