Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (13): 2642-2650.doi: 10.3864/j.issn.0578-1752.2016.13.020

• RESEARCH NOTES • Previous Articles    

The Gene Expression Assays and Protein Interactions of MdJAZ1 in Apple

AN Xiu-hong, LI En-mao, LI Min, LI Zhuang, ZHANG Xiu-de, CHENG Cun-gang   

  1. Institute of Pomology, Chinese Academy of Agricultural Sciences/Key Laboratory of Fruit Germplasm Resources Utilization, Ministry of Agriculture, Xingcheng 125100, Liaoning
  • Received:2016-02-03 Online:2016-07-01 Published:2016-07-01

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Abstract: 【Objective】MdJAZ1 was isolated from apple which encoded a repressor protein in jasmonate signaling pathway. The expression of MdJAZ1 and the interactions of MdJAZ1 with other JAZ proteins or F-box protein were determined in order to lay a foundation for further functional analysis of this gene.【Method】The MdJAZ1 was selected among multiple genes which were obtained through blasting in the apple genome database using the TIFY and Jas domains. The molecular weight and isoelectric point of MdJAZ1 were analyzed by the DNAMan software, and the functional domain was analyzed using the SMART soft. The phylogenetic tree of the proteins, including MdJAZ1 and the JAZ family proteins from Arabidopsis, was constructed using the neighbor-joining (NJ) method of MEGA5.0 soft. In addition, the expression of MdJAZ1 was detected in different tissues of apple and with MeJA or wounding treatment using qPCR. The cis-acting regulatory elements of the promoter of MdJAZ1 were analyzed through PlantCARE database. Finally, the interactions of MdJAZ1 with other JAZ proteins and AtCOI1 were detected through the yeast two-hybrid assays. 【Result】The open reading frame (ORF) of MdJAZ1 was 1 149 bp in length and encoded 382 amino acids. The calculated molecular mass of MdJAZ1 was 40.536 kDa, and the isoelectric point was 9. Sequence analysis showed that MdJAZ1 protein contained the typical TIFY domain and C-terminal Jas domain. The phylogenetic relationship of MdJAZ1 was clustered with the Arabidopsis JAZ family members AtJAZ3 and AtJAZ4. qPCR analysis revealed that MdJAZ1 was expressed in all apple tissues, including roots, stems, leaves, flowers and fruits, however, the transcriptional level was significantly different in the detected tissues, with the highest expression in roots and the lowest expression in fruits. In addition, MdJAZ1 was induced by MeJA and wounding treatments, with the highest expression at 1 h. Promoter analysis showed that there were multiple putative cis-acting elements which are involved in abscisic acid response, ethylene response, and defense and stress responses; MYB binding sites were also found in the promoter of MdJAZ1 gene. The yeast two-hybrid assays showed that MdJAZ1 formed homo- and heterodimers and also interacted with AtCOI1 which encoded a F-box protein in Arabidopsis.【Conclusion】MdJAZ1 was induced by MeJA and wounding treatments. MdJAZ1 protein formed homo- and heterodimers and also interacted with AtCOI1 in the present of coronatine.

Key words: apple, jasmonate, MdJAZ1, expression, protein interaction

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