Back
 AJPS  Vol.9 No.11 , October 2018
Cloning and Expression Analysis of RrG-Beta1 Gene Related to Anthocyanin Biosynthesis in Rosa rugose
Abstract: As an important signal transduction protein, G protein beta subunit gene encoded by oligonucleotides plays an important role in many physiological, biochemical and environmental stresses in plants. In order to understand the action mode of G protein beta subunit gene, this paper cloned a Wd40 gene related to G protein beta subunit gene, named RrG-beta1, based on the R. rugose—transcriptome data, using Rosa rugose “Zi zhi” as experimental materials. The full length of cDNA of the gene was obtained by RT-PCR and RACE methods. The total length of this gene is 981 bp, and it encodes 326 amino acids. After bioinformatics analysis, the molecular formula C1601H2520N450O486S11 was predicted; the relative molecular weight was 36,201.00 Da; the theoretical isoelectric point PI value was 6.71; and its instability index was 30.44. The total average hydrophobic index was -0.847. In the secondary structure of RrG-beta1 protein, there are 17 α-helix, 131 Random coil, and 141 extended peptide chain. Gene Bank Blast results showed that the amino acid sequence encoded by RrG-beta1 was more than 90% homologous with the beta-like protein of Rosa chinensis, Fragaria, Malus, Pyrus, Prunus, Arabidopsis and tobacco, so it can be inferred that the RrG-beta1 Gene is guanine nucleotide-binding protein subunit beta-like protein. Fluorescence quantitative expression analysis of RrG-beta1 protein decreased with the development of flower color, and it was speculated that it could exert negative regulation effect on flower color. The leaf expression was highest in the tissue part, so it was inferred that the signal was transmitted through the stoma on the leaf.
Cite this paper: Wang, Y. , Zhao, M. , Han, X. , Zhao, L. and Xu, Z. (2018) Cloning and Expression Analysis of RrG-Beta1 Gene Related to Anthocyanin Biosynthesis in Rosa rugose. American Journal of Plant Sciences, 9, 2244-2255. doi: 10.4236/ajps.2018.911162.
References

[1]   Buer, C.S., Imin, N., Djordjevic, M.A. and Lucas, W.J. (2010) Flavonoids: New Roles for Old Molecules. Journal of Integrative Plant Biology, 52, 98-111.
https://doi.org/10.1111/j.1744-7909.2010.00905.x

[2]   Wenjian, Y.U., Gao, P., Xiong, Y. and Wang, J. (2017) Evaluation and Selection of Adaptability of Twelve Climbing Rose Varieties. Northern Horticulture, 15, 79-83.

[3]   Ben-Simhon, Z., Judeinstein, S., Nadler-Hassar, T., Trainin, T., Bar-Ya’Akov, I., Borochov-Neori, H., et al. (2011) A Pomegranate (Punica granatum L.) WD40-Repeat Gene Is a Functional Homologue of Arabidopsis TTG1, and Is Involved in the Regulation of Anthocyanin Biosynthesis during Pomegranate Fruit Development. Planta, 234, 865-881.
https://doi.org/10.1007/s00425-011-1438-4

[4]   Lloyd, A., Brockman, A., Aguirre, L., Campbell, A., Bean, A., Cantero, A., et al. (2017) Advances in the MYB-BHLH-WD Repeat (MBW) Pigment Regulatory Model: Addition of a WRKY Factor and Co-Option of an Anthocyanin MYB for Betalain Regulation. Plant & Cell Physiology, 58, 1431-1441.
https://doi.org/10.1093/pcp/pcx075

[5]   Islasflores, T., Guillén, G., Islasflores, I., San, R.C., Sánchez, F., Lozatavera, H., et al. (2009) Germination Behavior, Biochemical Features and Sequence Analysis of the RACK1/arcA Homolog from Phaseolus vulgaris. Physiologia Plantarum, 137, 264-280.
https://doi.org/10.1111/j.1399-3054.2009.01280.x

[6]   Sethi, V.P. and Dubey, R.K. (2007) Multi-Rack Tray System for Off-Season Greenhouse Nursery Raising. Journal of Research, 357-363.

[7]   Nitta, Y., Ding, P. and Zhang, Y. (2015) Heterotrimeric G Proteins in Plant Defense against Pathogens and ABA Signaling. Environmental & Experimental Botany, 114, 153-158.
https://doi.org/10.1016/j.envexpbot.2014.06.011

[8]   Chakravorty, D., Trusov, Y. and Botella, J.R. (2012) Site-Directed Mutagenesis of the Arabidopsis Heterotrimeric G Protein β Subunit Suggests Divergent Mechanisms of Effector Activation between Plant and Animal G Proteins. Planta, 235, 615-627.
https://doi.org/10.1007/s00425-011-1526-5

[9]   Urano, D., Maruta, N., Trusov, Y., Stoian, R., Wu, Q., Liang, Y., et al. (2016) Saltational Evolution of the Heterotrimeric G Protein Signaling Mechanisms in the Plant Kingdom. Science Signaling, 9, ra93.
https://doi.org/10.1126/scisignal.aaf9558

[10]   Zhang, H., Wang, M., Wang, W., Li, D., Huang, Q., Wang, Y., et al. (2012) Silencing of G Proteins Uncovers Diversified Plant Responses When Challenged by Three Elicitors in Nicotiana benthamiana. Plant Cell & Environment, 35, 72-85.
https://doi.org/10.1111/j.1365-3040.2011.02417.x

[11]   Gookin, T.E. and Bendtsen, J.D. (2013). Topology Assessment, G Proein-Coupled Receptor (GPCR) Prediction, and in Vivo Interaction Assays to Identify Plant Candidate GPCRS. G Protein-Coupled Receptor Signaling in Plants, 1043, 1-12.
https://doi.org/10.1007/978-1-62703-532-3_1

[12]   Hao, Z.M., Li, Z.Y., Dong, J.G., et al. (2008) Cloning and Expression of the Gene Encoding the G Protein of Maize Big Spotted Bacteria. Journal of Agricultural Biotechnology, 16, 1025-1030.

[13]   Jones, A.M. (2002) G-Protein-Coupled Signaling in Arabidopsis. Current Opinion in Plant Biology, 5, 402-407.
https://doi.org/10.1016/S1369-5266(02)00288-1

[14]   Lein, W. and Saalbach, G. (2001) Cloning and Direct G-Protein Regulation of Phospholipase D from Tobacco. BBA—Molecular and Cell Biology of Lipids, 1530, 172-183.
https://doi.org/10.1016/S1388-1981(00)00182-7

[15]   Ishikawa, A., Iwasaki, Y. and Asahi, T. (1995) Molecular Cloning and Characterization of a cDNA for the α Subunit of a G Protein from Rice. Plant & Cell Physiology, 36, 353-359.
https://doi.org/10.1093/oxfordjournals.pcp.a078767

[16]   Kaydamov, C., Tewes, A., Adler, K. and Manteuffel, R. (2000) Molecular Characterization of Cdnas Encoding G Protein α and β Subunits and Study of Their Temporal and Spatial Expression Patterns in Nicotiana Plumbaginifolia Viv. Biochimica et Biophysica Acta (BBA)—Gene Structure and Expression, 1491, 143-160.
ttps://doi.org/10.1016/S0167-4781(00)00039-7

[17]   Lease, K.A., Wen, J., Li, J., Doke, J.T., Liscum, E. and Walker, J.C. (2001) A Mutant Arabidopsis Heterotrimeric g-Protein Beta Subunit Affects Leaf, Flower, and Fruit Development. Plant Cell, 13, 2631.
https://doi.org/10.1105/tpc.13.12.2631

[18]   Trusov, Y., Rookes, J.E., Tilbrook, K., Chakravorty, D., Mason, M.G., Anderson, D., et al. (2007) Heterotrimeric G Protein γ Subunits Provide Functional Selectivity in g β γ Dimer Signaling in Arabidopsis. Plant Cell, 19, 1235-1250.
https://doi.org/10.1105/tpc.107.050096

[19]   Rothe, C. and Lehle, L. (1998) Sorting of Invertase Signal Peptide Mutants in Yeast Dependent and Independent on the Signal-Recognition Particle. FEBS Journal, 252, 16-24.

[20]   Dodd, I.C. (2005) Root-to-Shoot Signalling: Assessing the Roles of “Up” in the up and down World of Long-Distance Signalling in Planta. Plant & Soil, 274, 251-270.
https://doi.org/10.1007/s11104-004-0966-0

[21]   Ullah, H., Temple, B., Alonso, J.M., Ecker, J.R. and Jones, A.M. (2006) Rack1 Mediates Multiple Hormone Responsiveness and Developmental Processes in Arabidopsis. Journal of Experimental Botany, 57, 2697-2708.
https://doi.org/10.1093/jxb/erl035

[22]   Clarke, J.M. and Mccaig, T.N. (1982) Evaluation of Techniques for Screening for Drought Resistance in Wheat. Crop Science, 22, 503-506.
https://doi.org/10.2135/cropsci1982.0011183X002200030015x

[23]   Winter, S.R. (1988) Evaluation of Screening Techniques for Breeding Drought-Resistant Winter Wheat. Crop Science, 28, 512-516.
https://doi.org/10.2135/cropsci1988.0011183X002800030018x

[24]   Rahman, S., Shaheen, M.S., Rahman, M. and Malik, T.A. (2000) Evaluation of Excised Leaf Water Loss and Relative Water Content, as Screening Techniques for Breeding Drought Resistant Wheat. Pakistan Journal of Biological Sciences, 3, 663-665.
https://doi.org/10.3923/pjbs.2000.663.665

[25]   Tachibana, M., Wilcox, E., Yokotani, N., Schneider, M. and Fex, J. (1992) Selective Amplification and Partial Sequencing of Cdnas Encoding G Protein Alpha Subunits from Cochlear Tissues. Hearing Research, 62, 82.
https://doi.org/10.1016/0378-5955(92)90204-Z

 
 
Top