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Báo cáo khoa học: "propagation of several Betula species representing various ploidy levels"

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In vitro propagation of several Betula species representing various ploidy levels E. Särkilahti Department of Biology University of Turku, SF-20500 Turku, Finland and multiplication, was studied to Introduction improve survival in a greenhouse. One of the most important applications of in vitro culture is micropropagation, which enables vegetative propagation of adult Materials and Methods trees, not usually achieved with traditional methods. Genotypic variability in response In vitro cultures were initiated from dormant api- to successful in vitro propagation within a cal and axillary buds of 6 birch genotypes species has been a difficulty in finding uni- representing different species and ploidy levels versal propagation methods for some spe- (Table I). cies. This paper introduces the esta- Cultures were grown on MS medium (Mura- blishment of sterile shoot culture and shige and Skoog, 1962) having a modified macromineral composition (Simola, 1985), sup- micropropagation of 4 different mature plemented with 2.0 mg/l benzylaminopurine birch species having tetraploid and penta- (BAP), 0.01 mg,’I a-naphthalene acetic acid ploid chromosome sets. The effect of a (NAA) and casein hydrolysate (CH) under lanolin layer on relative humidity (RH) in a growth conditions described in detail by Sarki- lahti (1988). The same medium was used for culture vessel, as well as on shoot growth
both induction of adventitious buds and culture senting different Betula species and dif- of new shoots. Rooting medium had the same ploidy levels. New buds and shoots ferent composition but with 0.1 mg/i NAA as the sole were produced in a multiplication cycle of growth regulator. Rooted plantlets were trans- 3-4 wk from the cut ends of shoots trans- ferred into a peat/soil mixture (1:1) and acclima- ferred into fresh medium (Fig. 1Shoot tized in a greenhouse. Chromosome counts determined from root tips of 5 cultures of the tetraploid ssp. tortuosa and were plantlets/genotype to check the ploidy level. the tetraploid variety subcordata were lost The effects of a lanolin layer and lack of CH because of bacterial contamination. the multiplication rate and growth of the on Adventitious shoot induction and shoot shoots were studied separately. In the lanolin were accelerated on the medium growth experiment, culture media were covered with a lanolin layer of 1-2 mm. Shoots measuring 5 CH (Fig. 2). Rooting ability of the lacking mm with 2 nodes from sterile shoot cultures of genotypes cloned in vitro varied from 50 to the colchicine-tetraploid and irradiation-mutant 100% and the survival rate after transfer B. pendula were used to start the experiments. into soil was poor, around 50%. Lanolin Both the experiments and the controls con- sisted of 60 shoots in 10 culture vessels. After 4 did not reduce the RH, furthermore, it wk, the number of regenerated shoots/original severely retarded both shoot growth and shoot, their lengths and number of nodes were multiplication. RH was 97.1 % with and evaluated. 96.8% without lanolin at the beginning of the experiment. After 4 wk, RH was 95.1% with and 99.3% without lanolin. The tem- perature was 26 ± 1 °C during the experi- Results ment. All the genotypes, except the moun- tain birch hybrid, had the original ploidy level in the in vitro culture. Plantlets of the The medium (Sdrkilahti, 1988) allowed mountain birch hybrid showed variability in both the induction of adventitious buds their ploidy level: 2 tetraploids, 2 aneu- and the development of new shoots from ploids and 1 pentaploid were recorded. in vitro culture of 6 genotypes repre-
Discussion and Conclusion Acknowledgments The author thanks Dr. Terho Valanne for his Both genetic variation and environmental advice in preparing the manuscript and B.Sc. conditions are known to have an effect on Soile Kiilunen for technical assistance. This the responses of different genotypes to in study was financially supported by the Founda- vitro culture. The present results with posi- tion for Research of Natural Resources in Fin- tive responses of different Betula geno- land. types in a single culture medium would support the major role of environmental conditions and the minor one of genotype. References Thus, it should be possible to develop a universal in vitro culture medium for the genus Betula. Growth and multiplication T. & Skoog F. (1962) A revised Murashige can be further improved, as shown by medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473- omitting CH from the culture medium. 497 Three genotypes retained their ploidy Simola L.K. (1985) Propagation of plantlets levels during several subcultures. This from leaf callus of Betula pendula f. purpurea. suggests that the instability of chromo- Sci. Hortic. 26, 77-85 some number in plantlets of the mountain Sdrkilahti E. (1988) Micropropagation of a birch was due to the hybrid nature of the mature colchicine-polyploid and irradiation- mother tree rather than the culture condi- mutant of Betula pendula Roth. Tree PhysioL 4, 173-179 tions.

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