Cloning and Expression Analysis of AcWRKY70 Gene Sequence under Psa and Hormone Treatment in Kiwifruit

doi:10.7668/hbnxb.20192987

Abstract

Abstract:

In order to explore the mechanism of AcWRKY70 transcription factor in response to kiwifruit canker stress in different resistant varieties,AcWRKY70 gene was cloned from the leaf cDNA of resistant variety Xuxiang and highly susceptible variety Hongyang kiwifruit.The sequence structure characteristics of Hongyang AcWRKY70 gene,subcellular localization and evolutionary relationship of Hongyang AcWRKY70 protein were analyzed. The expression patterns of AcWRKY70 gene,including the tissue expression and different expression in resistant kiwifruit varietie and susceptible variety under Pseudomonas syringae pv. Actinidiae(Psa),salicylic acid(SA)and methyl jasmonate(MeJA)treatment were analyzed by RT-qPCR. Results showed that the full length of the Hongyang AcWRKY70 gene was 906 bp(GenBank accession number was MW881147). AcWRKY70 gene contained 885 bp of open reading frame(ORF)and encoded 294 amino acids. AcWRKY70 protein had typical WRKYGQK domain and the zinc finger structure was C2-HC. It belonged to class Ⅲ group of WRKY family and islocated in the nucleus. Phylogenetic tree analysis showed that it had the closest genetic relationship with tea plant. Both Hongyang and Xuxiang AcWRKY70 genes had the highest expression level in leaves. Under Psa treatment, the highest expression of AcWRKY70 gene reached at 12 h in Xuxiang, the expression of Hongyang reached the maximum at 48 h.Under the co-treatment of salicylic acid and Psa(SA+Psa),Methyl jasmonate and Psa(MeJA+Psa),the highest expression of AcWRKY70 gene reached at 12 h in Xuxiang. Under SA+Psa and MeJA+Psa treatment,the expression of Hongyang reached the maximum at 72,24 h,respectively. AcWRKY70 gene played a certain role in resistance stress of kiwifruit,and the response mechanism to pathogens in different resistance kiwifruit varieties may vary considerably.

Key words: Kiwifruit, AcWRKY70, Expression analysis, Kiwifruit canker, Salicylic acid, Methyl jasmonate

QU Dong, YAN Fei, LIU Xinrui, KANG Xue, ZENG Haitao, ZHANG Yu. Cloning and Expression Analysis of AcWRKY70 Gene Sequence under Psa and Hormone Treatment in Kiwifruit[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 166-173. doi: 10.7668/hbnxb.20192987.

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Figures/Tables 7

Fig.1 The PCR amplified product of AcWRKY70 gene(A)and the conserved domain of AcWRKY70 transcription factor(B) M.GL DNA Marker 2000 Ladder;1.PCR product of Hongyang AcWRKY70 gene.

Fig.2 Multiple alignment of amino acid sequences of AcWRKY70 and WRKY transcription factors in different species Red box.The conserved domain of WRKYGQK;Red arrows.The zinc finger domain of C2-HC.

Fig.3 Phylogenetic tree analysis of AcWRKY70 protein in Hongyang The AcWRKY70 protein sequence of kiwifruit is located in the red frame.

Fig.4 The phosphorylation sites prediction of AcWRKY70 protein

Fig.5 The tertiary structure prediction of AcWRKY70 protein

Fig.6 Expression difference of AcWRKY70 gene in different tissues of Xuxiang and Hongyang Different lowercase indicate significant difference in different treatments, P<0.05.The same as Fig.7.

Fig.7 Different expression of AcWRKY70 gene under Psa(A),SA+Psa(B)and MeJA+Psa(C)treatment

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