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Journal of Zhejiang University SCIENCE B 2008 Vol.9 No.10 P.823-828

http://doi.org/10.1631/jzus.B0860006


Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum


Author(s):  Ji-ye WANG, Hong-ye LI

Affiliation(s):  Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China

Corresponding email(s):   hyli@zju.edu.cn

Key Words:  Penicillium digitatum, Agrobacterium tumefaciens-mediated transformation (ATMT), Hygromycin B resistance gene, Insertional mutagenesis


Ji-ye WANG, Hong-ye LI. Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum[J]. Journal of Zhejiang University Science B, 2008, 9(10): 823-828.

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Abstract: 
Agrobacterium tumefaciens-mediated transformation (ATMT) system was assessed for conducting insertional mutagenesis in Penicillium digitatum, a major fungal pathogen infecting post-harvest citrus fruits. A transformation efficiency of up to 60 transformants per 106 conidia was achieved by this system. The integration of the hph gene into the fungal genome was verified by polymerase chain reaction (PCR) amplification and sequencing. These transformants tested were also shown to be mitotically stable. Southern blot analysis of 14 randomly selected transformants showed that the hph gene was randomly integrated as single copy into the fungal genome of P. digitatum. Thus, we conclude that ATMT of P. digitatum could be used as an alternatively practical genetic tool for conducting insertional mutagenesis in P. digitatum to study functional genomics.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1] An, G., Ebert, P.R., Mitra, A., Ha, S.B., 1988. Binary Vectors. In: Gelvin, S.B., Schilperoort, R.A. (Eds.), Plant Molecular Biology Manual. Kluwer Academic Publishers, Great Britain, p.1-19.

[2] Bowyer, P., 2001. DNA-mediated Transformation of Fungi. In: Talbot, N. (Ed.), Molecular and Cellular Biology of Filamentous Fungi. Oxford Univ. Press, Oxford, p.33-46.

[3] Bundock, P., Dulk-Ras, A., Beijersbergen, A.G.M., Hooykaas, P.J.J., 1995. Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Ssaccharomyces cerevisiae. EMBO J., 14(13):3206-3214.

[4] Bush, D.A., Codner, R.C., 1968. The nature of macerating factor of Penicillium digitatum saccardo. Phytochemistry, 7(5):863-869.

[5] Hamamoto, H., Nawata, O., Hasegawa, K., Nakaune, R., Lee Y.J., Makizumi, Y., Akutsu, K., Hibi, T., 2001. The role of the ABC transporter gene PMR1 in demethylation inhibitor resistance in Penicillium digitatum. Pest. Biochem. Physiol., 70(1):19-26.

[6] Holmes, G.J., Eckert, J.W., 1999. Sensitivity of Penicilium digitatum and P. italicum to postharvest citrus fungicides in California. Phytopathology, 89(9):716-721.

[7] Hooykaas, P.J.J., Roobol, C., Schilperoort, R.A., 1979. Regulation of the transfer of Ti-plasmids of Agrobacterium tumefaciens. J. Gen. Microbiol., 110(1):99-109.

[8] Irie, T., Honda, Y., Watanabe, T., Kuwahara, M., 2001. Efficient transformation of the filamentous fungus Pleurotus ostreatus using single-strand carrier DNA. Appl. Microbiol. Biotechnol., 55(5):563-565.

[9] Malonek, S., Meinhardt, F., 2001. Agrobacterium tumefaciens mediated genetic transformation of the phytopathogenic ascomycete Calonectria morganii. Curr. Genet., 40(2):152-155.

[10] Morioka, L.R.I., Furlaneto, M.C., Bogas, A.C., Pompermayer, P., Duarte, R.T.D., Vieira, M.L.C., Watanabe, M.A.E., Fungaro, M.H.P., 2006. Efficient genetic transformation system for the ochratoxigenic fungus Aspergillus carbonarius. Curr. Microbiol., 52(6):469-472.

[11] Mullins, E.D., Chen, X., Romaine, P., Raina, R., Geiser, D.M., Kang, S., 2001. Agrobacterium tumefaciens-mediated transformation of Fusarium oxysporum: an efficient tool for insertional mutagenesis and gene transfer. Phytopathology, 91(2):173-180.

[12] Nakaune, R., Adachi, K., Nawata, O., Tomiyama, M., Akutsu, K., Hibi, T., 1998. A novel ATP-binding cassette transporter involved in multidrug resistance in the phytopathogenic fungus Penicillium digitatum. Appl. Environ. Microbiol., 64(10):3983-3988.

[13] Nakaune, R., Hamamoto, H., Imada, J., Akutsu, K., Hibi, T., 2002. A novel ABC transporter gene, PMR5, is involved in multidrug resistance in the phytopathogenic fungus Penicillium digitatum. Mol. Genet. Genomics, 267(2):179-185.

[14] Raeder, U., Broda, P., 1985. Rapid prepraration of DNA from filamentous fungi. Lett. Appl. Microbiol., 1(1):17-20.

[15] Rooney, P.J., Sullivan, T.D., Klein, B.S., 2001. Selective expression of the virulence factor BAD1 upon morphogenesis to the pathogenic yeast form of Blastomyces dermatitidis: evidence for transcriptional regulation by a conserved mechanism. Mol. Microbiol., 39(4):875-889.

[16] Sambrock, J., Fritsch, E.F., Maniatis, T., 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

[17] Smilanick, J.L., Mansour, M.F., Sorensen, D., 2006. Pre- and postharvest treatments to control green mold of citrus fruit during ethylene degreening. Plant Dis., 90(1):89-96.

[18] Song, A.H., Li, H.Y., Liu, X.H., 2004. Isolation and regeneration of protoplasts from Penicillium digitatum. Chin. J. Agric. Biotechnol., 1(3):197-202.

[19] Zhang, J.X., 2007. The potential of a new fungicide fludioxonil for stem-end rot and green mold control on Florida citrus fruit. Postharvest Biol. Technol., 46(3):262-270.

[20] Zhu, J.W., Xie, Q.Y., Li, H.Y., 2006. Occurrence of imazalil-resistant biotype of Penicillium digitatum in China and the resistant molecular mechanism. J. Zhejiang Univ. Sci. A, 7(2):362-365.

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