Abstract
Popular is not only an important tree species economically, but is also a model system for plant biology. Marssonina leaf spot is a serious leaf disease caused by Marssonina brunnea, which can result in significant economic losses. M. brunnea is considered to be a hemibiotrophic pathogen but its pathogenesis remains unclear. Full length cDNA of 24 LysM effector genes were obtained based on the sequencing of the M. brunnea genome. All of these genes belonged to a single gene family for which the characteristics of the amino acid sequences and protein domains were analyzed. Further detailed analysis was carried out on two representative gene family members. These two genes were up-regulated during the infection process and, when transformed into Arabidopsis thaliana, reduced the plant’s sensitivity to chitin thus might compromised its immune system. LysM effectors appear to show a specific affinity for chitin since the expressed proteins co-precipitated with insoluble chitin. We performed a series of studies into the molecular mechanism of LysM effectors from M. brunnea and carried out a preliminary analysis into the interaction between the pathogen and its host. This research lays a foundation for further investigation into forest tree resistance breeding.
Similar content being viewed by others
References
Bi DL, Cheng YT, Li X, Zhang YL (2010) Activation of Plant Immune Responses by a Gain-of-Function Mutation in an Atypical Receptor-Like Kinase. Plant Physiol 153:1771–1779
Buist G, Steen A, Kok J, Kuipers OP (2008) LysM, a widely distributed protein motif for binding to (peptido) glycans. Mol Microbiol 68:838–847
Burnette WN (1981) “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203
Cheng Q, Cao Y, Jiang C, Xu L, Wang M, Zhang S, Huang M (2010) Identifying secreted proteins of Marssonina brunnea by degenerate PCR. Proteomics 10:2406–2417
Chisholm ST, Coaker G, Day B, Staskawicz BJ (2006) Host-microbe interactions: shaping the evolution of the plant immune response. Cell 124:803–814
Clough SJ (2005) Floral dip: agrobacterium-mediated germ line transformation. Methods Mol Biol 286:91–102
Czechowski T, Bari RP, Stitt M et al (2004) Real-time RT-PCR profiling of over 1400 Arabidopsis transcription factors: unprecedented sensitivity reveals novel root- and shoot- pecific genes. The Plant J 38:366–379
de Jonge R, van Esse HP, Kombrink A, Shinya T, Desaki Y, Bours R, van der Krol S, Shibuya N, Joosten MH, Thomma BP (2010) Conserved fungal LysM effector Ecp6 prevents chitin-triggered immunity in plants. Science 329:953–955
Galletti R, Ferrari S, De Lorenzo G (2011) Arabidopsis MPK3 and MPK6 play different roles in basal and oligogalacturonide-or flagellin-induced resistance against Botrytis cinerea. Plant Physiol 157:804–814
Giraldo MC, Valent B (2013) Filamentous plant pathogen effectors in action. Nat Rev Microbiol 11:800–814
Hamamouch N, Li C, Seo PJ, Park CM, Davis EL (2011) Expression of Arabidopsis pathogenesis-related genes during nematode infection. Mol Plant Pathol 12:355–364
Hayafunea M, Berisiob R, Marchettic R, Silipoc A, Kayamaa M, Desakia Y, Arimaa S, Squegliab F, Ruggierob A, Tokuyasud K, Molinaroc A, Kakua H, Shibuyaa N (2014) Chitin-induced activation of immune signaling by the rice receptor CEBiP relies on a unique sandwich-type dimerization. Proc Natl Acad Sci 10:1073
Jiang C, Wen Q, Chen Y, Xu L, Huang M (2011) Efficient extraction of RNA from various Camellia species rich in secondary metabolites for deep transcriptome sequencing and gene expression analysis. Afr J Biotechnol 10:16769–16773
Jones JDG, Dangl JL (2006) The plant immune system. Nature 444:323–329
Lee SJ, Rose JKC (2010) Mediation of the transition from biotrophy to necrotrophy in hemibiotrophic plant pathogens by secreted effector proteins. Plant Signal Behav 5:769–772
Marshall R, Kombrink A, Motteram J, Loza-Reyes E, Lucas J, Hammond-Kosack KE, Thomma BP, Rudd JJ (2011) Analysis of two in planta expressed LysM effector homologs from the fungus Mycosphaerella graminicola reveals novel functional properties and varying contributions to virulence on wheat. Plant Physiol 156:756–769
Mentlak TA, Kombrink A, Shinya T, Ryder LS, Otomo I, Saitoh H, Terauchi R, Nishizawa Y, Shibuya N, Thomma BP, Talbot NJ (2012) Effector-mediated suppression of chitin-triggered immunity by Magnaporthe oryzae is necessary for rice blast disease. The Plant Cell 24:322–335
Mitsuhara I, Matsufuru H, Ohshima M, Kaku H, Nakajima Y, Murai N, Natori S, Ohashi Y (2000) Induced expression of sarcotoxin IA enhanced host resistance against both bacterial and fungal pathogens in transgenic tobacco. Mol Plant-Microbe In 13:860–868
Morris K, Thornber S, Codrai L, Richardson C, Craig A, Sadanandom A, Thomas B, Jackson S (2010) DAY NEUTRAL FLOWERING represses CONSTANS to prevent Arabidopsis flowering early in short days. The Plant Cell 22:1118–1128
Shimizu T, Nakano T, Takamizawa D, Desaki Y, Ishii-Minami N, Nishizawa Y, Minami E, Okada K, Yamane H, Kaku H, Shibuya N (2010) Two LysM receptor molecules, CEBiP and OsCERK1, cooperatively regulate chitin elicitor signaling in rice. Plant J 64:204–214
Spiers AG (1983) Host range and pathogenicity studies of Marsonnina brunnea to poplars. Eur J For Pathol 13:181–196
Wu R, Wang M, Huang M (1992) Quantitative genetics of yield breeding for Populus short rotation culture. I. Dynamics of genetic control and selection model of yield traits. Can J For Res 22:175–182
Xu M, Zhang B, Yao H, Huang M (2009) Isolation of high quality RNA and molecular manipulations with various tissues of Populus. Rus J Plant Physiol 56:716–719
Zhu S, Cao Y, Jiang C, Tan B, Wang Z, Feng S, Zhang L, Su X, Brejova B, Vinar T, Xu M, Wang M, Zhang S, Huang M, Wu R, Zhou Y (2012) Sequencing the genome of Marssonina brunnea reveals fungus-poplar co-evolution. BMC Genomics 13:382
Acknowledgements
This research was supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) and the Collaborative Innovation Plan of Jiangsu Higher Education. We acknowledge Professor Dacheng Tian and Yonghua Yang from Nanjing University who provide us A. thaliana Col-0.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Table S1
PCR primers used in the real-time PCR assay. (DOCX 14 kb)
Fig. S1
Sequence analysis of LysM genes and LysM domains. (A). Multiple sequence alignment of LysM genes in Marssonina.brunnea; (B). Consensus sequence of LysM domain (GIF 432 kb)
Rights and permissions
About this article
Cite this article
Jiang, C., He, B., Huang, R. et al. Identification and functional analysis of LysM effectors from Marssonina brunnea . Australasian Plant Pathol. 43, 615–622 (2014). https://doi.org/10.1007/s13313-014-0316-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13313-014-0316-5