苹果MdMYB9、MdMYB11表达及其蛋白互作分析

doi:10.3864/j.issn.0578-1752.2015.11.012

Abstract

Abstract: 【Objective】 This study is aiming at isolating MdMYB9 and MdMYB11 genes that are homologous to TT2 from Arabidopsis, detecting their expression in different tissues and under light treatment, identifying the interaction of MdMYB9 and MdMYB11 with MdbHLH33, and laying the foundation for further function analysis of this two genes. 【Method】 MdMYB9 the protein molecular weight, isoelectric point and the full length amino acid sequences were analyzed using DNAMAN software. In addition, the phylogenetic tree of the MYB proteins, including MdMYB9, MdMYB11 and the R2R2-MYB family proteins from Arabidopsis, was constructed using the neighbor-joining (NJ) method of MEGA 4.0 software. The transcriptional levels of MdMYB9 and MdMYB11 genes were detected in different tissues, fruit skins and seeds at different developmental stages and under light treatment using qPCR. Finally, the interaction of MdMYB9 and MdMYB11with MdbHLH33 protein was detected by Y2H assay.【Result】Sequence analysis showed that the open reading frame (ORF) of MdMYB9 was 873 bp in length, encoding 290 amino acids and sequence alignment revealed 38.13% identity with TT2 in Arabidopsis. The ORF of MdMYB11 gene was 861 bp in length, encoding 286 amino acids and sequence alignment revealed 32.44% identity with TT2. Both the proteins contained the typical R2R3 domain in the N-terminal. qPCR analysis indicated that MdMYB9 and MdMYB11 were expressed in all organs, including roots, stems, leaves and flowers, but their expressions exhibited different levels in the detected tissues. Both genes expressed at different developmental stages of seeds and skins, and the highest expression appeared at the middle stages; however, the expression of MdMYB9 and MdMYB11 were not obviously different under light treatment. Yeast two-hybrid assay showed that both MdMYB9 and MdMYB11 interacted with MdbHLH33.【Conclusion】MdMYB9 and MdMYB11 were expressed in different tissues, developmental fruit skins and seeds, and interacted with MdbHLH33 protein. and MdMYB11 genes associated with proanthocyanin biosynthesis were isolated using homology-based cloning method, and the primers were designed to amplify these two genes. Subsequently,

Key words: apple, proanthocyanin, MdMYB9, MdMYB11, expression analysis, yeast two-hybrid

AN Xiu-hong, ZHANG Xiu-de, CHEN Ke-qin, LIU Xiao-juan, HAO Yu-jin, CHENG Cun-gang. Expression and Protein Interaction Analysis of MdMYB9 and MdMYB11 in Apple[J].Scientia Agricultura Sinica, 2015, 48(11): 2208-2216.

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Expression and Protein Interaction Analysis of MdMYB9 and MdMYB11 in Apple

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