黑殭菌 (Metarhizium anisopliae (Metsch.) Sorokin ) 是一種昆蟲病原真菌，寄主範圍極廣，可感染 200 種以上的昆蟲，深具經濟及研究的價值。本研究收集國內外共計 32 株供試黑殭菌菌株，配合使用分類上常利用之核糖體核酸內轉錄間隔區 (rDNA-ITS) 定序法直接比對各菌株間核酸序列的差異度，並以鄰近接連法 (Neighbor-Joining , NJ) 及最大簡約法 (Maximum-Parsimony, MP) 建構出親緣關係樹 （phylogenetic tree）。同時本研究更進一步分析黑殭菌各菌株的二次代謝物 (secondary metabolites)，檢測目標以黑殭菌素 (destruxins, DTX) 和細胞鬆弛素 (cytochalasins) 為主，而此二類化合物皆各有其生物活性及醫學上之利用價值。本研究室在以往曾自南投採集所得之 Ma0407 菌株發現其可產生類細胞鬆弛素之新化合物，而購自食品工業發展研究所之菌株 BCRC35507 可產生黑殭菌素，因此由 Ma0407 及 BCRC35507 兩菌株產生之已知代謝物作為對照組，將供試菌株代謝萃取物經由高效液相層析儀 (high-performance liquid chromatography; HPLC) 分析後結果顯示 ML 13 可產生和 Ma0407 相同的類細胞鬆弛素，而 Ma0319及 BCRC 35520 和 ARSEF 488、7488、5369 則可比對出和 BCRC 35507 相同的黑殭菌素，但各菌株間所產之黑疆菌素種類及含量均有差異，而黑疆菌素A、B、B2/E2 和 C 則普遍存在於個菌株間。經由 rDNA-ITS 定序法探討 Metarhizium 種間及變種間之親緣關係及了解變種間二次代謝物之化學多樣性 （chemodiversity），發現縱使 ITS 序列完全相同、親緣關係相近者也未必會產生完全相同之代謝物。

Metarhizium anisopliae (Metsch.) Sorokin, a fungal entomopathogen, can infect more than 200 kinds of insect; the it is a value fungus for extensive studies both in industry and medicine. In the present study, 32 isolates of M. anisopliae collected from domestic and abroad were analyzed for ribosomal DNA internal transcribed spacer (rDNA-ITS) and secondary metabolites. The results of the difference between varieties were subjected the collected species. The difference of DNA sequence were analyzed by method to construct Neighbor-Joining (NJ) and Maximum-Parsimony (MP) phylogeny tree. The differences of secondary metabolites, mainly destruxins and cytochalasins of M. anisopliae varieties were also to be subjected to statistically analysis further. Ourrently. We use Ma0407, which collected from Nanto with analogs of cytochalasin, and BCRC 35507 purchased from Food Industry Research and Development Institute with destruxins. I employ Ma0407 and BCRC 35507 as reference group for chemodiversity study. After high-performance liquid chromatography (HPLC) analysis, the data reveals that ML13 and Ma0407 could produce the identical analogs of cytochalasins, in addition, BCRC 35507, Ma0319, BCRC 35520, and ARSEF 488, 7488, 5369 have the same destruxins; however, the classification and that quantity of destruxins in M. anisopliae strains are generally different, but destruxins A, B, B2/E2 and C exist commonly in every M. anisopliae isolate. In the present study, we explore the phylogeny of Metarhizium species and of Metarhizium varieties through rDNA-ITS sequencing method, and then investigate the chemodiversity of secondary metabolites between varieties. Results show that even if different M. anisopliae strains that possess identical rDNA-ITS sequence, meaning they have close phylogenetic relationship, cannot produce completely identical metabolites.

摘要..........................................II
Abstract ....................................III
第一章、 前言 .............................1
第二章、 文獻回顧 .............................3
一、 黑殭菌 (M. anisoplia) 之介紹..........3
二、 黑殭菌屬的分類沿革及特性 ..........4
三、 黑殭菌系統分類的研究進展 ..........6
1. 同功酵素分析 ...................6
2. RFLP 和 RAPD 技術的研究 ..........7
3. rDNA 序列分析 ...................8
四、 黑殭菌所產之代謝物 ..................10
1. 黑殭菌素 ...........................11
2. 細胞鬆弛素 ..................13
第三章、 材料與方法 ..................15
一、 供試菌株的來源、保存與製備 .........15
二、 黑殭菌親緣關係之建立 .........15
(一)、 DNA 萃取 ...........................15
(二)、 PCR 與 ITS 定序 ..................16
(三)、 序列排列及親緣圖建立 .........18
三、 黑殭菌代謝物質分析 ..................18
(一)、 樣品萃取 ...........................18
(二)、 HPLC分析 ...........................18
第四章、 結果 ...........................20
一、 黑殭菌親緣關係之建立 .........20
二、 黑殭菌代謝產物之分析 .........22
第五章、 討論 ...........................23
第六章、 參考文獻 ...........................28
附錄、 ...........................65

古欣平。2004。黑殭菌素B (Destruxin B) 抑制人類血癌細胞之探討。私立朝陽科技大學應用化學系碩士論文。94頁。
匡治州、許楊。2004。核醣體 rDNA ITS 序列在真菌學研究中的應用。生命的化學。24： 120-122。
高穗生、蔡勇勝。2007。真菌殺蟲劑研發現況、潛力和展望。生物性肥料與農藥。12： 21-27。
曹暉、劉玉萍。1999。DNA 分子標記技術：一種新的中藥分析方法。藥物分析雜誌。19： 355-360。
黃勃、樊美珍、李增智。2002。綠殭菌系統分類的研究進展 (綜述)。安徽農業大學學報。29： 169-172。
梁宗琦、劉愛英、劉杰麟。1991。蟲草新種及其綠殭菌無性型。真菌學報。10： 257-262。
曾敏南。2001。黑殭菌(Metarhizium anisopliae var. anisopliae)耐高溫突變菌株之篩選及其生理、病原性之探討。國立台灣大學植物病理學系碩士論文。106頁。
趙國柱、張天宇、張猛。2002。核醣體基因簇在真菌系統學研究中的意義。生命的化學。22： 13-15。
黎培琪。2004。黑殭菌素B (Destruxin B) 對 DBA/2 老鼠 L5178Y 淋巴癌細胞之抑制效果及其機制。私立台北醫學大學生物醫學技術研究所碩士論文。81頁。
劉顯達、林秀芬。2007。黑殭菌 MA126 菌株 DNA 探針製備及田間偵測應用。植物保護學會會刊。49： 347-354。
蔣盛岩、張志光。2002。真菌的分子生物學鑑定方法研究進展。生物學通報。37：4-6。
Aldridge, D. C., and Turner, W. B. 1969. Structures of cytochalasin C andcytochalasin D from Metarhizium anisopliae. J. Chem. Soc. C. 6: 923-928.
Amiri, B., Ibrahim, L., and Butt, T. M. 1999. Antifeedant Properties of Destruxins and their Potential Use with the Entomogenous Fungus Metarhizium anisopliae for Improved Control of Crucifer Pests. Biocontrol Sci. Technol. 9: 487-498.
Amiri-Besheli, B., Khambay, B. Cameron, S., Deadman, M. L., and Butt, T. M. 2000. Iner-and intra-specific variation in destruxin production by insect pathogenic Metarhizium spp. and its significance to phthogenesis. Mycol. Res. 104: 447-452.
Ayer, W. A., and Pena-Rodriguez, L. M. 1987. Metabolites produced by Alternaria brassicae, the black spot pathogen of Canola. Part 1, the phytotoxic compoments. J. Nat. Prod. 50: 400-407.
Bains, P. S., and Tewari, J. P. 1987. Purification, chemical characterization and host-specificity of the toxin produced by Alternaria brassicae. Physiol. Mol. Plant P. 30: 259-271.
Barrett, J. 1986. Molecular variation and evolution. pp. 83-95. Evolution Biology of the Fungi: Symposium of the British Mycological Society Held at the University of Bristol. Rayner, A. D., Brasier, C. M., and Moore, D. eds. Cambridge University Press, New York.
Berreta, M. F., Lecuona, R. E., Zandomeni, R. O., and Grau, O. 1998. Genotyping isolates of the entomopathogenic fungus Beauveria bassiana by RAPD with fluorescent labels. J. Invertebr. Pathol. 71: 145-150.
Bischoff, J. F., Rehner, S. A., and Humber, R. A. 2006. Metarhizium frigidum sp. Nov.: a cryptic species of M. anisopliae and a member of the M. flavoviride complex. Mycologia 98: 737-745.
Bräse, S., Encinas, A., Keck, J., and Nising, C. F. 2009. Chemistry and Biology of Mycotoxins and Related Fungal Metabolites. Chem. Rev. 109: A-CJ.
Brousseau, G., Charpentier, G., and Belloncik, S. 1996. Susceptibility of spruce budworm, Choristoneura fumiferana Clemens, to destruxins, cyclodepsipeptidic mycotoxin of Metarhizium anisopliae. J. Invertebr. Pathol. 68: 180 -182.
Bruns, T. D., White, T. J., and Taylor, J. W. 1991. Fungal molecular systematic. Annu. Rev. Ecol. Syst. 55: 525-564.
Buchwaldt, L., and Jensen, J.S. 1991. HPLC purication of destruxins produced by Alternaria brassicae in culture and leaves of Brassica napus. Phytochemistry 30: 2311-2316.
Cai, P., Smith, D., Katz, B., Pearce, C., Venables, D., and Houck, D. 1998. Destruxin-A4 chlorohydrin, a novel destruxin from fungus OS-F68576: isolation, structure determination, and biological activity as an inducer of erythropoietin. J. Nat. Prod. 61: 290-293.
Calmes, M., Cavelier-Frontin, F., Jacquier, R., Mercadier, J. L., Sabil, S., Quiot, J. M., and Vey, A. 1993. Synthesis and biological activity of a destruxin analogue:D-Lac-6 destruxin E. Int. J. Pept. Prot. Res. 41: 528-535.
Carter, S. B. Effects of cytochalasins on mammalian cells. 1967. Nature 213: 261-264.
Chen, H. C., Chou, C. K., Sun, C. M., and Yeh, S. F. 1997. Suppressive effects of destruxin B on hepatitis B virus surface antigen gene expression in human hepatoma cells. Antiviral Res. 34: 137-144.
Clarkson, J. M., and Charnley, A. K. 1996. New insights into the mechanisms of fungal pathogenesis in insects. Trends Microbiol. 4: 197-203.
Cobb, B. D., and Clarkson, J. M. 1993. Detection of molecular variation in the insect pathogenic fungus Metarhizium using RAPD-PCR. FEMS Micro. Biol. Lett. 112: 319-324.
Curtis, M. D., Gore, J., and Oliver, R. P. 1994. The phylogeny of the tomato leaf mould fungus Cladosporium fulvum syn. Fulvia fulva by analysis of rDNA sequences. Curr. Genet. 25: 318-322.
David, J. R. 1991. Past and future for taxomony. Molecular Techniques in Taxomony: Proceedings of the NATO Advanced Study Institute on Molecular Techniques in Taxonomy Held in Norwich (England). pp. 1-4. Hewitt, G. M., Johnston, A. W. B., and Young, J. P. W. eds. Springer-Verlag Berlin Heidelbera, Germany.
D'Ovidio, R. 1992. Nucleotide sequence of a 5.8S rDNA gene and of the internal transcribed spacers from Populus deltoides. Plant Mol. Biol. 19: 1069-1072.
Driver, F., Milner, R.J., and Trueman, W. H. J. 2000. Taxonomic revision of Metarhizium based on a phylogenetic analysis of rDNA sequence data. Mycol. Res. 104: 134-150.
Espada, A., and Dreyfuss, M. M. 1997. Effect of the cyclopeptolide 90-215 on the production of destruxins and helvolic acid by Metarhizium anisopliae. J. Ind. Microbiol. Biotechnol. 19: 7-11.
Fegan, M., Manners, J.M., Maclean, D.J., Irwin, J.A., Samuels, K.D., Holdom, D.G., and Li, D.P. 1993. Random amplified polymorphic DNA markers reveal a high degree of genetic diversity in the entomopathogenic fungus Metarhizium anisopliae var. anisopliae. J. Gen. Microbiol. 139: 2075-2081.
Felsenstein, J. 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39: 783-791.
Fujii, Y. Tani, H. Ichinoe, M., and Nakajima, H. 2000. Zygosporin D and Two New Cytochalasins Produced by the Fungus Metarrhizium anisopliae. J. Nat. Prod. 63: 132-135
Gams, W., and Rozsypal, J. 1973. Metarrhizium flavoviride n. sp. isolated from insects and from soil. Acta Bot. Neerl. 22: 518-521.
Gardes, M., White, T. J., Fortin, J. A., Bruns, T. D., and Taylor, J. W. 1991. Identification of indigenous and introduced symbiotic fungi in ectomycorrhizae by amplification of nuclear and mitochondrial ribosomal DNA. Can. J. Bot. 6: 180-190.
Gerbi, S. A. 1985. Evolution of ribosomal DNA. pp. 419-517. In: Molecular Evolutionary Genetics. Maclntype R. J. ed. Plenum, New York.
Grajal-Martin, M. J., C. J. Simon, and F. J. Muehlbauer. 1993. Use of random amplified polymorphic DNA (RAPD) to characterize race 2 of Fusarium oxysporum f.sp. pisi. Phytopathology 83: 612-614.
Gross, M. L., McCrery, D., Grow, F., and Tomer, K. B. 1982. The structure of the toxin from Metarhizium anisopliae. Tetrahedron Lett. 23: 5381-5384.
Guo, H. L., Ye, B. L., Yue, Y. Y., Chen, Q. T., and Fu, C. S. 1986. Three new species of Metarhizium. Acta Mycol. Sinica. 5: 185-190.
Gupta, S., Krasnoff, S. B. Renwick, J. A. A. Roberts, D. W. Steiner, J. R. and Clardy, J. 1993. Viridoxins A and B: novel toxins from the fungus Metarhizium avoviride. J. Org. Chem. 58: 1062–1067.
Gupta, S., Roberts, D. W., and Renwick, J. A. A. 1989. Preparative isolation of destruxins from Metarhizium anisopliae by high perfermance liquid chromatography. J. Liquid Chromatogr. R. T. 12: 383-395.
Henrion, B., Chevalier, G., and Martin, F. 1994. Typing truffle species by PCR amplification of the ribosomal DNA spacer. Mycol. Res. 98: 37-43.
Hibbert, D. S., Fukumasa-Nakai, Y., Tsuneda, A., and Donoghue, M. J. 1995. Phylogenetic diversity in shiitake inferred from nuclear ribosomal DNA sequences. Mycologia 87: 618-638.
Huang, B., Li, C., Humber, R. A., Hodge, K. T., Fan, M., and Li, Z. 2005. Molecular evidence for the taxonomic status of Metarhizium taii and its teleomorph, Cordyceps taii (Hypocreales, Clavicipitaceae). Mycotaxon. 94: 137-147.
Huxham, I.M., Lackie, A.M., and McCorkindale, N.J., 1989. Inhibitory effects of cyclodepsipeptides, destruxins, from the fungus Metarhizium anisopliae on cellular immunity in insects. J. Insect Physiol. 35: 97-105.
Iijima, M., Masuda, T. Nakamura, H. Naganawa, H. Kurosawa, S. Okami, Y. Ishizuka, M. Takeuchi, T., and Iitaka, Y. 1992. Metacytolin, a novel immunomodulator produced by Metarhizium sp. TA2759. J. Antibiot. 45: 1553-1556.
Ito, Y., and Hirano, T. 1997. The detremination of partial 18S ribosomal DNA sequence of Cordyceps species. Lett. Appl. Microbiol. 25: 239-242.
Iwen, P. C., Hinrichs, S. H., Rupp, M. E. 2002. Utilization of the internal transcribed spacer regions as molecular targets to detect and identify human fungal pathogens. Med. Mycol. 40: 87-109.
James, P. J., Kershaw, M. J., Reynolds, S. E., and Charnley, A. K. 1993. Inhibition of desert locust (Schistocerca gregaria) Malpighian tubule fluid secretion by destruxins, cyclic peptide toxins from the insect pathogenic fungus Metarhizium anisopliae. J. Insect Physiol. 39: 797-807.
Kasuga, T., Woods, C. Woodward, S., and Mitchelson, K. 1993. Heterobasidion annosum 5.8S ribosomal DNA and internal transcribed spacer sequence: Rapid identification of European intersterility groups by ribosomal DNA restriction polymorphism. Curr. Genet. 24: 433-436.
Kato, A., Nakajima, T., Yamashita, J., Yakura, K., and Tanifuji, S. 1990. The structure of the large spacer region of the rDNA in Vicia faba and Pisum sativum. Plant Mol. Biol. 14: 983-993.
Kershaw, M. J., Moorhouse, E. R., Bateman, R., Reynolds, S. E., and Charnley, A. K. 1999. The role of destruxins in the pathogenicity of Metarhizium anisopliae for three species of insect. J. Invertebr. Pathol. 74: 213-223.
Kodaria, Y. 1961. Toxic substances to insects, produced by Aspergillus ochraceus and Oopsra destructor. Agric. Biol. Chem. 25: 261-262.
Kohn, L. M. 1992. Developing new characters for fungal systematics: An experimental approach for determining the rank of resolution. Mycologia 84: 139-153.
Kondo, S. Meguriya, N. Mogi, H. Aota, T., and Miura, K. 1980. K-582, a new peptide antibiotic. I. J. Antibiot. 33: 533-542.
Kopecky, J., Matha, V., and Jegorov, A. 1992. The inhibitory effect of destruxin A on the replication of arboviruses in Aedes albopictus C6/36 cell line. Comp. Biochem. Physiol. 103C: 23-25.
Krasnoff, S. B., Keresztes, I., Gillilan, R. E., Szebenyi, D. M. E., Donzelli, B. G. G., Churchill, A. C. L., and Gibson, D. M. 2007. Serinocyclins A and B, Cyclic Heptapeptides from Metarhizium anisopliae. J. Nat. Prod. 70: 1919-1924
Krasnoff, S. B., Sommers, C. H., Moon, Y-S, Donzelli, B. G. G., Vandenberg, J. D., Churchill, A. C. L., and Gibson, D. M. 2006. Production of Mutagenic Metabolites by Metarhizium anisopliae. J. Agric. Food Chem. 54: 7083-7088.
Kuboki, H., Tsuchida, T. Wakazono, K. Isshiki, K. Kumagai, H., and Yoshioka, T. 1999. Mer-f3, 12-hydroxy-ovalicin, produced by Metarhizium sp. f3. J. Antibiot. 52: 590-593.
Kumar, S., Tamura, K., and Nei, M. 2004. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief. Bioinform. 5: 150-163.
Latch, G. C. M. 1965. Metarhizium anisopliae (Metschnikoff) Sorokin strains in New Zealand and their possible use for controlling pasture inhabiting insects. N. Z. J. Agric. Res. 8: 384-396.
Liu, A. Y., Liang, Z. Q., and Cao, L. 1989. Isolation and identication for Metarhizium biformisporae. J. Guizhou Agric. Coll. 2: 27-31.
Liu, Z. Y., Liang, Z. Q., and Liu, A. Y. 1995. Analysis of esterase isozyme bands and protein bands of Metarhizium spp. pp. 131-135. In: Advanced Research of Chinese Young Mycologists. Tang, W. Z. Ed. Southwest Normal Univ. Press. Chongqing, China
Liu, Z. W., Guo, H. L., and Kurtzman, C. P. 1994. Phylogeny of Metarhizium species as determined by partial ribosomal RNA sequence. Acta Mycol. Sinica. 13: 139-151.
Liu, Z. Y., Liang, Z. Q., Whalley, A. J. S. Yao, Y. -J., and Liu, A. Y. 2001. Cordyceps brittlebankisoides, a New Pathogen of Grubs and Its Anamorph, Metarhizium anisopliae var. majus. J. Invertebr. Pathol. 78: 178-182.
Liu, S. D., and Lin, S. C. 1989. Microbial control of coconut leaf beetle (Brontispa longissima) with green muscardine fungus, Metarhizium anisopliae var. anisopliae. J. Invertebr. Pathol. 53: 307-314.
Maclean, D. J., Braithwaite, K. S., Manners, J. M., and Irwin, J. A. G. 1993. How do we identify and classify fungal plant pathogens in the era of DNA analysis? Adv. Plant Pathol. 10: 207-244.
Micales, J. A., Bonde, M. R., and Peterson, G. L. 1986. The use of isozyme analysis in fungal taxonomy and genetics. Mycotaxon 27: 405-449.
Moon, Y-S., Donzelli, B. G. G., Krasnoff, S. B. McLane, H., Griggs, M. H. Cooke, P., Vandenberg, J. D., Gibson, D. M., and Churchill1, A. C. L. 2008. Agrobacterium-Mediated Disruption of a Nonribosomal Peptide Synthetase Gene in the Invertebrate Pathogen Metarhizium anisopliae Reveals a Peptide Spore Factor. Appl. environ. microbiol. 74: 4366-4380.
Moore, D., and Prior, C. 1993. The potential of my coinsecticides. 1993. Biocontrol News Info. 14: 31 -40.
Mugnai, L., Bridge, P. D., and Evans, H. C. 1989. A chemotaxonomic evaluation of the genus Beauveria. Mycol. Res. 92: 199-209.
Odier, F., Vey, A., and Bureau, J. P. 1992. In vitro effect fungal cyclodepsipeptides on leukemic cells : study of destruxins A、B and E. Biocell. 74: 267-271.
Patrick, M., Adlard, M. W., and Keshavarz, T. 1993. Production of anindolizidine alkaloid, swainsonine by the filamentous fungus, Metarhizium anisopliae. Biotechnol. Lett. 15: 997-1000.
Prasain, J. K., Ueki, M., Stefanowicz, P., and Osada, H. 2002. Rapid screening and identication of cytochalasins by electrospray tandem mass spectrometry. J. Mass Spectrom. 37: 283-291.
Pedras, M. S. C., Zaharia, L. I., and Ward, D. E. 2002. The destruxins: synthesis, biotransformation, and biological activity. Phytochemsitry. 59: 579-796.
Rakotonirainy, M.S., Cariou, M. L., Brygoo, Y., and Riba, G. 1994. Phylogenetic relationships within the genus Metarhizium based on 28S rRNA sequences and isozyme comparison. Mycol. Res. 98: 225-230.
Ritland, C. and Straus, N. A. 1993. High evolutionary divergence of the 5.8S ribosomal DNA in Mimulus glaucescens (Scrophulariaceae). Plant Mol. Biol. 22: 691-696.
Roberts, D. W., and Humber, R. A. 1981. Entomogenous fungi. pp.201-236. In: Biology of Conidial Fungi Volume 2. Cole, G. T. ed. Academic press, New York.
Rombach, M. C., Humber, R. A., and Roberts, D.W. 1986. Metarhizium flavoviride var. minus var. nov., a pathogen of plant-and leafhoppers on rice in the Philippines and Solomon Islands. Mycotaxon 27: 87-92.
Rombach, M. C., Humber, R. A., and Evans, H. C. 1987. Metarhizium album, a fungal pathogen of leaf- and planthoppers of rice. Trans. Br. Mycol. Soc. 88: 451-459.
Saitou, U., and Nei, M. 1987. The neighbor-joining methods: A new method for reconstructing phylogenetics tree. Mol. Biol. Evol. 4: 406-425.
Samuels, R. I., Reynold, S. E. and Charnley, A. K. 1992. Calcium channel activation of insect muscle by destruxins, insecticidal compounds produced by the entomopathogenic fungus Metarhizium anisopliae. Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology 90: 403-412.
Sanger, F., Nicklen, S., and Coulson, A. R. 1977. DNA sequencing with chain-termination inhibitors. Proc. Natl. Acad. Sci. 74: 5463-5467.
Schultz, N., Őnfelt, A. 2000. Sensitivity of cytokinesis to hydrophobic interactions. Chemical induction of bi-and multinucleated cells. Chem. Biol. Interact. 126: 97-123.
Springer, J. P., Cole, R. J., Dorner, J. W., Cox, R. H., Richard, J. L., Barnes, C. L., and van der Helm, D. 1984. Structure and conformation of roseiotoxin B. J. Am. Chem. Soc. 106: 2388-2392.
Steyn, P. S. 1977. Some new mycotoxins. Pure & Appl. Chem. 49: 1971-1978.
St Leger, R. J., May, B., Allee, L. L., Frank, D. C., Staples, R. C., and Roberts, D. W. 1992. Genetic differences in allozymes and in formation of infection structures among isolates of the entomopathogenic fungus Metarhizium anisopliae. J. Invertebr. Pathol. 60: 89-101.
Strasser, H., Vey, A., and Butt, T. M. 2000. Are there any risks in using entomopathogenic fungi for pest control, with particular reference to the bioactive metabolites of Metarhizium, Tolypocladium and Beauveria species? Biocontrol Sci. Technol. 10: 717-735.
Suzuki, A., Taguchi, H., and Tamura, S. 1970. Isolation and structure elucidation of three new insecticidal cyclodepsipeptides, Destruxins C and D and desmethyldestruxin B, produced by Metarhizium anisopliae. Agri. Biol. Chem. 34: 813-816.
Taira, T., Kato, A., and Tanifuji, S. 1988. Difference between two major size classes of carrot rDNA repeating units is due to reiteration of sequences of about 460 bp in the large spacer. Mol. Gen. Genet. 213: 170-174.
Thomsen, J., and Eilenberg, J. 2000. Time-concentration mortality of Pieris brassicae and Agrotis segetum larvae from different destruxins. Environ. Eentomol. 29: 1041-1047.
Tulloch, M. 1976. The genus Metarhizium. Trans. Brit. Mycol. Soc. 66: 407-411.
Tzean, S. S., Hsieh, L. S., Chen, J.L., and Wu, W. J. 1993. Nomuraea cylindrospora comb. nov. Mycologia 85: 514-519.
Uchida, R., Imasoto, R. Yamaguchi, Y. Masuma, R. Shiomi, K. Tomoda, H., and Ōmura , S. 2005. New insecticidal antibiotics, hydroxyfungerins A and B, produced by Metarhizium sp. FKI-1079. J. Antibiot. 58: 804-809.
Veen, K. H. 1968. Recherches sur la maladie, due à Metarhizium anisopliae chez le criquet pélerin. PhD-thesis, Landbouwhogeschool Wageningen 68-5, 77 pp.
Venkatsubbaiah, P., Tisserat, N. A., and Chilton, W. S. 1994. Metabolites of Ophiosphaerella herpotricha. Mycopathologia 128: 155-159.
Vey, A. and Quiot, J. M. 1989. Effet cytotoxique in vitro et chez l'insecte hôte des destruxines, toxines cyclodepsipeptidiques produites par le champignon entomopathogène Metarhizium anisopliae. Can. J. Microbiol. 35: 1000-1008.
White, T. J., Bruns, T., Lee, S., and Taylor, J. 1990. Amplification and direct..sequencing of fungal ribosomal RNA genes for phylogenetics. pp. 315-322. In PCR Protocols: A Guide to Methods and Applications, Innis, M. A., Gelfand, D. H., Sninsky, J. J., and White, T. J. eds. San Diego, Academic Press.
Williams, J. G. K., Kubelik, A. R., Rafalski . J. A., and Tingey, S. V. 1990. DNA polymorphisms amplified by arbitrary primers are useful genetic markers. Nucleic Acid Res. 18: 6531-6535.
Yahara, I., Harada, F., Sekita, S., Yoshihira, K., and Natori, S. 1982. Correlation between Effects of 24 Different Cytochalasins on Cellular Structures and Cellular Events and Those on Actin In Vitro. J. Cell Biol. 92: 69-78.
Yip, H. I., Rath, A. C., and Koen, T. B. 1992. Characterization of Metarhizium anisopliae isolates from Tasmanian pasture soil and their pathogenicity to redheaded cockchafer (Coleoptera: Scarabaeidae: Adoryphorus couloni). Mycol. Res. 96: 92-96.
Zimmermann, G. 1993. The entomopathogenic fungus Metarhizium anisopliae and its potential as a biocontrol agent. Pestic. Sci. 37: 375-379.