Volume 3, Issue 2, April 2015, Page: 85-91
Molecular Cloning, Characterization and Expression Analysis of MhRAR1 Gene from Malus Hupehensis
Zhang Ji-Yu, Institute of Botany, Jiangsu Province and the Chinese Academy of Sciences, Nanjing, China
Guo Zhong-Ren, Institute of Botany, Jiangsu Province and the Chinese Academy of Sciences, Nanjing, China
Received: Mar. 9, 2015;       Accepted: Mar. 22, 2015;       Published: Mar. 26, 2015
DOI: 10.11648/j.jps.20150302.17      View  2707      Downloads  155
Abstract
A novel RAR1 gene, designated MhRAR1, was cloned by the methods of RT-PCR and RACE from Malus hupehensis. The full length sequence of MhRAR1 is 1065 bp with an open reading frame of 678 bp, encoding a protein of 225 amino acids. As found in other plant RAR1 proteins, sequence alignment showed that MhRAR1 protein contains two CHORD domains and one plant-specific CCCH domain. In addition, the MhRAR1 contains conserved strings of invariant cysteine and histidine residues within the CHORD domains and CCCH domain. These results suggested that MhRAR1 protein from M. hupehensis might share the similar function with the Arabidopsis thaliana RAR1 and Hordeum vulgare RAR1, and is an important component of R gene–mediated disease resistance. Phylogenetic analysis revealed that MhRAR1 was closely related to Ricinus communis RAR1. The analysis by qRT-PCR revealed that the expression of MhRAR1 gene was higher in leaves than that in stems and roots. SA, MeJA and ACC treatment induced MhRAR1 expression in stems and roots, but not in leaves. Expression of MhRAR1 was weakly induced in M. hupehensis after infection with Botryosphaeria berengeriana. The cloning and characterization of the MhRAR1 gene will be useful for further studies of biological roles of MhRAR1 in plants.
Keywords
Malus hupehensis, MhRAR1, cDNA Cloning, Expression Pattern
To cite this article
Zhang Ji-Yu, Guo Zhong-Ren, Molecular Cloning, Characterization and Expression Analysis of MhRAR1 Gene from Malus Hupehensis, Journal of Plant Sciences. Vol. 3, No. 2, 2015, pp. 85-91. doi: 10.11648/j.jps.20150302.17
Reference
[1]
Shirasu K., Schulze-Lefert P. 2000. Regulators of cell death in disease resistance. Plant Molecular Biology 44: 371–385
[2]
Azevedo C., Sadanandom A., Kitagawa K., Freialdenhoven K., Shirasu A., Schulze-Lefert P. 2002. The RAR1 Interactor SGT1, an Essential Component of R Gene-Triggered Disease Resistance. Science 295: 2073-2076
[3]
Bieri S., Mauch S., Shen Q.H., Peart J., Devoto A., Casais C., Francesca C., Schulze S., Steinbiß H-H., Shirasu K., Schulze-Leferta P. 2004. RAR1 Positively Controls Steady State Levels of Barley MLA Resistance Proteins and Enables Sufficient MLA6 Accumulation for Effective Resistance. The Plant Cell Online 16: 3480-3495
[4]
Muskett P.R., Kahn K., Austin M.J., Moisan L.J., Sadanandom A., Shirasu K., Jones J.D.G., Parker J.E. 2002. Arabidopsis RAR1 Exerts Rate-Limiting Control of R Gene-Mediated Defenses against Multiple Pathogens. The Plant Cell Online 14: 979-992
[5]
Tornero P., Merritt P., Sadanandom A., Shirasu K., nnes R.W.I., Dangl J.L. 2002. RAR1 and NDR1 Contribute Quantitatively to Disease Resistance in Arabidopsis, and Their Relative Contributions Are Dependent on the R Gene Assayed. The Plant Cell Online 14: 1005-1015
[6]
Shirasu K., Lahaye T., Tan M-W., Zhou F., Azevedo C., Schulze-Lefert P. 1999. A novel class of eukaryotic zinc-binding proteins is required for disease resistance signaling in barley and development in C. elegans. Cell 99: 355-366
[7]
Shang Y., Li X., Cui H., He P., Thilmony R., Chintamanani S., Zwiesler-Vollick J., Gopalan S., Tang X., Zhou J.M. 2006. RAR1, a central player in plant immunity, is targeted by Pseudomonas syringae effector AvrB. Proceedings of the National Academy of Sciences 103: 19200-19205
[8]
Takahashi A., Casais C., Ichimura K., Shirasu K. 2003. HSP90 interacts with RAR1 and SGT1 and is essential for RPS2-mediated disease resistance in Arabidopsis. Proceedings of the National Academy of Sciences 100: 11777-11782
[9]
Tai Y.S. 2007. Interactome of signaling networks in wheat: the protein–protein interaction between TaRAR1 and TaSGT1. Molecular Biology Reports 35: 337-343
[10]
Zellerhoff N., Jansen M., Schaffrath U. 2008. Barley Rom1 antagonizes Rar1 function in Magnaporthe oryzae-infected leaves by enhancing epidermal and diminishing mesophyll defence. New Phytologist 180: 702-710
[11]
Zhang L., Yang W., Liu D. 2011. TaRAR1 is Required for Lr24-Mediated Wheat Leaf Rust Resistance. Agricultural Sciences in China 10: 1732-1738
[12]
Liu Y., Schiff M., Marathe R., Dinesh-Kumar S.P. 2002. Tobacco Rar1, EDS1 and NPR1 NIM1 like genes are required for N mediated resistance to tobacco mosaic virus. The Plant Journal 30: 415-429
[13]
Glazebrook J. 2001. Genes controlling expression of defence responses in Arabidopsis- 2001 status. Curr Opin Plant Biol 4:301–308
[14]
Kim J.C., Lee S.H., Cheong Y.H., Yoo C.M., Lee S.I., Chun H.J., Yun D.J., Hong J.C., Lee S.Y., Lim C.O., Cho M.J. 2001. A novel cold-inducible zinc finger protein from soybean, SCOF-1, enhances cold tolerance in transgenic plants. Plant J 25:247–251
[15]
Dong X. 1998. SA, JA, ethylene, and disease resistance in plants. Curr.Opin. Plant Biol 1:316–323
[16]
Spoel S.H., Koornneef A., Claessens S.M., Korzelius J.P., Pelt J.A., Mueller M.J., Buchala A.J., Metraux J.P., Brown R., Kazan K., Loon L.C., Dong X.N., Pieterse C.M. 2003. NPR1 modulates cross-talk between salicylate- and jasmonate-dependent defense pathways through a novel function in the cytosol. The Plant Cell 15:760-770
[17]
Leon-Reyes A., Spoel S.H., Lange E.S., Abe H., Kobayashi M., Tsuda S., Millenaar F.F., Welschen R.A., Ritsema T., Pieterse C.M. 2009. Ethylene modulates the role of nonexpressor of pathogenesis-related genes1 in cross talk between salicylate and jasmonate signaling. Plant Physiology 149:1797–1809
[18]
Lu Q.N., Jia D.X. 1999. China Fruit Records: Apple Volume, First Ed., China’s Agricultural Science & Technology Press, Beijing
[19]
Zhang J.Y., Qu S.C., Dong C., Gao Z.H., Qiao Y.S., Zhang Z. 2010. Utility and construction of full-length cDNA library of Malus hupehensis post-introduced with salicylic acid. Acta Bot Boreal -Occident. Sin 30 (8):1527-1533
[20]
Tong Z.G., Wang F.R., Zhang Z., Zhao J.B., Zhang K.C., Yan G.H., Zhou Y., Jiang L.J. 2008. A method for DNA extraction from mature leaves of fruit trees. J. Fruit Sci. 25, 122-125
[21]
Tamura K., Dudley J., Nei M., Kumar S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599
[22]
Page R.D. 1996. Tree view: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358
[23]
Cai B.H., Zhang J.Y., Gao Z.H., Qu S.C., Tong Z.G., Mi L., Qiao Y.S., Zhang Z. 2008. An improved method for isolation of total RNA from the leaves of Fragaria spp. Jiangsu Journal of Agricultural Sciences. 24, 875–877
[24]
Livak K.J., Schmittgen T.D. 2001. Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method. Methods 25: 402-408
[25]
Wang Q.J., Xu K.Y., Tong Z.G., Wang S.H., Gao Z.H., Zhang J.Y., Zong C.W., Qiao Y.S., Zhang Z. 2010. Characterization of a new dehydration responsive element binding factor in central arctic cowberry. Plant Cell, Tissue and Organ Culture (PCTOC) 101: 211-219
[26]
Wu J., Luo S., Jiang H., Li H. 2005. Mammalian CHORD-containing protein 1 is a novel heat shock protein 90-interacting protein. FEBS Letters 579: 421-426
[27]
Thao N.P., Chen L., Nakashima A., Hara S., Umemura K., Takahashi A., Shirasu K., Kawasaki T., Shimamoto K. 2007. RAR1 and HSP90 Form a Complex with Rac/Rop GTPase and Function in Innate-Immune Responses in Rice. The Plant Cell Online 19: 4035-4045
[28]
Zhang M., Kadota Y., Prodromou C., Shirasu K., Pearl L.H. 2010. Structural Basis for Assembly of Hsp90-Sgt1-CHORD Protein Complexes: Implications for Chaperoning of NLR Innate Immunity Receptors. Molecular Cell 39: 269-281
[29]
Eitas T.K., Nimchuk Z.L., Dangl J.L. 2008. Arabidopsis TAO1 is a TIR-NB-LRR protein that contributes to disease resistance induced by the Pseudomonas syringae effector AvrB. Proceedings of the National Academy of Sciences of the USA, 105: 6475-6480
[30]
Hubert D.A., He Y., McNulty B.C., Tornero P., Dangl J.L. 2009. Specific Arabidopsis HSP90.2 alleles recapitulate RAR1 cochaperone function in plant NB-LRR disease resistance protein regulation. Proceedings of the National Academy of Sciences 106: 9556-9563
[31]
Dangl J.L., Jones J.D. 2001. Plant pathogens and integrated defence responses to infection. Nature 411: 826-833
[32]
Austin M.J., Muskett P., Kahn K., Fey B.J., Jones J.D.G., Parker J.E. .2002. Regulatory Role of SGT1 in Early R Gene-Mediated Plant Defenses. Science 295: 2077-2080
Browse journals by subject