SUMMARYFrom cellulose and cellobiose the formation of sophorose, laminaribiose, and gentiobiose was catalyzed by ~'r~chode~a reesei culture filtrale containing exo-and endoglucanase and B-glucosidase activity and from cellobiose by a broken cell suspension from T. reesei with B-glucosidase activity. The results indicate that B-glucosidase is the component responsible for transglycosylation reaction catalyzed by T. reesei cellulase enz~ne complex.The normal enzymatic degradation products of cellulose by T~choderma ree~ei are glucose and cellobiose. However, the cellulase enzyme complex has been shown to posses also transglycosylation activity. Toda et ,:~i.(1968) have found that a mixture of pentasaccharides and higher oligosacchavides were formed from cellotetraose by purified cellulase preparation.According to Okada et al. (1975)

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Transgenic, fertile barley (Hordeum vulgare L.) from the Finnish elite cultivar Kymppi was obtained by particle bombardment of immature embryos. Immature embryos were bombarded to the embryonic axis side and grown to plants without selection. Neomycin phosphotransferase II (NPTII) activity was screened in small plantlets. One out of a total of 227 plants expressed the transferred nptII gene. This plant has until now produced 98 fertile spikes (T0), and four of the 90 T0 spikes analyzed to date contained the nptII gene. These shoots were further analyzed and they expressed the transferred gene. From green grains, embryos were isolated and grown to plantlets (T1). The four transgenic shoots of Toivo (the T0 plant) produced 25 plantlets as T1 progeny. Altogether fifteen of these T1 plants carried the transferred nptII gene as detected with the PCR technique, fourteen of which expressed the nptII gene. The integration and inheritance of the transferred nptII gene was confirmed by Southern blot hybridization. Although present as several copies, the transferred gene was inherited as a single Mendelian locus into the T2 progeny.

Somatic embryogenesis was induced in cell cultures of birch (Betula pendula Roth.) derived from juvenile tissue of seed embryos and from mature leaf tissue. Embryos were formed in liquid and on solidified medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-furfurylaminopurine (kinetin). Sometimes somatic embryos formed only after transfer to medium devoid of growth regulators. The embryos germinated on hormone-free medium and were potted in soil and grown in the greenhouse.

must be based on adequate knowledge of their population genetic parameters, especially gene flow between Genetic engineering is becoming a useful tool in the improvement different populations in cultivation. of plants and plant-based raw materials. Varieties with value-added traits are developed for nonfood use in industrial and medical produc-The first transgenic barley plants were produced by tion, and different production lines must be kept separate. For good particle bombardment (Hagio et al., 1995; Jä hne et al., management practices, knowledge of relevant gene flow parameters 1994;Wan and Lemaux, 1994). Later is required. In the present study, pollen-mediated dispersal of transon other techniques have also resulted in transgenic genes via cross-fertilization was examined. A transgenic barley (Horbarley plants (Funatsuki et al., 1995; Matthews et al., deum vulgare L.) line carrying a marker gene coding for neomycin 1997; Nobre et al., 2000; Salmenkallio-Marttila et al., phosphotransferase II (nptII ) was used as a pollen donor. For maxi-1995; Tingay et al., 1997; Zhang et al., 1999). The most mum resolution, a cytoplasmically male-sterile barley line was utilized reproducible gene transfer method in barley has been as recipient and the flow of nptII transgene was monitored at distances the bombardment of embryos of the variety Golden of 1, 2, 3, 6, 12, 25, 50, and 100 m from the donor plots of 225 and Promise (Wan and Lemaux, 1994). The majority of gene 2000 m 2 . Male-fertile plots at a distance of 1 m were included to measure the transgene flow in normal barley. The number of seeds transfers aiming at commercial applications have been obtained from male-sterile heads diminished rapidly with distance carried out by this method (e.g., Jensen et al., 1996; and only a few seeds were found at distances of 50 and 100 m. Molecu-Nuutila et al., 1999). lar genetic analysis (polymerase chain reaction-PCR) revealed that When varieties with novel traits are being developed, all seeds obtained from male-sterile heads at a distance of 1 m were safety questions should also be considered. In the retransgenic, as anticipated. However, only 3% of the distant seeds lease of transgenic plants, the effect of the gene is of (50 m) actually carried the transgene, whereas most of them resulted prime importance, not the way in which it was introfrom fertilization with nontransgenic background pollen. This backduced into the genome (EUCARPIA, 1989). The safety ground pollen was mainly due to pollen leakage in some male-sterile assessment should cover the whole chain from research heads. In normal male-fertile barley, the cross-fertilization frequency and production of transgenic plants to cultivation, pro-A. Ritala, A.M. Nuutila and V. Kauppinen, VTT Biotechnology, P.O. uted to the evolution of new cultivated plant species,