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An X.-H.,Shandong Agricultural University | Tian Y.,Shandong Agricultural University | Tian Y.,Research Institute of Pomology | Chen K.-Q.,Shandong Agricultural University | And 2 more authors.
Journal of Plant Physiology | Year: 2012

The abundance of anthocyanins and proanthocyanins in apples is tightly regulated by three classes of regulatory factors, MYB, bHLH and WD40 proteins, only some of which have been previously identified. In this study, we identified an apple WD40 protein (MdTTG1) that promotes the accumulation of anthocyanins. The biosynthetic genes required downstream in the flavonoid pathway were up-regulated when MdTTG1 was over-expressed in Arabidopsis. Consistent with its role as a transcriptional regulator, an MdTTG1-GFP fusion protein was observed only in the nucleus. We assayed the expression patterns of this gene in different organs and found that they were positively correlated with anthocyanin accumulation in the apple. Yeast two-hybrid and bimolecular fluorescence complementation assays demonstrated that MdTTG1 interacted with bHLH transcription factors (TFs) but not MYB protein, whereas bHLH was known to interact with MYB in apples. However, based on a ChIP assay, MdTTG1 does not appear to bind to the promoter of the anthocyanin biosynthetic genes MdDFR and MdUFGT. Taken together, these results suggest that the apple WD40 protein MdTTG1 interacts with bHLH but not MYB proteins to regulate anthocyanin accumulation. © 2012 Elsevier GmbH. Source

An X.-H.,Shandong Agricultural University | An X.-H.,Research Institute of Pomology | Tian Y.,Shandong Agricultural University | Tian Y.,Research Institute of Pomology | And 7 more authors.
Plant and Cell Physiology | Year: 2015

Anthocyanin and proanthocyanidin (PA) are important secondary metabolites and beneficial to human health. Their biosynthesis is induced by jasmonate (JA) treatment and regulated by MYB transcription factors (TFs). However, which and how MYB TFs regulate this process is largely unknown in apple. In this study, MdMYB9 and MdMYB11 which were induced by methyl jasmonate (MeJA) were functionally characterized. Overexpression of MdMYB9 or MdMYB11 promoted not only anthocyanin but also PA accumulation in apple calluses, and the accumulation was further enhanced by MeJA. Subsequently, yeast two-hybrid, pull-down and bimolecular fluorescence complementation assays showed that both MYB proteins interact with MdbHLH3. Moreover, Jasmonate ZIM-domain (MdJAZ) proteins interact with MdbHLH3. Furthermore, chromatin immunoprecipitation- quantitative PCR and yeast one-hybrid assays demonstrated that both MdMYB9 and MdMYB11 bind to the promoters of ANS, ANR and LAR, whereas MdbHLH3 is recruited to the promoters of MdMYB9 and MdMYB11 and regulates their transcription. In addition, transient expression assays indicated that overexpression of MdJAZ2 inhibits the recruitment of MdbHLH3 to the promoters of MdMYB9 and MdMYB11. Our findings provide new insight into the mechanism of how MeJA regulates anthocyanin and PA accumulation in apple. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. Source

Tian L.,Research Institute of Pomology | Gao Y.,Research Institute of Pomology | Cao Y.,Research Institute of Pomology | Liu F.,Research Institute of Pomology | Yang J.,Research Institute of Pomology
Genetic Resources and Crop Evolution | Year: 2012

109 Pyrus accessions including 92 local Chinese accessions of P. bretschneideri were identified genetically using nine microsatellite loci developed from apple and pear. The nine SSR loci revealed 129 alleles in 109 pear accessions and 114 alleles in 92 Chinese white pears. Among the 92 local Chinese accessions of P. bretschneideri, 70 could be differentiated successfully except for 10 sets of synonymous or mutants. For the 92 accessions, the number of putative alleles per locus ranged from seven to 18, with an average of 12.67; the average values of observed heterozygosity and Shannon's Information index were 0.60 and 1.85, respectively. A phenogram based on the SSR (simple sequence repeat) genotypes was obtained. The 109 accessions clustered into 11 groups based on geographical origin. The European pears and the Asian pears did not form independent two groups, but three P. communis cultivars grouped together independently. The Japanese P. pyrifolia cultivars mingled together with the Chinese P. bretschneideri cultivars, but four P. ussuriensis cultivars except for one (Jianbali) grouped together independently. The results indicated that the relationship of P. bretschneideri cultivars and P. pyrifolia cultivars was much closer than others. © 2012 Springer Science+Business Media B.V. Source

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