The Botrytis cinerea effector BcXYG1 suppresses immunity in Fragaria vesca by targeting FvBPL4 and FvACD11

Abstract

Botrytis cinerea is one of the most destructive pathogens in strawberry cultivation. Successful infection by B. cinerea requires releasing a large number of effectors that interfere with the plant’s immune system. One of the effectors required by B. cinerea for optimal virulence is the secreted protein BcXYG1, which is thought to associate with proteins near the plasma membrane of the host plant to induce necrosis. However, the host proteins that associate with BcXYG1 at the plasma membrane are currently unknown. We found that BcXYG1 binds to FvBPL4 and FvACD11 at the plasma membrane. Both FvBPL4 and FvACD11 are negative regulators of plant immunity in strawberry. Our results demonstrate that degradation of FvBPL4 by BcXYG1 promotes disease resistance while stabilization of FvACD11 by BcXYG1 suppresses the immune response. These findings suggest that BcXYG1 suppresses plant immunity and promotes B. cinerea infection by regulating FvBPL4 and FvACD11 protein levels.

Acknowledgements

This research was supported in part by the Priority Academic Program Development of Jiangsu Higher Education Institutions and the State Key Laboratory of Crop Genetics and Germplasm Enhancement (ZW201813). This study was also supported by the high-performance computing platform of the Bioinformatics Cen-ter, Nanjing Agricultural University. We would like to thank A&L Scientific Editing (www.alpublish.com) for its linguistic assistance during the preparation of this manuscript.

Author contributions

Z.-M. and L.S. designed the research plan. L.S., T.Z. and B.Y. performed the experiments. L.S. performed the data analyses and wrote the manuscript. Y.B., W.F. and J.X. were involved in manuscript editing. All authors read and approved the final manuscript.

Data availability

The authors confirm that all data from this study are avail-able and can be found in this article and its supplementary information.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary data

Supplementary data is available at Horticulture Research online.

References

[1] Zipfel C. Pattern-recognition receptors in plant innate immunity. Curr Opin Immunol. 2008;20:10-6

[2] Wu S, Shan L, He P. Microbial signature-triggered plant defense responses and early signaling mechanisms. Plant Sci. 2014;228: 118-26

[3] Couto D, Zipfel C. Regulation of pattern recognition receptor signalling in plants. Nat Rev Immunol. 2016;16:537-52

[4] Houterman PM, Cornelissen B, Rep M. Suppression of plant resis-tance gene-based immunity by a fungal effector. PLoS Pathog. 2008;4:e1000061

[5] Cui H, Tsuda K, Parker JE. Effector-triggered immunity: from pathogen perception to robust defense. Annu Rev Plant Biol. 2015;66:487-511

[6] Prins TW, Tudzynski P, Tiedemann AV. et al. Infection strate-gies of Botrytis cinerea and related necrotrophic pathogens. In: Kronstad JW (ed.), Fungal Pathology. Dordrecht: Springer, 2000, 33-64.

[7] Kan J. Licensed to kill: the lifestyle of a necrotrophic plant pathogen. Trends Plant Sci. 2006;11:247-53

[8] Williamson B, Tudzynski B, Tudzynski P. et al. Botrytis cinerea:the cause of grey mould disease. Mol Plant Pathol. 2007;8:561-80

[9] Soares F, Pimentel D, Erban A. et al. Virulence-related metabolism is activated in Botrytis cinerea mostly in the interaction with tolerant green grapes that remain largely unaffected in contrast with susceptible green grapes. Hortic Res. 2022;9:c217

[10] Cantarel BL, Coutinho PM, Corinne R. et al. The Carbohydrate-Active EnZymes database (CAZy): an expert resource for glycogenomics. Nucleic Acids Res. 2009;37:D233-8

[11] Kubicek CP, Trevor LS, Glass NL. Plant cell wall-degrading enzymes and their secretion in plant-pathogenic fungi. Annu Rev Phytopathol. 2014;52:427-51

[12] Noda J, Brito N, González C. The Botrytis cinerea xylanase Xyn11A contributes to virulence with its necrotizing activity, not with its catalytic activity. BMC Plant Biol. 2010;10:38

[13] Yang Y, Yang X, Dong Y. et al. The Botrytis cinerea xylanase BcXyl1 modulates plant immunity. Front Microbiol. 2018;9:9

[14] Kars I, Krooshof GH, Wagemakers L. et al. Necrotizing activity of five Botrytis cinerea endopolygalacturonases produced in Pichia pastoris. Plant J. 2005;43:213-25

[15] Zhang L, Kars I, Essenstam B. et al. Fungal endopolygalactur-onases are recognized as microbe-associated molecular pat-terns by the Arabidopsis receptor-like protein RESPONSIVENESS TO BOTRYTIS POLYGALACTURONASES1. Plant Physiol. 2014;164: 352-64

[16] Frías M, González M, González C. et al. BcIEB1, a Botrytis cinerea secreted protein, elicits a defense response in plants. Plant Sci. 2016;250:115-24

[17] Arenas YC, Kalkman E, Schouten A. et al. Functional analysis and mode of action of phytotoxic Nep1-like proteins of Botrytis cinerea. Physiol Mol Plant Pathol. 2010;74:376-86

[18] Frías M, González C, Brito N. BcSpl1, a cerato-platanin family protein, contributes to Botrytis cinerea virulence and elicits the hypersensitive response in the host. New Phytol. 2011;192:483-95

[19] Zhu WJ, Ronen M, Gur Y. et al. BcXYG1, a secreted xyloglucanase from Botrytis cinerea, triggers both cell death and plant immune responses. Plant Physiol. 2017;175:438-56

[20] Brodersen P, Petersen M, Pike HM. et al. Knockout of Arabidopsis ACCELERATED-CELL-DEATH11 encoding a sphingosine trans-fer protein causes activation of programmed cell death and defense. Genes Dev. 2002;16:490-502

[21] Li Q, Ai G, Shen D. et al. A Phytophthora capsici effector targets ACD11 binding partners that regulate ROS-mediated defense response in Arabidopsis. Mol Plant. 2019;12:565-81

[22] Petersen NHT, Joensen J, McKinney LV. et al. Identification of pro-teins interacting with Arabidopsis ACD11. J Plant Physiol. 2009;166: 661-6

[23] Liu H, Ravichandran S, Teh O. et al. The RING-type E3 ligase XBAT35.2 is involved in cell death induction and pathogen response. Plant Physiol. 2017;175:1469-83

[24] Martínez-Rivas FJ, Blanco-Portales R, Moyano E. et al. Strawberry fruit FanCXE1 carboxylesterase is involved in the catabolism of volatile esters during the ripening process. Hortic Res. 2022;9:uhac95

[25] YinX FuQ, Shang B. et al. An RxLR effector from Plasmopara viticola suppresses plant immunity in grapevine by targeting and stabilizing VpBPA1. Plant J. 2022;112:104-14

[26] Zhang X, Ai G, Wang X. et al. Genome-wide identification and molecular evolution analysis of BPA genes in green plants. Phy-topathol Res. 2020;2:6

[27] Simanshu D, Zhai X, Munch D. et al. Arabidopsis accelerated cell death 11, ACD11, is a ceramide-1-phosphate transfer protein and intermediary regulator of phytoceramide levels. Cell Rep. 2014;6:388-99

[28] Williams B, Kabbage M, Kim HJ. et al. Tipping the balance: Sclero-tinia sclerotiorum secreted oxalic acid suppresses host defenses by manipulating the host redox environment. PLoS Pathog. 2011;7:e1002107

[29] Kabbage M, Williams B, Dickman MB. et al. Cell death control: the interplay of apoptosis and autophagy in the pathogenicity of Sclerotinia sclerotiorum. PLoS Pathog. 2013;9:e1003287

[30] van Kan JAL, Shaw MW, Grant-Downton RT. Botrytis species: relentless necrotrophic thugs or endophytes gone rogue? Mol Plant Pathol. 2014;15:957-61

[31] Islam MN, Jacquemot M, Coursol S. et al. Sphingosine in plants -more riddles from the sphinx? New Phytol. 2012;193:51-7

[32] Markham JE, Lynch DV, Napier JA. et al. Plant sphingolipids: function follows form. Curr Opin Plant Biol. 2013;16:350-7

[33] Bi FC, Liu Z, Wu JX. et al. Loss of ceramide kinase in Arabidopsis impairs defenses and promotes ceramide accumulation and mitochondrial H2O2 bursts. Plant Cell. 2014;26:3449-67