Stable transformation of embryogenic tissues of Pinus nigra Arn. using a biolistic method

Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademická 2, P.O. Box 39 A, 950 07, Nitra 1, Slovak Republic.
Biotechnology Letters (Impact Factor: 1.59). 08/2005; 27(13):899-903. DOI: 10.1007/s10529-005-7178-4
Source: PubMed


The stable transformation of embryogenic tissues of Pinus nigra Arn., cell line E104, has been achieved using a biolistic approach. The introduced DNA consisted of the uidA reporter gene under the control of the double CaMV 35S promoter and the nptII selection gene controlled by the single CaMV 35S promoter. Three days after bombardment, putative transformed tissues were selected for continued proliferation on a medium containing 20 mg geneticin l(-1). Resistant embryogenic tissue recovery required 10-12 weeks. The integration of the nptII and uidA genes was confirmed by both histochemical/fluorimetric GUS assays and PCR amplification of the inserts in the five geneticin resistant sub-lines of line E104. The activity of the uidA reporter gene in transgenic, embryogenic tissue lines was stable and could be detected after one year of culture. Somatic embryo maturation was, however, poor and no plantlet regeneration could be obtained.

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    • "AT, LISW, PB S Stomp et al. 1990; Newton et al. 1993; Tang et al. 2006b griffithii PB T Fernando et al. 2000 halepensis AR S Tzfira et al. 1996 jeffreyi AT Gall Stomp et al. 1990 lambertiana AT S Loopstra et al. 1990 maximartinezii AT S Villalobos-Amador et al. 2002 monticola AR, PB T McAfee et al. 1993; Fernando et al. 2000 nigra AR, AT, EP, PB S Mihaljevic et al. 1996; Lopez et al. 2000a; Salaj et al. 2005 palustris AR, AT, PB T Diner 1999 patula PB SP Nigro et al. 2004 pinaster AT, EP, PB SP Gomez-Maldonado et al. 2001; Trontin et al. 2002 pinceana AT S Villalobos-Amador et al. 2002 pinea AT, PB, SAAT T Humara et al. 1999a, 1999b ponderosa AT Gall Morris et al. 1989; Stomp et al. 1990 radiata AT, EP, PB SP Stomp et al. 1990; Campbell et al. 1992; Walter et al. 1998; Charity et al. 2005 rigida x taeda AT, PB SP Connett-Porceddu et al. 2003, 2007 roxburghii PB SP Parasharami et al. 2006 strobus AT, PB SP Tian et al. 1997; Levée et al. 1999 sylvestris AT, AR, PB SP Stomp et al. 1990; Aronen et al. 2003 taeda AT, PB, SAAT SP Sederoff et al. 1986; Tang et al. 2001; Connett-Porceddu et al. 2003, 2007 taeda x elliottii AT Gall Huang and Tauer 1994 thunbergii PB T Taniguchi et al. 2004 virginiana AT, PB SP Stomp et al. 1990; Tang and Newton 2004b, 2005 the genome by a variety of recombination events. In the AT method, the gene(s) of interest are inserted into the transfer- DNA (T-DNA) region of tumor-inducing plasmid (pTi) from a disarmed A. tumefaciens strain. "
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