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http://dx.doi.org/10.4489/KJM.2010.39.3.235

Agrobacterium-Mediated Transformation of Flammulina velutipes with NaOH Treatment  

Shin, Dong-Il (Department of Biotechnology, Catholic University of Daegu, Kyungsan)
Park, Hee-Sung (Department of Biotechnology, Catholic University of Daegu, Kyungsan)
Publication Information
The Korean Journal of Mycology / v.39, no.3, 2011 , pp. 235-238 More about this Journal
Abstract
Agrobacterium harboring vector pCHBs with hygromycin phosphotransferase(hph) and hepatitis B virus surface antigen(HBsAg)gene was transformed into the mycelial culture of Flammulina velutipes. In particular, mild NaOH solution was treated to the mycelia before Agrobacterium infection step. This was purposed to generate putative surface wounds in the mycelial cell walls. The results showed that hygromycin-resistant($hyg^r$) mycelia could be obtained only from NaOH-treated mycelia but not from intact mycelia. The integration of $hyg^r$ gene in fungal genome was confirmed by PCR. In addition, a single transgene integration and heterologous protein expression in F. velutipes could be verified by Southern blot hybridization and western blot analysis, respectively. This study demonstrated an efficient tool for the Agrobacterium-mediated transformation of F. velutipes mycelia.
Keywords
Agrobacterium; Flammulina velutipes; Mycelium; NaOH; Transformation;
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1 Shin, D.-I. and H.-S. Park. 2008. Expression of recombinant protein from Oenanthe javanic DC. leaf tissues as a biofactory. Kor. J. Biotechnol. Bioeng. 23:554-556.   과학기술학회마을
2 Singh, N. and Chawla, H. S. 1999. Use of silicon carbide fibers for Agrobacterium-mediated transformation in wheat. Curr. Sci. 76:1483-1485.
3 Uze, M., Wunn, J., Pounti-Kaelas, J., Potrykus, I. and Sauter, C. 1997. Plasmolysis of precultured immature embryos improves Agrobacterium mediated gene transfer to rice (Oryza sativa L.). Plant Sci. 130:87-95.   DOI   ScienceOn
4 Wasser, S. P. and Weis, A. L. 1999. Medicinal properties of substances occurring in higher basidiomycetes mushrooms: Current perspectives (review). Int. J. Med. Mushr. 1:31-62.   DOI
5 Kilaru, S. and Kes, U. 2005. Comparison of gpd genes and their protein products in basidiomycetes. Fungal Genet. Newslett. 52:18-23.
6 Kuo, C. Y., Chou, S. Y., Hseu, R. S. and Huang, C. T. 2010. Heterologous expression of EGFP in enoki mushroom Flammulina velutipes. Bot. Studies. 51:303-309.
7 Maehara T., Yoshida, M., Ito, Y., Tomita, S., Takabatake, K., Ichinose, H. and Kaneko, S. 2010. Development of a gene transfer system for the mycelia of Flammulina velutipes Fv-1 strain. Biosci. Biotechnol. Biochem. 74:1126-1128.   DOI   ScienceOn
8 Meyer, V. 2008. Genetic engineering of filamentous fungi - progress, obstacles and future trends. Biotech. Adv. 26:177-185.   DOI   ScienceOn
9 Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15:473-497.   DOI
10 Olhoft, P. M. and Somers, D. A. 2001. L-Cysteine increases Agrobacterium-mediated T-DNA delivery into soybean cotyledonary-node cells. Plant Cell Rep. 20:706-711.   DOI
11 Bechtold, N. and Pelletier, G. 1998. In planta Agrobacteriummediated gene transfer of adult Arabidopsis thaliana plants by vacuum infiltration. Meth. Mol. Biol. 82:259-266.
12 Pan, S. Q., Charles, T., Jin, S., Wu, Z. L. and Nester, E. W. 1993. Preformed dimeric state of the sensor protein VirA is involved in plant-Agrobacterium signal transduction. Proc. Natl. Acad. Sci. USA 90: 9939-9943.   DOI
13 Park, S.-Y., van Peer, A. F., Jang, K.-Y., Shin, P.-G., Park, Y., Yoo, Y.-B., Park, K.-M. and Kong, W.-S. 2010. Agrobacterium-mediated transformation using gill tissue of Flammulina velutipes. Kor. J. Mycol. 38:48-53.   과학기술학회마을   DOI   ScienceOn
14 Sharma, K. K. and Kuhad, R. C. 2010. Genetic transformation of lignin degrading fungi facilitated by Agrobacterium tumefaciens. BMC Biotechnol. 10:67-74.   DOI   ScienceOn
15 Cheng, M., Fry, J. E., Pang, S., Zhou, I., Hironaka, C., Duncan, D. R. I., Conner, T. W. L. and Wang, Y. 1997. Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol. 115:971-980.   DOI
16 Cheng, M., Hu, T., Layton, J. L., Liu, C. N. and Fry, J. E. 2003. Desiccation of plant tissues post-Agrobacterium infection enhances T-DNA delivery and increases stable transformation efficiency in wheat. In Vitro Cell. Dev. Biol. Plant 39:595-604l.   DOI   ScienceOn
17 Kim, S., Shin, D.-I. and Park, H.-S. 2007. Transient ${\beta}-glucuronidase$ expression in lily (Lilium longiflorum L.) pollen via woundingassisted Agrobacterium-mediated transformation. Biotech. Lett. 29:965-969.   DOI
18 Enriquez-Obregon, G. A., Vazquez-Padron, R. I., Prieto-Samsonov, D. L. de la Riva, G. A. and Selman-Housein, G. 1998. Herbicideresistant sugarcane (Saccharum officinarum L.) plants by Agrobacterium-mediated transformation. Planta 205:20-27.
19 Flores Sols, J. L., Mlejnek, P., Studen, K. and Prochzka, S. 2003. Application of sonication-assisted Agrobacterium-mediated transformation in Chenopodium rubrum L. Plant Soil Environ. 49:255-260.   DOI
20 Floros, J. D., Wetzstein, H. Y. and Chinnan, M. S. 1987. Chemical (NaOH) peeling as viewed by scanning electron microscopy: pimiento peppers as a case study. J. Food Sci. 52:1312-1316.   DOI