Chinese Journal OF Rice Science >
Signal Peptide Validation and Expression Analysis of Multiple Effectors from Ustilaginoidea virens
Received date: 2023-10-30
Revised date: 2023-12-12
Online published: 2024-05-13
【Objective】Signal peptides are crucial for effector secretion and affect the pathogenic process of pathogenic microorganisms. Previously, ten effector genes were identified to be significantly up-regulated during Ustilaginoidea virens infection in rice spikelets. However, it remains unclear whether these effectors contain functional signal peptides and how they express.【Method】The signal peptides of these effectors were predicted and verified by using SignaIP-5.0 database, Agrobacterium-mediated transient expression in Nicotiana benthamiana, and yeast invertase assay. Besides, the expression patterns of these effectors were analyzed by using RT-qPCR.【Results】All the ten effectors, including UV_44, UV_1548, UV_1567, etc., were predicted to contain a signal peptide. The transient expression method showed that eYFP fusion proteins containing signal peptides were successfully secreted into the extracellular space after plasmolysis in N. benthamiana. Yeast invertase assay showed that the yeast strains harboring signal peptides could grow on the medium with raffinose as the only carbon source with the occurrence of a chromogenic reaction using TTC. All the effector genes had distinct expression patterns, except for UV_6754. Seven of them showed significant up-regulation after inoculation, including UV_44, UV_1567, UV_3667, UV_4213, UV_4989, UV_5215, and UV_8184.【Conclusion】The signal peptides of these effectors had secretory functions, and the effector genes have different expression patterns during U. virens infection. These findings lay a foundation for further research on the functions of the above-mentioned effectors from U. virens.
Key words： rice; Ustilaginoidea virens; effector; signal peptide
YIN Xiaoxiao, ZHANG Zhihan, YAN Xiulian, LIAO Rong, YANG Sijia, Beenish HASSAN, GUO Daiming, FAN Jing, ZHAO Zhixue, WANG Wenming . Signal Peptide Validation and Expression Analysis of Multiple Effectors from Ustilaginoidea virens[J]. Chinese Journal OF Rice Science, 2024 , 38(3) : 256 -265 . DOI: 10.16819/j.1001-7216.2024.231012
| [1] | Brooks S A, Anders M M, Yeater K M. Effect of cultural management practices on the severity of false smut and kernel smut of rice[J]. Plant Disease, 2009, 93(11): 1202-1208. |
| [2] | Tang Y X, Jin J, Hu D W, Yong M L, Xu Y, He L P. Elucidation of the infection process of Ustilaginoidea virens (teleomorph: Villosiclava virens) in rice spikelets[J]. Plant Pathology, 2013, 62(1): 1-8. |
| [3] | Cooke M C. Some extra-european fungi[J]. Grevillea, 1878, 7: 13-15. |
| [4] | Sun W, Fan J, Fang A, Li Y, Tariqjaveed M, Li D, Hu D, Wang W M. Ustilaginoidea virens: Insights into an emerging rice pathogen[J]. Annual Review of Phytopathology, 2020, 58(1): 363-385. |
| [5] | Fan J, Liu J, Gong Z Y, Xu P Z, Hu X H, Wu J L, Li G B, Yang J, Wang Y Q, Zhou Y F, Li S C, Wang L, Chen X Q. The false smut pathogen Ustilaginoidea virens requires rice stamens for false smut ball formation[J]. Environmental Microbiology, 2020, 22(2): 646-659. |
| [6] | Fan J, Guo X Y, Li L, Huang F, Sun W X, Li Y, Huang Y Y, Xu Y J, Shi J, Zheng A P, Wang W M. Infection of Ustilaginoidea virens intercepts rice seed formation but activates grain-filling-related genes[J]. Journal of Integrative Plant Biology, 2015, 57(6): 577-590. |
| [7] | Fan J, Yang J, Wang Y Q, Li G B, Li Y, Wang W M. Current understanding on Villosiclava virens, a unique flower-infecting fungus causing rice false smut disease[J]. Molecular Plant Pathology, 2016, 17(9): 1321-1330. |
| [8] | Wang B, Liu L, Li Y, Zou J, Li D, Zhao D, Li W, Sun W. Ustilaginoidin D induces hepatotoxicity and behaviour aberrations in zebrafish larvae[J]. Toxicology, 2021, 456: 152786. |
| [9] | Lin X, Bian Y, Mou R, Cao Z, Cao Z, Zhu Z, Chen M. Isolation, identification, and characterization of Ustilaginoidea virens from rice false smut balls with high ustilotoxin production potential[J]. Journal of Basic Microbiology, 2018, 58(8): 670-678. |
| [10] | Rafiqi M, Ellis J G, Ludowici V A, Hardham A R, Dodds P N. Challenges and progress towards understanding the role of effectors in plant-fungal interactions[J]. Current Opinion in Plant Biology, 2012, 15(4): 477-482. |
| [11] | Zhang K, Zhao Z, Zhang Z, Li Y, Li S, Yao N, Hsiang T. Insights into genomic evolution from chromosomal and mitochondrial genomes of Ustilaginoidea virens[J]. Phytopathology Research, 2021, 3(1): 9. |
| [12] | Zhang N, Yang J, Fang A, Wang J, Li D, Li Y, Wang S, Cui F, Yu J, Liu Y, Peng Y L, Sun W. The essential effector SCRE1 in Ustilaginoidea virens suppresses rice immunity via a small peptide region[J]. Molecular Plant Pathology, 2020, 21(4): 445-459. |
| [13] | Zheng X, Fang A, Qiu S, Zhao G, Wang J, Wang S, Wei J, Gao H, Yang J, Mou B, Cui F, Zhang J, Liu J, Sun W. Ustilaginoidea virens secretes a family of phosphatases that stabilize the negative immune regulator OsMPK6 and suppress plant immunity[J]. The Plant Cell, 2022, 34(8): 3088-3109. |
| [14] | Chen X, Duan Y, Qiao F, Liu H, Huang J, Luo C, Chen X, Li G, Xie K, Hsiang T, Zheng L. A secreted fungal effector suppresses rice immunity through host histone hypoacetylation[J]. New Phytologist, 2022, 235(5): 1977. |
| [15] | Li G B, He J X, Wu J L, Wang H, Zhang X, Liu J, Hu X H, Zhu Y, Shen S, Bai Y F, Wang W M, Fan J. Overproduction of OsRACK1A, an effector-targeted scaffold protein promoting OsRBOHB-mediated ROS production, confers rice floral resistance to false smut disease without yield penalty[J]. Molecular Plant, 2022, 15(11): 1790-1806. |
| [16] | Chen X, Li X, Duan Y, Pei Z, Liu H, Yin W, Huang J, Luo C, Chen X, Li G, Xie K, Hsiang T, Zheng L. A secreted fungal subtilase interferes with rice immunity via degradation of SUPPRESSOR OF G2 ALLELE OF skp1[J]. Plant Physiology, 2022, 190(2): 1474-1489. |
| [17] | Owji H, Nezafat N, Negahdaripour M, Hajiebrahimi A, Ghasemi Y. A comprehensive review of signal peptides: Structure, roles, and applications[J]. European Journal of Cell Biology, 2018, 97(6): 422-441. |
| [18] | von Heijne G. Signal sequences: Limits of variation[J]. Journal of Molecular Biology, 1985, 184(1): 99-105. |
| [19] | Rafiqi M, Gan P H P, Ravensdale M, Lawrence G J, Ellis J G, Jones D A, Hardham A R, Dodds P N. Internalization of flax rust avirulence proteins into flax and tobacco cells can occur in the absence of the pathogen[J]. The Plant Cell, 2010, 22(6): 2017-2032. |
| [20] | Yin W, Wang Y, Chen T, Lin Y, Luo C. Functional evaluation of the signal peptides of secreted proteins[J]. Bio-protocol, 2018, 8(9): e2839. |
| [21] | Gu B, Kale S D, Wang Q, Wang D, Pan Q, Cao H, Meng Y, Kang Z, Tyler B M, Shan W. Rust secreted protein Ps87 is conserved in diverse fungal pathogens and contains a RXLR-like motif sufficient for translocation into plant cells[J]. PLOS ONE, 2011, 6(11): e27217. |
| [22] | Bao J, Wang R, Gao S, Wang Z, Fang Y, Wu L, Wang M. High-Quality genome sequence resource of a rice false smut fungus Ustilaginoidea virens isolate, UV-FJ-1[J]. Phytopathology, 2021, 111(10): 1889-1892. |
| [23] | Zhang Y, Zhang K, Fang A, Han Y, Yang J, Xue M, Bao J, Hu D, Zhou B, Sun X, Li S, Wen M, Yao N, Ma L J, Liu Y, Zhang M, Huang F, Luo C, Zhou L, Li J, Chen Z. Specific adaptation of Ustilaginoidea virens in occupying host florets revealed by comparative and functional genomics[J]. Nature Communications, 2014, 5(1): 3849. |
| [24] | Wang H, Yang X, Wei S, Wang Y. Proteomic analysis of mycelial exudates of Ustilaginoidea virens[J]. Pathogens, 2021, 10(3): 364. |
| [25] | Fang A, Han Y, Zhang N, Zhang M, Liu L, Li S, Lu F, Sun W. Identification and characterization of plant cell death-inducing secreted proteins from Ustilaginoidea virens[J]. Molecular Plant-Microbe Interactions, 2016, 29(5): 405-416. |
| [26] | Wei S, Wang Y, Zhou J, Xiang S, Sun W, Peng X, Li J, Hai Y, Wang Y, Li S. The conserved effector UvHrip1 interacts with OsHGW and infection of Ustilaginoidea virens regulates defense- and heading date-related signaling pathway[J]. International Journal of Molecular Sciences, 2020, 21(9): 3376. |
| [27] | Qiu S, Fang A, Zheng X, Wang S, Wang J, Fan J, Sun Z, Gao H, Yang J, Zeng Q, Cui F, Wang W M, Chen J, Sun W. Ustilaginoidea virens nuclear effector SCRE4 suppresses rice immunity via inhibiting expression of a positive immune regulator OsARF17[J]. International Journal of Molecular Sciences, 2022, 23(18): 10527. |
| [28] | Fang A, Gao H, Zhang N, Zheng X, Qiu S, Li Y, Zhou S, Cui F, Sun W. A novel effector gene SCRE2 contributes to full virulence of Ustilaginoidea virens to rice[J]. Frontiers in Microbiology, 2019, 10. |
| [29] | Presti L L, Lanver D, Schweizer G, Tanaka S, Liang L, Tollot M, Zuccaro A, Reissmann S, Kahmann R. Fungal Effectors and plant susceptibility[J]. Annual Review of Plant Biology, 2015, 66(1): 513-545. |
| [30] | Fan J, Du N, Li L, Li G B, Wang Y Q, Zhou Y F, Hu X H, Liu J, Zhao J Q, Li Y, Huang F, Wang W M. A core effector UV_1261 promotes Ustilaginoidea virens infection via spatiotemporally suppressing plant defense[J]. Phytopathology Research, 2019, 1(1): 11. |
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