The obtaining of transgenic potato Solanum tubersosum L. with high productivity by the
transfer of the gene ugt/iaglu for Zea mays L.
Salyaev, R.K.1, Rekoslavskaya, N.I.1, Mapelli, S.2, Korneva, A.V.1, Stepanova, E.G.1, Chepinoga,
A.V.1, and A.A. Truchin1
1Siberian Institute of Plant Physiology and Biochemistry, Siberian Branch of RAS, PO Box 1243,
Irkutsk, 664033, Russia, e-mail: phytolab@sifibr.irk.ru
2Istituto Biologia Biotecnologia Agraria, C.N.R., via Bassini 15, 20133 Milan, Italy, e-mail:
mapelli@ibba.cnr.it
Introduction
Potato is a very valuable food source for the Siberian region with short summer and
continental climate. To obtain transgenic plants with enhanced productivity, the target gene with
predictable significance such as ugt(iaglu) from Zea mays L. was used. The gene ugt is encoding
the synthesis of UDPG-transferase (IAA-glucose synthase) from maturing corn endosperm thereby
the stored form of the main phytohormone indoleacetic acid (IAA) accumulates in kernels and is
employed in growth stimulation during germination and growth of seedlings.
The goal of transgenesis was to create transgenic potato plants with high energy of growth,
enhanced productivity and new valuable features.
Materials and Methods
The gene ugt cloned under pT3 promoter in pBluescript of E. coli DH5a
was introduced by
the use of triparental mating with disarmed strain of A. tumefaciens 699 (EHA105 on the base C58)
with pCNL30 (nptII, gus under nos and 35S promoters, respectively) and E. coli K802 with pRT104
(Kozak sequence for improving eukaryotic translation, gus under 35S promoter). The presence of
marker gene nptII and target gene ugt in transconjugant cells was demonstrated by PCR with
suitable primers (not shown). For conjugation, fresh cultures of bacteria were placed on YPD agar
medium and kept for 3 days at 26° in an incubator. Then transconjugants were selected on the YPD
medium with 50 mg/l of kanamycin and 50 mg/l of ampicillin. Selected colonies were used for
transformation of potato by pricking of tubers with a needle coated with bacterial slurry.
Potato cv. Borodyanski was used for transformation in planta. Potato tubers infected in planta
were placed in greenhouse, phytotrone, or in field plot beds. The expression of the reporter gene
gus was confirmed by determination of specific activity of GUS (1). The integration and expression
of the selective gene nptII was determined by PCR (not shown) and by checking of chlorophyll after
treating of leaves with kanamycin solution (200 mg/l). To check the integration and expression of
target gene ugt, Southern blot hybridization, PCR and RT-PCR were employed. Southern blot
hybridization was carried out with ECL system Gene Image Random Prime Labeling Module and
Gene Images CDP-Star Detection Module (Amersham, UK)
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