Agrobacterium rhizogenes-mediated transformation of a high oil-producing filamentous fungus Umbelopsis isabellina

• Autorzy: Wei D-S , Zhang Y-H , Xing L-J , Li M-C
• Języki publikacji: EN

Abstrakty

EN

The Agrobacterium rhizogenes-mediated transformation procedure was developed by using the hygromycin B phosphotransferase gene (hph) as a selective marker for the oil-producing fungus Umbelopsis isabellina. Different conditions were combined to increase the transformation efficiency. The highest efficiency was obtained by using A. rhizogenes strain R105 and a vector with zygomycete promoter. Southern blot analysis demonstrated that 71% of transformants contained random integrations of T-DNA sequences under optimal conditions. We randomly selected 115 positive transformants resistant to hygromycin to analyze the amount of total fatty acid and gamma-linolenic acid (GLA). Six transformants produced a higher amount of total fatty acids than the wild strain, and one transformant also produced a higher level of GLA than the wild strain in gas chromatography analysis. This is the first report about using A. rhizogenes strain R105 and germinated conidia to transform successfully the recalcitrant zygomycetes and to obtain transformants with a stable phenotype.

Słowa kluczowe

PL

Agrobacterium rhizogenes; conidium; Umbelopsis isabellina; fungi; filamentous fungi; plasmid construction; fungal transformation; molecular analysis; functional analysis; total fatty acid; wild strain

• Wydawca: -
• Czasopismo: Journal of Applied Genetics
• Rocznik: 2010
• Tom: 51
• Numer: 2
• Opis fizyczny: p.225-232,fig.,ref.

Twórcy

autor

Wei D-S

• Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin, 300071, PR China

autor

Zhang Y-H

• Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin, 300071, PR China

autor

Xing L-J

• Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin, 300071, PR China

autor

Li M-C

• Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin, 300071, PR China

Bibliografia

• Ando A, Sumida Y, Negoro H, Suroto DA, Ogawa J, Sakuradani E, Shimizu S, 2009. Establishment of Agrobacterium tumefaciens-mediated transformation of an oleaginous fungus Mortierella alpina 1S-4 and its application for eicosapentaenoic acid-producer breeding. Appl Environ Microbiol (in press), Doi: 10.1128/AEM.00648-09.
• Bundock P, Dulk-Ras A, Beijersbergen A, Hooykaas PJ, 1995. Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae. EMBO J 14: 3206-3214.
• Combier JP, Melayah D, Raffier C, Gay G, Marmeisse R, 2003. Agrobacterium tumefaciens-mediated transformation as a tool for insertional mutagenesis in the symbiotic ectomycorrhizal fungus Hebeloma cylindrosporum. FEMS Microbiol Lett 220: 141-148.
• Czapek F, 1902. Untersuchungen über die Stickstoffgewinnung und Eiweißbildung der Pflanzen. Beitr Chem Physiol u Pahtol 1: 540-560.
• De Groot MJA, Bundock P, Hooykaas PJJ, Beijersbergen AGM, 1998. Agrobacterium tumefaciens-mediated transformation of filamentous fungi. Nat Biotechnol 16: 839-842.
• Dox AW, 1910. The intracellular enzymes of Penicillium and Aspergillus with special references to those of P. camenberti. US Dept Agr Bur Anim Ind Bull 120: 170.
• Fang G, Hammar S, Grumet R, 1992. A quick and inexpensive method for removing polysaccharides from plant genomic DNA. BioTechniques 13: 52-57.
• Fang W-G, Zhang Y-J, Yang X-Y, Zheng X-L, Duan H, Li Y, Pei Y, 2004. Agrobacterium tumefaciens-mediated transformation of Beanveria bassiana using an herbicide resistance gene as a selection marker. J Invertebr Pathol 85: 18-24.
• Gardiner DM, Jarvis RS. Howlett BJ, 2005. The ABC transporter gene in the sirodesmin biosynthetic gene cluster of Leptosphaeria maculans is not essential for sirodesmin production but facilitates self-protection. Fungal Genet Biol 42: 257-263.
• Gill I, Valivety R, 1997. Polyunsaturated fatty acids, part 1: Occurrence, biological activities and application. Trends Biotechnol 15: 401-409. Hoekema A, Hirsch PR, Hooyka PK, Schilperoort RA, 1983. A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303: 179-180.
• Hood EE, Gelvin SB, Melchers LS, Hoekema A, 1993. New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res 2: 208-218.
• Horrobin DF, 1992. Nutritional and medical importance of gamma-linolenic acid. Prog Lipid Res 31: 163-194.
• Leclerque A, Wan H, Abschütz A, Chen S, Mitina GV, Zimmermann G, Schairer HU, 2004. Agrobacterium-mediated insertional mutagenesis (AIM) of the entomopathogenic fungus Beauveria bassiana. Curr Genet 45: 111-119.
• Li W, Guo G-Q, Zheng G-Ch, 2000. Agrobacterium- mediated transformation: state of the art and future prospect. Chin. Science Bull 45: 1537-1546.
• Mackenzie DA, Wongwathanarat P, Carter AT, Archer DB, 2000. Isolation and use of a homologous histone H4 promoter and a ribosomal DNA region in a transformation vector for the oil- producing fungus Mortierella alpina. Appl Environ Microbiol 66: 4655-4661.
• Meyer V, Mueller D, Strowig T, Stahl U, 2003. Comparison of different transformation methods for Aspergillus giganteus. Curr Genet 43: 371-377.
• Miao X-L, Wu Q-Y, 2006. Biodiesel production from heterotrophic microalgal oil. Bioresour Technol 97: 841-846.
• Michielse CB, Hooykaas PJJ, van den Hondel CAMJJ, Ram AFJ, 2005. Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Curr Genet 48: 1-17.
• Michielse CB, Salim K, Ragas P, Ram AFJ, Kudla B, Jarry B, et al. 2004. Development of asystem for integrative and stable transformation of the zygomycete Rhizopus oryzae by Agrobacterium-mediated DNA transfer. Mol Genet Genomics 271: 499-510.
• Monfort A, Cordero L, Maicas S, Polaina J, 2003. Transformation of Mucor miehei results in plasmid deletion and phenotypic instability. FEMS Microbiol Lett 224: 101-106.
• Mullaney EJ, Hamer JE, Roberti KA, Yelton MM, Timberlake WE, 1985. Primary structure of the trpC gene from Aspergillus nidulans. Mol Gen Genet 199: 37-45.
• Mullins ED, Chen X, Romaine P, Raina R, Geiser DM, Kang S, 2001. Agrobacterium-mediated transformation of Fusarium oxysporum: an efficient tool for insertional mutagenesis and gene transfer. Phytopathology 91: 173-180.
• Mullins ED, Kang S, 2001. Transformation: a tool for studying fungal pathogens of plants. Cell Mol Life Sci 58: 2043-2052.
• Nyilasi I, Papp T, Csernetics Á, Vágvölgyi C, 2008. Agrobacterium tumefaciens-mediated transformation of the zygomycete fungus Backusella lamprospora. J Basic Microbiol 48: 59-64.
• Pahl G, McKibben B, 2005. Biodiesel: growing a new energy economy. White River Junction, Vermont: Chelsea Green.
• Papanikolaou S, Sarantou S, Komaitis M, 2004. Repression of reserve lipid turnover in Cunninghamella echinulata and Mortierella isabellina cultivated in multiple limited media. J Appl Microbiol 97: 867-75.
• Rho H, Kang S, Lee Y, 2001. Agrobacterium tumefaciens-mediated transformation of the plant pathogenic fungus Magnaporthe grisea. Mol cells 12: 407-411.
• Stafford HA, 2000. Crown gall disease and Agrobacterium tumefaciens: a study of the history, present knowledge, missing information and impact on molecular genetics. Bot Rev 66: 101-118.
• Sugui JA, Chang YC, Kwon-Chung KJ, 2005. Agrobacterium tumefaciens-mediated transformation of Aspergillus fumigatus: an efficient tool for insertional mutagenesis and targeted gene disruption. Appl Environ Microbiol 71: 1798-1802.
• Takahara H, Tsuji G, Kubo Y, Yamamoto M, Toyoda K, Inagaki Y, et al. 2004. Agrobacterium tumefaciens-mediated transformation as a tool for random mutagenesis of Colletotrichum trifolii. J Gen Plant Pathol 70: 93-96.
• Yang X-H,Sun Zh-H, Tong R-J, 2006. Optimizing culture system of Ri T-DNA transformed roots for Citrus grandis cv. Changshou Shatian You. Agrie Sci China 5: 90-97.
• Yan Z, Chen J, 2003. Research advance on microbial oils and their exploitation and utilization. Journal of Cereals & Oils 7: 13-15.
• Zhang P-Y, Xu B, Wang Y-Zh, Li Y-Q, Qian Zh, Tang Sh-R, et al. 2008a. Agrobacterium tumefaciens-mediated transformation as a tool for insertional mutagenesis in the fungus Penicillium marneffei. Mycol Res 112: 943-949.
• Zhang X-W, Li M-Ch, Wei D-Sh, Wang X-M, Chen X, Xing L-J, 2007. Disruption of the fatty acid delta6-desaturase gene in the oil-producing fungus Mortierella isabellina by homologous recombination. Curr Microbiol 55: 128-134.
• Zhang Y-H, Wei D-Sh, Xing L-J, Li M-Ch, 2008b. A modified method for isolating DNA from fungus. Microbiology 35: 466-469 (In Chinese).