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PLANT GENETIC TRANSFORMATION, CELLSELECTION AND SOMATICHYBRIDIZATION
GENETICTRANSFORMATIONOF
TRITICUM AESTIVUM L .
BYUSE
HAPLOIDCELLS
B. Anapiayev, O.Blokhina,N.Yevdakova, A.Kaliyev, S.Zairov
TheM.A . Aytkhohzin Institute o fmolecular biology & biochemistry, the Republic o f
Kazakhstan, Almaty, 480012, Dosmukhamedov st., 86.
The creation o f transgenic plants is a widespread method fo r getting
the plant sorts which are the resistant forms to an unfavourable environmental
factors as well as the highly productive sorts and forms o f the important
agricultural plants. Formerly we have obtained the transgenic plants of
hexaploid wheat which were transformed using the binary vector system of
the
Agrobacterium
plasmids, when we have demonstrated the integration of
a foreign genes into genom of the transformed plants (Kaliyev, Kornienko
1991,1992).
Howeverw e th in k that using the haploid cells, which are obtained from
the culture of the
in vitro
isolated microspores, is more interesting and more
perspectively fo r the theoretical and practical aspects o f the genetic
transformation by use a foreign genes. A fter getting the transgenic plants
we'll research the reveal of the influence o f a gen engineering manipulations
on a phenotype, on a molecular-biochemistry indexes, on a level of harvest
and on a technologycal quality of grain.
The haploid cells have been obtained from the culture of the isolated
microspores on the modificated by us culture media N6 (1 mg/l 2,4-D). The
generative cells were cultivated on the stage of the vesicular microspore
after the precultivation by cold (4-7C) in the temperature of 27C on dark. As
far as appearance o f a androclinic structures (embryoedes, calluses, globules)
they were transplanted to the modificated cultural media MS (2mg/l 2,4-D or
0,5 mg/l IAA) and were cultivated on light. The obtained regenerates have
been used for the genetictransformation.
The basal part of regenerates have been plased in the solid nourishing
cultural media that contains the MS salts, vitam ines, the 3% sugar,
acetoseringon (AS) and 2-4 mg/l 2,4-D, pH 5,5 - 5,7 and they have been
cultivating for 2 days in the temperature of 25C. After this the regenerates
were inoculated with the cells suspension o f
Agrobacterium
and were plased
on the same cultural media and they were incubating for 2 days. The obtained
sprouts have been transplanted to the fresh nourishing media without AS,
but with the addition of the kanamicin antibiotik (the concentration of 50 mg/
I) and klaforan (300 mg/l).
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