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Regeneration and Agrobacterium-mediated
transformation studies in carnation (Dianthus
caryophyllus L. cv. Turbo)
Ş. E. Arici1* and N. K. Koç2
1University
of Süleyman Demirel, Faculty of Agriculture, Department of
Plant Protection, Çünür/Isparta.
2University
of Çukurova, Faculty of Agriculture, Department of Plant
Protection Balcalı/Adana.
*Corresponding author. E-mail:
evrima@ziraat.sdu.edu.tr.
Abbreviations:
BA,
benzyladenine; GUS, β-glucuronidase; IBA,
indole-3-butyric acid; MS, Murashige and Skoog
nutrient medium; NAA, naftalin acedic acid,
npt II,
neomycin phosphotansferase II,
PCR,
polymerase chain reaction;
SPSS,
Statistical Package for the Social Sciences,
X-gluc,
5-bromo-4-chloro-3-indolyl-D-glucuronide.
Accepted 28 August, 2009 |
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Leaf
explants of carnation (Dianthus caryophyllus L. cv.
Turbo) were used for the transformation of gene performed by
the EHA 105 strain of Agrobacterium tumefaciens
harboring the binary vector, pGA482GG. This vector carries
the marker genes, neomycin phosphotansferase II (npt II)
that determine resistance to kanamycin and
β-glucoronidase
(GUS). Leaf segments of carnation plants were cultured on MS
medium containing different combinations and concentrations
of hormones. The best shoot regeneration was observed on MS
medium with 1 mg/L BAP+ 0.05 mg/L NAA. Initiation of root
formation was performed on MS medium containing 0.5 mg /L
IBA. Transformation was done by the EHA 105 strain of A.
tumefaciens, after determining the appropriate medium
and the plant tissue for the transformation. Plant tissues,
selected on MS medium containing kanamycin, were tested by
isolating them from transgenic plant tissues and shoots
regenerated from these transgenic plants. PCR were done to
indicate transformed GUS gene; 660 bp specific DNA bands of
GUS were observed. In this work, an appropriate regeneration
system for later studies on carnation and an efficient
technique for the transformation of important genes that can
resist diseases, using A. tumefaciens were developed.
Key words:
Agrobacterium tumefaciens, carnation (Dianthus
caryophyllus L. cv. Turbo), gene transformation, plant
regeneration. |
