Identification of Key Ester Substances and Screening of Enzyme Gene Family Members for Aroma Differences in Fruits of Oriental Melons

doi:10.7668/hbnxb.20195590

Abstract

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

CLC Number:

S652.2

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.

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Figures/Tables 12

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

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

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

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

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

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

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.

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

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.

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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