乙烯利和1-MCP对菠萝蜜果实中AheAAT和 AheERF表达的影响

doi:10.3864/j.issn.0578-1752.2019.21.017

Abstract

Abstract:

【Objective】 The effects of Ethephon (ETH) and 1-methylcyclopropene (1-MCP) on AAT activities, the expression of AheAAT gene and AheERF1/2 transcription factor were studied during ripening of jackfruit to provide a theoretical basis for further understanding the role of AATS in the synthesis of esters and its regulation. 【Method】 The test material was jackfruit of Haida2. During the full bloom period, the fruit with the representative flowering period was labeled and harvested about 150 days after flowering. The fruits with the same size and maturity and no disease and insect damage were selected to divide into three groups for treatment. The first group was immersed in 1 000 mg?L ^-1 ETH solution for 2-3 min; the second group was fumigated with 0.5 mg?L ^-1 1-MCP for 15 h; the third group, as a control group, was naturally matured at 22℃ and 90% relative humidity conditions. During fruit ripening, samples were taken periodically with 3-5 fruits per times. The flesh of the jackfruit was frozen in liquid nitrogen and then stored in an ultra-low temperature refrigerator at -80℃ for determination of AAT activities, the expression of AheAAT gene and AheERF1/2 of jackfruit.【Result】 By analyzing the activity of AAT enzyme, it was found that ETH treatment promoted the ripening and senescence of jackfruit, advanced the peak of the activity peak of AAT, but reduced AAT enzyme activity. 1-MCP treatment inhibited the ripening and senescence of fruits by inhibiting the AAT activity in the pre-storage period, delaying the time of peak AAT activity, and significantly reducing the activity of AAT enzyme. Sequence analysis of AheAAT gene showed that the total length of AheAAT gene was 1 380 bp, encoding 459 amino acids. AheAAT had a conserved domain H-x-x-x-D and an L-x-x-YYPLAGR active site motif, a D(N)F(V) GWG add-on motif. The AheAAT gene belonged to the BAHD alcohol acyltransferase family, and its amino acid sequence had the highest similarity to apples and pears. At the same time, two ERF transcription factors, named AheERF1/2, were cloned from the fruit of jackfruit, of which AheERF1 was 648 bp in length, encoding 215 amino acids, and had the highest homology with apple and kidney bean. AheERF2 was 657 bp in length, encoding 218 amino acids, with the highest homology with apples, papaya. The amino acid sequence of AheERF1/2 contained an AP2/ERF conserved sequence consisting of 64/65 amino acid residues and had a characteristic sequence of an ERF family transcription factor. ETH treatment advanced the expression peak of AheAAT gene in jackfruits and down-regulated its expression. The effects of ETH treatment on the expression of AheERF1/2 transcription factor were identical to that of AheAAT gene. ETH treatment decreased the correlation between AheAAT gene and AheERF2 transcription factor. 1-MCP treatment prolonged the storage period of jackfruit fruit. 1-MCP treatment prolonged the storage period of jackfruit fruit. The expression of AheAAT gene and AheERF1/2 transcription factors were significantly reduced during the pre-storage period after 1-MCP treatment. 1-MCP down-regulated the expression levels of AheAAT and AheERF1/2 transcription factor at the peak of their expressions. However, 1-MCP had little effect on the correlations of the AheERF1/2 transcription factor with the AheAAT gene. 【Conclusion】 ETH treatment advanced the peak activity of AAT enzyme, the peak expression of AheAAT and AheERF1/2 transcription factors, down-regulated the expression of AheAAT and AheERF1/2 transcription factors, and decreased the activity of AAT enzyme and the correlation between AheAAT and AheERF2 transcription factors. Compared with the control group, 1-MCP treatment inhibited the activity of AAT enzyme, the expression of AheAAT gene and AheERF1/2 transcription factors in the pre-storage phase, decreased the activity of AAT enzyme and down-regulated the expression levels of AheAAT and AheERF1/2 transcription factors during the late storage period.

Key words: jackfruit, ETH, 1-MCP, AheAAT gene

REN XueYan,LIU GuangCai,LI GuoPeng,YE ChunHai,FENG Feng,WANG JunNing. Effects of Ethephon and 1-MCP on the Expression of AheAAT Gene and AheERF Transcription Factors in Jackfruit Fruit[J].Scientia Agricultura Sinica, 2019, 52(21): 3890-3902.

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References 33

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引物名称 Primers name	引物序列 Primer sequence
AheAAT -F AheAAT -R	5′- ATGGCATCAACGCCATCCTCATCGTTAG-3′ 5′- TTAGAGCGAGGATGAAATCTTAAAGCCCTTC-3′
AheERF1 -F AheERF1 -R	5′- ATGACCCGACCGCAACAGCGATATC-3′ 5′- TGGGTACTAGTAGTATTCCTAAAGGAGTG-3′
AheERF2 -F AheERF2 -R	5′- ATGGAATCCCCTGTTTTCCACTCCCAAACT-3′ 5′- TTAGATTGAAGTGGTTAAAAGTTCTTCTAA-3′

引物名称 Primers name	引物序列 Primer sequence
AheAAT-qup	5′- CATCAACGCCATCCTCAT -3′
AheAAT-qdw	5′-TGAAACCTAAGCCCTCCT -3′
AheERF1-qup	5′-CCCGCACCAACTTCCCTT -3′
AheERF1-qdw	5′-GTTGGGTTTCGTGAGAATGCC -3′
AheERF2-qup	5′-CGAACCAGGCGATAAAGGA -3′
AheERF2-qdw	5′-TGCCGAGCCACACCCTTA -3′
AheGAPDH-qup	5′-TTGAAGGGTGGNGCNAARAARG -3′
AheGAPDH-qdw	5′-ATAACCCCAYTCRTTRTCRTAC -3′

	AheAAT
	CK	ETH	1-MCP
AheERF1转录因子	0.993**	0.965**	0.968**
AheERF2转录因子	0.956**	0.915*	0.817**

Effects of Ethephon and 1-MCP on the Expression of AheAAT Gene and AheERF Transcription Factors in Jackfruit Fruit

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