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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳昭瑩 | |
dc.contributor.author | Wei-Hsiang Tsai | en |
dc.contributor.author | 蔡瑋祥 | zh_TW |
dc.date.accessioned | 2021-06-17T02:14:08Z | - |
dc.date.available | 2020-01-04 | |
dc.date.copyright | 2018-01-04 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-11-16 | |
dc.identifier.citation | 1. 王清玲、邱一中。2007。作物蟲害之非農藥防治技術。利用天敵防治害蟲。行政院農業委員會農業試驗所。1-18 頁。
2. 林玉儒。2008。臘狀芽孢桿菌 C1L 菌株在臺灣百合群聚能力之探討。國立臺灣大學植物病理與微生物學系碩士論文。台北。台灣。88頁。 3. 陳榮五。2007。台灣有機農業發展之瓶頸。行政院農業委員會臺中區農業改良場。9 頁。 4. 陳鈺婷。2013。草莓灰黴病生物防治之應用研究。國立臺灣大學植物病理與微生物學系碩士論文。台北。台灣。62頁。 5. 曾安慈。2010。臘狀芽孢桿菌於玉米根圈群聚與誘導抗病能力相關性探討。國立臺灣大學植物病理與微生物學系碩士論文。台北。台灣。62頁。 6. 劉益宏。2004。臺灣百合根圈細菌之篩選及灰黴病菌防治應用研究。國立臺灣大學植物病理與微生物學系碩士論文。台北。台灣。68頁。 7. Abraham, N. M., Liu, L., Jutras, B. L., Murfin, K., Acar, A., Yarovinsky, T. O., Sutton, E., Heisig, M., Jacobs-Wagner, C., and Fikrig, E. 2017. A tick antivirulence protein potentiates antibiotics against Staphylococcus aureus. Antimicrob. Agents Chemother. 61(7): 113-117. 8. Ahemad, M. and Kibret, M. 2014. Mechanisms and applications of plant growth promoting rhizobacteria: Current perspective. J. King Saud Univ. Sci. 26(1): 1-20. 9. Asai, S., Mase, K., and Yoshioka, H. 2010. Role of nitric oxide and reactive oxide species in disease resistance to necrotrophic pathogens. Plant Signal. Behav. 5(7): 872-874. 10. Asselbergh, B., Curvers, K., França, S. C., Audenaert, K., Vuylsteke, M., Breusegem, F. V., and Höfte, M. 2007. Resistance to Botrytis cinerea in sitiens, an abscisic acid-deficient tomato mutant, involves timely production of hydrogen peroxide and cell wall modifications in the epidermis. Plant Physiol. 144: 1863-1877. 11. Bailey, M. J., Koronakis, V., Schmoll, T., and Hughes, C. 1992. Escherichia coli HlyT protein, a transcriptional activator of haemolysin synthesis and secretion is encoded by the rfaH (sfrB) locus required for expression of sex factor and lipopolysaccharide gene. Mol. Microbiol. 6: 1003-1012. 12. Beneduzi, A., Ambrosini, A., and Passaglia, L. M. P. 2012. Plant growth-promoting rhizobacteria (PGPR): Their potential as antagonists and biocontrol agents. Genet. Mol. Biol. 35(4): 1044-1051. 13. Bhattacharyya, P. H. and Jha, D. K. 2012. Plant growth-promoting rhizobacteria (PGPR): emergence in agriculture. World J. Microbiol. Biotechnol. 28(4): 1327-1350. 14. Chang, L. K., Chen, C. L., Chang, Y. S., Tschen, J. S. M., Chen, Y. M., and Liu, S. T. 1994. Construction of Tn917ac1, a transposon useful for mutagenesis and cloning of Bacillus subtilis genes. Gene 150(1): 129-134. 15. Choudhary, D. K., Prakash, A., and Johri, B. N. 2007. Induced systemic resistance (ISR) in plants: mechanism of action. Indian J. Microbiol. 47(4): 289-297. 16. Cook, R. J. 2002. Advances in plant health management in the twentieth century. Annu. Rev. Phytopathol. 38(1): 95-116. 17. De Torres Zabela, M., Fernandez-Delmond, I., Niittyla, T., Sanchez, P., and Grant, M. 2002. Differential expression of genes encoding Arabidopsis phospholipases after challenge with virulent or avirulent Pseudomonas isolates. Mol. Plant-Microbe Interact. 15: 808-816. 18. De Vleesschauwer, D. and Höfte, M. 2009. Bacterial determinants and host defense responses underpinning rhizobacteria-mediated systemic resistance in rice. In: Wang GL., Valent B. (eds) Advances in Genetics, Genomics and Control of Rice Blast Disease. Springer, Dordrecht, Holland. 19. Doll, V. M., Ehling-Schulz, M., and Vogelmann, R. 2013. Concerted action of sphingomyelinase and non-hemolytic enterotoxin in pathogenic Bacillus cereus. PLoS ONE 8(4): 1-13. 20. Dutta, S., Rani, T. S., and Podile, A. R. 2013. Root exudate-induced alterations in Bacillus cereus cell wall contribute to root colonization and plant growth promotion. PLoS ONE 8(10): 1-12. 21. Elsharkawy, M. M., Shivanna, M. B., Meera, M. S., and Hyakumachi, M. 2015. Mechanism of induced systemic resistance against anthracnose disease in cucumber by plant growth-promoting fungi. Acta Agriculturae Scandinavica, Section B. Soil Plant Sci. 65(4): 287-299. 22. Engebrecht, J., Brent, R., and Kaderbhai, M. A. 1991. Minipreps of Plasmid DNA. Curr. Protoc. Mol. Biol. 66: 1.6.1- 1.6.10. 23. Engebrecht, J., Brent, R., and Kaderbhai, M. A. 2001. Preparation genomic DNA from bacteria. Curr. Protoc. Mol. Biol. 66: 2.4.1. 24. Espinosa-Urgel, M., Salido, A., and Ramos J-L. 2000. Genetic analysis of functions involved in adhesion of Pseudomonas putida to seeds. J. Bacteriol. 182(9): 2363–2369. 25. Evans, K. J. 2010. Botrytis Management. Integrated Management of Botrytis Bunch Rot Fact Sheet. Tasmania Institute of Agriculture Science, University of Tasmania, Australia. 26. Fan, B., Carvalhais, L. C., Becker, A., Fedoseyenko, D., von Wirén, N., and Borriss, R. 2012. Transcriptomic profiling of Bacillus amyloliquefaciens FZB42 in response to maize root exudates. BMC Microbiol. 12: 116-129. 27. Hartmann, A., Rothballer, M., and Schmid, M. 2008. Lorenz Hiltner, a pioneer in rhizosphere microbial ecology and soil bacteriology research. Plant Soil 312(7): 7-14. 28. He, Q. F., Chen, J. X., and Li, C. F. 2011. An extracellular oligopeptide permease may be a potential virulence factor of Vibrio harveyi. J. Ocean Univ. 10(4): 343-350. 29. Hiltner, L. 1904. Über neuere Erfahrungen und Probleme auf dem Gebiete der Bodenbakteriologie unter besonderer Berücksichtigung der Gründüngung und Brache. Arb DLG 98: 59-78. 30. Hobbs, M. and Reeves, P. R. 1997. The JUMPstart sequence: a 39 bp element common to several polysaccharide gene clusters. Mol. Microbiol. 12(5): 855-856. 31. Huang, C. J., Liu, Y. H., Yang, K. H., and Chen, C. Y. 2012. Physiological response of Bacillus cereus C1L-induced systemic resistance in lily against Botrytis leaf blight. Eur. J. Plant Pathol. 134: 1-12. 32. Huang, C. J., Tsay, J. F., Chang, S. Y., Yang, H. P., Wu, W. S., and Chen, C. Y. 2013. Dimethyl disulfide is an induced systemic resistance elicitor produced by Bacillus cereus C1L. Pest Manag. Sci. 68(9): 1306-1310. 33. Jarvis, W.R. 1962. The dispersal of spores of Botrytis cinerea Fr. in a raspberry plantation. Trans. Br. Mycol. Soc. 45(4): 549-559. 34. Jumpertz, T., Chervaux, C., Racher, K., Zouhair, M., Blight, M. A., Holland, I. B., and Schmitt, L. 2010. Mutations affecting the extreme C terminus of Escherichia coli haemolysin A reduce haemolytic activity by altering the folding of the toxin. Microbiology 156(8): 2495-2505. 35. Kamilova, F., Kravchenko, L. V., Shaposhnikov, A. I., Azarova, T., Makarova, N., and Lugtenberg, B. 2006. Organic acids, sugars, and L-tryptophane in exudates of vegetables growing on stonewool and their effects on activities of rhizosphere bacteria. Mol. Plant-Microbe Interact.19(3): 250-256. 36. Kloepper, J. W. and Schroth, M. N. 1978. Plant growth-promoting rhizobacteria on radishes. In: Proceedings of the 4th international conference on plant pathogenic bacteria. Gilbert-Clarey. Tours. France. pp 879-882. 37. Laxalt, A. M., ter Riet, B., Verdonk, J. C., Parigi, L., Tameling, W. I., Vossen, J., Haring, M., Musgrave, A., and Munnik, T. 2001. Characterization of five tomato phospholipase D cDNAs: Rapid and specific expression of LePLDβ1 on elicitation with xylanase. Plant J. 26: 237-247. 38. Lee, J., Klessig, D. F., and Nürnberger, T. 2001. A harpin binding site in tobacco plasma membranes mediates activation of the pathogenesis-related gene HIN1 independent of extracellular calcium but dependent on mitogen-activated protein kinase activity. Plant Cell 13(5): 1079-1093. 39. Leeds, J. A. and Welch, R. A. 1997. Enhancing transcription through the Escherichia coli hemolysin operon, hlyCABD: RfaH and upstream JUMPStart DNA sequences function together via a postinitiation mechanism. J. Bacteriol. 179(11): 3519-3527. 40. Leroux, P. 2007. Chemical control of Botrytis and its resistance to chemical fungicides. pp. 195-222 in: Botrytis: Biology, Pathology and Control. Williamson, B., Tudzynski, P., Nafiz Delen Fillinger, S., and Elad, Y. Springer. France. 41. Li, X. Z. and Nikaido, H. 2016. Antimicrobial Drug Efflux Pumps in Escherichia coli. In: Li XZ., Elkins C., Zgurskaya H. (eds) Efflux-Mediated Antimicrobial Resistance in Bacteria. Adis, Cham. 42. Lippolis, R., Siciliano, R. A., Mazzeo, M. F., Abbrescia, A., Gnoni, A., Sardanelli A. M., and Papa, S. 2013. Comparative secretome analysis of four isogenic Bacillus clausii probiotic strains. Proteome Sci. 11(28): 1-14. 43. Liu, Y. H., Huang, C. J., and Chen, C. Y. 2008. Evidence of induced systemic resistance against Botrytis elliptica in lily. Phytopathology 98(7): 830-836. 44. Mark, G. L., Dow, J. M., Kiely, P. D., Higgins, H., Haynes, J., and Baysse, C. 2005. Transcriptome profiling of bacterial responses to root exudates identifies genes involved in microbe-plant interactions. Proc. Natl. Acad. Sci. USA. 102: 17454-17459. 45. Marquis, H., Goldfine, H., and Portnoy, D. A. 1997. Proteolytic pathways of activation and degradation of a bacterial phospholipase C during intracellular infection by Listeria monocytogenes. J. Cell Biol. 137(6): 1381-1392. 46. Marschner, H. 1995. Mineral nutrition of higher plant. Annals. of Botany. Volume 78(4): 527-528. 47. Newman, M. A., Sundelin, T., Nielsen, J. T., and Erbs, G. 2013. MAMP (microbe-associated molecular pattern) triggered immunity in plants. Front. Plant Sci. 4(139): 1-14. 48. Pieterse, C. M., Zamioudis, C., Berendsen, R. L., Weller, D. M., Van Wees, S. C., and Bakker, P. A. 2014. Induced systemic resistance by beneficial microbes. Annu. Rev. Phytopathol. 52: 347-75. 49. Pizzuti, F. and Daroda, L. 2008. Investigating recombinant protein exudation from roots of transgenic tobacco. Environ. Biosafety Res. 7(4): 219-226. 50. Profotová, B., Burketová, L., Novotná, Z., Martinec, J., and Valentová, O. 2006. Involvement of phospholipases C and D in early response to SAR and ISR inducers in Brassica napus plants. Plant Physiol. Biochem. 44(2-3): 143-151. 51. Qiu, M., Xu, Z., Li, X., Li, Q., Zhang, N., Shen, Q., and Zhang, R. 2014. Comparative proteomics analysis of Bacillus amyloliquefaciens SQR9 revealed the key proteins involved in in situ root colonization. J. Proteome Res. 13(12): 5581-5591. 52. Quentin, Y., Fichant, Q., and Denizot, F. 1999. Inventory, assembly and analysis of Bacillus subtilis ABC transport systems. J. Mol. Biol. 287(3): 467-484. 53. Ramarao, N. and Sanchis, V. 2013. The pore-forming haemolysins of Bacillus cereus: a review. Toxins 5(6): 1119-1139. 54. Rebecchi M. J. and Pentyal, S. N. 2000. Structure, function, and control of phosphoinositide-specific phospholipase C. Physiol. Rev. 80(4): 1292-1324. 55. Redmon. J. W., Batley, M., Djordjevic, M. A., Innes, R. W., Kuempel P. L., and Rolfe, B. G. 1986. Flavones induce expression of nodulation genes in Rhizobium. Nature 323: 632-635. 56. Roberts, H. M. 2007. Does GABA act as a signal in plants? Plant Signal. Behav. 2(5): 408-409. 57. Romanazzi, G., Smilanick, J. L., Feliziania, E., and Droby, S. 2016. Integrated management of postharvest gray mold on fruit crops. Postharvest Biol. Technol. 113(16): 69-76. 58. Rossier, O. and Ciancitto, N. P. 2005. The Legionella pneumophila tatB gene facilitates secretion of phospholipase C, growth under iron-limiting conditions, and intracellular infection. Infect. Immun. 73(4): 2020-2032. 59. Rudrappa, T., Czymmek, K. J., Paré, P. W., and Bais, H. P. 2008. Root-secreted malic acid recruits beneficial soil bacteria. Plant Physiol. 148: 1547-1556. 60. Sharma, A., Jansen, R., Nimtz, M., Johri, B. N., and Wray, V. 2007. Rhamnolipids from the rhizosphere bacterium Pseudomonas sp. GRP3 that reduces damping-off disease in Chilli and tomato nurseries. J. Nat. Prod. 70(6): 941-947. 61. Sivasakthi, S., Usharani, G., and Saranraj, P. 2014. Biocontrol potentiality of plant growth promoting bacteria (PGPR) - Pseudomonas fluorescent and Bacillus subtilis: A review. Afr. J. Agri. Res. 9(16): 1265-1277. 62. Shi, S., Richardson, A. E., O'Callaghan, M., DeAngelis, K. M., Jones, E. E., Stewart, A., Firestone, M. K., and Condron, L. M. 2011. Effects of selected root exudate components on soil bacterial communities. FEMS Microbiol. Ecol. 77(3): 600-610. 63. Song, F. and Goodman, R. M. 2002. Molecular cloning and characterization of a rice phosphoinositide-specific phospholipase C gene, OsPI-PLC1, that is activated in systemic acquired resistance. Physiol. Mol. Plant Pathol. 61(1): 31-40. 64. Sousa, C. S., Soares, A. C. F., and Garrido, M. S. 2008. Characterization of streptomycetes with potential to promote plant growth and biocontrol. Sci. Agric. 65(1): 50-55. 65. Streb, H., Irvine, R. F., Berridge, M. J., and Schulz, I. 1983. Release of Ca2+ from a nonmitochondrial intracellular store in pancreatic acinar cells by inositol-1,4,5-trisphosphate. Nature 306(5938): 67-69. 66. Taghavi, S., Lelie, D. van der, Hoffman, A., Zhang, Y-B., Walla, M. D., Vangronsveld, J., Newman, L., and Monchy, S. 2010. Genome Sequence of the plant growth promoting endophytic bacterium Enterobacter sp. 638. PLoS Genet. 6(5): 1-15. 67. Thomas, S., Holland, I. B., and Schmitt, L. 2014. The Type 1 secretion pathway- The hemolysin system and beyond. Biochim. Biophys. Acta 1843 1629–1641. 68. Torres, M. A., Jones, J. D. G., and Dangl, J. L. 2006. Reactive oxygen species signaling in response to pathogens. Plant Physiol. 141(2): 373-378. 69. Vicedo, B., Flors, V., Leyva, M. de la O., Finiti, I., Kravchuk, Z., Real, M. D., García-Agustín, P., and González-Bosch, C. 2009. Hexanoic acid-induced resistance against Botrytis cinerea in tomato plants. Mol. Plant Microbe Interact. 22(11): 1455-1465. 70. Wang, P., Yu, Z., Li, B., Cai, X., Zeng, Z., Chen, X., and Wang, X. 2015. Development of an efficient conjugation-based genetic manipulation system for Pseudoalteromonas. Microb. Cell Fact. 14(11): 1-11. 71. Wiesel, L., Newton, A. C., Elliott, I., Booty, D., Gilroy, E. M., Birch, P. R. J., and Hein, I. 2014. Molecular effects of resistance elicitors from biological origin and their potential for crop protection. Front. Plant Sci. 5(655): 1-13. 72. Williamson, B., Bettina, T., Tudzynski, P., and Van Kan, J. A. L. 2007. Botrytis cinerea: the cause of grey mould disease. Mol. Plant Pathol. 8(5): 561-580. 73. Yang, C. K., Ewis, H. E., Zhang, X. Z., Lu, C. D., Hu, H. J., Pan, Y., Abdelal, A. T., and Tai1 P. C. 2011. Nonclassical protein secretion by Bacillus subtilis in the stationary phase is not due to cell lysis. J. Bacteriol. 193(20): 5607-5615. 74. Yi, H-S., Ahn, Y-R., Song, G. C., Ghim, S-Y., Lee, S., Lee, G., and Ryu, C-M. 2016. Impact of a bacterial volatile 2,3-butanediol on Bacillus subtilis rhizosphere robustness. Front. Microbiol. 7(993): 1-11. 75. Zheng, J., Guan, Z., Cao, S., Peng, D., Ruan, L., Jiang, D., and Sun, M. 2015. Plasmids are vectors for redundant chromosomal genes in the Bacillus cereus group. BMC Genomics 16(1): 6-16. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68179 | - |
dc.description.abstract | 施用化學農藥是常見的高效植物病害管理策略之一,但過度施用化學農藥可能造成環境污染以及抗藥性病原菌的出現,因此,研發生物性農藥及其活性成份作為替代方案顯得特別重要。一類常用的生物性農藥來自植物促生根圈細菌,它擁有群聚於植物根圈的能力並有利於植物生長。部份植物促生根圈細菌亦可增強植物全體抵抗病原微生物的危害,此現象稱為誘導性系統抗病。前人研究顯示由臺灣花蓮太魯閣國家公園的臺灣百合根圈分離出之植物促生根圈細菌臘狀芽孢桿菌 (Bacillus cereus) 菌株 C1L, 被證實有降低百合灰黴病及南方玉米葉枯病罹病嚴重度的功效。本研究自臘狀芽孢桿菌C1L跳躍子突變庫,篩選誘導圓葉菸草對灰黴病菌 (Botrytis cinerea) 系統性抗病能力減弱之突變株,結果顯示突變株 M177經由定序與南方雜合分析確認跳躍子之單插入位置為巨環內酯通透酶基因 (macrolide permease),且經反轉錄聚合酶連鎖反應確定無法表現巨環內酯通透酶之訊息核酸。隨後藉由液相層析串連質譜儀分析臘狀芽孢桿菌野生株C1L與突變株M177的外泌液差異性累積物質,得知臘狀芽孢桿菌 C1L 與誘導系統性抗病能力相關之細菌產物可能有flagellin、hemolysin 與 phospholipase C等,而這些候選因子的外泌與巨環內酯通透酶的關係,以及其對菸草誘導抗病性的促進作用尚待進一步研究。綜合上述,本研究增進對臘狀芽孢桿菌 C1L 介導系統性抗病之瞭解,且提供進一步探視的相關線索。 | zh_TW |
dc.description.abstract | Applying chemical pesticides is one of the common and efficient plant disease management measures. However, overuse of chemical pesticides causes environmental pollution and emergence of antibiotic-resistant strains of pathogens. Therefore, developing active ingredients of biopesticides has become an important issue. Several kinds of the popular biopesticides are developed based on plant growth-promoting rhizobacteria (PGPR) which are capable of colonizing plant rhizosphere and beneficial for plant growth. Some PGPR also increase whole-plant resistance against biotic and abiotic stresses, a phenomenon of induced systemic resistance (ISR). A PGPR strain named Bacillus cereus C1L, isolated from the rhizosphere of Lilium formosanum in Taroko National Park, Hualien County, Taiwan, was proved able to reduce disease severity of lily gray mold and southern corn leaf blight after soil-drench application. The aim of this study was to identify the bacterial genes involved in the systemic disease resistance induced by B. cereus C1L. To find the related genes, a transposon insertion library of B. cereus C1L was used and screened by the phenotype of decreased ISR on Nicotiana benthamiana against fungal pathogen Botrytis cinerea. In the mutant strain M177 of B. cereua C1L, a single transposon insertion was located in the gene encoding macrolide permease as confirmed by sequencing and Southern blot hybridization. The failure to express macrolide permease gene was demonstrated by reverse transcription-polymerase chain reaction. Presumably, a lack of macrolide permease function in M177 would retard the secretion of products required for ISR elicitation. LC-MS/MS analysis of the compounds differentially present in the secretions of B. cereus C1L wild-type strain and mutant M177 showed that flagellin, hemolysin and phospholipase C might be involved in the ISR elicitation of B. cereus C1L in N. benthamiana. The implication of permease involving in the secretion of candidate factors for eliciting effect of candidate factors on systemic resistance needs to be further confirmed. In this study, the mechanisms of B. cereus C1L-mediated ISR are better understood and give some clues for further investigations. | en |
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dc.description.tableofcontents | 壹、中⽂摘要 ...................................... 5
貳、英⽂摘要 ........................................ 6 參、前⾔ ................................................ 8 肆、前⼈研究 ....................................... 10 ⼀、植物促⽣根圈細菌 (Plant growth-promoting rhizobacteria, PGPR) .................... 10 ⼆、植物防禦機制 ................................ 11 三、植物根分泌物 ......................................... 12 四、已知之 ISR 誘導因⼦ .......................... 13 五、溶⾎素 (Hemolysin) ........................... 14 六、磷脂酶 C (Phospholipase C) ..................... 15 七、灰黴病菌 (Botrytis cinerea) .................... 17 伍、材料⽅法 ..................................... 19 ⼀、供試菌株與植株 ................................ 19 1. 細菌菌株培養及保存 ........................... 19 2. 真菌菌株培養及保存 ........................... 19 3. 圓葉菸草 (Nicotiana benthamiana) 栽培....................... 19 ⼆、臘狀芽孢桿菌 C1L 突變基因分析 ............... 20 1. 臘狀芽孢桿菌 C1L 突變菌株篩選 ......................... 20 2. C1L 突變株基因體 DNA 抽取 ..................... 20 3. ⼤腸桿菌質體萃取 ............................... 21 4. ⼤腸桿菌勝任細胞 DH5α製備 ................. 21 5. ⼤腸桿菌勝任細胞熱休克轉形法 ................ 21 6. 南⽅雜合法分析 .................................. 22 7. 臘狀芽孢桿菌C1L 突變株基因定序 ..................... 23 三、突變株 M177 之突變基因及其上下游基因表現檢視................. 23 1. 細菌 RNA 萃取 ................................... 23 2. 反轉錄-聚合酶連鎖反應 ....................... 24 四、突變株 M177 特性分析 ....................... 24 1. 突變株 M177 ⽣⾧勢檢視 .................... 24 2. 突變株 M177 抗紅黴素測試 ................. 24 3. 突變株 M177 外泌液及純菌體誘導圓葉菸草抗灰黴病之能⼒測試........ 24 五、 突變株M177 外泌液蛋⽩質分析 ................. 25 1. 菸草根分泌物收集 ............................ 25 2. 細菌外泌液收集及蛋⽩質電泳分析 .............. 25 六、突變株 M177 誘導植物系統性抗病能⼒之檢視 .............. 25 1. 活性氧化物染⾊ ................................ 25 2. 癒傷葡聚糖染⾊ ................................ 26 3. 灰黴病菌核酸萃取與基因定量 ............................... 26 七、突變株 M177 在菸草根部群聚與內⽣情形檢視 ......... 27 陸、結 果 ................... 28 ⼀、臘狀芽孢桿菌C1L 突變株篩選結果 ............... 28 1. 依據菸草病徵篩選突變株 ....................... 28 2. Tn917ac1 周邊序列分析 ....................... 28 3. 跳躍⼦ Tn917ac1 插⼊數⽬確認 .................. 28 ⼆、突變株基因分析 ................................. 29 1. 突變株基因表現分析 ......................... 29 2. 啟動⼦及終⽌⼦預測分析 ................. 29 3. Tn917ac1 插⼊位置繪圖 .................... 30 三、突變株 M177 對接種灰黴病菌之菸草葉⽚防禦反應之影響 .. 30 1. 癒傷葡聚糖 (callose) 表現量⽐較 ................ 30 2. 活性氧物質誘發時間⽐較 ................................ 31 3. 灰黴病菌菌量差異 ............................ 31 四、M177 突變株之特性 ........................... 31 1. ⽣⾧趨勢⽐較 .................................. 31 2. 突變株 M177 對紅黴素的抗性較野⽣株 C1L 弱 ............................. 31 3. 突變株 M177 菸草根部群聚與內⽣菌量較野⽣株 C1L 低 ......... 32 五、突變株 M177 外泌液組成分析 ......................... 32 1. 突變株 M177 外泌液之誘導系統性抗病能⼒漸弱 .......... 32 2. 菸草根分泌物濃度差異對突變株 M177 之外泌液組成影響 ....... 32 3. 突變株M177 外泌液之溶⾎素及磷脂酶 C 含量減少 ...... 33 4. 臘狀芽孢桿菌 C1L 之溶⾎素與磷脂酶相關基因 ......... 33 柒、討 論 ................... 34 ㄧ、ISR 減弱之臘狀芽孢桿菌突變株之突變基因功能 ............ 34 ⼆、三磷酸腺苷結合傳送蛋⽩系統之通透酶突變株病徵差異探討 ..... 35 三、突變株M177 誘導植物抗性的表徵改變 .................... 36 1. 突變株M177 造成植物活性氧物質累積增加與癒傷葡聚糖沉積減少 .................. 36 2. 前處理突變株M177 之菸草感染灰黴病菌菌量增加 ............ 36 四、突變株 M177 環境⽣存機率與寄主附著的能⼒較低 ......... 37 1. 突變株 M177 抗紅黴素測試 .............. 37 2. 臘狀芽孢桿菌 C1L 及突變株 M177 根部群聚與內⽣情形 ................................... 37 五、臘狀芽孢桿菌 C1L 與突變株M177 外泌液組成⽐較 ................. 37 1. 突變株 M177 之溶⾎素參與植物 ISR 表現 ............. 37 2. 突變株 M177 之磷脂酶 C 參與植物 ISR 表現 ..................................................... 38 3. 臘狀芽孢桿菌 C1L 之溶⾎素相關基因與磷脂酶相關基因 ................................... 38 捌、參考⽂獻 ........................... 40 玖、圖表集 ............................... 49 表ㄧ、供試菌株 .................................. 50 表⼆、引⼦ .................... 51 表三、臘狀芽孢桿菌C1L 誘導植物系統性抗病⼒弱化突變株的對應病徵與跳躍⼦插⼊ 位置 ................. 52 表四、經 LC-MS/MS 鑑定之臘狀芽孢桿菌 C1L 與突變株M177 的差異性外泌候選蛋 ⽩ .................................. 53 表五、候選外泌蛋⽩之基因位置 .......................... 54 圖⼀、臘狀芽孢桿菌C1L 野⽣株與突變株之誘導植物系統性抗病能⼒試驗。 ......... 55 圖⼆、臘狀芽孢桿菌C1L 突變株基因組中Tn917ac1 插⼊數⽬確認。 ...................... 56 圖三、以反轉錄聚合酶連鎖反應檢測 Tn917ac1 對插⼊基因與鄰近基因表現之影響。 ..................................... 57 圖四、臘狀芽孢桿菌 C1L 之突變株染⾊體上 Tn917ac1 跳躍⼦插⼊位置圖。 ........... 58 圖五、臘狀芽孢桿菌C1L 野⽣株與M177 突變株於圓葉菸草上誘發抗灰黴病菌侵染之 癒傷葡聚糖沉積。 ................... 59 圖六、臘狀芽孢桿菌C1L 野⽣株與M177 突變株於圓葉菸草上所誘發的癒傷葡聚糖相對累積量。 ............................ 60 圖七、⽐較臘狀芽孢桿菌C1L 野⽣株與M177 突變株於圓葉菸草上誘發抗灰黴病菌侵染的活性氧化物累積情形。 ................. 61 圖⼋、以定量 PCR 偵測經臘狀芽孢桿菌C1L 野⽣株與M177 突變株處理之菸草葉⽚中灰黴病菌的相對累積量。 .................. 62 圖九、臘狀芽孢桿菌C1L 野⽣株與M177 突變株之⽣⾧曲線。 ................ 63 圖⼗、臘狀芽孢桿菌C1L 野⽣株與M177 突變株抗紅黴素能⼒測試。 .................... 64 圖⼗⼀、分析臘狀芽孢桿菌C1L 與M177 突變株於圓葉菸草根的群聚量。 ............. 65 圖⼗⼆、臘狀芽孢桿菌C1L 野⽣株與M177 突變株的上清液及菌體處理菸草對灰黴病菌之系統性抗病試驗。 ........................ 66 圖⼗三、⽐較經不同濃度之菸草根分泌物刺激後的臘狀芽孢桿菌C1L 野⽣株與 M177突變株之外泌性蛋⽩。 ......................... 67 圖⼗四、以 SDS-PAGE ⽐較分析菸草根分泌物預處理之臘狀芽孢桿菌野⽣株 C1L 與突變株 M177 的外泌性蛋⽩。 .................. 68 | |
dc.language.iso | zh-TW | |
dc.title | 誘導圓葉菸草系統性抗病之細菌通透酶相關外泌因子鑑定 | zh_TW |
dc.title | Identification of bacterial permease-related secreted factors involved in the induction of systemic disease resistance in Nicotiana benthamiana | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 沈偉強,廖秀娟,吳蕙芬,黃健瑞 | |
dc.subject.keyword | 植物促生根圈細菌,誘導系統性抗病,臘狀芽孢桿菌,圓葉菸草,巨環內脂通透?, | zh_TW |
dc.subject.keyword | Plant growth-promoting rhizobacteria,induced systemic resistance,Bacillus cereus,Nicotiana benthamiana,macrolide permease, | en |
dc.relation.page | 68 | |
dc.identifier.doi | 10.6342/NTU201704381 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-11-17 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 植物病理與微生物學研究所 | zh_TW |
顯示於系所單位: | 植物病理與微生物學系 |
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