AS  Vol.9 No.3 , March 2018
Cloning and Expression of Anthocyanin Biosynthesis Related Gene RrMYB6 in Rosa rugosa
Abstract: R2R3-MYB transcription factor plays an important role in plant anthocyanin synthesis. Based on the transcriptional database of Rosa rugosa, one MYB transcription factor related to floral color, RrMYB6, was cloned. By using bioinformatics analysis method, cloning MYB gene and analyzing its function in anthocyanin biosynthesis regulation, we hope to lay a solid foundation for new color variety breeding of R. rugosa. Using the R. rugosa “Zi zhi” as the material, we obtained the total length of cDNA of RrMYB6 by RT-PCR and RACE. By analyzing its bioinformatics, we found that the formula of the protein was C1491H2368N452O470S17, molecular weight was 34690.97 Da, the theoretical pI was 8.74. In addition, it belonged to unstable protein with an unstable index at 50.59, and it was also a hydrophilic protein with the total average hydrophobic index at -0.847. In the secondary structure of RrMYB6 protein, the Alpha helix accounted for 32.35%, random coil was 47.39%, extended strand was 11.11%, and beta turn was 9.15%. The sequence analysis showed that RrMYB6 had a typical R2R3-MYB domain and bHLH binding domain, and it also had an N1, C1, C2 inhibitory motif, belonging to the Sg4 subfamily MYB protein. What’s more, evolutionary analysis indicated that the RrMYB6 protein was closely related with the MYB protein in Rosacea family, while it was far from those in other families. The expression analysis showed that RrMYB6 protein decreased with the color of petals deeping, and its expression was the lowest in the petals while the highest in stamens. According to the above results, it was speculated that RrMYB6 was involved in regulating the anthocyanin synthesis of R. rugosa, which belonged to negative regulatory mechanism.
Cite this paper: Zou, K. , Wang, Y. , Zhao, M. , Zhao, L. and Xu, Z. (2018) Cloning and Expression of Anthocyanin Biosynthesis Related Gene RrMYB6 in Rosa rugosa. Agricultural Sciences, 9, 374-383. doi: 10.4236/as.2018.93026.

[1]   Buer, C.S., Imin, N. and Djordjevic, M.A. (2010) Flavonoids: New Roles for Old Molecules. Journal of Integrative Plant Biology, 52, 98-111.

[2]   Schwinn, K.E., Boase, M.R., Bradley, J.M., et al. (2014) MYB and bHLH transcription Factor Transgenes Increase Anthocyanin Pigmentation in Petunia and Lisianthus Plants, and the Petunia Phenotypes Are Strongly Enhanced under Field Conditions. Frontiers in Plant Science, 5, 603.

[3]   Ibraheem, F., Gaffoor, I., Tan, Q., et al. (2015) A Sorghum MYB Transcription Factor Induces 3-deoxyanthocyanidins and Enhances Resistance against Leaf Blights in Maize. Molecules, 20, 2388-2404.

[4]   Schwinn, K., Venail, J., Shang, Y., et al. (2006) A Small Family of MYB-Regulatory Genes Controls Floral Pigmentation Intensity and Patterning in the Genus Antirrhinum. The Plant Cell, 18, 831-851.

[5]   Ravaglia, D., Espley, R., Henry, K.R., et al. (2013) Transcriptional Regulation of Flavonoid Biosynthesis in Nectarine (Prunus persica) by a Set of R2R3-MYB Transcription Factors. BMC Plant Biology, 13, 68.

[6]   Li, L., Ban, Z.J., Li, X.H., et al. (2012) Differential Expression of Anthocyanin Biosynthetic Genes and Transcription Factor PcMYB10 in Pears (Pyrus communis L.). Plos One, 7, e46070.

[7]   Huang, W., Sun, W., Lv, H., et al. (2013) A R2R3-MYB Transcription Factor from Epimedium sagittatum Regulates the Flavonoid Biosynthetic Pathway. PLoS ONE, 8, e70778.

[8]   Feng, S., Wang, Y., Yang, S., et al. (2010) Anthocyanin Biosynthesis in Pears Is Regulated by a R2R3-MYB Transcription Factor PyMYB10. Planta, 232, 245-255.

[9]   Starkevic, P., Paukstyte, J., Kazanaviciute, V., et al. (2015) Expression and Anthocyanin Biosynthesis-Modulating Potential of Sweet Cherry (Prunus avium L.) MYB10 and bHLH Genes. PLoS ONE, 10, e0126991.

[10]   Zhao, J., Liu, R., Yang, F., et al. (2015) Cloning and Expression Analyses of R2R3-MYB Genes Related to Anthocyanin Biosynthesis in Rose. Scientia Agricultura Sinica, 48, 1392-1404.

[11]   Xu, Z.S., Feng, K., Que, F., et al. (2017) A MYB Transcription Factor, DcMYB6, Is Involved in Regulating Anthocyanin Biosynthesis in Purple Carrot Taproots. Scientific Reports 7, Article ID: 45324.

[12]   Butelli, E., Licciardello, C., Zhang, Y., Liu, J., et al. (2012) Retrotransposons Control Fruit-Specific, Cold-Dependent Accumulation of Anthocyanins in Blood Oranges. The Plant Cell, 24, 1242-1255.

[13]   Yamagishi, M., Shimoyamada, Y., Nakatsuka, T., et al. (2010) Two R2R3-MYB Genes, Homologs of Petunia AN2, Regulate Anthocyanin Biosyntheses in Flower Tepals, Tepal Spots and Leaves of Asiatic Hybrid Lily. Plant and Cell Physiology, 51, 463-474.

[14]   Xu, Z.S., et al. (2014) Transcript Profiling of Structural Genes Involved in Cyanidin-Based Anthocyanin Biosynthesis between Purple and Non-Purple Carrot (Daucus carota L.) Cultivars Reveals Distinct Patterns. BMC Plant Biology, 14, 1583-1588.

[15]   Wang, H.Z., Qu, H.Y., Zhou, T.T., et al. (2017) Cloning and Expression Analysis of Anthocyanin Biosynthesis-Associated DFR and MYB Genes in Calyx of Eggplant (Solanum melongena L.). Scientia Agricultura Sinica, 50, 2781-2792.

[16]   Jin, H., Cominelli, E., Bailey, P., et al. (2000) Transcriptional Repression by AtMYB4 Controls Production of UV-Protecting Sunscreens in Arabidopsis. The EMBO Journal, 19, 6150-6161.

[17]   Lin, W.K., Bolitho, K., Grafton, K., et al. (2010) An R2R3-MYB Transcription Factor Associated with Regulation of the Anthocyanin Biosynthetic Pathway in Rosaceae. BMC Plant Biology, 10, 50.

[18]   Vimolmangkang, S., Han, Y., Wei, G., et al. (2013) An Apple MYB Transcription Factor, MdMYB3, Is Involved in Regulation of Anthocyanin Biosynthesis and Flower Development. BMC Plant Biology, 13, 176.

[19]   Aharoni, A., De, V.C.H., Wein, M., et al. (2001) The Strawberry FaMYB1 Transcription Factor Suppresses Anthocyanin and Flavonol Accumulation in Transgenic Tobacco. The Plant Journal, 28, 319-332.

[20]   Wang, L., Han, W., Xie, C., et al. (2013) Comparing the Acceptor Promiscuity of a Rosa hybrida Glucosyltransferase RhGT1 and an Engineered Microbial Glucosyltransferase OleD (PSA) toward a Small Flavonoid Library. Carbohydrate Research, 368, 73-77.

[21]   Liu, X.F., Li, F., Yin, X.R., et al. (2013) Recent Advances in the Transcriptional Regulation of Anthocyanin Biosynthesis. Acta Horticulturae Sinica, 40, 2295-2306.

[22]   Bendahmane, M., Dubois, A., Raymond, O., et al. (2013) Genetics and Genomics of Flower Initiation and Development in Roses. Journal of Experimental Botany, 64, 847-857.

[23]   Albert, N.W., Davies, K.M., Lewis, D.H., et al. (2014) A Conserved Network of Transcriptional Activators and Repressors Regulates Anthocyanin Pigmentation in Eudicots. The Plant Cell, 26, 962-980.

[24]   Takos, A.M., Jaffe, F.W., Jacob, S.R., et al. (2006) Light-Induced Expression of a MYB Gene Regulates Anthocyanin Biosynthesis in Red Apples. Plant Physiology, 142, 1216-1232.

[25]   Lin, W.K., Micheletti, D., Palmer, J., et al. (2011) High Temperature Reduces Apple Fruit Colour via Modulation of the Anthocyanin Regulatory Complex. Plant, Cell & Environment, 34, 1176-1190.