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Although seeds are mainly used for food or feed, they are also used as raw materials for various other industrial applications. Improvement of the nutritional quality of protein in legumes and cereals has been the prime aim of plant breeders. Crop breeding has undergone major changes during the last decades and new selection strategies are based on sophisticated genetics, the use of computational power, and biometric methods in field trials. On the other hand, the processing quality of seeds can be modified by altering the structural grain constituents or the enzyme activities that mobilize storage reserves of the seeds. Recent developments in the molecular biology of seed proteins offer breeders new tools for this work.

The use of recombinant DNA-based protein production using genetically modified plants could provide a reproducible, consistent quality, safe, animal-component free, origin-traceable, and cost-effective source for industrial proteins required in large amounts (1000s of metric tons) and at low cost (below US$100/Kg). The aim of this work was to demonstrate the feasibility of using barley suspension cell culture to support timely testing of the genetic constructs and early product characterization to detect for example post-translational modifications within the industrial protein caused by the selected recombinant system. For this study the human Collagen I alpha 1 (CIa1) chain gene encoding the complete helical region of CIa1 optimized for monocot expression was fused to its N- and C-terminal telopeptide and to a bacteriophage T4 fibritin foldon peptide encoding sequences. The CIa1 accumulation was targeted to the endoplasmic reticulum (ER) by fusing the CIa1 gene to an ER-directing signal peptide sequence and an ER retention signal HDEL. The construct containing the CIa1 gene was then introduced into immature barley half embryos or barley cells by particle bombardment. Transgenic barley cells resulting from these transformations were grown as suspension cultures in flasks and in a Wave bioreactor producing CIa1 similar to CIa1 purified from the yeast Pichia pastoris based on Western blotting, pepsin resistance, and mass spectroscopy analysis. The barley cell culture derived-CIa1 intracellular accumulation levels ranged from 2 to 9 μg/l illustrating the need for further process improvement in order to use this technology to supply material for product development activities.

The F4-positive enterotoxigenic Escherichia coli (ETEC) strains are a frequent cause of porcine post-weaning diarrhea. Orally administered F4 fimbriae or FaeG, the major subunit and adhesin of F4, induce a protective mucosal immune response in F4 receptor-positive piglets. Feed plants carrying immunogenic subunit proteins can offer great advantages for oral vaccination of domestic animals. Here, we describe high-level endosperm-specific production (1% of total soluble proteins) of FaeG in the crop plant barley. The endoplasmic reticulum-targeted recombinant endospermic FaeG (erFaeG) was shown to be heterogeneously glycosylated. The erFaeG showed resistance at digestive conditions simulating piglet gastric fluid. Glycosylation did not abolish the immunogenic character of the FaeG protein, since erFaeG was able to induce F4 fimbria-specific antibodies in mice. Biological activity of these anti-F4 antibodies was demonstrated in vitro by blocking the attachment of the F4+ ETEC to the F4 receptors present on porcine intestinal enterocytes.

Homozygosity was induced in transgenic barley by microspore culture. Spikes of transgenic barley plants carrying microspores in the late uni-nucleate stage were cold pretreated. Teflon rod maceration and a density of 100 000 viable micropores per plate were used. The developed calli were regenerated and plantlets were treated with colchicine. The microspore culture of 16 mother plants (three transgenic lines) resulted in 927 green regenerants. Of these plants, 476 were transferred to soil, 380 were transgenic, 358 reached maturity and 350 were fertile with a normal seed-set carrying a yield of 6.9 kg. A production efficiency of 0.8 fertile transgenic doubled haploid barley plants per spike used for microspore isolation was recorded. The produced transgenic seeds were used in malting experiments.

An extensive study on the effects of blanching/freezing and long-term freezer storage on various bioactive compounds of more than 20 commonly used vegetables was performed. Effects were strongly plant species-dependent. Contents of dietary fibre components either were not affected or increased slightly. Minerals in general were also stable, but some losses of soluble minerals by leaching were observed. Phenolic antioxidants and vitamins were clearly more sensitive. Significant losses (20-30%) of antioxidant activity and total phenolics were detected in many vegetables. A qualitative HPLC profiling method for phenolic antioxidants was developed which proved to be very useful when evaluating the complex behaviour of phenolics during food processing. Up to one-third of vitamin C contents were lost during blanching, and further slight losses were detected during storage. Folic acid turned out to be very sensitive to blanching, with more than half of the vitamin being lost, but was stable during freezer storage. Carotenoids and sterols were not affected by blanching or freezer storage. The usefulness of the applied screening methods for evaluation of the effects of processing on vegetables is shown.  2003 Society of Chemical Industry

For the quantitative determination of individual flavonoid glycosides in plant materials, the glycosides are normally hydrolysed and the resulting aglycones are identified and quantified. However, the hydrolysis conditions which result in optimal breakdown of glycosides are too harsh for some of the other phenolic compounds present in the same plant material. Therefore, the effects of different hydrolysis conditions and different antioxidants on pure flavonoid glycones and aglycones were studied. On the basis of the results obtained with standards, suitable hydrolysis methods for red spring onion and spinach were developed. The best results from these vegetables were obtained by refluxing at 80 °C for 2 h with 1.2 M HCl in 50% aqueous methanol with addition of 2 mg ascorbic acid as an antioxidant. The method developed in this study is suited to the screening of flavonoids in vegetables and leafy vegetables.

The processing quality of cereals can be modified by altering the structural grain constituents or the enzyme activities that mobilize storage reserves of the seeds. In order to complement the malt enzyme spectrum, a gene encoding for a thermotolerant fungal endo-(1,4)-ß-glucanase was introduced into two barley cultivars, Kymppi and Golden Promise. The gene was expressed in the seeds during germination, thus providing a thermotolerant enzyme that is active under mashing conditions. The amount of thermotolerant ß-glucanase produced by the seeds (ca. 0.025% soluble seed protein) has been shown to be sufficient to reduce wort viscosity by decreasing the soluble ß-glucan content. For the safe commercial cultivation of transgenic plants risk assessment of their cultivation is needed. In our study experimental estimates of the transgene flow from transgenic barley by pollen dispersal were produced. Field trials were conducted during the summers of 1996 and 1997. A transgenic barley line homozygous for the gene encoding for neomycin phosphotransferase was used as a source of pollen and male-sterile barley lines as recipients. In order to be able to transform the cross-fertilization frequencies to corresponding values of normal male-fertile barley, plots of normal barley were also included in the experimental plan. On the basis of our study, cross-fertilization in male-sterile recipient barley is possible with very low frequency up to 50 meters from the donor area. However, the frequency dramatically decreases with distance and due to self-pollination the possibility of cross-fertilization remains very low in normal cultivated barley.

By using a batch in vitro anaerobic fecal fermentation model, we have shown that the fecal microflora can rapidly deconjugate rutin, isoquercitrin, and a mixture of quercetin glucuronides. High levels of beta,D-glucosidase, alpha,L-rhamnosidase, and beta,D-glucuronidase were present. Rutin underwent deglycosylation, ring fission, and dehydroxylation. The main metabolite, 3,4-dihydroxyphenylacetic acid, appeared rapidly (2 h) and was dehydroxylated to 3-hydroxyphenylacetic acid within 8 h. The pattern of in vitro fermentation of rutin was not changed by changing the pH (6.0 or 6.9), fermentation scale (10 or 1000 mL), or donors of the inoculum. Hydroxyphenylacetic acids were not methylated by colon flora in vitro. The colonic microflora has enormous potential to transform flavonoids into lower molecular weight phenolics, and these might have protective biological activities in the colon. The site of absorption of flavonoids and the form in which they are absorbed are critical for determining their metabolic pathway and consequent biological activities in vivo.

Genetic engineering is becoming a useful tool in the improvement of plants and plant-based raw materials. Varieties with value-added traits are developed for nonfood use in industrial and medical production, and different production lines must be kept separate. For good management practices, knowledge of relevant gene flow parameters is required. In the present study, pollen-mediated dispersal of transgenes via cross-fertilization was examined. A transgenic barley (Hordeum vulgare L.) line carrying a marker gene coding for neomycin phosphotransferase II (nptII) was used as a pollen donor. For maximum resolution, a cytoplasmically male-sterile barley line was utilized as recipient and the flow of nptII transgene was monitored at distances of 1, 2, 3, 6, 12, 25, 50, and 100 m from the donor plots of 225 and 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 obtained from male-sterile heads diminished rapidly with distance and only a few seeds were found at distances of 50 and 100 m. Molecular genetic analysis (polymerase chain reaction-PCR) revealed that all seeds obtained from male-sterile heads at a distance of 1 m were transgenic, as anticipated. However, only 3% of the distant seeds (50 m) actually carried the transgene, whereas most of them resulted from fertilization with nontransgenic background pollen. This background pollen was mainly due to pollen leakage in some male-sterile heads. In normal male-fertile barley, the cross-fertilization frequency with transgenic pollen varied from 0 to 7% at a distance of 1 m, depending on weather conditions on the heading day. We conclude that, because of competing self-produced and nontransgenic background pollen, the possibility of cross-pollination is very low between a transgenic barley field and an adjacent field cultivated with normal barley. However, adequate isolation distances and best management practices are needed for cultivation of transgenic barley.

In recent years particle bombardment has become the most used method for gene transfer to barley (Hordeum vulgare L.). Transformation efficiency depends greatly on the ability of the target material to regenerate into green plants. In this work we improved the regeneration efficiency of polyembryogenic cultures of barley (cv. Kymppi) by optimizing the nitrogen and copper concentrations in the media.

The malting quality of two barley cultivars, Kymppi and Golden Promise, was modified to better meet the requirements of the brewing process. The egl1 gene, coding for fungal thermotolerant endo-1,4-beta-glucanase (EGI, cellulase), was transferred to the cultivars using particle bombardment, and transgenic plants were regenerated on bialaphos selection. Integration of the egl1 gene was confirmed by Southern blot hybridization. The transgenic seeds were screened for the expression of the heterologous EGI. Under the high-pI alpha-amylase promoter, the egl1 gene was expressed during germination. The heterologous enzyme was thermotolerant at 65 degrees C for 2 h, thus being suitable for mashing conditions. The amount of heterologous EGI produced by the seeds (ca. 0.025% of soluble seed protein), has been shown to be sufficient to reduce wort viscosity by decreasing the soluble beta-glucan content. A decrease in the soluble beta-glucan content in the wort improves the filtration rate of beer.

The effects of salinity, nitrate concentration, pH and temperature on eicosapentaenoic acid (EPA) production by Porphyridium purpureum were studied. Both systematic and stepwise optimization were used in order to determine the best conditions for production of EPA. An analysis method for fatty acids was also developed. The highest EPA production per cell was achieved at pH 7.6 with 41.8 g l −1 NaCl and 12.1 mM nitrate in the production medium and using a growth temperature of 8°C.

A high reproducibility of batch growth of Catharanthus roseus could only be achieved by using identical inoculum material. It was found that the initial growth rate of the cells increased with increasing inoculation density, whereas the maximum growth rate was not affected. Withdrawal of the conditioned medium from the inoculum resulted in a strong decline in the initial growth rate, indicating that conditioning factors affected the initial growth rate. Neither increasing the air flow rate from 0.5 to 1.5 volume of air per volume of broth-min−1 (vvm) nor CO2 enrichment of the aeration gas had any effect on the growth of the cells.

The nutrient uptake of an embryogenic and of a non-embryogenic cell line of birch (Betula pendula Roth.) during cell growth and embryo production was studied in suspension culture. The embryogenic and non-embryogenic cell suspensions grew differently in the same medium. The non-embryogenic cell line started to grow without any lag period after the inoculation. It rapidly hydrolyzed sucrose in the medium to glucose and fructose and consumed the glucose as carbon source. The concentration of fructose in the medium decreased only after the depletion of glucose. The embryogenic cell line also rapidly hydrolyzed the sucrose to glucose and fructose, but the monosaccharides were consumed only after the embryos started to germinate after three weeks of culture. Both monosaccharides were then taken up at the same rate.

The vhb gene encoding Vitreoscilla haemoglobin (VHb) was transferred to barley with the aim of studying the role of oxygen availability in germination and growth. Previous findings indicate that VHb expression improves the efficiency of energy generation during oxygen-limited growth, and germination is known to be an energy demanding growth stage during which the embryos also suffer from oxygen deficiency. When subjected to oxygen deficiency, the roots of vhb-expressing barley plants showed a smaller increase in alcohol dehydrogenase (ADH) activity than those of the control plants. This indicates that VHb plants experienced less severe oxygen deficiency than the control plants, possibly due to the ability of VHb to substitute ADH for recycling NADH and maintaining glycolysis. In contrast to previous findings, we found that constitutive vhb expression did not improve the germination rate of barley kernels in any of the conditions studied. In some cases, vhb expression even slowed down germination slightly. VHb production also appeared to restrict root formation in young seedlings. The adverse effects of VHb on germination and root growth may be related to its ability to scavenge nitric oxide (NO), an important signal molecule in both seed germination and root formation. Because NO has both cytotoxic and stimulating properties, the effect of vhb expression in plants may depend on the level and role of endogenous NO in the conditions studied. VHb production also affected the levels of endogenous barley haemoglobin, which may explain the relatively moderate effects of VHb in this study.

Transgenic plants are attractive bioreactors to large-scale production of recombinant proteins because of their relatively low cost. This study reports for the first time the use of transgenic plants to reduce enterotoxigenic Escherichia coli (ETEC) excretion in its natural host species. The DNA sequence encoding the major subunit and adhesin FaeG of F4+ ETEC was transformed into edible alfalfa plants. Targeting of FaeG production to chloroplasts led to FaeG levels of up to 1% of the total soluble protein fraction of the transgenic alfalfa. Recombinant plant-produced FaeG (pFaeG) remained stable for 2 years when the plant material was dried and stored at room temperature. Intragastric immunization of piglets with pFaeG induced a weak F4-specific humoral response. Co-administration of pFaeG and the mucosal adjuvant cholera toxin (CT) enhanced the immune response against FaeG, reflected a better induction of an F4-specific immune response. In addition, the intragastric co-administration of CT with pFaeG significantly reduced F4+ E. coli excretion following F4+ ETEC challenge as compared with pigs that had received nontransgenic plant material. In conclusion, transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against F4+ ETEC infections.

The Vitreoscilla hemoglobin gene (vhb) was introduced into Hyoscyamus muticus with the aim of investigating its effect on growth and alkaloid production of Agrobacterium rhizogenes-induced hairy root cultures. We were able to generate several VHb-expressing hairy root lines with different integration patterns. Substantial somaclonal variation was observed in growth and hyoscyamine production amongst both VHb-expressing lines and controls. Despite this variation, the growth properties of single lines remained stable over time. Expression of VHb was found to improve growth of H. muticus hairy roots in shake-flask cultures. The dry weights of the root cultures expressing Vitreoscilla hemoglobin were on average 18 % higher than those of the controls. VHb expression also increased the volumetric hyoscyamine production, mainly due to the improved growth properties. However, this difference was not statistically significant due to the wide somaclonal variation and fluctuations over time in both VHb and control hairy root lines.

The health benefits of oat (Avena sativa L.) are mainly associated with its mixed-linked beta-glucan. Mixed-linked beta-glucan is not metabolised by digestive enzymes. It lowers the cholesterol levels of blood and balances the glucose and insulin contents of serum after meals. These physiological effects reduce the risks of cardiovascular diseases. Our aim is to increase the beta-glucan content of Finnish oat cultivars through genetic engineering. The ultimate aim is to use plant-derived genes to elevate the beta-glucan content of oat to levels not obtainable through traditional plant breeding methods. Embryogenic cell cultures were started from mature embryos of oat cultivars ‘Aslak’, ‘Veli’ and ‘Kolbu’. Microscopic- and HPLC-analysis of beta-glucan of seeds, apical meristems and cell cultures were carried out. Gene transfer of microbial 1,3-beta-glucan synthases by particle bombardment was performed. The beta-glucan was mainly localized in subaleurone layers of oat seeds by Calcofluor staining. The molecular weight of oat seed beta-glucan was ca. 2 000 000 and the amount varied from 40 to 60 g/kg. In apical meristems trace amounts of beta-glucan were observed. In cell cultures the molecular weight of the beta-glucan was ca. 200 000 and the amount varied from 2 to 3 g/kg. Gene transfer experiments with microbial 1,3-beta-glucan synthase genes have been started in order to evaluate their effect on beta-glucan contents of oat cell lines. The cloning of plant beta-glucan synthase genes is on the way. References Salmenkallio-Marttila, M., K. Katina & K. Autio, 2002. Effects of bran fermentation on quality and microstructure of high-fiber wheat bread. Cereal Chem. 78: 429-435. Suortti, P. 1993. Size-exclusion chromatographic determination of beta-glucan with postcolumn reaction detection. J. Chrom. 632: 105-110.

Anaerobiosis occurs in seeds of many higher plants during imbibition. After successful germination followed by penetration of the enclosing structures by the radicle, the embryo switches from fermentative to aerobic energy metabolism. The microbial population, which is naturally present on the covering layers of barley grains, may lead to oxygen deficiency by competing for oxygen with the grain tissues. Although imbibed barley grains can survive several days under anoxia, they require oxygen to complete germination. Therefore, novel methods are useful to improve oxygen transfer and enhance germination of seeds. The advances in genetic engineering have opened new possibilities to study oxygen limitation and other critical mechanisms of germination. Previously, heterologous expression of Vitreoscilla hemoglobin (VHb) has been shown to improve either growth properties or productivity of various microbes and plants. Therefore, our metabolic engineering approach was to express the Vitreoscilla hemoglobin gene (vhb) in germinating barley to alleviate adverse effects of inadequate oxygen availability. However, our results show that VHb expression did not improve germination or growth of barley.

The antioxidant activities of the methanol extracts of selected varieties and parts of garlic and onion were determined by two methods: inhibition of lipid peroxidation induced by tert-butyl hydroperoxide in isolated rat hepatocytes and scavenging activity against diphenylpicrylhydrazyl radical. The total phenolics and the main flavonoids of the hydrolysed onion and garlic samples were also analysed. The antioxidant activities obtained by the two methods were compared. Both methods gave similar antioxidant activities for pure compounds and Allium extracts. However, the radical scavenging method had many benefits compared to the lipid peroxidation method, being easier, cheaper, more specific and reproducible. The radical scavenging activities also correlated positively with the total phenolics of the extracts. Onions had clearly higher radical scavenging activities than garlic, red onion being more active than yellow onion. The skin extracts of onion possessed the highest activities.

Many important pharmaceutical compounds are still isolated from plants. Due to their complex structures the chemical synthesis of these secondary metabolites is usually not applied. Plant cell cultures would offer an attractive alternative production system. However, the yields in cell cultures have only in few cases been commercially feasible. Futhermore, due to the lack of understanding of the biosynthetic pathways, genetic engineering has not had much success until now. It is expected that the application of genomics, proteomics and metabolomics tools will create a paradigm shift in our ability to engineer the often complex biosynthetic pathways of plant secondary metabolites. To illustrate the power of this approach, we present some data on the genome-wide transcript cDNA-AFLP profiling in combination with a GC-MS-SIM alkaloid analysis of methyl jasmonate elicited BY-2 tobacco cells used as a model system.

Despite the tremendous importance of secondary metabolites for humans as for the plant itself, plant secondary metabolism remains poorly characterized. Here, we present an experimental approach, based on functional genomics, to facilitate gene discovery in plant secondary metabolism. Targeted metabolite analysis was combined with cDNA-amplified fragment length polymorphism-based transcript profiling of jasmonate-elicited tobacco Bright yellow 2 cells. Transcriptome analysis suggested an extensive jasmonate-mediated genetic reprogramming of metabolism, which correlated well with the observed shifts in the biosynthesis of the metabolites investigated. This method, which in addition to transcriptome data also generates gene tags, in the future might lead to the creation of novel tools for metabolic engineering of medicinal plant systems in general.

New selectable markers and selection systems are needed to increase the efficiency and flexibility of plant transformation. The objective of this research was to determine if the green fluorescent protein (gfp) gene could be utilized as a visual selectable marker for transformation of oat (Avena sativa L.). A modified gfp gene was delivered into oat cells by microprojectile bombardment. Cell clusters expressing gfp were visually identified using fluorescence microscopy and physically isolated at each subculture. Eleven independent transgenic cell lines were obtained, and fertile plants regenerated from all lines. Transgene integration and expression were confirmed in transgenic plants and progeny. Transgene expression segregated in a 3 : 1 ratio in progeny of the majority of the transgenic lines.

Hairy root cultures of strawberry (Fragaria x ananassa Duch.) were induced with the Agrobacterium rhizogenes strain A4. Cultures were maintained on B50 medium but could also grow on a minimal medium, which did not inhibit the growth of arbuscular mycorrhizal fungi. The growth and nutrient uptake were characterized in shake flasks and in a bioreactor. Spores of the native Finnish arbuscular mycorrhizal fungus Glomus fistulosum V128 were used to infect strawberry (Fragaria x ananassa Duch. 'Senga Sengana') hairy roots in vitro. During cultivation, vegetative spore formation was observed. At the end of the cultivation, hyphae and arbuscules were observed in the stained roots.

Members of the genus Betula are deciduous trees, bushes, or shrubs with brown branches and a trunk, the upper part of which is covered with smooth, silver-white to white bark and the base with blackish, fissured bark. Birches are monoecious: the purple-brown male flowers are housed is drooping catkins and the green female catkins are small and erect but hang when mature. The fruit is a winged achene (Jury 1978).

Hairy roots of Catharanthus roseus were cultivated in three different types of bioreactors. The best growth and indole alkaloid production was achieved in an airsparged bioreactor with no other mixing. In the stirred bioreactor or in the bioreactor with medium circulation the roots did not grow, suggesting that hairy roots of C. roseus are more sensitive to stress than root cultures of many other plant species.

The effect of scaleup on he production of ajmalicine by a Catharanthus roseus cell suspension culture in a selected induction medium were studied. In preliminary experiments it was observed that the culture turned brown and the production was inhibited upon transfer from a shake flask to a stirred bioreactor with forced aeration. Two factors were recognized as the potential origin of the differences between shake flask and bioreactor cultures: gas composition and mechanical shear forces. These factors were studied separately. By recirculating a large part of the exhaust gas, a comparable gas regime was obtained in a bioreactor as occurred in a shake flask cultures. This resulted in the absence of browning and a similar pattern of ajmalicine production as observed in shake flasks. The effect of shear forces could not be demonstrated. However, the experiments showed that the culture may be very sensitive to liquid phase concentrations of gaseous compounds. The effects of kLa, aeration rate, CO2 production rate, and influent gas phase CO2 concentration on the liquid phase CO2 concentration are discussed.

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.

Among the cereal crops cultivated in Finland oat (Avena saliva L.) ranks second, after barley. Finland is one of the major oat producers in the world and a considerable part of the harvest is exported. The Finnish oat cultivars are generally well adapted to the humid and cool growth conditions in Finland. In order to improve the Finnish oat cultivars better to meet the requirements of the food industry, modern biotechnical methods are being used. Our first aim was to develop oat varieties with resistance to BYDV (Barley Yellow Dwarf Virus) through gene transfer. Six Finnish oat cultivars were used. The genotypes Kolbu, Rhiannon, GP-1 and Avena sterilis were used as controls. Cell cultures were started from mature embryos of all of these cultivars. From two Finnish cultivars leaf bases were also used. Embryogenic tissue cultures or oat leaf bases were used as targets for gene transfer, which was accomplished using particle bombardment. The transformants from the gene transfer experiments with the BYDV resistance gene were regenerated and rooted on selection media and transferred in soil in the greenhouse. Transgenic plants carrying the BYDV resistance gene were obtained only from those embryogenic cell cultures, which were started from mature embryos of oat. The progeny of these transgenic plants was germinated and screened for the inheritance of the transgene. The germination frequency of the seeds has been good, for example ranging from 92 to 100 % in TI and in T2, respectively. The progeny carrying the BYDV resistance gene was submitted to viral resistance testing.

Finland is one of the major oat (Avena sativa L.) producers, holding approximately a 20% share of the worldwide oat trade. The health benefits of oats are mainly associated with its mixed beta-glucan. Mixed beta-glucan is not metabolised by digestive enzymes and lowers the cholesterol levels of blood and balances the glucose and insulin contents of serum after meals. These physiological effects reduce the risks of cardiovascular diseases. Our aim is to increase the beta-glucan content of Finnish oats through genetic engineering. The ultimate aim is to use plant-derived genes to elevate the beta-glucan content of oats to levels not obtainable through traditional plant breeding methods. The induction of embryogenic cell cultures from mature embryos of oat cultivars Aslak, Veli and Kolbu was performed according to Somers et al. (1992). Calcofluor staining (Salmenkallio-Marttila et al. 2002) and HPLC-analysis (Suortti 1993) of beta-glucan of seeds, apical meristems and cell cultures were carried out. Gene transfer of microbial 1,3-beta-glucan synthases (Inoue et al. 1995) was accomplished by using particle bombardment (Bio-Rad, PDS-1000/He) according to Wan & Lemaux (1994). The beta-glucan was mainly localized in subaleurone layers of oat seeds by Calcofluor staining. The molecular weight of oat seed 13-glucan was ca. 2 000 000 and the amount varied from 40 to 60 g/kg. In apical meristems trace amounts of beta-glucan were observed. In cell cultures the molecular weight of the beta-glucan was ca. 200 000 and the amount varied from 2 to 3 g/kg. Gene transfer experiments with microbial 1,3-beta-glucan synthase genes have been started in order to evaluate their effect on beta-glucan contents of oat cell lines. The cloning of plant beta-glucan synthase genes is on the way. Inoue et al. 1995. European Journal of Biochemistry 231: 845-854; Salmenkallio-Marttila et al. 2002. Cereal Chemistry 78: 429-435.; Somers et al. 1992. Bio/Technology 10: 1589-1594; Suortti 1993. Journal of Chromatography 632: 105-110; Wan & Lemaux 1994. Plant Physiology 104: 37-48.