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H E A L T H Y P L A N T S A N D M IC R O P R O P A G A T IO N

PCR-ANALYSISOFGENOMEOFPOTATE,CULTIVATED

IN VITRO

FORA

LONGTIME

G.A.Gerashenkov, A.G.Mardamshin, G.G.Sharafutdinova

Biochemistry andCytochemistryDepartment, Ufa Science Center, RussianAcademy o f

Sciences, Prospect Octyabrya 69, Ufa, Russia, 450054, E-Mail:

The problem o f genome stability in the process of prolonged clonal

micropropagation o f plants is insufficiently studied. PCR-analysis is one of

the perspective approaches in study of plant genome polymorphism. RAPD-

method based on the use of arbitrary primers is a variant of PCR-analysis.

The essential moment for polymorphism detection is the choice of optimal

primers. It is known that short oligonucleotides 8-12 bp long may give artefacts

owing to the excess o f regions of non-specific binding.

More extended gene specific primers 19-30 bp long don’t have this

shortcoming, however, the polymorphism revealed in this case is insignificant

as a rule that is why the primers 15-18 bp long are optimal fo r such research.

The aim of the given work is the comparative analysis of amplification

products of DNA total preparation of potate plants o f Nevsky variety cultured

in vitro

fo r 9 years and non-cultured

in vitro

potate plants o f the same variety.

DNA was isolated according to Graham method, the following primers were

u sed

R A P D -P C R

re a c tio n

G AG G G TG G CG G T TC T ,

GAGGGAGAGAGGGAG , TAGGGAACGTGAGCTGGG . Amplification

products were fractionated in 0,8% agarose gel and analysed after staining

w ith ethidium bromide. The obtained results showed polymorphism in the

compared samples. It may be the result o f somatic mutations in some loci

o f the studied genome which took place in the process of cultivation

in vitro

and/or autoselection processes which lead to the selection of definite lines

different from the initial forms.

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