Identification,Cloning and Expression Analysis of PavJAZ1 in the Fruit of Prunus avium L.

doi:10.7668/hbnxb.20195818

Abstract

Abstract:

Jasmonates play a crucial role in regulating fruit ripening and quality formation.The JAZ proteins are key repressors of the jasmonic acid signaling pathway.To investigate the function of PavJAZ1 in sweet cherry fruit ripening and quality regulation,the PavJAZ1 gene was identified and cloned from the sweet cherry genome.Bioinformatics analysis was performed to explore its physicochemical properties,protein structure,conserved domains,and cis-acting elements in the promoter region.The gene expression pattern of PavJAZ1 in fruits was analyzed by Quantitative Real-time PCR.The subcellular localization of the PavJAZ1 protein was observed,and the interaction between PavJAZ1 and PavMYC2 was verified using the bimolecular fluorescence complementation(BiFC).The results of gene identification and cloning showed that the open reading frame of the PavJAZ1 gene was 837 bp in length,encoding 278 amino acids.The molecular weight of the PavJAZ1 was 30.09 ku,the theoretical isoelectric point was 9.21,the instability index was 54.14,and the grand average of hydropathicity was -0.564,indicating that it was an alkaline hydrophilic and unstable properties.Protein structure analysis revealed that PavJAZ1 was mainly composed of random coils and α-helices,containing two conserved domains,the Tify domain and the Jas domain.It also exhibited high homology with proteins from other species,among which it showed the highest homology with that of peach,a plant that also belonged to the genus Prunus of the Rosaceae family.Subcellular localization analysis showed that the PavJAZ1 protein was localized in the nucleus.The expression level of PavJAZ1 increased significantly in the early stages of fruit development and then gradually decreased.This result suggested that PavJAZ1 may play different roles at different developmental stages.In addition,the BiFC results showed that PavJAZ1 interacted with PavMYC2,confirming that PavJAZ1 participates in the jasmonic acid signaling pathway.

Key words: Sweet cherry, JAZ, Fruit ripening, Expression pattern, Fruit quality

CLC Number:

WANG Wei, RAN Jianglin, ZHANG Xiaoming, YAN Guohua, WANG Jing, ZHOU Yu, WU Chuanbao, FENG Chen, ZHANG Kaichun, DUAN Xuwei. Identification,Cloning and Expression Analysis of PavJAZ1 in the Fruit of Prunus avium L.[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(2): 28-35. doi: 10.7668/hbnxb.20195818.

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URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20195818

http://www.hbnxb.net/EN/Y2026/V41/I2/28

Figures/Tables 9

Fig.1 PCR amplification of PavJAZ1 gene M.DNA marker;1,2.Target fragment.

Fig.2 Phylogenetic analysis of PavJAZ1 and JAZ1 in other species

Fig.3 Conserved domain analysis and sequence alignment of PavJAZ1 A.Conserved domain analysis of PavJAZ1; B.Conserved domain sequence alignment of PavJAZ1.

Fig.4 Structure analysis of PavJAZ1 and prediction of the structure of the encoded protein A.Gene structure analysis of PavJAZ1;B.Secondary structure prediction of PavJAZ1;C.Tertiary structure prediction of PavJAZ1.

Tab.1 PavJAZ1 gene promoter cis-acting elements

分类 Type	名称 Name	序列 Sequence	个数 Number	功能 Function
胁迫应答元件	DRE core	GCCGAC	1	干旱应答元件
Stress related element	3-AF1 binding site	TAAGAGAGGAA	1	光应答元件
	Box 4	ATTAAT	4	光应答元件
	G-Box	CACGTG	5	光应答元件
	GATA-motif	AAGGATAAGG	1	光应答元件
	Sp1	GGGCGG	2	光应答元件
	TCCC-motif	TCTCCCT	1	光应答元件
	GC-motif	CCCCCG	1	缺氧特异性诱导元件
	TC-rich repeats	ATTCTCTAAC	1	防御和应激调控元件
	WUN-motif	CCATTTCAA	1	创伤响应元件
激素应答元件	ABRE	ACGTG	4	脱落酸应答元件
Hormone response element	TGACG-motif	TGACG	1	茉莉酸应答元件
	CGTCA-motif	CGTCA	1	茉莉酸应答元件
	P-box	CCTTTTG	1	赤霉素应答元件
	TATC-box	TATCCCA	1	赤霉素应答元件
	TCA-element	CCATCTTTTT	1	水杨酸应答元件
	TGA-element	AACGAC	1	生长素应答元件
	ERE	ATTTCATA	2	乙烯应答元件

Fig.5 Subcellular localization of PavJAZ1 protein

Fig.6 Expression analysis of PavJAZ1 in different fruit developmental stage Different lowercase letters indicate significant differences(P<0.05).

Fig.7 Predicted proteins interacting with PavJAZ1 A.Prediction based on protein-protein interactions in peach;B.Prediction based on protein-protein interactions in Arabidopsis.

Fig.8 Analysis of the interaction between PavJAZ1 and PavMYC2

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