請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/95780
標題: 小麥赤黴病生物防治策略之研發
Development of Biocontrol Strategies for Fusarium Head Blight of Wheat
作者: 楊翰祁
Han-Chi Yang
關鍵字: 小麥赤黴病
Fusarium head blight
biological control
response surface methodology
trichothecene biosynthesis
引用: 沈勳。2011。淺談小麥與在麵粉的應用。台中區農業改良場特刊105:240。 郭建志、林訓仕、廖君達、黃冬青。2013。小麥赤黴病之病原鑑定及防治藥劑篩選。台中區農業改良場101年度科技計畫研究成果發表會論文輯。169-175。趙佳鴻主編。台中:農委會台中區農業改良場。 Abadias, M., Benabarre, A., Teixido, N., Usall, J., Vinas, I. 2001. Effect of freeze drying and protectants on viability of the biocontrol yeast Candida sake. International Journal of Food Microbiology 65, 173-182. Ahlem, H., Mohammed, E., Badoc, A., Ahmed, L. 2012. Effect of pH, temperature and water activity on the inhibition of Botrytis cinerea by Bacillus amyloliquefaciens isolates. African Journal of Biotechnology 11, 2210-2217. Alina, S. O., Constantinscu, F., Petruţa, C. C. 2015. Biodiversity of Bacillus subtilis group and beneficial traits of Bacillus species useful in plant protection. Romanian Biotechnological Letters 20, 10737. Amarasinghe, C., Wang, J. H., Liao, Y. C., Fernando, W. G. D. 2011. Difference in Tri13 gene sequences between the 3-acetyldeoxynivalenol producing Fusarium graminearum Chemotypes from Canada and China. International Journal of Molecular Sciences 12, 6164-6175. Bai, G., Shaner, G. 2004. Management and resistance in wheat and barley to Fusarium head blight. Annual Review of Phytopathology 42, 135-161. Brady, C., Cleenwerck, I., Venter, S., Vancanneyt, M., Swings, J., Coutinho, T. 2008. Phylogeny and identification of Pantoea species associated with plants, humans and the natural environment based on multilocus sequence analysis (MLSA). Systematic and Applied Microbiology 31, 447-460. Brady, C. L., Venter, S. N., Cleenwerck, I., Engelbeen, K., Vancanneyt, M., Swings, J., Coutinho, T. A. 2009. Pantoea vagans sp. nov., Pantoea eucalypti sp. nov., Pantoea deleyi sp. nov. and Pantoea anthophila sp. nov. International Journal of Systematic and Evolutionary Microbiology 59, 2339-2345. Campestre, M. P., Castagno, L. N., Estrella, M. J., Ruiz, O. A. 2016. Lotus japonicus plants of the Gifu B-129 ecotype subjected to alkaline stress improve their Fe2+ bio-availability through inoculation with Pantoea eucalypti M91. Journal of Plant Physiology 192, 47-55. Cao, F., Shen, D., Li, J., Guan, D., Jiang, X., Li, L., Feng, R., Yang, X., Chen, H., Ge, Y. 2008. Multiplex-PCR approach to identify Bacillus species applied in microbial fertilizers. Acta microbiologica Sinica 48, 651-656. Castagno, L. N., Garcia, I. V., Sannazzaro, A. I., Bailleres, M., Ruiz, O. A., Mendoza, R. E., Estrella, M. J. 2014. Growth, nutrient uptake and symbiosis with rhizobia and arbuscular mycorrhizal fungi in Lotus tenuis plants fertilized with different phosphate sources and inoculated with the phosphate-solubilizing bacterium Pantoea eucalypti M91. Plant and Soil 385, 357-371. Chen Y. C., Su W. T., Liao C. D., Lin H. Y., Chiueh L. C. and Shih Y. C. 2012. A survey of deoxynivalenol and zearalenone in cereal products. Annual Report of Food and Drug Research 3, 159-164. Chen, Z. M., Li, Q., Liu, H. M., Yu, N., Xie, T. J., Yang, M. Y., Shen, P., Chen, X. D. 2010. Greater enhancement of Bacillus subtilis spore yields in submerged cultures by optimization of medium composition through statistical experimental designs. Applied Microbiology and Biotechnology 85, 1353-1360. Choi, D., Park, S. S., Ahn, B. K., Lim, D. H., Lee, Y. W., Moon, J. H., Shin, D. Y. 2008. Studies on production of gentamicin from Micromonosporas purpurea using crude vegetable oils. Process Biochemistry 43, 835-841. Chowdhury, S. P., Dietel, K., Rändler, M., Schmid, M., Junge, H., Borriss, R., Hartmann, A., Grosch, R. 2013. Effects of Bacillus amyloliquefaciens FZB42 on lettuce growth and health under pathogen pressure and its impact on the rhizosphere bacterial community. PLoS ONE, e68818. Compant, S., Duffy, B., Nowak, J., Clement, C., Barka, E. A. 2005. Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Applied and Environmental Microbiology 71, 4951-4959. Costa, E., Teixido, N., Usall, J., Atares, E., Vinas, I. 2001. Production of the biocontrol agent Pantoea agglomerans strain CPA-2 using commercial products and by-products. Applied Microbiology and Biotechnology 56, 367-371. Costa, E., Teixido, N., Usall, J., Atares, E., Vinas, I. 2002. The effect of nitrogen and carbon sources on growth of the biocontrol agent Pantoea agglomerans strain CPA-2. Letters in Applied Microbiology 35, 117-120. de Almeida, F. C. G., e Silva, T. A. D. L., Garrard, I., Asfora, L., Sarubbo, L. A., de Campos-Takaki, G. M., Tambourgi, E. B. 2015. Optimization and evaluation of biosurfactant produced by Pantoea sp. using pineapple peel residue, vegetable fat and corn steep liquor. Journal of Chemistry and Chemical Engineering 9, 269-279. Esitken, A., Yildiz, H. E., Ercisli, S., Donmez, M. F., Turan, M., Gunes, A. 2010. Effects of plant growth promoting bacteria (PGPB) on yield, growth and nutrient contents of organically grown strawberry. Scientia Horticulturae 124, 62-66. Gardiner, D. M., Kazan, K., Manners, J. M. 2009a. Nutrient profiling reveals potent inducers of trichothecene biosynthesis in Fusarium graminearum. Fungal Genetics Biology 46, 604-613. Gardiner, D. M., Osborne, S., Kazan, K., Manners, J. M. 2009b. Low pH regulates the production of deoxynivalenol by Fusarium graminearum. Microbiology 155, 3149-3156. Gillis, A., Guo, S. X., Bolotin, A., Makart, L., Sorokin, A., Mahillon, J. 2017. Detection of the cryptic prophage-like molecule pBtic235 in Bacillus thuringiensis subsp. israelensis. Research in Microbiology 168, 319-330. Hammond, T. M., Xiao, H., Rehard, D. G., Boone, E. C., Perdue, T. D., Pukkila, P. J., Shiu, P. K. T. 2011. Fluorescent and bimolecular-fluorescent protein tagging of genes at their native loci in Neurospora crassa using specialized double-joint PCR plasmids. Fungal Genetics and Biology 48, 866-873. He, J., Boland, G. J., Zhou, T. 2009. Concurrent selection for microbial suppression of Fusarium graminearum, Fusarium head blight and deoxynivalenol in wheat. Journal of Applied Microbiology 106, 1805-1817. Herrera, S. D., Grossi, C., Zawoznik, M., Groppa, M. D. 2016. Wheat seeds harbour bacterial endophytes with potential as plant growth promoters and biocontrol agents of Fusarium graminearum. Microbiological Research 186, 37-43. Holtmann, G., Bremer, E. 2004. Thermoprotection of Bacillus subtilis by exogenously provided glycine betaine and structurally related compatible solutes: Involvement of Opu transporters. Journal of Bacteriology 186, 1683-1693. Hu, W., Gao, Q., Hamada, M. S., Dawood, D. H., Zheng, J., Chen, Y., Ma, Z. 2014. Potential of Pseudomonas chlororaphis subsp. aurantiaca strain Pcho10 as a biocontrol agent against Fusarium graminearum. Phytopathology 104, 1289-1297. Huang, C. W. 2008. Potato common scab caused by Streptomyces scabies in Taiwan-biological characteristics of the pathogen and an atempted biocontrol by antagonistic Bacillus subtilis var. amyloliquefaciens WG6-14. Taichung, Taiwan: National Chung Hsing Unisversity. Master thesis. Ippolito, A., Nigro, F. 2000. Impact of preharvest application of biological control agents on postharvest diseases of fresh fruits and vegetables. Crop Protection 19, 715-723. Jung, B., Park, S. Y., Lee, Y. W., Lee, J. 2013. Biological efficacy of Streptomyces sp. strain BN1 against the cereal head blight pathogen Fusarium graminearum. Plant Pathology Journal 29, 52-58. Khan, N. I., Schisler, D. A., Boehm, M. J., Slininger, P. J., Bothast, R. J. 2001. Selection and evaluation of microorganisms for biocontrol of Fusarium head blight of wheat incited by Gibberella zeae. Plant Disease 85, 1253-1258. Kimura, M., Tokai, T., Takahashi-Ando, N., Ohsato, S., Fujimura, M. 2007. Molecular and genetic studies of Fusarium trichothecene biosynthesis: pathways, genes, and evolution. Bioscience, Biotechnology and Biochemistry 71, 2105-2123. Kluiver, J., Gibcus, J. H., Hettinga, C., Adema, A., Richter, M. K. S., Halsema, N., Slezak P. I., Ding Y., Kroesen B. J., Berg A. 2012. Rapid generation of microRNA sponges for microRNA inhibition. PLoS ONE, e29275. Kucherlapati, R. S., Eves, E. M., Song, K.-Y., Morse, B. S., Smithies, O. 1984. Homologous recombination between plasmids in mammalian cells can be enhanced by treatment of input DNA. Proceedings of the National Academy of Sciences 81, 3153-3157. Kulik, T., Łojko M., Jestoi M., Perkowski J. 2012. Sublethal concentrations of azoles induce tri transcript levels and trichothecene production in Fusarium graminearum. FEMS Microbiol Letter 335, 58-67. Landschoot, S., Audenaert, K., Waegeman, W., De Baets, B., Haesaert, G. 2013. Influence of maize–wheat rotation systems on Fusarium head blight infection and deoxynivalenol content in wheat under low versus high disease pressure. Crop Protection 52, 14-21. Landschoot, S., Audenaert, K., Waegeman, W., Pycke, B., Bekaert, B., De Baets, B., Haesaert, G. 2011. Connection between primary Fusarium inoculum on gramineous weeds, crop residues and soil samples and the final population on wheat ears in Flanders, Belgium. Crop Protection 30, 1297-1305. Lee, T., Han, Y. K., Kim, K. H., Yun, S. H., Lee, Y. W. 2002. Tri13 and Tri7 determine deoxynivalenol- and nivalenol- producing chemotypes of Gibberella zeae. Applied and Environmental Microbiology 68, 2148-2154. Lyznik, L. A., Mcgee, J. D., Tung, P.-Y., Bennetzen, J. L., Hodges, T. K. 1991. Homologous recombination between plasmid DNA molecules in maize protoplasts. Molecular and General Genetics 230, 209-218. Mishra, P., Kshirsagar, P., Nilegaonkar, S., Singh, S. 2012. Statistical optimization of medium components for production of extracellular chitinase by Basidiobolus ranarum: a novel biocontrol agent against plant pathogenic fungi. Journal of Basic Microbiology 52, 539-548. O''Donnell, K., Kistler, H. C., Tacke, B. K., Casper, H. H. 2000. Gene genealogies reveal global phylogeographic structure and reproductive isolation among lineages of Fusarium graminearum, the fungus causing wheat scab. Proceedings of the National Academy of Science 97, 7905-7910. O''Donnell, K., Ward, T. J., Aberra, D., Kistler, H. C., Aoki, T., Orwig, N., Kimura, M., Bjornstad, S., Klemsdal, S. S. 2008. Multilocus genotyping and molecular phylogenetics resolve a novel head blight pathogen within the Fusarium graminearum species complex from Ethiopia. Fungal Genetics and Biology 45, 1514-1522. Ohta, N., Park, Y. S., Yahiro, K., Okabe, M. 1995. Comparison of neomycin production from streptomyces fradiae cultivation using soybean oil as the sole carbon source in an airlift bioreactor and a stirred-tank reactor. Journal of Fermentation and Bioengineering 79, 443-448. Palazzini, J. M., Alberione, E., Torres, A., Donat, C., Kohl, J., Chulze, S. 2016. Biological control of Fusarium graminearum sensu stricto, causal agent of Fusarium head blight of wheat, using formulated antagonists under field conditions in Argentina. Biological Control 94, 56-61. Palazzini, J. M., Ramirez, M. L., Torres, A. M., Chulze, S. N. 2007. Potential biocontrol agents for Fusarium head blight and deoxynivalenol production in wheat. Crop Protection 26, 1702-1710. Pastor, J. M., Salvador, M., Argandoña, M., Bernal, V., Reina-Bueno, M., Csonka, L. N., Iborra, J. L., Vargas, C., Nieto, J. J., Cánovas, M. 2010. Ectoines in cell stress protection: uses and biotechnological production. Biotechnology Advances 28, 782-801. Pestka, J. J. 2007. Deoxynivalenol: toxicity, mechanisms and animal health risks. Animal Feed Science and Technology 137, 283-298. Pirgozliev, S. R., Edwards, S. G., Hare, M. C., Jenkinson, P. 2003. Strategies for the control of Fusarium head blight in cereals. European Journal of Plant Pathology 109, 731-742. Proctor, R. H., Hohn, T. M., Mccormick, S. P. 1995. Reduced virulence of Gibberella zeae caused by disruption of a trichothecene toxin biosynthetic gene. Molecular Plant-Microbe Interactions 8, 593-601. Raaijmakers, J. M., De Bruijn, I., Nybroe, O. Ongena, M., 2010. Natural functions of lipopeptides from Bacillus and Pseudomonas: more than surfactants and antibiotics. FEMS Microbiology Reviews 34, 1037-1062. Reva, O. N., Dixelius, C., Meijer, J., Priest, F. G. 2004. Taxonomic characterization and plant colonizing abilities of some bacteria related to Bacillus amyloliquefaciens and Bacillus subtilis. FEMS Microbiology Ecology 48, 249-259. Rong, C. B., Ma, Y. W., Wang, S. X., Liu, Y., Chen, S. F., Huang, B., Wang, J., Xu, F. 2016. Pantoea hericii sp. nov., isolated from the fruiting bodies of Hericium erinaceus. Current Microbiology 72, 738-743. Sarver, B. A., Ward, T. J., Gale, L. R., Broz, K., Kistler, H. C., Aoki, T., Nicholson, P., Carter, J., O’Donnell, K. 2011. Novel Fusarium head blight pathogens from Nepal and Louisiana revealed by multilocus genealogical concordance. Fungal Genetics and Biology 48, 1096-1107. Schisler, D. A., Slininger, P., Behle, R., Jackson, M. 2004. Formulation of Bacillus spp. for biological control of plant diseases. Phytopathology 94, 1267-1271. Schisler, D. A., Khan, N. I., Boehm, M. J., Lipps, P. E., Slininger, P. J., Zhang, S. 2006. Selection and evaluation of the potential of choline-metabolizing microbial strains to reduce Fusarium head blight. Biological Control 39, 497-506. Singh, R. P., Singh, P. K., Rutkoski, J., Hodson, D. P., He, X., J?rgensen, L. N., Hovm?ller, M. S., Huerta-Espino, J. 2016. Disease impact on wheat yield potential and prospects of genetic control. Annual Review of Phytopathology 54, 303-322. Stockwell, V. O., Johnson, K. B., Sugar, D., Loper, J. E. 2010. Control of fire blight by Pseudomonas fluorescens A506 and Pantoea vagans C9-1applied as single strains and mixed inocula. Phytopathology 100, 1330-1339. Tamura, K., Stecher, G., Peterson, D., Filipski, A., Kumar, S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30, 2725-2729. Teixidó, N., Cañamás, T., Usall, J., Torres, R., Magan, N., Viñas, I. 2005. Accumulation of the compatible solutes, glycine–betaine and ectoine, in osmotic stress adaptation and heat shock cross‐protection in the biocontrol agent Pantoea agglomerans CPA‐2. Letters in Applied Microbiology 41, 248-252. Vinas, I., Usall, J., Teixidó, N., Sanchis, V. 1998. Biological control of major postharvest pathogens on apple with Candida sake. International Journal of Food Microbiology 40, 9-16. Walkowiak, S., Rowland, O., Rodrigue, N., Subramaniam, R. 2016. Whole genome sequencing and comparative genomics of closely related Fusarium Head Blight fungi: Fusarium graminearum, F. meridionale and F. asiaticum. BMC Genomics 17, 1014. Walterson, A. M., Stavrinides, J. 2015. Pantoea: insights into a highly versatile and diverse genus within the Enterobacteriaceae. FEMS Microbiology Reviews 39, 968-984. Wang, C. L., Cheng, Y. H. 2017. Identification and trichothecene genotypes of Fusarium graminearum species complex from wheat in Taiwan. Botanical Studies 58, 4. Wang, C. L., Shim, W. B., Shaw, B. D. 2016. The Colletotrichum graminicola striatin orthologue Str1 is necessary for anastomosis and is a virulence factor. Molecular plant pathology 17, 931-942. Wang, L. Y., Xie, Y. S., Cui, Y. Y., Xu, J. J., He, W., Chen, H. G., Guo, J. H. 2015. Conjunctively screening of biocontrol agents (BCAs) against Fusarium root rot and Fusarium head blight caused by Fusarium graminearum. Microbiological Research 177, 34-42. Wang, Y. Z., Miller, J. D. 1988. Effects of Fusarium graminearum metabolites on wheat tissue in relation to Fusarium head blight resistance. Journal of Phytopathology 122, 118-125. Wu, A. B., Li, H. P., Zhao, C. S., Liao, Y. C. 2005. Comparative pathogenicity of Fusarium graminearum isolates from China revealed by wheat coleoptile and floret inoculations. Mycopathologia 160, 75-83. Yánez‐Mendizábal, V., Viñas, I., Usall, J., Torres, R., Solsona, C., Abadias, M., Teixidó, N. 2012. Formulation development of the biocontrol agent Bacillus subtilis strain CPA‐8 by spray‐drying. Journal of Applied Microbiology 112, 954-965. Yang, J. Y. 2016. Efficacy evaluation for tobacco debris combined with microorganisms on control of cruciferous vegetable anthracnose. Taichung, Taiwan: National Chung Hsing Unisversity. Master thesis. Yu, J. H., Hamari, Z., Han, K. H., Seo, J. A., Reyes-Dominguez, Y., Scazzocchio, C. 2004. Double-joint PCR: a PCR-based molecular tool for gene manipulations in filamentous fungi. Fungal Genetics and Biology 41, 973-981. Zhao, Y., Selvaraj, J. N., Xing, F., Zhou L., Wang, Y., Song, H., Tan, X., Sun, L., Sangare, L., Folly, Y. M. E., Liu, Y. 2014. Antagonistic action of Bacillus subtilis strain SG6 on Fusarium graminearum. PLoS ONE, 92486.
摘要: 小麥赤黴病在世界各地的產區造成嚴重的危害,除了造成直接的經濟損失外,赤黴病菌另會產生新月毒素污染穀粒。許多台灣農民以友善環境的耕作方式生產小麥,但缺乏有效防治小麥赤黴病的方法,本研究欲開發生物防治策略,包含拮抗微生物之開發及建立可篩選降低新月毒素生合成之拮抗微生物的平台,期融入台灣小麥的生產體系。本研究拮抗微生物有二,一來自台灣農業試驗所提供之保存菌株,另分離小麥花藥之拮抗微生物,依菌落型態又可分為類芽孢桿菌型與黃色黏稠型,經過孢子發芽抑制測試後分別選擇黃色黏稠型菌株I-1-4,與類芽孢桿菌型菌株I-2-9進行後續試驗,分別以PDB及LB作為基礎培養基,進行工業生產原料的替換與篩選。為鑑定2菌株的種名,菌株I-1-4以多位點基因分析法鑑定為Pantoea sp.;菌株I-2-9則以專一性引子對鑑定為Bacillus amyloliquefaciens。經一系列之營養源篩選,菌株I-1-4以蔗糖、馬鈴薯萃取物與葵花油組成的配方有較佳的拮抗能力;菌株I-2-9以脫脂奶粉、酵母粉、蔗糖與氯化鈉組成之配方有較佳的拮抗效果。為瞭解合適的培養條件與成分配置,進行環境因子與培養基成分影響之分析,經過二因子水準分析發現,菌株I-1-4的配方中,酵母萃取物與馬鈴薯萃取物顯著影響拮抗能力;菌株I-2-9的配方則以綠豆粉顯著影響拮抗能力。以二因子水準分析的配方進行溫室試驗,結果顯示在接種病原菌20天後,施用菌株I-2-9的10倍稀釋培養液,可降低58%的罹病度以及40%的罹病率,而施用菌株I-1-4的25倍稀釋培養液則在接種病原菌20天後,可降低67%的罹病度以及73%的罹病率,顯示其具有開發為生物製劑之潛力,因此進一步以最陡上升分析及中央合成設計分析優化上述顯著影響成分,中央合成設計分析獲得之迴歸方程式顯示,菌株I-1-4稀釋5倍之濾液可達到83.3%孢子發芽抑制率;菌株I-2-9以20倍稀釋的培養液可達到77.95%的孢子發芽抑制率。在建立偵測新月毒素生合成的平台方面,利用新月毒素生合成路徑第一個酵素TRI5的啟動子與GFP基因結合後之載體,進行PEG媒介基因轉殖後,產生9個轉殖株,南方墨點分析證實上述載體均成功插入轉殖株的TRI5基因座,並於誘導TRI5表現之培養基中,可見到明顯的GFP表現,於即時聚合酶鏈鎖反應顯示,野生型r-p-1的TRI5基因表現與轉殖株TPG-1及TPG-6的GFP表現,分別於新月毒素誘導及非誘導培養基中,有相同趨勢的表現情形,期未來可以轉殖株作為新月毒素生合成的偵測平台,篩選能夠降低新月毒素產生之拮抗微生物。
Fusarium head blight (FHB) is a devastating disease worldwide. Besides the direct economic loss, the pathogens produce trichothecene that contaminates kernels. The environmentally-friendly cultural practice is used to produce wheat in Taiwan, which lacks of an effective method in the control of FHB. This study aimed to develop a biocontrol agent and establish a platform to screen antagonists which can reduce trichothecene biosynthesis. The sources of antagonistic strains were the collection of Taiwan Agricultural Research Institute and the isolates from wheat anther. The antagonists were divided into two colony types, Bacillus-like colony and yellow mucoid colony. The strain I-2-9 with Bacillus-like colony and strain I-1-4 with yellow mucoid colony highly inhibited the spore germination of FHB pathogens in LB and PDB, respectively. Strain I-1-4 was identified as Pantoea sp. by multilocus sequence analysis while strain I-2-9 was identified as Bacillus amyloliquefaciens by species-specific primers. The strains and media were used as the basis for nutrient screening. The formula of strain I-1-4 composed of sucrose, potato extract and sunflower oil displayed a high antagonistic ability. The formula of strain I-2-9 composed of skim milk, yeast powder, sucrose and NaCl had a better antagonistic ability. For optimizing the formula, the environmental conditions and significance of ingredients were evaluated. The two-level factorial designed assay showed that yeast extract and potato extract in I-1-4 medium, and mung bean in I-2-9 medium significantly influenced antagonistic ability. A greenhouse test was carried out with the best medium composition in the two-level factorial design assay. The 10-fold dilution of I-2-9 reduced the disease severity by 58% and disease incidence by 40%. The 25-fold dilution of I-1-4 reduced the disease severity by 67% and disease incidence by 73%. The two strains showed the potential to develop as biocontrol agents. Moreover, the steepest ascent assay and central composited design were used to optimize the formula. Accroding to the regression equation obtained from central composite design. The 5-fold dilution filtrate of the strain I-1-4 could reach 83.3% spore inhibition rate. The strain I-2-9 could reach 77.95% spore inhibition rate with 20-fold dilution culture. On the other hand, to select antagonists that can suppress trichothecene biosynthesis, 9 fungal transformants were gernerated by PEG-mediated transformation, which contained the TRI5 promoter::GFP construct and significantly expressed GFP in a TRI5-inducing condition. The real-time PCR showed that TRI5 of wild type r-p-1 had a corresponding effect on GFP expression of transformants TPG-1 and TPG-6. The transformants can be used as a detection platform for screening antagonists which reduce the biosynthesis of trichothecene.
URI: http://hdl.handle.net/11455/95780
文章公開時間: 2020-08-23

檔案 大小格式 
nchu-106-7103035012-1.pdf2.07 MBAdobe PDF 請求副本

在 DSpace 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。