biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 49:593-598, 2005 | DOI: 10.1007/s10535-005-0054-1

Transgenic tobacco plants carrying the non-structural P3 gene of potato virus A

S. Nováková1, Ą. Mazúrová1, N. Čeřovská2, Z. W. ©ubr1,*
1 Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovak Republic
2 Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Praha, Czech Republic

Transgenic tobacco (Nicotiana tobacum) plants carrying the gene coding for potato virus A (PVA) non-structural P3 protein were prepared by inoculation with Agrobacterium tumefaciens. Seeds from self-pollinated flowers (T1 generation) were collected. To estimate the effectiveness of vertical transfer of the introduced gene and usefulness of respective plant lines for further experiments, the T1 generation was characterized by testing its ability to grow in the presence of kanamycin (Km) and by PCR of both neomycin phosphotransferase (nptII) and PVA P3 genes. Eight and ten of 29 lines showed Mendelian segregation of Km-resistant phenotype 3:1 and ≥15:1, respectively, the T1 of eleven lines showed low Km resistance. Selected PCR-positive lines were tested for the presence of P3 mRNA. In most cases the transgene transcription was dependent on the presence or absence of Km in the plant growth medium. Prepared transgenic plants were furthermore tested for sensitivity to PVA and potato virus Y (PVY) infection. All of them showed identical symptom development as the non-transgenic control plants.

Keywords: potyvirus; Agrobacterium tumefaciens; Nicotiana tabacum; PCR; P3 protein; transcription
Subjects: Agrobacterium tumefaciens; germination; Nicotiana tabacum; nutrient medium MS (Murashige and Skoog); polymerase chain reaction (PCR); potato virus A, non-structural P3 gene; potyvirus; protein, P3; tobacco, transgenic plants; transgenic plants; virus

Received: September 10, 2004; Accepted: April 19, 2005; Published: December 1, 2005  Show citation

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Nováková, S., Mazúrová, Ą., Čeřovská, N., & ©ubr, Z.W. (2005). Transgenic tobacco plants carrying the non-structural P3 gene of potato virus A. Biologia plantarum, 49(4), 593-598. doi: 10.1007/s10535-005-0054-1
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