請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/95772
標題: 辣椒炭疽病之候選效應蛋白功能性分析
Functional Analysis of Candidate Effector Proteins of Colletotrichum acutatum strain 524
作者: 江政霖
Chen-Lin Chiang
關鍵字: 辣椒炭疽病
Colletotrichum acutatum
效應蛋白
LysM domain
Colletotrichum acutatum
effector protein
hypothetical protein
LysM effector
virulence
引用: 王添成、許宗銘。2013。番椒炭疽病的發生、病原鑑定與病害管理。農藝之友 159:31-38。 Azmi NSA, Singkaravanit-Ogawa S, Ikeda K, et al., 2018. Inappropriate expression of an NLP effector in Colletotrichum orbiculare impairs infection on Cucurbitaceae cultivars via plant recognition of the C-terminal region. Mol Plant Microbe Interact 31, 101-11. Bombarely A, Rosli HG, Vrebalov J, Moffett P, Mueller LA, Martin GB, 2012. A draft genome sequence of Nicotiana benthamiana to enhance molecular plant-microbe biology research. Mol Plant Microbe Interact 25, 1523-30. Chen M.Y., 2012. Induction and regulation of highly branched penetration structure of Colletotrichum acutatum. National Chung Hsing University, Master Thesis. Chuang S.C.,Lee M.H., Shih M.C., 2014. Genomic and transcriptomic analyses unravel effector repertoire of Colletotrichum acutatum during the infection on Capsicum annuum. . 12th European Conference on Fungal Genetics. Spanish, 175. Chung K.R., Lee M.H., 2015. Split-marker-mediated transformation and targeted gene disruption in filamentous fungi. Fungal Biology 2, 175-180. Cruz CLaSS, 1999. Bax-induced cell death in tobacco is similar to the hypersensitive response. Proc Natl Acad Sci 96, 7956-61. De Jonge R, Bolton MD, Thomma BP, 2011. How filamentous pathogens co-opt plants: the ins and outs of fungal effectors. Curr Opin Plant Biol 14, 400-6. De Wit PJ, 2016. Apoplastic fungal effectors in historic perspective: a personal view. New Phytol 212, 805-13. Dou D, Kale SD, Wang X, et al., 2008. Conserved C-terminal motifs required for avirulence and suppression of cell death by Phytophthora sojae effector Avr1b. Plant Cell 20, 1118-33. El Oirdi M, Bouarab K, 2007. Plant signalling components EDS1 and SGT1 enhance disease caused by the necrotrophic pathogen Botrytis cinerea. New Phytol 175, 131-9. Fu ZQ, Dong X, 2013. Systemic acquired resistance: turning local infection into global defense. Annu Rev Plant Biol 64, 839-63. Gawehns F, Houterman PM, Ichou FA, et al., 2014. The Fusarium oxysporum effector Six6 contributes to virulence and suppresses I-2-mediated cell death. Mol Plant Microbe Interact 27, 336-48. Jeff H. Chang, John P. Rathjen, Adriana J. Bernal, And BJS, Michelmore RW, 2000. AvrPto enhances growth and necrosis caused by Pseudomonas syringae pv. tomato in tomato lines lacking either Pto or Prf. Molecular Plant-Microbe Interactions 13, 568-71. Jing M, Guo B, Li H, et al., 2016. A Phytophthora sojae effector suppresses endoplasmic reticulum stress-mediated immunity by stabilizing plant Binding immunoglobulin Proteins. Nat Commun 7, 11685. Kombrink A, Rovenich H, Shi-Kunne X, et al., 2017. Verticillium dahliae LysM effectors differentially contribute to virulence on plant hosts. Mol Plant Pathol 18, 596-608. Kuo C-C, 2016. Functional analysis of genes involved in vegetative growth and virulence of Colletotrichum acutatum, a causing agent of chili pepper anthracnose. National Chung Hsing University, Master Thesis, 22-3. Li Z, Yin Z, Fan Y, Xu M, Kang Z, Huang L, 2015. Candidate effector proteins of the necrotrophic apple canker pathogen Valsa mali can suppress BAX-induced PCD. Front Plant Sci 6, 579. Liao C.Y., Chen M.Y., Chen Y.K., et al., 2012. Characterization of three Colletotrichum acutatum isolates from Capsicum spp. European Journal of Plant Pathology 133, 599-608. Liao C.Y., Chen M.Y., Chen Y.K., Kuo K.C.,Chung K.R. And Lee M.H., 2012. Formation of highly branched hyphae by Colletotrichum acutatum within the fruit cuticles of Capsicum spp. Plant pathology 61, 262-70. Lin H-C, 2016. The study on histidine kinase gene group of Colletotrichum gloeosporioides, a causal agent of mango anthracnose. National Chung Hsing University, Master Thesis, 94. Marina Pais JW, Kentaro Yoshida, Graham J Etherington, Liliana M Cano, Sylvain Raffaele, Mark J Banfield, Alex Jones, Sophien Kamoun and Diane Go Saunders, 2013. From pathogen genomes to host plant processes the power of plant parasitic oomycetes. Genome Biol 14. Melanie L. Lewis Ivey CN-D, And Sally A. Miller, 2004. Identification and management of Colletotrichum acutatum on immature bell peppers. Plant disease 88, 1198-204. Mentlak TA, Kombrink A, Shinya T, et al., 2012. Effector-mediated suppression of chitin-triggered immunity by Magnaporthe oryzae is necessary for rice blast disease. Plant Cell 24, 322-35. Nguyen QB, Kadotani N, Kasahara S, Tosa Y, Mayama S, Nakayashiki H, 2008. Systematic functional analysis of calcium-signalling proteins in the genome of the rice-blast fungus, Magnaporthe oryzae, using a high-throughput RNA-silencing system. Mol Microbiol 68, 1348-65. Oliveira-Garcia E, Deising HB, 2013. Infection structure-specific expression of beta-1,3-glucan synthase is essential for pathogenicity of Colletotrichum graminicola and evasion of beta-glucan-triggered immunity in maize. Plant Cell 25, 2356-78. Petre B, Saunders DG, Sklenar J, et al., 2016. Heterologous expression screens in Nicotiana benthamiana identify a candidate effector of the wheat yellow rust pathogen that associates with processing bodies. PLoS One 11, e0149035. Plissonneau C, Benevenuto J, Mohd-Assaad N, Fouche S, Hartmann FE, Croll D, 2017. Using population and comparative genomics to understand the genetic basis of effector-driven fungal pathogen evolution. Front Plant Sci 8, 119. Rovenich H, Boshoven JC, Thomma BP, 2014. Filamentous pathogen effector functions: of pathogens, hosts and microbiomes. Curr Opin Plant Biol 20, 96-103. Selin C, De Kievit TR, Belmonte MF, Fernando WG, 2016. Elucidating the role of effectors in plant-fungal interactions: progress and challenges. Front Microbiol 7, 600. Shah J, Zeier J, 2013. Long-distance communication and signal amplification in systemic acquired resistance. Front Plant Sci 4, 30. Sharpee WC, Dean RA, 2016. Form and function of fungal and oomycete effectors. Fungal Biology Reviews 30, 62-73. Sheu ZMaW, Tien Cheng 2005. Identification and management of Colletotrichum acutatum on immature bell peppers. The Second Asian Conference on Plant Pathology, 25-8. Sonah H, Deshmukh RK, Belanger RR, 2016. Computational prediction of effector proteins in fungi: opportunities and challenges. Front Plant Sci 7, 126. Sperschneider J, Dodds PN, Gardiner DM, Manners JM, Singh KB, Taylor JM, 2015. Advances and challenges in computational prediction of effectors from plant pathogenic fungi. PLoS Pathog 11, e1004806. Sperschneider J, Gardiner DM, Dodds PN, et al., 2016. EffectorP: predicting fungal effector proteins from secretomes using machine learning. New Phytol 210, 743-61. Surico G, 2013. The concepts of plant pathogenicity, virulence, avirulence and effector proteins by a teacher of plant pathology. Phytopathologia Mediterranea 52, 399-417. Takahara H, Hacquard S, Kombrink A, et al., 2016. Colletotrichum higginsianum extracellular LysM proteins play dual roles in appressorial function and suppression of chitin-triggered plant immunity. New Phytol 211, 1323-37. Than P JR, Hyde K, Pongsupasamit S, Mongkolporn O, Taylor P, 2008. Characterization and pathogenicity of Colletotrichum species associated with anthracnose on chili (Capsicum spp.) in Thailand. Plant pathology 57, 562-72. Thomma BP, Nurnberger T, Joosten MH, 2011. Of PAMPs and effectors: the blurred PTI-ETI dichotomy. Plant Cell 23, 4-15. Vega-Arreguin JC, Jalloh A, Bos JI, Moffett P, 2014. Recognition of an Avr3a homologue plays a major role in mediating nonhost resistance to Phytophthora capsici in Nicotiana species. Mol Plant Microbe Interact 27, 770-80. Wang Y, Wu J, Kim SG, et al., 2016. Magnaporthe oryzae-secreted protein MSP1 induces cell death and elicits defense responses in rice. Mol Plant Microbe Interact 29, 299-312.
摘要: 辣椒炭疽病菌Colletotrichum acutatum在台灣危害辣椒及多種重要經濟作物。目前已知此病原菌以半活體營養生長策略來獲取寄主養分,但詳細的致病機制則很少有相關的探討。近年來我們已建立辣椒炭疽病菌強毒力菌株C. acutatum Coll-524侵染辣椒果實過程之轉錄體。於轉錄體分析的結果發現Coll-524在感染辣椒的過程中,會外泌一群小分子的蛋白質,這些小分子蛋白可能參與Coll-524於辣椒果實之半活體營養生長。我們從中挑選出13個Candidate Effector Proteins (CEPs),這13個CEPs蛋白質長度小於300個胺基酸、帶有訊息傳遞胜肽、功能尚未被研究及感染過程中於特定時期表現量最高。本研究進行其中2個CEPs功能的探討,為功能未知的CEP002以及帶有LysM domain的CEP012;另外也同時研究CEP012的旁系同源基因Ca_5620的功能。本研究使用基因剔除、過量表現及於菸草葉片中短暫大量表現等策略進行基因功能分析。將CEP002基因剔除後,進行致病力、生長與孢子發芽率等試驗分析,結果顯示CEP002 並未影響Coll-524 之上述功能,但若將CEP002持續大量表現於Coll-524,則可提升Coll-524對辣椒的致病力;在圓葉菸草葉片中表現CEP002進行誘導細胞死亡及抑制BAX所誘導的細胞死亡分析,並未觀察到顯著的差異。在CEP012與Ca_5620的功能研究,剔除CEP012或Ca_5620後,發現皆可造成Coll-524對辣椒之致病力下降。未來可針對持續表現CEP002是如何提升Coll-524致病力以及CEP012與Ca_5620同時存在於Coll-524基因體中的意義等方面進行探討。
Colletotrichum acutatum dramatically diminishes yield rates of chili peppers and other crops in Taiwan. This pathogen uses hemibiotrophic strategy for nutrient acquisition but how pathogen effectors alter plant immunity or responses remains unknown. In our lab, transcriptomic analysis of Coll-524 during pathogenesis on chili pepper fruit showed that a group of genes encoding small and secreted proteins expressed specifically during infection processes, called candidate effector proteins (CEPs). In this study, we analyzed the function of unknown effector CEP002 and two LysM domain-containing effector, CEP012 and its paralogous gene Ca_5620, by gene knockout and overexpression in Coll-524, and transient expression in Nicotiana benthamiana. Here, we found that knocking out CEP002 in Coll-524 genome did not affect on pathogenicity, mycelia growth and spore germination, but constitutive expression of CEP002-gfp in Coll-524 could increase fungal virulence to chili peppers. However, transient expression of CEP002 in N. benthamiana didn’t show any notable phenotype. For the functional analysis of CEP012 and Ca_5620, we found that knocking out eitheir CEP012 or Ca_5620 in Coll-524 genome would reduce the virulence of Coll-524 when inoculated on chili peppers. In the future, the reason that the constitutive expression of CEP002-gfp increases virulence of Coll-524 on chili peppers and the biological meaning of the two LysM effectors existing in Coll-524 genome remain to be investigated.
URI: http://hdl.handle.net/11455/95772
文章公開時間: 10000-01-01
顯示於類別:植物病理學系

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