ABA and Ethylene Enhance the Expression VlMybA1 and VlMybA2 and Promote Pigmentation in the Berry Skin via Their Interaction

doi:10.16420/j.issn.0513-353x.2022-0919

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (11): 2323-2336.doi: 10.16420/j.issn.0513-353x.2022-0919

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

ABA and Ethylene Enhance the Expression VlMybA1 and VlMybA2 and Promote Pigmentation in the Berry Skin via Their Interaction

SUN Yushuai¹, WANG Fei¹, GUAN Xueqiang², CHI Huiru¹, YAO Yuxin¹^,^*()

¹ State Key Laboratory of Crop Biology，Shandong Collaborative Innovation Center for Fruit & Vegetable Production with High Quality and Efficiency，College of Horticultural Science and Engineering，Shandong Agricultural Univerity，Tai’an，Shandong 271018，China
² Shandong Academy of Grape，Jinan 250100，China

Received:2023-04-21 Revised:2023-08-13 Online:2023-11-25 Published:2023-11-28
Contact: YAO Yuxin

Abstract

Abstract:

The patterns of anthocyanin and ABA accumulation and ethylene production as well as the changes in expression of the key genes associated with their synthesis or regulation during berry development were studied using Kyoho grapes（Vitis vinifera × V. labrusca‘Kyoho’）. Analysis of gene expression，promoter response and relative mRNA abundance showed that exogenous ABA treatment upregulated the expression level of VlMybA1 and VlMybA2，in which VlMybA1 was significantly expressed and promoted berry skin pigmentation；In contrast，exogenous ACC treatment increased the VlMybA2 expression and anthocyanin accumulation. The interaction between ABA and ethylene was found，100 μmol · L^-1ABA largely induced the production of endogenous ethylene，and 500 μmol · L^-1ACC significantly increased ABA content. Additionally，exogenous ABA treatment increased the relative mRNA abundance of VlMybA2 via ethylene. Collectively，ABA promoted berry skin pigmentation by upregulating the VlMybA1 expression directly and the VlMybA2 expression indirectly via ethylene.

Key words: grape, ABA, ethylene, VlMybA1, VlMybA2, anthocyanins

SUN Yushuai, WANG Fei, GUAN Xueqiang, CHI Huiru, YAO Yuxin. ABA and Ethylene Enhance the Expression VlMybA1 and VlMybA2 and Promote Pigmentation in the Berry Skin via Their Interaction[J]. Acta Horticulturae Sinica, 2023, 50(11): 2323-2336.

Figures/Tables 7

Table 1 Primer sequence used in this study

目的基因 Target gene	登录号 Accession number	上游引物（5′-3′） Forward primer sequence	下游引物（5′-3′） Reverse primer sequence
VlMybA1，VlMybA2，VlMybA3（qRT-PCR）	AB427165，AB073013， AB073012	GAGGGTGATTTTCCATTTGAT	CAAGAACAACTTTTGAACTTAAACAT
VlMybA2（qRT-PCR）	AB073013	GTTCCAAAGTTGTTTAATCTTAG	ATCCACTGCTCACATCCAT
VlUFGT（qRT-PCR）	AF000372^*	GGGATGGTAATGGCTGTGG	ACATGGGTGGAGAGTGAGTT
VlNCED1（qRT-PCR）	XM_002277318^*	ATACATAGCTGCTTGGTGTCAGA	CTGTAGAGAGTTTGGAGATTCCG
VlNCED4（qRT-PCR）	XM_002270125^*	GGCTTCCATGGATTGTTTGT	CCTAATTCGAGCTTGCTTTCC
VlACS（qRT-PCR）	XM_002278453^*	CTCAGGTAATTGTCGGATGTG	GATCAAAACGAGAATCCGGT
VlACO（qRT-PCR）	XM_002273394^*	CTGTTGTCGTTCTAGTGTTGGTA	CATCAAACCATACCTTAGGAGCT
VlUbiquitin1（qRT-PCR）	BN000705^*	GTGGTATTATTGAGCCATCCTT	AACCTCCAATCCAGTTATCTAC
VlMybA1（promoter）	AB427165	TGCCATCTCCATGCAGC	GAGTCAACTCAACACAAGAGACC
VlMybA2（promoter）	AB073013	GCGATAAATTCACAAATAGAGGA	GAGTCAACTCAACACAAGAGACC

Fig. 1 Alignment of VlMybA1，VlMybA2和VlMybA3 fragments The region used for PCR amplification is indicated with red letters，the primers are highlighted with underlined letters，asterisk means the same letter for the three genes，tga is the stop codonh，the blue letters represent the bases that were different between the three VlMybAs and were utilized to differentiate VlMybA1，VlMybA2和VlMybA3.

Fig. 2 Changes in grape berry ripening-related parameters（A，B），ABA and ethylene（C），and the expression of the genes（D，E） involved in biosynthesis of ABA and ethylene

Fig 3 Effects of ABA and ACC treatment time on anthocyanin and anthocyanin-related gene expression The significant differences was analyzed between each treatment and control group was analyzed，* indicates a significant at the 0.05 level，**indicates a significant difference at the 0.01 level. The same below.

Fig. 4 Promoter analysis of VlMybA1 and VlMybA2 genes（A），GUS staining and GUS activity measurement of VlMybA1/2 promoter:: GUS overexpressed by ABA or ACC treatment（B，C）

Fig. 5 Effects of treatment with ABA or different ACC concentrations on ABA and ethylene release rates and corresponding gene expression

Fig. 6 Effects of ABA or ACC treatment time on the relative abundance of VlMybA2 Figure C，D significance analysis shows the comparison of the same gene among different treatments. Different lowercase letters indicate significant differences at 0.05 level.

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ABA and Ethylene Enhance the Expression VlMybA1 and VlMybA2 and Promote Pigmentation in the Berry Skin via Their Interaction

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