Please use this identifier to cite or link to this item:
標題: 具有防治十字花科蔬菜根瘤病潛力的技術開發
Development of the Potential Techniques for Control of Cruciferous Clubroot Disease
作者: 鄭倩芸
Cheng, Chien-Yun
關鍵字: Plasmodiophora brassicae
Plasmodiophora brassicae
fast analysis technique
biological control
Bacillus mycoides
出版社: 植物病理學系所
引用: 行政院農業委員會。2010。99年農業統計年報。行政院農委會出版。115 頁。 陳俊欽。1996。利用拮抗細菌防治十字花科蔬菜根瘤病。國立中興大學植物病理研究所第二十六屆碩士論文。57頁。 郭魁士。1983。土壤學。中國書局。801頁。 黃一修。1988。台灣十字花科蔬菜根瘤病菌菌系之調查與土壤酸鹼值及鈣離子對病原菌之影響。國立中興大學植物病理研究所第十八屆碩士論文。53頁。 黃美茹。2010。甜瓜萎凋病的病原菌鑑定與其基本生物特性及生物防治菌篩選。國立中興大學植物病理研究所第四十屆碩士論文。51頁。 黃勝智。1986。育苗土壤添加鈣化物防治十字花科蔬菜根瘤病之研究。國立中興大學植物病理研究所第十六屆碩士論文。45頁。 楊慶鴻。1984。土壤添加物防治十字花科蔬菜根瘤病之研究。國立中興大學植物病理研究所第十四屆碩士論文。58頁。 澤田兼吉。1933。台灣產菌類調查報告第六篇。台灣總督府農業試驗所報告第61號。 謝文瑞。1992a。十字花科蔬菜根瘤病之防治。中華植物保護學會出版。病蟲害非農藥防治技術研討會專刊:293-299。 謝文瑞。1992b。十字花科蔬菜根瘤病菌之生態。中華植物保護學會出版。作物絕對寄生真菌性病害研討會刊:145-156。 謝文瑞。1993。十字花科蔬菜根瘤病。中華植物保護學會出版。蔬菜保護研討會專刊 1:257- 272。 Aist, J. R. and Williams, P. H. 1971. The cytology kinetics of cabbage root hair penetration by Plasmodiophora brasicae. Can. J. Bot. 49: 2023-2024. Alexopoulos, C. J. 1952. Introductory Mycology (3th ed.). New York. John Wiley & Sons., Inc. 482pp. Anon. 1987. Insect pests of economic significance affecting major crops of countries in Asia and the Pacific region. APPPC Technical Document No. 135. Bangkok and Thailand. Regional FAO office for Asia and the Pacific (RAPA). Anon. 1996. Plant Quarantine Report (PQR) Database. Paris and France. European Plant Protection Organisation (EPPO). Appel, O. and Werth, E. 1910. Infektionsversuche mit Plasmodiophora brassicae Woronin. Mitteilungen aus der Kaiserlichen Bioloischen Anstalt fiir Land-und Forstwirtschaft. 10: 17. Arie T., Kobayashi Y., Okada G., Kono Y. and Yamaguchi, I. 1998. Control of soilborne clubroot disease of cruciferous plants by epoxydon from Phoma glomerata. Plant Pathol. 47: 743-748. Arie, T., Namba, S., Yamashita, S. and Doi, Y. 1988. Detection of resting spores of Plasmodiophora brassicae Woron. from soil and root by fluorescent antibody technique. Ann. Phytopathol. Soc. Jpn. 54: 242-245. Asano, T., Kodama, A. and Kageyama, K. 2006. Susceptibility of hairy root lines of Brassica species to Plasmodiophora brassicae and in an in vitro subculture system. J. Gen. Plant Pathol. 72: 85-91. Baker, K. F. and Cook, R. J. 1974. Biological Control of Plant Pathogens. W. H. Freeman Co., San Francisco, California. 433pp. Buczacki, S. T., Toxopeus, H., Mattusch, P., Johnston, T. D., Dixon, G. D. and Hobolth, L. A. 1975. Study of physiologic specialization in Plasmodiophora brassicae: proposals for rationalization through an international approach. Trans. Br. Mycol. Soc. 65: 295-303. Conway, W. S. and Sams, C. E. 1984. Possible mechanisms by which postharvest calcium treatment reduces decay in apples. Phytopathology 74: 208-210. Cook, W. R. I. and Swartz, E. J. 1930. The life history, cytology and method of infection of Plasmodiophora brassicae Woron., the cause of finger and toe disease of cabbage and other crucifers. Phil. Trans. R. Soc. Lond. B. 218: 283-314. Crête, R. 1981. Worldwide importance of clubroot. Clubroot Newslett. 11: 6-7. Dixon, G. R. 2009. The Occurrence and economic impact of Plasmodiophora brassicae and clubroot disease. J. Plant Growth Regul. 28: 194-202. Dixon, G. R. and Robinson, D. L. 1986. The susceptibility of Brassica oleracea cultivars to Plasmodiophora brassicae (clubroot). Plant Pathol. 35: 101-107. Dobson, R. L., Gabrielson, R. L., Baker and A. S. 1982. Soil water matric potential requirements for root-hair and cortical infection of Chinese cabbage by Plasmodiophora brassicae. Phytopathology 72: 1598-1600. Dobson, R. L., Gabrielson, R. L., Baker, A. S. and Bennett, L. 1983. Effects of lime particle size and distribution and fertilizer formulation on clubroot disease caused by Plasmodiophora brassicae. Plant Dis. 67: 50-52. Donald, F. M. and Campbell, R. N. 1985. Lime and the control of clubroot of crucifers: Effects of pH, calcium, magnesium, and their interactions. Phytopathology 75: 670-673. Faggian, R., Bulman, S. R., Lawrie, A. C. and Porter, I. J. 1999. Specific polymerase chain reaction primers for the detection of Plasmodiophora brassicae in soil and water. Phytopathology 89: 392-397. Fletcher, J. T., Hims, M. J., Archer, F. C. and Brown, A. 1982. Effects of adding calcium and sodium salts to field soils on the incidence of clubroot. Ann. Appl. Biol. : 245-251 Gleisberg, W. 1923. Plasmodiophora brassicae Woron: Zur Auswertung von Kruciferen- Infektionsreihen. Nachrichtenblatt für den Deutschen Pflanzenschutzdienst. 3: 10-12. Honig, F. 1931. Der Kohklorpferreger. (Plasmoidophora brassicae Wor.) Cartenbauwissen-schaft. 5: 116-225. Höstermann, G. 1922. Versuche zur Bekämpfung der Kohlhernie (Plasmoidophora brassicae). Landwirtschaftliche Jahrbücher, Ergänzungsband. 1: 100-103. Hsieh, W. H. and Yang, C. H. 1985. Investigations on the clubroot and susceptibility of crucifers in Taiwan. Plant Prot. Bull. 27: 3-10. Hsieh, W. H., Yang, C. H. and Huang, S. C. 1987. Effects of soil amendments on root hair infection, zoosporangium formation and the incidence of clubroot disease caused by Plasmodiophora brassicae. Plant Prot. Bull. 29: 117-122. Huang, S. C., and Hsieh, W. H. 1987. Studies on nursery soil amended with calcium compounds for controlling clubroot of Chineses cabbage. Plant Prot. Bull. 29: 25-32. Hugh, R. and Leifson, E. 1953. The taxonomic significance of fermentative versus oxidative metabolism of carbohydrates by various Gram negative bacteria. J. Bacteriol. 66: 24-26. Ingram, D. S. and Tommerup, I. C. 1972. The life history of Plasmodiophora brassicae Woron. Proc. R. Soc. Lond. B. 180: 103-112. Jäschke D., Dugassa-Gobena D., Karlovsky P., Vidal S., and Ludwig-Müller J. 2010. Suppression of clubroot (Plasmodiophora brassicae) development in Arabidopsis thaliana by the endophytic fungus Acremonium alternatum. Plant Pathol. 59: 100-111. Jones, D. R., and Ingram, D. S. 1982. Factors affecting test for differential pathogenicity in populations of Plasmodiophora brassicae. Plant Pathol. 31: 229-238. Kageyama, K., Komatsu, T., and Suga, H. 2003. Refined PCR protocol for detection of plant pathogens in soil. J. Gen. Plant Pathol. 69(3): 153-160. Karling, J. 1968. The Plasmodiophorales 2nd ed. Hafner Publishing Company. London and New York. 256 pp. Lange, L., Heide, M., Hobolth, L., and Olsen, L. W. 1989. Serological detection of Plasmodiophora brassicae by dot immunobinding and visualization of the serological reaction by scanning electron microscopy. Phytopathology 79: 1066-1071. Macfarlane, I. 1952. Factors affecting the survival of Plasmodiophora brassicae Wor. in the soil and its assessment by a host test. Ann. Appl. Biol. 39: 239-256. Macfarlane, I. 1955. Variation in Plasmodiophora brassicae. Ann. Appl. Biol. 43: 297-306. Melville, I. E., and Hawken, R. W. 1967. Soil testing for club root in Devon and Cornwall. Plant Pathol. 16: 145-147. Mitani, S., Sugimoto, K., Hayashi, H., Takii, Y., Ohshima, T., and Matsuo, N. 2003. Effects of cyazofamid against Plasmodiophora brassicae Woronin on Chinese cabbage. Pest. Manag. Sci. 59: 287-293. Morre, D. J., and Bracker, C. E. 1976. Ultrastructural alteration of plant plasma membrane induced by auxin and calcium ions. Plant Physiol. 58: 544-547. Nakamura, L. K., and Jackson, M. A. 1995. Clarification of Taxonomy of Bacillus mycoides. Int. J. Syst. Bacteriol. Vol. 45, 1: 46-49. Narisawa, K., Tokumasu, S. and Hashiba, T., 1998. Suppression of clubroot formation in Chinese cabbage by the root endophytic fungus, Heteroconium chaetospira. Plant Pathol. 47: 206-210. Orihara, S. and Yamamoto, T. 1997. Specific detection of resting spores of Plasmodiophora brassicae in soil and plant tissue by enzyme immunoassay. Page 220 in: Programme and Summaries. Biennial Conf., 11th . Australasian Plant Pathology Society, Perth. Parry, J. M., Turnbull, P. C. B. and Gibson, J. R. 1983. A Color Atlas of Bacillus Species. Wolfe Medical Publications Ltd, London. 272 pp. Samuel, G., and Garrett, S. D. 1945. The infected root hair count for estimating the activity of Plasmodiophora brassicae Woron. in the soil. Ann. Appl. Biol. 32: 96-101. Sasser, M. 1990. Identification of bacteria by gas chromatography of celluar fatty acids. Technical Note 101. Microbial ID, Int., Newark, Del. Schaad, N. W., Jones, J. B. and Chun, W. 2001. Laboratory Guide for Identification of Plant Pathogenic Bacteria (3rd ed.). The American Phytopathological Society. 373pp. Scheijgrond, W. and Vos, H. 1954. Investigation on the susceptibility to clubroot. Euphytica 3: 125-139. Schofield,R.K. and Taylor, A.W. 1955. The measurement of soil pH. Soil Sci. Soc. Amer. Proc. 19: 164-167. Simmons, J. S. 1926. A culture medium for differentiating organisms of typhoid-colon aeorgenes groups and for isolation of certain fungi. J. Inf. Dis. 39:209-214. Sneath, P. H. A. 1986. Endospore-forming gram-positive rods and cocci. Pages 1104-1139 in: Bergey’s Manual of Systematic Bacteriology. Vol. 2. Sneath, P. H. A. (2nd ed.). Williams and Wilkins, Baltimore. 2898 pp. Sundelin, T. 2008. Identification of Expressed Plasmodiophora brassicae Genes During Plant Infection and Molecular Diagnosis of Plant Pathogens. Ph. D. thesis, University of Copenhagen, Copenhagen. Suwabe, K., Tsukazaki, H., Hatakeyama., Fujimura, M., Nunome, T., Fukuoka, H., Matsumoto, S. and Hirai, M. 2003. Identification of two loci for resistance to clubroot (Plasmodiophora brassicae Woronin) in Brassica rapa L. Theor. Appl. Genet. 107: 997-1002. Takahashi, K. 1991. Correlation between fluorescent staining reaction and germination in resting spores of Plasmodiophora brassicae. Ann. Phytopathol. Soc. Jpn. 57: 160-164. Takahashi, K. and Yamaguchi, T. 1989. Assessment of pathogenicity of resting spores of Plasmodiophora brassicae in soil by fluorescence microscopy. Ann. Phytopathol. Soc. Jpn. 55: 621-628. Tanaka S., Kochi S., Kunita H., Ito S. and Kameya-Iwaki M. 1999. Biological mode of action of fungicide, flusulfamide, against Plasmodiophora brassicae (clubroot). Eur. J Plant Pathol. 105: 577-584. Tanaka, S., Sakamoto, Y., Kajima, K., Fujieda, K. Katumoto, K. and Nishi, Y. 1991. Pathogenicity of three isolates of clubroot fungus attacking clubroot-resistant cultivars of Chinese cabbage. Bull. Fac. Agric. 39: 113-122. Wakeham, A. J. and White, J. G. 1996. Serological detection in soil of Plasmodiophora brassicae resting spores. Physiol. Mol. Plant Pathol. 48: 289-303. Wallenhammar, A. C. and Arwidsson, O. 2002. Detection of Plasmodiophora brassicae by PCR in naturally infested soils. Eur. J. Plant Pathol. 107: 313-321. Wallenhammar, A. C., Johnsson, L. and Gerhardson, B. 2000. Agronomic performance of partly clubroot-resistant spring oilseed turnip rape lines. J. Phytopathol. 148: 495-499. Williams, P. H. and Mcnabola, S. S. 1967. Fine structure of Plasmodiophora brassicae in sporogensis. Can. J. Bot. 49: 41-47. Williams, P. H., and Mcnabola, S. S. 1970. Fine stracture of the host-parasite interface of Plasmodiophora brassicae in cabbage. Phytopathology 60: 1557-166.
摘要: 西元1933年,台灣新竹與桃園一帶發現十字花科蔬菜發生葉片失水萎凋與根部畸形腫大之病徵,研究證實係由Plasmodiophora brassicae Woronin引起之根瘤病。近年來,十字花科根瘤病廣泛發生於台灣新竹、桃園、台中及宜蘭等地區,造成十字花科蔬菜的收益鉅減。田間P. brassicae從感染甘藍植株直到出現顯著病徵約需二至三個月,若要調查其各種防治成效卻頗為耗時,因此本文的主要目的即在建立一套快速且能精準評估十字花科蔬菜根瘤病危害的分析技術與標準流程,並利用此技術篩選拮抗微生物與土壤添加物之防病潛力。首先利用甘藍種子播種於病土中8天後進行根部染色,並調查根毛受感染之百分率,結果發現在24 oC接種病原菌濃度108 spores/g dry cultural medium時,主根在2-4公分有最高的感染率;將此區間分成4等分所計算出之數據,證實其與主根2-4公分間所有根毛的感染率相近。利用主根2-4公分間所調查的結果,進行統計分析發現接種濃度與感染率 (Y = 1.4773 + 12.3707 X – 0.4161 X2 , r2 = 0.98)、罹病度 (Y = 2.9764 + 2.3577 X + 1.3008 X2 , r2 = 0.97) 及植株鮮重 (Y = 8.2314 – 0.6148 X , r2 = 0.91 ; Y = 2.2850 – 0.1840 X , r2 = 0.95) 間均具有高度的相關性;至於感染率與罹病度 (Y = 3.0320 – 0.1336 X + 0.0193 X2 , r2 = 0.94) 間亦具有頗高的相關性。進一步採取此評估方法篩選有效防治十字花科根瘤病的微生物和土壤添加物,結果發現拮抗微生物Wt15及Wt16兩菌株可有效降低64.0 %和56.4 %的根毛感染率,並可使罹病度分別減少44.4 %與40.8 %,兩菌株經由生理生化及分子鑑定等結果證明均為蕈狀芽孢桿菌Bacillus mycoides。將兩拮抗菌株以種子粉衣、混拌入介質及澆灌等三種不同施用方式處理甘藍幼苗,結果以B. mycoides Wt15菌株粉衣種子之處理效果最為顯著,不僅較接種根瘤病菌之對照組延遲兩週產生病徵,罹病度亦由100 %降至48.15 %,且分別可提高甘藍地上部與地下部植體的鮮重達5.4與2.3倍;進一步分析整體根系結瘤比例後,結果顯示B. mycoides Wt15菌株粉衣種子之處理組植株的結瘤比為22.57 %,而對照組則為69.53 %。利用B. mycoides Wt15菌株處理本病原菌的休眠孢子36小時後,結果顯示孢子發生縊縮變形的現象,孢子處理48小時後活性由66.3 %降至44.7 %。至於土壤添加物方面,中興一百衍生物雖亦可有效降低53 %的根毛感染率,惟其防治根瘤病的效果與拮抗菌B. mycoides Wt15菌株的處理間並無顯著的協力加成效果。
The clubroot disease of cruciferous crops caused by Plasmodiophora brassicae, is one of the most important soilborne diseases. Infected plants typically exhibit abnormal growth, foliar wilting and swollen roots. In Taiwan, the disease was seriously found in Taoyuan, Hsinshu, Taichung and Ilan, causing huge ecomonic losses in agricultural production. For surveying the clubroot disease severity of cabbage plants in the field, it takes long time and heavy work. Therefore, the main purpose of this study is to establish a fast analysis technique for developing and evaluating the control measures of cruciferous clubroot disease in early stage of the crop. In this study, we found the highest infection occurred within 2-4 cm of the main root at the 8th day of inoculation under 24℃ . Besides, examining the infection of 40 root hairs within 2-4 cm region could stand for the whole infection. On the other hand, we also found that relationships among inoculum density, infection, fresh wieght, root wieght and disease severity were highly correlative. By using the fast analysis technique, we screened the antagonistic bacteria and soil amendments for the disease control. The results showed that Bacillus mycoides Wt15 delayed the disease development over two weeks which could reduce 52 % and 47 % of disease severity and clubbed root ratio. Moreover, B. mycoides Wt15 could also increase respectively 5.4 and 2.3 fold of fresh weight and root weight by seed coating method. The observation under the fluorescence microscopy indicated that B. mycoides Wt15 could reduce 21 % viability of resting spores compared with the control and result in shrinking shape of germinated resting spores. The CH100 derivative (250X) was able to reduce 53 % of root hairs infection, however, it did not show the synergistic effect with B. mycoides Wt15 on reducing disease severity.
其他識別: U0005-1408201204461400
Appears in Collections:植物病理學系



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