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M O L E C U L A R B IO L O G Y OF P L A N T C E L L S IN V IT R O

M O L E C U L A R A N A L Y SIS O F G E N O M E V A R IA B IL IT Y IN S O M A T IC H Y B R ID S

T.P.Suprunova, E.Z.Kochieva*, M.K.Lapteva, G.E.Pozmogova**, D.B.Dorokhov

Instilute o f VegetableBreeding and SeedProduction, VNIISSOK, MoscowDistrict,

Russia. E-Mail vniissok@cea. tv.

*Center "Bioengineering", Moscow, Russia

*TimiryazevAgriculturalAcademy, Russia.

Recently, we applied the rapid random amplified polymorphic DNA

polymerase chain reaction (RAPD) technique to fingerprint genomes of various

varieties and wild plants (D. Dorokhov and E. Klocke 1997, Rus. J. Genetics).

This technique we successful used for genome analysis of wide number of

different somatic hybrids obtained in various research groups: interspecific

somatic hybrids of brassicas (Institute for Breeding of Vegetable, Medicinal and

Spice Plants, Quedlinburg, Germany), interspecific somatic hybrids of potatoes

(Institute for Breeding of Crop Plants, Gross Lusewits, Germany), asymmetric

interspecific somatic hybrids of tomatoes (Institute of Cell Biology and Genetic

Engineering o fNASU, Kiev, Ukraine), intergeneric somatic hybrids of

Lycopersicon

and nontuberous potato species (N.l. Vavilov Institute of Plant Industry,

St.Petersburg, Russia).

Comparative RAPD analysis of the hybridization partners and somatic

hybrids has revealed the following, nearly all somatic hybrids reveal partial loss

of amplified DNA fragments typical for RAPD patterns of parental plants and

induction of new bands, not typical for hybridization partners ("non parental"

bands).

For more detail investigation phenomena of "non parental" bands and to

find out a nature of new bands in RAPDspectra of plant DNA, we conducted a

number of experiments with fertile asymmetric interspecific somatic hybrid

Lycopersicon esculentum Mill.

Lycopersicon peruvianum var. dentatum Dun.

and F1-F2 hybrid progeny (Ratushnyake tal. 1993, MGG; Cherepe tal. 1995,

Rus.

J. Genetics). Formation of new "non parental" bands in RAPD spectra is

Induced by fivefromsixprimers tested. One ofthe "non parental" band (580 b.p.),

which observed inR0 and

F1 RAPD

spectra and non detected in

and parental

RAPD

patterns, was cloning in pGem-T and sequenced. The determined

sequence was compared with the

EMBL

database. The deduced sequence has

high

level of homology to catehepsin

B-like

cysteine proteinase from

Nicotiana

rustica.

the base of Southern blotting experimentswe found that, "non parental"

band (580

b.p.) did not cross-hybridizewith

and parental

RAPD

patterns,

but

hybridize

with genomic

DNA of

parental

plants,

somatic hybrid and F1 -

hybrid

progeny. Others

"parental"

and

"non parental"

bands was investigated too. The

hypothesis

of correlation

"non parental’ bands

with process of reparation and

recombination are discussed.

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