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P L A N T C E L L , O R G A N , A N D T IS S U E C O L L E C T IO N S , T H E M Е Т Н О D S О F

G E R M P L A SM P R E S E R V A T IO N

CRYOPRESERVATION:COMBINEDAPLICATIONOFMANNITOLAND

ABAFORPRE-GROWTHOFPLANTCELLSTRAINS

O.U.Baskakova, T.V.Nikishina (Korneeva), S.E.Zorinyants, A.S.Popov,

S.M.Adueva, L.N.Kulikova, E.A .Zhivukhina

К .A.Timiryazev Institute o fPlant Physiology, Russian Acad. ofSci.; V.l.LeninMoscow

State Theachers University, Moscow.

The most o f long term cultivated plant cell strains had loosed their

morpho- and embryogenetic capacities and their cryopreservation demanded

a specia l p re lim ina ry cu ltiva tion . The Rhs-2 and Rhs-8 cell lines o f

Rhaponticum carthamoides

were not alike a Rhs-1 strain that had been

successful cryopreserved early (Volkova L.A., Popov A.S.) with 7% DMSO

(dimethilsulfoxide) only without any special pre-growth. The Rhs-2 and Rhs-

8 strains are more sensitive to DMSO. They were successful cryopreserved

w ith cryoprotector m ixture: 2% DMSO + 5% trehalose + 4% glycerol after

prelim inary cultivation fo r 7 days with 6% mannitol and 7.5x10-5 M ABA in

combination. The ir survival was rised up to 44% and the growth o f these

strains was recovered a fte rthawing from liquid nitrogen. Pre-growth with the

mannitol only w ithout ABA was not effective fo r re-growth.

But the same precultivation with mannitol and ABA was not effective for

cell strain SAS-2n o f Beta vulgaris. Also cold hardening before deep freezing

was not effective. M itotic sinchronization o f SAS-2n cell culture have not

produced good results too. A vitrification (an ultra-rapid freezing) with PVS-

3 solution of Japan investigators after the above prelim inary cultivation and

cryoprotection did not provide any advantage over slowly program freezing:

the best survival was 1 5 -1 7% only. At the slow freezing the survival was 20

- 30% but the growth of this culture afterthaw ing from liquid nitrogen was not

renewed.

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