“blank” bacterial vector alone. PCR product of whole sequence of the gene ugt was about of 1800
bp (Figure 1, D).
Figure 1. PCR (A,D), Southern blot hybridization (B) and RT-PCR (C) of the gene ugt on potato
genomic DNA and total RNA.
A – 1 - standard DNA (100 bp), 2 – initial nontransformed tuber, 3 – nontransformed sprout, 4 –
sprout from transformed tuber, 5 – pBluescript with the gene ugt (234 bp fragment from 567 to 801
nucleotides);
B - 2 – initial nontransformed tuber, 3 – nontransformed sprout, 4 – sprout from transformed tuber,
5 – pBluescript with the gene ugt (234 bp fragment from 567 to 801 nucleotides);
C - RT-PCR with primers to the whole sequence of the gene ugt (1800 bp),
1,2 and 3 - independent transgenic potato plants, 4 – nontransformed plant, 5 – transformed with
blank A.t. vector C58;
D – PCR on plasmid DNA of pBluescript harboring the gene ugt with primers to the whole sequence
of 1800 bp of the gene ugt.
By using developed methods of plant infection with transconjugants created from triparental
mating we obtained the transgenic potato with increased growth and increased productivity. The
presence in transconjugant plasmid, the integration and the expression of marker and target genes
in transgenic potato were confirmed by PCR, Southern blot hybridization and RT-PCR (Figure 1)
and by measuring of the activity of GUS and NPTII (Table 1). Transgenic potato revealed high
energy of growth because of elevated levels of free and bound IAA which was due to UDPG-
transferase (IAA-glucose synthase) encoded by gene ugt transferred from corn (Table 4).
Transgenic plants developed large total leaf surface (Tables 2 and 4) because of high level of IAA.
Transpiration and photosynthesis were higher in transgenic plants (Table 5) which correlated with
the harvest of transgenic potato, the highest per plant in most cases in comparison to
nontransformed in 2000-2001 years (Tables 6).
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