|
|
|
|
|
| Molecular cloning and functional identification of an apple flagellin receptor MdFLS2 gene |
| QI Chen-hui1, ZHAO Xian-yan1, JIANG Han1, LIU Hai-tao2, WANG Yong-xu1, HU Da-gang1, HAO Yu-jin1 |
1 National Key Laboratory of Crop Biology/Key Laboratory of Horticultural Crop Biology and Germplasm Innovation, Ministry of Agriculture and Rural Affairs of China/College of Horticulture Science and Engineering, Shandong Agricultural University, Tai’an 271018, P.R.China
2 Shandong Yihui Detection Technology Co., Ltd., Tai’an 271000, P.R.China |
|
|
|
|
Abstract The leucine-rich repeat receptor kinase flagellin-sensing 2 gene (MdFLS2; Gene ID: MDP0000254112) was cloned from Royal Gala apple (Malus×domestica Borkh.). This gene contained a complete open reading frame of 3 474 bp that encoded 1 158 amino acids. The phylogenetic tree indicated that Prunus persica FLS2 exhibited the highest sequence similarity to MdFLS2. The PlantCare database suggests that the promoter sequence of MdFLS2 contains several typical cis-acting elements, including ethylene-, gibberellin-, salicylic acid-, and drought-responsive elements. Quantitative real-time PCR analysis showed that MdFLS2 was widely expressed in the different tissues of the apple and most highly expressed in the leaves. Furthermore, MdFLS2 was significantly induced by the flagellin elicitor peptide flg22. Treatment of the apple seedling leaves with flg22 resulted in an increase in leaf callose levels with increased treatment duration. An increase in the production of O2– along with the expression of disease-related genes was also observed. An oxidative burst was detected in the treated seedlings, but not in the control seedlings, indicating that flg22 had stimulated the expression of the MdFLS2 gene and its downstream target genes. Furthermore, the ectopic expression of MdFLS2 complemented the function of the Arabidopsis fls2 mutant and conferred enhanced flg22 tolerance to the transgenic Arabidopsis, suggesting that MdFLS2 acts as a positive regulator in the response to pathogens in apple.
|
|
Received: 11 January 2018
Accepted:
|
| Fund: This work was supported by the National Natural Science Foundation of China (31601728 and 31430074), the Ministry of Education of China (IRT15R42), the Natural Science Foundation of Shandong Province, China (ZR2016CQ13 and SDAIT-06-03), the Young Scientists Funds of Shandong Agricultural University, China (564024 and 24024). |
|
Corresponding Authors:
CorrespondenceHUD a-gang, Tel/Fax: +86-538-8246151,E-mail:fap_296566@163.com; HAO Yu-jin, E-mail: haoyujin@sdau.edu.cn
|
| About author: QI Chen-hui, E-mail: 18763823286@163.com; |
Cite this article:
QI Chen-hui, ZHAO Xian-yan, JIANG Han, LIU Hai-tao, WANG Yong-xu, HU Da-gang, HAO Yu-jin.
2018.
Molecular cloning and functional identification of an apple flagellin receptor MdFLS2 gene. Journal of Integrative Agriculture, 17(12): 2694-2703.
|
Albert M, Jehle A K, Fürst U, Chinchilla D, Boller T, Felix G. 2013. A two-hybrid-receptor assay demonstrates heteromer formation as switch-on for plant immune receptors. Plant Physiology, 163, 1504–1509.
Böhm H, Albert I, Fan L, Reinhard A, Nürnberger T. 2014. Immune receptor complexes at the plant cell surface. Current Opinion in Plant Biology, 20, 47–54.
Boller T, Felix G. 2009. A renaissance of elicitors: Perception of microbe-associated molecular patterns and danger signals by pattern-recognition receptors. Annual Review of Plant Biology, 60, 379–406.
Cai R, Lewis J, Yan S, Liu H, Clarke C R, Campanile F, Almeida N F, Studholme D J, Lindeberg M, Schneider D, Zaccardelli M, Setubal J C, Morales-Lizcano N P, Bernal A, Coaker G, Baker C, Bender C L, Leman S, Vinatzer B A. 2011. The plant pathogen Pseudomonas syringae pv. tomato is genetically monomorphic and under strong selection to evade tomato immunity. PLoS Pathogens, 7, e1002130.
Chinchilla D, Bauer Z, Regenass M, Boller T, Felix G. 2006. The Arabidopsis receptor kinase FLS2 binds flg22 and determines the specificity of flagellin perception. The Plant Cell, 18, 465–476.
Chinchilla D, Shan L, He P, de Vries S, Kemmerling B. 2009. One for all: The receptor-associated kinase BAK1. Trends in Plant Science, 14, 535–541.
Clarke C R, Chinchilla D, Hind S R, Taguchi F, Miki R, Ichinose Y, Martin G B, Leman S, Felix G, Vinatzer B A. 2013. Allelic variation in two distinct Pseudomonas syringae flagellin epitopes modulates the strength of plant immune responses but not bacterial motility. New Phytologist, 200, 847–860.
Dou D, Zhou J M. 2012. Phytopathogen effectors subverting host immunity: Different foes, similar battleground. Cell Host & Microbe, 12, 484–495.
Felix G, Duran J D, Volko S, Boller T. 1999. Plants have a sensitive perception system for the most conserved domain of bacterial flagellin. The Plant Journal, 18, 265–276.
Gómez-Gómez L, Boller T. 2000. FLS2: An LRR receptor-like kinase involved in the perception of the bacterial elicitor flagellin in Arabidopsis. Molecular Cell, 5, 1003–1011.
Gómez-Gómez L, Felix G, and Boller T. 1999. A single locus determines sensitivity to bacterial flagellin in Arabidopsis thaliana. The Plant Journal, 18, 277–284.
Gust A A, Biswas R, Lenz H D, Rauhut T, Ranf S, Kemmerling B, Götz F, Glawischnig E, Lee J, Felix G, Nürnberger T. 2007. Bacteria-derived peptidoglycans constitute pathogen-associated molecular patterns triggering innate immunity in Arabidopsis. The Journal of Biological Chemistry, 282, 32338–32348.
Hao G, Pitino M, Duan Y, Stover E. 2016. Reduced susceptibility to Xanthomonas citri in transgenic citrus expressing the FLS2 receptor from Nicotiana benthamiana. Molecular Plant-Microbe Interactions, 29, 132–142.
Hu D G, Sun C H, Zhang Q Y, An J P, You C X, Hao Y J. 2016. Glucose sensor MdHXK1 phosphorylates and stabilizes MdbHLH3 to promote anthocyanin biosynthesis in apple. PLoS Genetics, 12, e1006273.
Kunze G, Zipfel C, Robatzek S, Niehaus K, Boller T, Felix G. 2004. The N terminus of bacterial elongation factor Tu elicits innate immunity in Arabidopsis plants. The Plant Cell, 16, 3496–3507.
Liu M, Lei L, Powers C, Liu Z, Campbell K G, Chen X, Bowden R L, Carver B F,Yan L. 2016. TaXA21-A1 on chromosome 5AL is associated with resistance to multiple pests in wheat. Theoretical and Applied Genetics, 129, 345–355.
Lu D, Lin W, Gao X, Wu S, Cheng C, Avila J, Heese A, Devarenne T P, He P, Shan L. 2011. Direct ubiquitination of pattern recognition receptor FLS2 attenuates plant innate immunity. Science, 332, 1439–1442.
Meyer A, Pühler A, Niehaus K. 2001. The lipopolysaccharides of the phytopathogen Xanthomonas campestris pv. campestris induce an oxidative burst reaction in cell cultures of Nicotiana tabacum. Planta, 213, 214–222.
Miya A, Albert P, Shinya T, Desaki Y, Ichimura K, Shirasu K, Narusaka Y, Kawakami N, Kaku H, Shibuya N. 2007. CERK1, a LysM receptor kinase, is essential for chitin elicitor signaling in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 104, 19613–19618.
Monaghan J, Zipfel C. 2012. Plant pattern recognition receptor complexes at the plasma membrane. Current Opinion in Plant Biology, 15, 349–357.
Ottmann C, Luberacki B, Küfner I, Koch K, Brunner F, Weyand M, Mattinen L, Pirhonen M, Anderluh G, Seitz H U, Nürnberger T, Oecking C. 2009. A common toxin fold mediates microbial attack and plant defense. Proceedings of the National Academy of Sciencesof the United States of America, 106, 10359–10364.
Takeuchi O, Akira S. 2010. Pattern recognition receptors and inflammation. Cell, 140, 805–820.
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731–2739.
Thomma B P, Nürnberger T, Joosten M H. 2011. Of PAMPs and effectors: The blurred PTI-ETI dichotomy. The Plant Cell, 23, 4–15.
Trdá L, Fernandez O, Boutrot F, Heloir M, Kelloniemi J, Daire X, Adrian M, Clement C, Zipfel C, Dorey S. 2013. The grapevine flagellin receptor VvFLS2 differentially recognizes flagellin derived epitopes from the endophytic growth-promoting bacterium Burkholderia phytofirmans and plant pathogenic bacteria. New Phytologist, 201, 1371–1384.
Win J, Chaparro-Garcia A, Belhaj K, Saunders D G O, Yoshida K, Dong S, Schornack S, Zipfel C, Robatzek S, Hogenhout S A, Kamoun S. 2012. Effector biology of plant-associated organisms: concepts and perspectives. Cold Spring Harbor Symposia on Quantitative Biology, 77, 235–247.
Zhao T, Jiang J, Liu G, He S, Zhang H, Chen X, Li J, Xu X. 2016. Mapping and candidate gene screening of tomato Cladosporium fulvum-resistant gene Cf-19, based on high-throughput sequencing technology. BMC Plant Biology, 16, 51.
Zipfel C, Oldroyd G E D. 2017. Plant signalling in symbiosis and immunity. Nature, 543, 328–336.
|
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
