Please use this identifier to cite or link to this item:
標題: The effect of NbSCL6 on Bamboo mosaic virus infection cycle in Nicotiana benthamiana
作者: 王斯遠
Ssu-yuan Wang
關鍵字: 菸草;竹嵌紋病毒;N. bentamiana;BaMV;GRAS;SCL-6;VIGS
引用: Bolle, C., 2004. The role of GRAS proteins in plant signal transduction and development. Planta 218, 683-692. Bolle, C., Koncz, C., Chua, N.H., 2000. PAT1, a new member of the GRAS family, is involved in phytochrome A signal transduction. Genes & development 14, 1269-1278. Brosche, M., Blomster, T., Salojarvi, J., Cui, F., Sipari, N., Leppala, J., Lamminmaki, A., Tomai, G., Narayanasamy, S., Reddy, R.A., Keinanen, M., Overmyer, K., Kangasjarvi, J., 2014. Transcriptomics and functional genomics of ROS-induced cell death regulation by RADICAL-INDUCED CELL DEATH1. PLoS genetics 10, e1004112. Cheng, S.F., Huang, Y.P., Wu, Z.R., Hu, C.C., Hsu, Y.H., Tsai, C.H., 2010. Identification of differentially expressed genes induced by Bamboo mosaic virus infection in Nicotiana benthamiana by cDNA-amplified fragment length polymorphism. BMC plant biology 10, 286. Day, R.B., Tanabe, S., Koshioka, M., Mitsui, T., Itoh, H., Ueguchi-Tanaka, M., Matsuoka, M., Kaku, H., Shibuya, N., Minami, E., 2004. Two rice GRAS family genes responsive to N -acetylchitooligosaccharide elicitor are induced by phytoactive gibberellins: evidence for cross-talk between elicitor and gibberellin signaling in rice cells. Plant molecular biology 54, 261-272. Desveaux, D., Subramaniam, R., Despres, C., Mess, J.N., Levesque, C., Fobert, P.R., Dangl, J.L., Brisson, N., 2004. A 'Whirly' transcription factor is required for salicylic acid-dependent disease resistance in Arabidopsis. Dev Cell 6, 229-240. Di Laurenzio, L., Wysocka-Diller, J., Malamy, J.E., Pysh, L., Helariutta, Y., Freshour, G., Hahn, M.G., Feldmann, K.A., Benfey, P.N., 1996. The SCARECROW gene regulates an asymmetric cell division that is essential for generating the radial organization of the Arabidopsis root. Cell 86, 423-433. Eulgem, T., 2005. Regulation of the Arabidopsis defense transcriptome. Trends in plant science 10, 71-78. Eulgem, T., Rushton, P.J., Schmelzer, E., Hahlbrock, K., Somssich, I.E., 1999. Early nuclear events in plant defence signalling: rapid gene activation by WRKY transcription factors. The EMBO journal 18, 4689-4699. Fu, X., Richards, D.E., Ait-Ali, T., Hynes, L.W., Ougham, H., Peng, J., Harberd, N.P., 2002. Gibberellin-mediated proteasome-dependent degradation of the barley DELLA protein SLN1 repressor. The Plant cell 14, 3191-3200. Greb, T., Clarenz, O., Schafer, E., Muller, D., Herrero, R., Schmitz, G., Theres, K., 2003. Molecular analysis of the LATERAL SUPPRESSOR gene in Arabidopsis reveals a conserved control mechanism for axillary meristem formation. Genes & development 17, 1175-1187. Helariutta, Y., Fukaki, H., Wysocka-Diller, J., Nakajima, K., Jung, J., Sena, G., Hauser, M.T., Benfey, P.N., 2000. The SHORT-ROOT gene controls radial patterning of the Arabidopsis root through radial signaling. Cell 101, 555-567. Hirsch, S., Oldroyd, G.E., 2009. GRAS-domain transcription factors that regulate plant development. Plant signaling & behavior 4, 698-700. Huang, Y.-L., Han, Y.-T., Chang, Y.-T., Hsu, Y.-H., Meng, M., 2004. Critical residues for GTP methylation and formation of the covalent m7GMP-enzyme intermediate in the capping enzyme domain of bamboo mosaic virus. Journal of virology 78, 1271-1280. Ikeda, A., Ueguchi-Tanaka, M., Sonoda, Y., Kitano, H., Koshioka, M., Futsuhara, Y., Matsuoka, M., Yamaguchi, J., 2001. slender rice, a constitutive gibberellin response mutant, is caused by a null mutation of the SLR1 gene, an ortholog of the height-regulating gene GAI/RGA/RHT/D8. The Plant cell 13, 999-1010. Itoh, H., Ueguchi-Tanaka, M., Sato, Y., Ashikari, M., Matsuoka, M., 2002. The gibberellin signaling pathway is regulated by the appearance and disappearance of SLENDER RICE1 in nuclei. The Plant cell 14, 57-70. Kalinina, N., Rakitina, D., Solovyev, A., Schiemann, J., Morozov, S.Y., 2002. RNA helicase activity of the plant virus movement proteins encoded by the first gene of the triple gene block. Virology 296, 321-329. Kasschau, K.D., Xie, Z., Allen, E., Llave, C., Chapman, E.J., Krizan, K.A., Carrington, J.C., 2003. P1/HC-Pro, a Viral Suppressor of RNA Silencing, Interferes with< i> Arabidopsis</i> Development and miRNA Function. Developmental cell 4, 205-217. Krishnamurthy, K., Heppler, M., Mitra, R., Blancaflor, E., Payton, M., Nelson, R.S., Verchot-Lubicz, J., 2003. The< i> Potato virus X</i> TGBp3 protein associates with the ER network for virus cell-to-cell movement. Virology 309, 135-151. Lee, C.-C., Ho, Y.-N., Hu, R.-H., Yen, Y.-T., Wang, Z.-C., Lee, Y.-C., Hsu, Y.-H., Meng, M., 2011. The interaction between bamboo mosaic virus replication protein and coat protein is critical for virus movement in plant hosts. Journal of virology 85, 12022-12031. Lee, M.H., Kim, B., Song, S.K., Heo, J.O., Yu, N.I., Lee, S.A., Kim, M., Kim, D.G., Sohn, S.O., Lim, C.E., Chang, K.S., Lee, M.M., Lim, J., 2008. Large-scale analysis of the GRAS gene family in Arabidopsis thaliana. Plant molecular biology 67, 659-670. Lee, Y.-S., Hsu, Y.-H., Lin, N.-S., 2000. Generation of subgenomic RNA directed by a satellite RNA associated with bamboo mosaic potexvirus: analyses of potexvirus subgenomic RNA promoter. Journal of virology 74, 10341-10348. Li, X., Qian, Q., Fu, Z., Wang, Y., Xiong, G., Zeng, D., Wang, X., Liu, X., Teng, S., Hiroshi, F., Yuan, M., Luo, D., Han, B., Li, J., 2003. Control of tillering in rice. Nature 422, 618-621. Li, Y.-I., Chen, Y.-J., Hsu, Y.-H., Meng, M., 2001a. Characterization of the AdoMet-dependent guanylyltransferase activity that is associated with the N terminus of bamboo mosaic virus replicase. Journal of virology 75, 782-788. Li, Y.-I., Cheng, Y.-M., Huang, Y.-L., Tsai, C.-H., Hsu, Y.-H., Meng, M., 1998. Identification and characterization of the Escherichia coli-expressed RNA-dependent RNA polymerase of bamboo mosaic virus. Journal of virology 72, 10093-10099. Li, Y.-I., Shih, T.-W., Hsu, Y.-H., Han, Y.-T., Huang, Y.-L., Meng, M., 2001b. The helicase-like domain of plant potexvirus replicase participates in formation of RNA 5′ cap structure by exhibiting RNA 5′-triphosphatase activity. Journal of virology 75, 12114-12120. Lin, M.-K., Chang, B.-Y., Liao, J.-T., Lin, N.-S., Hsu, Y.-H., 2004. Arg-16 and Arg-21 in the N-terminal region of the triple-gene-block protein 1 of Bamboo mosaic virus are essential for virus movement. Journal of general virology 85, 251-259. Lin, M.-K., Hu, C.-C., Lin, N.-S., Chang, B.-Y., Hsu, Y.-H., 2006. Movement of potexviruses requires species-specific interactions among the cognate triple gene block proteins, as revealed by a trans-complementation assay based on the bamboo mosaic virus satellite RNA-mediated expression system. Journal of general virology 87, 1357-1367. Lin, M., Kitajima, E., Cupertino, F., Costa, C., 1977. Partial purification and some properties of bamboo mosaic virus. Phytopathology 67, 1439-1443. Lin, N.-S., Chen, C.-C., 1991. Association of bamboo mosaic virus(BoMV) and BoMV-specific electron-dense crystalline bodies with chloroplasts. Phytopathology 81, 1551-1555. Lin, N.-S., Lin, F.-Z., Huang, T.-Y., Hsu, Y.-H., 1992. Genome properties of bamboo mosaic virus. Phytopathology 82, 731-734. Lin, N.S., Lin, B.Y., Lo, N.W., Hu, C.C., Chow, T.Y., Hsu, Y.H., 1994. Nucleotide sequence of the genomic RNA of bamboo mosaic potexvirus. The Journal of general virology 75 ( Pt 9), 2513-2518. Llave, C., Xie, Z., Kasschau, K.D., Carrington, J.C., 2002. Cleavage of Scarecrow-like mRNA targets directed by a class of Arabidopsis miRNA. Science (New York, N.Y.) 297, 2053-2056. Mitra, R., Krishnamurthy, K., Blancaflor, E., Payton, M., Nelson, R.S., Verchot-Lubicz, J., 2003. The< i> potato virus x</i> TGBp2 protein association with the endoplasmic reticulum plays a role in but is not sufficient for viral cell-to-cell movement. Virology 312, 35-48. Morohashi, K., Minami, M., Takase, H., Hotta, Y., Hiratsuka, K., 2003. Isolation and characterization of a novel GRAS gene that regulates meiosis-associated gene expression. The Journal of biological chemistry 278, 20865-20873. Palani, P.V., Kasiviswanathan, V., Chen, J.C.-F., Chen, W., Hsu, Y.-H., Lin, N.-S., 2006. The arginine-rich motif of Bamboo mosaic virus satellite RNA-encoded P20 mediates self-interaction, intracellular targeting, and cell-to-cell movement. Molecular plant-microbe interactions 19, 758-767. Pysh, L.D., Wysocka-Diller, J.W., Camilleri, C., Bouchez, D., Benfey, P.N., 1999. The GRAS gene family in Arabidopsis: sequence characterization and basic expression analysis of the SCARECROW-LIKE genes. Plant J 18, 111-119. Ruiz, M.T., Voinnet, O., Baulcombe, D.C., 1998. Initiation and maintenance of virus-induced gene silencing. The Plant cell 10, 937-946. Rushton, P.J., Reinstadler, A., Lipka, V., Lippok, B., Somssich, I.E., 2002. Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen- and wound-induced signaling. The Plant cell 14, 749-762. Rushton, P.J., Somssich, I.E., 1998. Transcriptional control of plant genes responsive to pathogens. Current opinion in plant biology 1, 311-315. Rushton, P.J., Torres, J.T., Parniske, M., Wernert, P., Hahlbrock, K., Somssich, I.E., 1996. Interaction of elicitor-induced DNA-binding proteins with elicitor response elements in the promoters of parsley PR1 genes. The EMBO journal 15, 5690-5700. Schumacher, K., Schmitt, T., Rossberg, M., Schmitz, G., Theres, K., 1999. The Lateral suppressor (Ls) gene of tomato encodes a new member of the VHIID protein family. Proceedings of the National Academy of Sciences of the United States of America 96, 290-295. Silverstone, A.L., Ciampaglio, C.N., Sun, T., 1998. The Arabidopsis RGA gene encodes a transcriptional regulator repressing the gibberellin signal transduction pathway. The Plant cell 10, 155-169. Singh, K.B., Foley, R.C., O?ate-S?nchez, L., 2002. Transcription factors in plant defense and stress responses. Current opinion in plant biology 5, 430-436. Stuurman, J., Jaggi, F., Kuhlemeier, C., 2002. Shoot meristem maintenance is controlled by a GRAS-gene mediated signal from differentiating cells. Genes & development 16, 2213-2218. Tian, C., Wan, P., Sun, S., Li, J., Chen, M., 2004. Genome-wide analysis of the GRAS gene family in rice and Arabidopsis. Plant molecular biology 54, 519-532. Van Verk, M.C., Gatz, C., Linthorst, H.J., 2009. Transcriptional regulation of plant defense responses. Advances in Botanical Research 51, 397-438. Voinnet, O., Lederer, C., Baulcombe, D.C., 2000. A Viral Movement Protein Prevents Spread of the Gene Silencing Signal in< i> Nicotiana benthamiana</i>. Cell 103, 157-167. Wen, C.-K., Chang, C., 2002. Arabidopsis RGL1 encodes a negative regulator of gibberellin responses. The Plant Cell Online 14, 87-100. Yang, Y., Klessig, D.F., 1996. Isolation and characterization of a tobacco mosaic virus-inducible myb oncogene homolog from tobacco. Proceedings of the National Academy of Sciences of the United States of America 93, 14972-14977. Zhou, J., Tang, X., Martin, G.B., 1997. The Pto kinase conferring resistance to tomato bacterial speck disease interacts with proteins that bind a cis-element of pathogenesis-related genes. The EMBO journal 16, 3207-3218.
竹嵌紋病毒(Bamboo mosaic virus; BaMV)為一外型絲狀的單一正意股RNA病毒,屬於Flexiviridae科,Potexvirus屬。本實驗主要是針對在病毒的感染週期中,菸草(Nicotiana benthamiana)基因差異性表現的基因對於BaMV的影響,並進而探討其影響病毒的機制。在初步的研究中,利用cDNA amplified fragment length polymorphism (cDNA-AFLP)的技術,將這些具差異性表現的基因片段篩選出來,目前已有90個基因片段被篩選出來,其中一個命名為ACTC7-1的基因片段被選擇出來進行後續的研究。菸草基因ACTC7-1在竹嵌紋病毒感染後,會有表現量增加的現象,表示它可能是為了防禦病毒或是被病毒利用而造成表現量增加。首先,我利用VIGS (virus-induced gene silencing)的技術將ACTC7-1的表現量進行靜默,然後在ACTC7-1基因靜默的菸草進行病毒的接種,發現竹嵌紋病毒的外鞘蛋白累積量有增加的現象。因此猜測ACTC7-1應該是屬於防禦病毒的相關基因。為了釐清這個基因的影響層次是作用在病毒的複製階段還是在病毒的移動階段,我在將病毒RNA接種在ACTC7-1基因靜默的原生質體中,發現病毒的外鞘蛋白累積量也有增加的現象。由此結果可以猜測ACTC7-1是作用在病毒的複製階段。最後,利用RACE的技術,我得到了ACTC7-1的全長基因,透過NCBI的資料庫比對,發現ACTC7-1跟SCL-6 (SCARECROW-LIKE-6) 蛋白的相似度很高。SCL屬於GRAS家族,是一種轉錄因子(transcription factor),對根部發育的調控有關。未來會大量表現ACTC7-1蛋白,判斷是否會對病毒的累積量有影響。並觀察ACTC7-1蛋白在菸草細胞中的位置,將可以更進一步的探討ACTC7-1對病毒影響的真正機制為何。

Bamboo mosaic virus ( BaMV) is a single-stranded positive-sense RNA virus with a 5¢cap and a 3¢ poly(A) tail. BaMV belongs to the genus Potexvirus and the family Flexiviridae. The gene expression profile in Nicotiana benthamiana may be altered after BaMV infection. To identify the possible host genes involving the infection cycle of BaMV, our lab used cDNA-AFLP technique to screen the differentially expressed genes in BaMV-inoculated N. benthamiana plants. ACTC7-1 is an upregulated gene when BaMV infects N. benthamiana. To characterize the function of ACTC7-1 involving in BaMV infection cycle, I used the virus-induced gene silencing (VIGS) technique to known down the expression of ACTC7-1 leveling N. benthamiana plant and then inoculated BaMV onto the knockdown leaves. The accumulation levels of BaMV coat protein was determined by Western blotting analysis. The accumulation of BaMV was enhanced when after the expression of ACTC7-1 was knocked down. To further analyze the effect of ACTC7-1 on BaMV infection is on viral RNA replication or virus movement, I infected BaMV RNA into the ACTC7-1-knockdown protoplasts which were derived from the knockdown plants. The result of the accumulation of BaMV coat protein in N. benthamiana protoplasts was also increased. Overall of these preliminary results suggest that ACTC7-1 is probably involved in the replication of BaMV. Furthermore, I used RACE technique to clone the full-length of ACTC7-1 and blasted the sequence to WORKBENCH. The identity of ACTC7-1 could be the SCARECROW-LIKE transcriotion factor in GRAS family. The future work of this research will focus on the transient expression the full-length of ACTC7-1 fused with Orange fluorescent protein in N. benthamiana to localize this protein and examine the effect on BaMV accumulation
Rights: 同意授權瀏覽/列印電子全文服務,2016-08-31起公開。
Appears in Collections:生物科技學研究所

Files in This Item:
File SizeFormat Existing users please Login
nchu-103-7099041008-1.pdf1.31 MBAdobe PDFThis file is only available in the university internal network    Request a copy
Show full item record

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.