Please use this identifier to cite or link to this item:
Molecular cloning and functional analysis of Papaya ringspot virus resistance genes in Cucumis metuliferus
|關鍵字:||刺角瓜;Cucumis metuliferus;木瓜輪點病毒;Papaya ringspot virus||出版社:||農藝學系所||引用:||Abalo G, Tongoona P, Derera J, Edema R (2009) A comparative analysis of conventional and marker-assisted selection methods in breeding Maize streak virus resistance in maize. Crop Sci 49:509-520. Adams SE, Jones RAC, Coutts RHA (1986) Expression of Potato virus X resistance gene Rx in potato leaf protoplasts. J Gen Virol 67:2341-2345. Ameline-Torregrosa C, Wang BB, O’Bleness MS, Deshpande S, Zhu H, Roe B, Young ND, Cannon SB (2008) Identification and characterization of nucleotide-binding site-leucine-rich repeat genes in the model plant Medicago truncatula. Plant Physiol 146:5-21. Asano M, Satoh R, Mochizuki A, Tsuda S, Yamanaka T, Nishiguchi M, Hirai K, Meshi T, Naito S, Ishikawa M (2005) Tobamovirus-resistant tobacco generated by RNA interference directed against host genes. FEBS Lett 579:4479-4484. Barker H, McGeachy KD, Ryabov EV, Commandeur U, Mayo MA, Taliansky M (2001) Evidence for RNA-mediated defence effects on the accumulation of Potato leafroll virus. J Gen Virol 82:3099-3106. Bendahmane A, Farnham G, Moffett P, Baulcombe DC (2002) Constitutive gain-of-function mutants in a nucleotide binding site-leucine rich repeat protein encoded at the Rx locus of potato. Plant J 32:195-204. Bendahmane A, Kanyuka K, Baulcombe DC (1999) The Rx gene from potato controls separate virus resistance and cell death responses. Plant Cell 11:781-792. Bendahmane A, Kohn BA, Dedi C, Baulcombe DC (1995) The coat protein of Potato virus X is a strain-specific elicitor of Rx1-mediated virus resistance in potato. Plant J 8:933-941. Bendahmane A, Querci M, Kanyuka K, Baulcombe DC (2000) Agrobacterium transient expression system as a tool for the isolation of disease resistance genes: application to the Rx2 locus in potato. Plant J 21:73-81. Bhattacharjee S, Zamora A, Azhar MT, Sacco MA, Lambert LH, Moffett P (2009) Virus resistance induced by NB-LRR proteins involves Argonaute4-dependent translational control. Plant J 58:940-951. Brommonschenkel SH, Frary A, Tanksley SD (2000) The broad-spectrum tospovirus resistance gene Sw-5 of tomato is a homolog of the root-knot nematode resistance gene Mi. Mol Plant-Microbe Interact 13:1130-1138. Browning KS (1996) The plant translational apparatus. Plant Mol Biol 32:107-144. Browning KS (2004) Plant translation initiation factors: it is not easy to be green. Biochem Soc Trans 32:589-591. Caplan JL, Mamillapalli P, Burch-Smith TM, Czymmek K, Dinesh-Kumar SP (2008) Chloroplastic protein NRIP1 mediates innate immune receptor recognition of a viral effector. Cell 132:449-462. Carrington JC, Kasschau KD, Mahajan SK, Schaad MC (1996) Cell-to-cell and long-distance transport of viruses in plants. Plant Cell 8:1669-1681. Chaddock JA, Lord JM, Hartley MR, Roberts LM (1994) Pokeweed antiviral protein (PAP) mutations which permit E. coli growth do not eliminate catalytic activity towards prokaryotic ribosomes. Nucleic Acids Res 22:1536-1540. Chen MH, Citovsky V (2003) Systemic movement of a tobamovirus requires host cell pectin methylesterase. Plant J 35:386-392. Chisholm ST, Mahajan SK, Whitham SA, Yamamoto ML, Carrington JC (2000) Cloning of the Arabidopsis RTM1 gene, which controls restriction of long-distance movement of Tobacco etch virus. Proc Natl Acad Sci USA 97:489-494. Clarke S (1992) Protein isoprenylation and methylation at carboxyl-terminal cysteine residues. Annu Rev Biochem 61:355-386. Cole AB, Kiraly L, Ross K, Schoelz JE (2001) Uncoupling resistance from cell death in the hypersensitive response of Nicotiana species to Cauliflower mosaic virus infection. Mol Plant-Microbe Interact 14:31-41. Cooley MB, Pathirana S, Wu HJ, Kachroo P, Klessig DF (2000) Members of the Arabidopsis HRT/RPP8 family of resistance genes confer resistance to both viral and oomycete pathogens. Plant Cell 12:663-676. Dai S, Wei X, Alfonso AA, Pei L, Duque UG, Zhang Z, Babb GM, Beachy RN (2008) Transgenic rice plants that overexpress transcription factors RF2a and RF2b are tolerant to Rice tungro virus replication and disease. Proc Natl Acad Sci USA 105:21012-21016. Dangl JL, Jones JD (2001) Plant pathogens and integrated defence responses to infection. Nature 411:826-833. Dinesh-Kumar SP, Baker BJ (2000) Alternatively spliced N resistance gene transcripts: their possible role in Tobacco mosaic virus resistance. Proc Natl Acad Sci USA 97:1908-1913. Dinesh-Kumar SP, Whitham S, Choi D, Hehl R, Corr C, Baker B (1995) Transposon tagging of Tobacco mosaic virus resistance gene N: its possible role in the TMV-N-mediated signal transduction pathway. Proc Natl Acad Sci USA 92:4175-4180. Flor HH (1971) Current status of the gene-for-gene concept. Annu Rev Phytopathol 9:275-296. Fraser RSS (1990) The genetics of resistance to plant viruses. Annu Rev Phytopathol 28:179-200. Fuchs M (2008) Plant resistance to viruses: Engineered resistance. Page 156-164. in: Encylopedia of Virology. May BWJ, Regenmortel MHV eds. 3rd edn., Elsevier, Maryland Heights, MO. Gallie DR, Browning KS (2001) eIF4G functionally differs from eIF(iso)4G in promoting internal initiation, cap-independent translation, and translation of structured mRNAs. J Biol Chem 276:36951-36960. Gassmann W, Hinsch ME, Staskawicz BJ (1999) The Arabidopsis RPS4 bacterial-resistance gene is a member of the TIR-NBS-LRR family of disease-resistance genes. Plant J 20:265-277. Gutierrez-Campos R, Torres-Acosta JA, Saucedo-Arias LJ, Gomez-Lim MA (1999) The use of cysteine proteinase inhibitors to engineer resistance against potyviruses in transgenic tobacco plants. Nat Biotechnol 17:1223-1226. Hagiwara Y, Komoda K, Yamanaka T, Tamai A, Meshi T, Funada R, Tsuchiya T, Naito S, Ishikawa M (2003) Subcellular localization of host and viral proteins associated with tobamovirus RNA replication. EMBO J 22:344-353. Hajimorad MR, Eggenberger AL, Hill JH (2005) Loss and gain of elicitor function of Soybean mosaic virus G7 provoking Rsv1- mediated lethal systemic hypersensitive response maps to P3. J Virol 79:1215-1222. Hajimorad MR, Hill JH (2001) Rsv1-mediated resistance against Soybean mosaic virus-N is hypersensitive response-independent at inoculation site, but has the potential to initiate a hypersensitive response-like mechanism. Mol Plant-Microbe Interact 14:587-598. Halterman DA, Wei FS, Wise RP (2003) Powdery mildew-induced Mla mRNAs are alternatively spliced and contain multiple upstream open reading frames. Plant Physiol 131:558-567. Hamalainen JH, Kekarainen T, Gebhardt C, Watanabe KN, Valkonen JP (2000) Recessive and dominant genes interfere with the vascular transport of Potato virus A in diploid potatoes. Mol Plant-Microbe Interact 13:402-412. Hong Y, Saunders K, Hartley MR, Stanley J (1996) Resistance to geminivirus infection by virus-induced expression of dianthin in transgenic plants. Virology 220:119-127. Hull R (2002) Economic losses due to plant viruses. in: Matthew’s Plant Virology. Hull R eds. Academic Press, New York, NY, USA. Ishikawa M, Naito S, Ohno T (1993) Effects on the tom1 mutation of Arabidopsis thaliana on the multiplication of Tobacco mosaic virus RNA in protoplasts. J Virol 67:5328-5338. Kachroo P, Yoshioka K, Shah J, Dooner HK, Klessig DF (2000) Resistance to Turnip crinkle virus in Arabidopsis is regulated by two host genes, is salicylic acid dependent but NPR1, ethylene and jasmonate independent. Plant Cell 12:677-690. Kaminaka H, Nake C, Epple P, Dittgen J, Schutze K, Chaban C, Holt III BF, Merkle T, Schafer E, Harter K, Dangl JL (2006) bZIP10-LSD1 antagonism modulates basal defense and cell death in Arabidopsis following infection. EMBO J 25:4400-4411. Kang BC, Yeam I, Frantz JD, Murphy JF, Jahn MM (2005a) The pvr1 locus in pepper encodes a translation initiation factor eIF4E that interacts with Tobacco etch virus VPg. Plant J 41:392-405. Kang BC, Yeam I, Jahn MM (2005b) Genetics of plant virus resistance. Annu Rev Phytopathol 43:581-621. Kneller EL, Rakotondrafara AM, Miller WA (2006) Cap-independent translation of plant viral RNAs. Virus Res 119:63-75. Kohm BA, Goulden MG, Gilbert JE, Kavanagh TA, Baulcombe DC (1993) A Potato virus X resistance gene mediates an induced, nonspecific resistance in protoplasts. Plant Cell 5:913-920. Kushner DB, Lindenbach BD, Grdzelishvili VZ, Noueiry AO, Paul SM, Ahlquist P (2003) Systematic, genome-wide identification of host genes affecting replication of a positive-strand RNA virus. Proc Natl Acad Sci USA 100:15764-15769. Lawrence GJ, Finnegan EJ, Ayliffe MA, Ellis JG (1995) The L6 gene for flax rust resistance is related to the Arabidopsis bacterial resistance gene RPS2 and the tobacco viral resistance gene N. Plant Cell 7:1195-1206. Lellis AD, Kasschau KD, Whitham SA, Carrington JC (2002) Loss-of susceptibility mutants of Arabidopsis thaliana reveal an essential role for eIF(iso)4E during potyvirus infection. Curr Biol 12:1046-1051. Leonard S, Plante D, Wittmann S, Daigneault N, Fortin MG, Laliberte J-F (2000) Complex formation between potyvirus VPg and translation eukaryotic initiation factor 4E correlates with virus infectivity. J Virol 74:7730-7737. Lin N-C, Martin GB (2007) Pto- and Prf-mediated recognition of AvrPto and AvrPtoB restricts the ability of diverse Pseudomonas syringae pathovars to infect tomato. Mol Plant-Microbe Interact 20:806-815. Liu Y, Schiff M, Marathe R, Dinesh-Kumar SP (2002a) Tobacco RAR1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to Tobacco mosaic virus. Plant J 30:415-429. Liu Y, Schiff M, Serino G, Deng XW, Dinesh-Kumar SP (2002b) Role of SCF ubiquitin-ligase and the COP9 signalosome in the N gene-mediated resistance response to Tobacco mosaic virus. Plant Cell 14:1483-1496. Lodge JK, Kaniewski WK, Tumer NE (1993) Broad-spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein. Proc Natl Acad Sci USA 90:7089-7093. Loebenstein G, Manadilova A (2003) Potatoes in the central Asian Republics. Page 195-222. in: Virus and Virus-like Disease of Major Crops in Developing Countries. Loebenstein G, Thottappilly G eds. Kluwer Academic Publishers, Dordrecht. Lucas WJ, Bouche-Pillon S, Jackson DP, Nguyen L, Baker L, Ding B, Hake S (1995) Selective trafficking of KNOTTED1 homeodomain protein and its mRNA through plasmodesmata. Science 270:1980-1983. Mackey D, Holt BF, Wiig A, Dangl JL (2002) RIN4 interacts with Pseudomonas syringae type III effector molecules and is required for RPM1- mediated resistance in Arabidopsis. Cell 108:743-754. Meshi T, Motoyoshi F, Adachi A, Watanabe H, Takamatsu N, Okada Y (1988) Two concomitant base substitutions in the putative replicase genes of Tobacco mosaic virus confer the ability to overcome the effects of a tomato resistance gene, Tm-1. EMBO J 7:1575-1581. Mestre P, Brigneti G, Baulcombe DC (2000) An Ry-mediated resistance response in potato requires the intact active site of the NIa proteinase from Potato virus Y. Plant J 23:653-661. Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW (2003) Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis. Plant Cell 15:809-834. Miller WA, White KA (2006) Long-distance RNA-RNA interactions in plant virus gene expression and replication. Annu Rev Phytopathol 44:447-467. Moffett P (2009) Mechanisms of recognition in dominant R gene mediated resistance. Adv Virus Res 75:1-33. Moreno IM, Thompson JR, Garcia-Arenal F (2004) Analysis of the systemic colonization of cucumber plant by Cucumber green mottle mosaic virus. J Gen Virol 85:749-759. Murphy JF, Blauth JR, Livingstone KD, Lackney VK, Jahn MK (1998) Genetic mapping of the pvr1 locus in Capsicum spp. and evidence that distinct potyvirus resistance loci control responses that differ at the whole plant and cellular levels. Mol Plant-Microbe Interact 11:943-951. Mysore KS, Ryu CM (2004) Nonhost resistance: How much do we know? Trends Plant Sci 9:97-104. Nagy PD (2008) Yeast as a model host to explore plant virus-host interactions. Annu Rev Phytopathol 46:217-242. Nagy PD, Pogany J (2006) Yeast as a model host to dissect functions of viral and host factors in tombusvirus replication. Virology 344:211-220. Nicaise V, German-Retana S, Sanjuan R, Dubrana MP., Mazier M, Maisonneuve B, Candresse T, Caranta C, LeGall O (2003) The eukaryotic translation initiation factor 4E controls lettuce susceptibility to the potyvirus Lettuce mosaic virus. Plant Physiol 132:1272-1282. Ohshima K, Taniyama T, Yamanaka T, Ishikawa M, Naito S (1998) Isolation of a mutant of Arabidopsis thaliana carrying two simultaneous mutations affecting Tobacco mosaic virus multiplication within a single cell. Virology 243:472-481. Parker JE, Coleman MJ, Szabo V, Frost LN, Schmidt R, van der Biezen E, Moores T, Dean C, Daniels M, Jones JDG (1997) The Arabidopsis downy mildew resistance gene RPP5 shares similarity to the Toll and interleukin-1 receptors with N and L6. Plant Cell 9:879-894. Parrella G, Gognalons P, Gebre-Selassie K, Vovlas C, Marchoux G (2003) An update of the host range of Tomato spotted wilt virus. J Plant Pathol 85:227-264. Ponz F, Russel ML, Rowhani A, Bruening G (1988) A cowpea line has distinct genes for resistance to Tobacco ringspot virus and Cowpea mosaic virus. Phytopathology 78:1124-1128. Porter BW, Paidi M, Ming R, Alam M, Nishijima WT, Zhu YJ (2009) Genome-wide analysis of Carica papaya reveals a small NBS resistance gene family. Mol Genet Genom 281:609-626. Provvidenti R, Robinson RW (1977) Inheritance of resistance to Watermelon mosaic virus 1 in Cucumis metuliferus (Naud.) May. J Hered 68:56-57. Provvidenti R, Gonsalves D (1982) Resistance to Papaya ringspot virus in Cucumis metuliferus and its relationship to resistance to Watermelon mosaic virus 1. J Hered 72:239-240. Querci M, Baulcombe DC, Goldbach RW, Salazar LF (1995) Analysis of the resistance-breaking determinant of Potato virus X (PVX) strain HB on different potato genotypes expressing extreme resistance to PVX. Phytopathology 85:1003-1010. Reddy DVR, Bragard C, Sreenivasulu P, Delfosse P (2008) Pecluvirus. Page 97-105. in: Encyclopedia of Virology. Mahy BWJ, van Regenmortel MHV eds. Elsevier, Maryland Heights, MO. Ren T, Qu F, Morris TJ (2000) HRT gene function requires interaction between a NAC protein and viral capsid protein to confer resistance to Turnip crinkle virus. Plant Cell 12:1917-1925. Rentel MC, Leonelli L, Dahlbeck D, Zhao B, Staskawicz BJ (2008) Recognition of the Hyaloperonospora parasitica effector ATR13 triggeres resistance against oomycete, bacterial, and viral pathogens. Proc Natl Acad Sci USA 105:1091-1096. Robaglia C, Caranta C (2006) Translation initiation factors: A weak link in plant RNA virus infection. Trends Plant Sci 11:40-45. Rogers GWJr, Komar AA, Merrick WC (2002) eIF4A: the godfather of the DEAD box helicases. Prog Nucleic Acid Res Mol Biol 72:307-331. Sacco MA, Mansoor S, Moffett P (2007) A RanGAP protein physically interacts with the NB-LRR protein Rx, and is required for Rx-mediated viral resistance. Plant J 52:82-93. Sacco MA, Moffett P (2009) Disease resistance genes: Form and function. Page 94-141. in: Molecular Plant-Microbe Interactions. Bouarab K, Brisson N, Daayf F eds. CABI, Wallingford, UK. Santa Cruz S (1999) Perspective: phloem transport of viruses and macromolecules- What goes in must come out. Trends Microbiol 7:237-241. Schaad MC, Anderberg RJ, Carrington JC (2000) Strain-specific interaction of the Tobacco etch virus NIa protein with the translation initiation factor eIF4E in the yeast two-hybrid system. Virology 273:300-306. Shao F, Golstein C, Ade J, Stoutemyer M, Dixon JE, Innes RW (2003) Cleavage of Arabidopsis PBS1 by a bacterial type III effector. Science 301:1230-1233. Shen QH, Saijo Y, Mauch S, Biskup C, Bieri S, Keller B, Seki H, Ulker B, Somssich IE, Schulze-Lefert P (2007) Nuclear activity of MLA immune receptors links isolate-specific and basal disease-resistance responses. Science 315:1098-1103. Singh DP, Moore CA, Gilliland A, Carr JP (2004) Activation of multiple antiviral defence mechanisms by salicylic acid. Mol Plant Pathol 5:57-63. Takahashi H, Miller J, Nozaki Y, Takeda M, Shah J, Hase S, Ikegami M, Ehara Y, Dinesh-Kumar SP (2002) RCY1, an Arabidopsis thaliana RPP8/ HRT family resistance gene, conferring resistance to Cucumber mosaic virus requires salicylic acid, ethylene and a novel signal transduction mechanism. Plant J 32:655-667. Tameling WIL, Nooijen C, Ludwig N, Boter M, Slootweg E, Goverse A, Shirasu K, Joosten MHAJ (2010) RanGAP2 mediates nucleocytoplasmic partitioning of the NB-LRR immune receptor Rx in the Solanaceae, thereby dictating Rx function. Plant Cell 22:4176-4194. Tornero P, Merritt P, Sadanandom A, Shirasu K, Innes RW, Dangl JL (2002) RAR1 and NDR1 contribute quantitatively to disease resistance in Arabidopsis, and their relative contributions are dependent on the R gene assayed. Plant Cell 14:1005-1015. Truniger V, Aranda MA (2009) Recessive resistance to plant viruses. Adv Virus Res 75:119-159. Tsujimoto Y, Numaga T, Ohshima K, Yano M-A, Ohsawa R, Goto DB, Naito S, Ishikawa, M (2003) Arabidopsis TOBAMOVIRUS MULTIPLICATION (TOM) 2 locus encodes a transmembrane protein that interacts with TOM1. EMBO J 22: 335-343. Tumer NE, Hwang DJ, Bonness M (1997) C-terminal deletion mutant of pokeweed antiviral protein inhibits viral infection but does not depurinate host ribosomes. Proc Natl Acad Sci USA 94:3866-3871. Uehara Y, Takahashi Y, Berberich T, Miyazaki A, Takahashi H, Matsui K, Ohme-Takagi M, Saitoh H, Terauchi R, Kusano T (2005) Tobacco ZFT1, a transcriptional repressor with a Cys2/His2-type zinc finger motif that functions in spermine-signaling pathway. Plant Mol Biol 59:435-448. van der Biezen EA, Jones JD (1998) Plant disease-resistance proteins and the gene-for-gene concept. Trends Biochem Sci 23:454-456. van der Hoorn RAL, Kamoun S (2008) From guard to decoy: A new model for perception of plant pathogen effectors. Plant Cell 20:2009-2017. van der Vossen EA, van der Voort JN, Kanyuka K, Bendahmane A, Sandbrink H, Baulcombe DC, Bakker J, Stiekema WJ, Klein-Lankhorst RM (2000) Homologues of a single resistance-gene cluster in potato confer resistance to distinct pathogens: A virus and a nematode. Plant J 23:567-576. Velasco R, Zharkikh A, Troggio M, Cartwright DA, Cestaro A, Pruss D, Pindo M, Fitzgerald LM, Vezzulli S, Reid J, Malacarne G, Iliev D, Coppola G, Wardell B, Micheletti D, Macalma T, Facci M, Mitchell JT, Perazzolli M, Eldredge G, Gatto P, Oyzerski R, Moretto M, Gutin N, Stefanini M, Chen Y, Segala C, Davenport C, Dematte L, Mraz A, Battilana J, Stormo K, Costa F, Tao Q, Si-Ammour A, Harkins T, Lackey A, Perbost C, Taillon B, Stella A, Solovyev V, Fawcett JA, Sterck L, Vandepoele K, Grando SM, Toppo S, Moser C, Lanchbury J, Bogden R, Skolnick M, Sgaramella V, Bhatnagar SK, Fontana P, Gutin A, van de Peer Y, Salamini F, Viola R (2007) A high quality draft consensus sequence of the genome of a heterozygous grapevine variety. PLoS ONE 2:e1326. Whitham SA, Anderberg RJ, Chisholm ST, Carrington JC (2000) Arabidopsis RTM2 gene is necessary for specific restriction of Tobacco etch virus and encodes an unusual small heat shock-like protein. Plant Cell 12:569-582. Whitham S, Dinesh-Kumar SP, Choi D, Hehl R, Corr C, Baker B (1994) The product of the Tobacco mosaic virus resistance gene N: similarity to Toll and the interleukin-1 receptor. Cell 78:1101-1115. Wiermer M, Feys BJ, Parker JE (2005) Plant immunity: the EDS1 regulatory node. Curr Opin Plant Biol 8:383-389. Xiang T, Zhong N, Zou Y, Wu Y, Zhang J, Xing W, Li Y, Tang X, Zhu L, Chai J, Zhou J-M (2008) Pseudomonas syringae effector AvrPto blocks innate immunity by targeting receptor kinases. Curr Biol 18:74-80. Xie Z, Fan B, Chen C, Chen Z (2001) An important role of an inducible RNA-dependent RNA polymerase in plant antiviral defense. Proc Natl Acad Sci USA 98:6516-6521. Xu L, Massague J (2004) Nucleocytoplasmic shuttling of signal transducers. Nat Rev Mol Cell Biol 5:209-219. Yoshii M, Shimizu T, Yamazaki M, Higashi T, Miyao A, Hirochika H, Omura T (2009) Disruption of a novel gene for a NAC-domain protein in rice confers resistance to Rice dwarf virus. Plant J 57:615-625. Yoshii M, Yamazaki M, Rakwal R, Kishi-Kaboshi M, Miyao A, Hirochika H (2010) The NAC transcription factor RIM1 of rice is a new regulator of jasmonate signaling. Plant J 61:804-815. Yoshii M, Yoshioka N, Ishikawa M, Naito S (1998a) Isolation of an Arabidopsis thaliana mutant in which accumulation of Cucumber mosaic virus coat protein is delayed. Plant J 13:211-219. Yoshii M, Yoshioka N, Ishikawa M, Naito S (1998b) Isolation of an Arabidopsis thaliana mutant in which the multiplication of both Cucumber mosaic virus and Turnip crinkle virus is affected. J Virol 72:8731-8737. Yu IC, Parker J, Bent AF (1998) Gene-for-gene disease resistance without the hypersensitive response in Arabidopsis dnd1 mutant. Proc Natl Acad Sci USA 95:7819-7824. Zhou JM, Chai J (2008) Plant pathogenic bacterial type III effectors subdue host responses. Curr Opin Microbiol 11:179-185. Zhou T, Wang Y, Chen JQ, Araki H, Jing Z, Jiang K, Shen J, Tian D (2004) Genome-wide identification of NBS genes in japonica rice reveals significant expansion of divergent non-TIR NBS-LRR genes. Mol Genet Genom 271:402-415. Zipfel C, Rathjen JP (2008) Plant Immunity: AvrPto targets the frontline. Curr Biol 18:R218-R22.||摘要:||
許多瓜類在熱帶及亞熱帶地區容易受到木瓜輪點病毒(Papaya ring spot virus, PRSV)侵害，造成植株生長受阻以及產量及品質下降，因此了解植物抗病毒機制與培育抗病之栽培品種為當前重要的工作。目前對於木瓜輪點病毒的防治策略，包括:交互保護方法和利用病毒誘導基因靜默方法來達到木瓜輪點病的防治。然而上述的方法雖然可以達到有效的成果，但是交互保護需要高度勞力，且病毒基因發生突變時，病毒誘導基因靜默的抗病方法就無法達到有效抑制病毒的效果。因此本研究的目地為探討木瓜輪點病毒與其寄主植物刺角瓜之間抗病反應的交互作用，並且從中選殖與PRSV抗病反應相關的候選基因，最後希望未來將該抗病基因應用在其它改善植物的病毒抗病育種。本實驗共分成三個部分，首先為了之後分析刺角瓜候選基因的功能，必須建立刺角瓜的再生與農桿菌轉殖系統。該系統主要是使用刺角瓜的子葉作為再生與農桿菌轉殖的培殖體，結合植物生長素(BA和NAA)的使用，最後只要6~8個星期的時間就可以成功得到刺角瓜轉殖株，目前轉殖系統的成功效率有2.5%。另外，為了探討刺角瓜在PRSV感染後，其基因的差異表現，本研究利用cDNA-AFLP方法針對兩個刺角瓜品系(分別為抗病品系PI 292190和感病品系Acc. 2459)進行多型性片段的分析。經由比較抗病品系PI 292190和感病品系Acc. 2459接種PRSV，以及對照組處理後，將差異表現的片段進行選殖。最後有139個候選片段可能與PRSV抗病的反應有關係，之後利用reverse northern blot和northern blot方法篩選出25個可能與PRSV抗病反應有關的候選基因。其中CmPI 1基因的表現分析在northern blot結果顯示，抗病品系PI 292190接種病毒後48小時，該基因的表現量快速提升並達到最高;反之感病品系Acc. 2459的基因表現需要到接種後21天才出現。經由序列比對CmPI 1基因可以得知其編碼出來的蛋白與potato I type蛋白酶抑制子有很高的相似。另外，PRSV屬於potyvirus家族，該家族的病毒須要利用自身的蛋白酶進行病毒的複製和感染反應。由這些證據顯示，CmPI 1基因的蛋白產物可能與病毒的蛋白酶有交互作用，進而干擾蛋白酶的功能。本研究因此將CmPI 1基因片段構築在可以產生siRNA的RNAi載體pEPJ86i，並利用農桿菌轉殖法進行刺角瓜抗病品系PI 292190轉殖，最後也獲得CmPI 1基因靜默的轉殖株。將CmPI 1基因靜默的轉殖株接種PRSV可以發現，轉殖株在接種後21天有病徵出現，證實抗病品系PI 292190的CmPI 1基因表現被抑制時，刺角瓜對PRSV的抗性會被破壞並且導致感病。此外，CmPI 1過量表現試驗的結果，證實感病品系Acc. 2459在持續表現CmPI 1基因的情形下，轉基因Acc. 2459可以抵抗PRSV感染。綜合上述結果，可以確認刺角瓜的CmPI 1基因參與PRSV抗病反應，並且可以提供刺角瓜抗PRSV的能力。本論文的結果有助於了解抗病相關的候選基因在植物與病毒抗病反應之間可能扮演的角色，另外利用植物蛋白酶抑制子提供植物在potyvirus屬病毒的抗病能力，也將是未來病毒防治上另一個可行的策略。
Papaya ringspot virus (PRSV) is Potyvirus genus and causes severe damages of crops especially papaya and cucurbits. It is a top priority to understand mechanisms of plant virus resistance to assist the breeding programs for developing resistant cultivars. Until now, the control of PRSV includes cross-protection and virus-induced gene silencing (VIGS). However, labor-intensive work and resistance broken have limited the application of these strategies. This study aims to investigate the interaction between Cucumis metuliferus and PRSV, and identify defense-related candidate genes. On top of that, the candidate genes could be used in the improvement of plant resistance breeding. This study contains three major parts of contribution. First, the regeneration and Agrobacterium-mediated transformation system of C. metuliferus were developed to assist functional characterization of candidate genes. The cotyledon of C. metuliferus was used as explants in the regeneration and Agrobacterium-mediated transformation experiments. By means of benzyl adenine (BA) (1 mg/l) and naphthaleneacetic acid (NAA) (0.02 mg/l), six to eight weeks are acquired to obtain transgenic C. metuliferus plants and the transformation efficiency was 2.5%. Second, the cDNA-AFLP method was used to analyze the differential displayed genes of C. metuliferus treated with or without PRSV-inoculation. The polymorphic fragments were identified by comparing the difference of fragments occurring in resistant line PI 292190 and susceptible line Acc. 2459 inoculated with PRSV or treated with mock. A total of 139 candidates were cloned and 25 candidates of them were selected based on further results of reverse northern blot and northern blot analysis. One of these candidates, CmPI 1 has shown the most possible candidate correlated with PRSV resistance. As the result of northern blot, the gene expression of CmPI 1 has been shown to increase quickly in resistant line PI 292190 at 48 hour-post inoculation (hpi), in contrast, express not until 21 day-post inoculation (dpi) in susceptible line Acc. 2459. CmPI 1 encodes a protein with homology to potato I type proteinase inhibitor by sequencing analysis. In addition, PRSV needs virus proteinases to complete its replication and infection cycle. This implicated that CmPI 1 could impede the function of virus proteinase by interacting with virus proteinases. To estimate this hypothesis, the fragment of CmPI 1 was constructed onto an RNAi vector, pEPJ86i. The resistant line PI 292190 was transferred with this RNAi construction using Agrobacterium-mediated transformation method. The transgenic plants were shown infectious and symptoms were observed after 21 dpi. In the over-expression CmPI 1, a resistant allele derived from resistant line PI 292190 was transformed into susceptible line Acc. 2459 to test the resistance ability of cloned CmPI 1 gene against PRSV. According to the above results, CmPI 1 were able to participate in PRSV resistance and contribute the resistance ability in C. metuliferus. Overall, the results in this dissertation provide a better understanding the roles of defense-related genes in host-virus interaction. The application of plant proteinase inhibitors in inducing resistance against potyviruses would also be another possible strategy for virus control.
|Appears in Collections:||農藝學系|
Show full item record
TAIR Related Article
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.