|
|
Study on the Function of TaGRF1-A in the Interaction Between Wheat (Triticum aestivum) and Puccinia triticina |
GU Jia, NIU Ze-Lin, WANG Qian, WANG Dong-Mei* |
State Key Laboratory of North China Crop Improvement and Regulation/Hebei Key Laboratory of Plant Physiology and Molecular Pathology/College of Life Sciences, Hebei Agricultural University, Baoding 071001, China |
|
|
Abstract The cloning of genes related to wheat (Triticum aestivum) resistance leaf rust and the in-depth study of their expression characteristics are of great significance for elucidating the molecular mechanisms of wheat resistance Puccinia triticina. By analyzing the RNA-seq transcriptome database obtained in the laboratory in the early stage, a gene belong to 14-3-3 family closely related to resistance to P. triticina infection was discovered, with significant differences in expression between compatible and incompatible combinations. In order to deeper understanding of the mechanism of this gene in wheat resistance to P. triticina infection, this study cloned the gene from wheat near isogenic lines TcLr26, the result of sequence alignment showed that the gene was TaGRF1-A in wheat. Preparation of polyclonal antibodies against TaGRF1-A protein, Western blot analysis showed that the antibody was able to bind specifically to TaGRF1-A protein in wheat, and showed an upregulation and then decrease expression trend induced by Puccinia triticina in the incompatible combinations. The virus induced gene silencing (VIGS) technology was used to reduce the expression of TaGRF1-A in incompatible combinations, inoculate with leaf rust showed a significant increase in the area of hypersensitive reaction (HR) and the number of haustorium mother cells (HMCs) at the site of P. triticina infection. After 15 d of inoculation with leaf rust, a spore of P. triticina appeared on the surface of the leaves, and the expression of pathogenesis-related (PR) genes were reduced. The results indicated that TaGRF1-A was positive regulating the wheat resistance to leaf rust infection. This study provides reference for further exploring the mechanism of TaGRF1-A in the interaction between wheat and leaf rust.
|
Received: 28 August 2023
|
|
Corresponding Authors:
*dongmeiwang63@126.com
|
|
|
|
[1] 刘娜, 乔妹, 孙嘉伟, 等. 2019. 叶锈菌侵染的小麦叶片转录组数据分析[J]. 植物遗传资源学报, 20(4): 991-1000. (Liu N, Qiao M, Sun J W, et al.2019. Transcriptome analysis of wheat leaves infected by Puccinia triticina[J]. Journal of Plant Genetic Resources, 20(4): 991-1000.) [2] 刘娜, 孙天杰, 陈琰等. 2020. 小麦与叶锈菌互作过程中TabZIP3的功能研究[J].农业生物技术学报, 28(02):201-210. (Liu N, Sun T J, Chen Y, et al.2020. Study on the function of TabZIP3 in the interaction between wheat (Triticum aestivum) and Puccinia triticina[J]. Journal of Agricultural Biotechnology, 28(02): 201-210.) [3] Aitken A.2006. 14-3-3 proteins: A historic overview[J]. Semin Cancer Biology, 16(3): 162-172. [4] Bian S, Jin D, Li R, et al.2017. Genome-wide analysis of CCA1-like proteins in soybean and functional characterization of GmMYB138a[J]. International Journal of Molecular Sciences, 18(10): 117-137. [5] Bihn E A, Paul A L, Wang S W, et al.1997. Localization of 14-3-3 proteins in the nuclei of Arabidopsis and maize[J]. Plant Journal, 12(6): 1439-1445. [6] Camoni L, Visconti S, Aducci P, et al.2018. 14-3-3 proteins in plant hormone signaling: Doing several things at once[J]. Front Plant Science, 9: 297-305. [7] Campo S, Peris-Peris C, Montesinos L, et al.2012. Expression of the maize ZmGF14-6 gene in rice confers tolerance to drought stress while enhancing susceptibility to pathogen infection[J]. Journal of Experimental Botany, 63(2): 983-999. [8] Cao A, Jain A, Baldwin J C, et al.2007. Phosphate differentially regulates 14-3-3 family members and GRF9 plays a role in Pi starvation induced responses[J]. Planta, 226: 1219-1230. [9] Chevalier D, Morris E R, Walker J C.2009. 14-3-3 and FHA domains mediate phosphoprotein interactions[J]. Annual Review of Plant Biology, 60: 67-91. [10] Chen F, Li Q, Sun L, et al.2006. The rice 14-3-3 gene family and its involvement in responses to biotic and abiotic stress[J]. DNA Research, 13(2): 53-63. [11] Deb S, Ghosh P, Patel H K, et al.2020. Interaction of the Xanthomonas effectors XopQ and XopX results in induction of rice immune responses[J]. Plant Journal, 104: 332-350. [12] Denison F C, Paul A L, Zupanska A K, et al.2011. 14-3-3 proteins in plant physiology[J]. Seminars in Cell & Developmental Biology, 22(7): 720-727. [13] Evangelisti E, Guyon A, Shenhav L, et al.2023. FIRE Mimics a 14-3-3-binding motif to promote Phytophthora palmivora infection[J]. Molecular Plant Microbe Interactions, 36(6): 315-322. [14] Faris J D, Haen K M, Gill B S.2000. Saturation mapping of a gene-rich recombination hot spot region in wheat[J]. Genetics, 154(2): 823-835. [15] Hermeking H, Benzinger A.2006. 14-3-3 proteins in cell cycle regulation[J]. Semin Cancer Biology, 16(3): 183-192. [16] Hirsch S, Aitken A, Bertsch U, et al.1992. A plant homologue to mammalian brain 14-3-3 protein and protein kinase C inhibitor[J]. FEBS Letter, 296(2): 222-224. [17] Huang Y, Wang W, Yu H, et al.2022. The role of 14-3-3 proteins in plant growth and response to abiotic stress[J]. Plant Cell Reports, 41(4): 833-852. [18] Konagaya K, Matsushita Y, Kasahara M, et al.2004. Members of 14-3-3 protein isoforms interacting with the resistance gene product N and the elicitor of Tobacco mosaic virus[J]. Journal of General Plant Pathology, 70: 221-231. [19] Li W, Yadeta K A, Elmore J M,et al.2013. The Pseudomonas syringae effector HopQ1 promotes bacterial virulence and interacts with tomato 14-3-3 proteins in a phosphorylation-dependent manner[J]. Plant Physiology,161(4): 2062-2074. [20] Liu Q, Zhang S, Liu B.2016. 14-3-3 proteins: Macro-regulators with great potential for improving abiotic stress tolerance in plants[J]. Biochemical and Biophysical Research Communications, 477(1): 9-13. [21] Manickavelu A, Kawaura K, Oishi K, et al.2010. Comparative gene expression analysis of susceptible and resistant near-isogenic lines in common wheat infected by Puccinia triticina[J]. DNA Research, 17(4): 211-222. [22] Manosalva P M, Bruce M, Leach J E.2011. Rice 14-3-3 protein (GF14e) negatively affects cell death and disease resistance[J]. The Plant Journal, 68(5): 777-787. [23] Nimchuk Z, Eulgem T, Holt B F, et al.2003. Recognition and response in the plant immune system[J]. Annual Review of Genetics, 37: 579-609. [24] Oh C S, Martin G B.2011. Tomato 14-3-3 protein TFT7 interacts with a MAP kinase kinase to regulate immunity-associated programmed cell death mediated by diverse disease resistance proteins[J]. Journal of Biological Chemistry, 286(16): 14129-14136. [25] Qiao M, Sun J W, Liu N, et al.2015. Changes of nitric oxide and its relationship with H2O2 and Ca2+ in defense interactions between wheat and Puccinia triticina[J]. PLOS ONE, 10(7): e0132265. [26] Ren YR, Yang YY, Zhang R, et al.2019. MdGRF11, an apple 14-3-3 protein, acts as a positive regulator of drought and salt tolerance[J]. Plant Science, 288: 110219-110230. [27] Robb J, Lee B, Nazar R N.2007. Gene suppression in a tolerant tomato-vascular pathogen interaction[J]. Planta, 226(2): 299-309. [28] Roberts M R, Bowles D J.1999. Fusicoccin, 14-3-3 proteins, and defense responses in tomato plants[J]. Plant Physiology, 119(4): 1243-1250. [29] Rohringer K, Kim W K.1977. An optical brighter for fluorescence microscopy of fungal plant parasites in leaves[J]. Phytopathology, 67(1): 808-810. [30] Rooney M F, Ferl R J.1995. Sequences of three Arabidopsis general regulatory factor genes encoding GF14 (14-3-3) proteins[J]. Plant Physiology, 107(1): 283-284. [31] Seo Y E, Yan X, Choi D, et al.2023. Phytophthora infestans RxLR effector PITG06478 hijacks 14-3-3 to suppress PMA activity leading to necrotrophic cell death[J]. Molecular Plant Microbe Interactions, 36(3):150-158. [32] Shao W, Chen W, Zhu X, et al.2021. Genome-wide identification and characterization of wheat 14-3-3 genes unravels the role of TaGRF6-A in salt stress tolerance by binding MYB transcription factor[J]. International Journal of Molecular Sciences, 22(4): 1904-1922. [33] Scofield S R, Huang L, Brandt A S, et al.2005. Development of a virus-induced gene-silencing system for hexaploid wheat and its use in functional analysis of the Lr21-mediated leaf rust resistance pathway[J]. Plant Physiology, 138(4): 2165-2173. [34] van Heusden G P, Griffiths D J, Ford J C, et al.1995. The 14-3-3 proteins encoded by the BMH1 and BMH2 genes are essential in the yeast Saccharomyces cerevisiae and can be replaced by a plant homologue[J]. European Journal of Biochemistry, 229(1): 45-53. [35] Wurtele M, Jelich-Ottmann C, Wittinghofer A, et al.2003. Structural view of a fungal toxin acting on a 14-3-3 regulatory complex[J]. EMBO Journal, 22(5): 987-994. [36] Yang X, Wang W, Coleman M, et al.2009. Arabidopsis 14-3-3 lambda is a positive regulator of RPW8-mediated disease resistance[J]. Plant Journal, 60(3): 539-550. [37] Yao Y, Du Y, Jiang L, et al.2007. Molecular analysis and expression patterns of the 14-3-3 gene family from Oryza sativa[J]. Journal of Biochemistry and Molecular Biology, 40(3): 349-357. [38] Yashvardhini N, Bhattacharya S, Chaudhuri S, et al.2018. Molecular characterization of the 14-3-3 gene family in rice and its expression studies under abiotic stress[J]. Planta, 247(1): 229-253. [39] Zhang Y, Zhao H, Zhou S, et al.2018. Expression of TaGF14b, a 14-3-3 adaptor protein gene from wheat, enhances drought and salt tolerance in transgenic tobacco[J]. Planta, 248(1): 117-137. [40] Zhao X, Li F, Li K.2021. The 14-3-3 proteins: Regulators of plant metabolism and stress responses[J]. Plant Biology , 23(4): 531-539. |
|
|
|