Vector Construction of Cis-acting Regulatory DNA Elements of Rice Blast Resistance Gene Pi63

doi:10.11924/j.issn.1000-6850.casb15060135

Abstract

Abstract: Rice blast resistance gene Pi63 has fine field resistance to the rice blast disease and its resistance levels are closely correlated with its gene expression levels. However, the mechanism of the expression of Pi63 has not been investigated. In this study, cis-acting regulatory DNA elements in promoter region of Pi63 were predicted by bioinformatics analysis. 4 constructs containing different elements were obtained using plant binary vector pCAMBIA1391Z. The positive clones were verified by colony PCR and restriction enzyme digestion and further confirmed by Sanger sequencing. The results showed that the 4 deletant vectors were successfully constructed, which laid the foundation for further research on the effect of different cis-acting regulatory DNA element on the resistance of Pi63 and the expression pattern of Pi63.

CLC Number:

S511

Liang Huabing,Wang Xiaowan,Hu Xiaolan,Zhao Feng,Zhang Chengguo,Han Shuhui,Yao Fucheng and Xu Xin. Vector Construction of Cis-acting Regulatory DNA Elements of Rice Blast Resistance Gene Pi63[J]. Chinese Agricultural Science Bulletin, 2015, 31(30): 220-224.

References

[1] Khush G S. What it will take to feed 5.0 billion rice consumers in 2030[J]. Plant Molecular Biology,2005,59(1):1-6.
[2] Khush G S, Jena K K. Current status and future prospects for research on blast resistance in rice (Oryza sativa L.)[M]. Advances in genetics, genomics and control of rice blast disease. Springer Netherlands,2009:1-10.
[3] McDowell J M, Woffenden B J. Plant disease resistance genes: recent insights and potential applications[J]. Trends in Biotechnology,2003,21(4):178-183.
[4] Koide Y, Telebanco-Yanoria M J, Fukuta Y, et al. Detection of novel blast resistance genes, Pi58 (t) and Pi59 (t), in a Myanmar rice landrace based on a standard differential system[J]. Molecular Breeding,2013,32(2):241-252.
[5] Liu J, Wang X, Mitchell T, et al. Recent progress and understanding of the molecular mechanisms of the rice-Magnaporthe oryzae interaction[J]. Molecular Plant Pathology,2010,11(3):419-427.
[6] Liu W, Liu J, Triplett L, et al. Novel insights into rice innate immunity against bacterial and fungal pathogens. Annual Review of Phytopathology,2014(52):213-241.
[7] Chen X, Shang J, Chen D, et al. A β-lectin receptor kinase gene conferring rice blast resistance[J]. The Plant Journal,2006,46(5):794-804.
[8] Fukuoka S, Saka N, Koga H, et al. Loss of function of a proline-containing protein confers durable disease resistance in rice. Science,2009,325(5943):998-1001.
[9] Inoue H, Hayashi N, Matsushita A, et al. Blast resistance of CC-NB-LRR protein Pb1 is mediated by WRKY45 through protein-protein interaction. Proceedings of the National Academy of Sciences,2013,110(23):9577-9582.
[10] Xu X, Chen H, Fujimura T, et al. Fine mapping of a strong QTL of field resistance against rice blast, Pikahei-1 (t), from upland rice Kahei, utilizing a novel resistance evaluation system in the greenhouse[J]. Theoretical and Applied Genetics,2008,117(6):997-1008.
[11] Xu X, Hayashi N, Wang C T, et al. Rice blast resistance gene Pikahei-1 (t), a member of a resistance gene cluster on chromosome 4, encodes a nucleotide-binding site and leucine-rich repeat protein[J]. Molecular Breeding,2014,34(2):691-700.
[12] Liu X, Lin F, Wang L, et al. The in silico map-based cloning of Pi36, a rice coiled-coilnucleotide-binding site leucine-rich repeat gene that confers race-specific resistance to the blast fungus[J]. Genetics,2007,176(4):2541-2549.
[13] Liu Y, Liu B, Zhu X, et al. Fine-mapping and molecular marker development for Pi56(t), a NBS-LRR gene conferring broad-spectrum resistance to Magnaporthe oryzae in rice[J]. Theoretical and Applied Genetics,2013,126(4):985-998.
[14] Hayashi N, Inoue H, Kato T, et al. Durable panicle blast-resistance gene Pb1 encodes an atypical CC-NBS-LRR protein and was generated by acquiring a promoter through local genome duplication[J]. The Plant Journal,2010,64(3):498-510.
[15] Eulgem T, Rushton P J, Schmelzer E, et al. Early nuclear events in plant defense signaling: rapid gene activation by WRKY transcription factors[J]. The EMBO Journal,1999,18(17):4689-4699.
[16] Eulgem T, Rushton P J, Robatzek S, et al. The WRKY superfamily of plant transcription factors[J]. Trends in Plant Science,2000,5(5):199-206.
[17] Shimono M, Koga H, Akagi A Y A, et al. Rice WRKY45 plays important roles in fungal and bacterial disease resistance[J]. Molecular Plant Pathology,2012,13(1):83-94.
[18] Brown R L, Kazan K, McGrath K C, et al. A role for the GCC-box in jasmonate-mediated activation of the PDF1. 2 gene of Arabidopsis[J]. Plant Physiology,2003,132(2):1020-1032.
[19] Luo H, Song F, Goodman R M, et al. Up-regulation of OsBIHD1, a rice gene encoding BELL homeodomain transcriptional factor, in disease resistance responses[J]. Plant Biology,2005,7(5):459-468.
[20] Fusada N, Masuda T, Kuroda H, et al. Identification of a novel cis-element exhibiting cytokinin-dependent protein binding in vitro in the 5′-region of NADPH-protochlorophyllide oxidoreductase gene in cucumber[J]. Plant Molecular Biology,2005,59(4):631-645.
[21] Choi J, Choi D, Lee S, et al. Cytokinins and plant immunity: old foes or new friends?[J]. Trends in Plant Science,2011,16(7):388-394.
[22] Xu N, Hagen G, Guilfoyle T. Multiple auxin response modules in the soybean SAUR 15A promoter[J]. Plant Science,1997,126(2):193-201.
[23] Chakravarthy S, Tuori R P, D''Ascenzo M D, et al. The tomato transcription factor Pti4 regulates defense-related gene expression via GCC box and non-GCC box cis elements[J]. The Plant Cell,2003,15(12):3033-3050.
[24] Wang D, Qin Y, Han J, et al. Expression analysis of innate immunity related genes in the true/field blast resistance gene-mediated defence response[J]. Biotechnology & Biotechnological Equipment,2014,28(6):999-1007.