Variation for winter hardiness generated by androgenesis from
Festuca pratensis × Lolium multiflorum amphidiploid cultivars with different winter susceptibility
Androgenic populations produced from three Festuca pratensis × Lolium multiflorum amphidiploid (2n = 4x = 28) cultivars show wide within population variation for winter hardiness. Populations comprising a total of 423 androgenic plants derived from 23 donor plants from 3 Polish F. pratensis × L. multiflorum cultivars Felopa, Sulino, and Rakopan were studied over 3 years (1999–2002) in a field experiment at Lopuszna, Poland (20∘08′E, 49∘28′N, altitude 568 m). The results indicate that despite differences in winter hardiness between the three amphidiploid cultivars, the mean winter hardiness of androgenic progeny from each cultivar was the same. Thus an androgenesis component in a plant breeding programme may provide an opportunity to recover winter hardy genotypes from high yielding cultivars that are themselves adapted poorly to stress conditions. Androgenesis also evoked variation in snow mould resistance. In this case, androgenic plants with the greatest snow mould resistance were recovered from the least winter hardy donor cultivars and plants. The results indicated the low importance of snow mould resistance as a component of winter hardiness under the field conditions used for these experiments.
Changes in protein abundance and activity involved in freezing tolerance acquisition in winter barley (Hordeum vulgare L.).
The changes in protein abundance induced by cold hardening were analysed by 2 DE in ten doubled haploid (DH) lines of winter barley, highly differentiated with respect to freezing tolerance level. Among 45 differential proteins identified by MALDI-TOF/TOF, the majority was classified as related to photosynthesis, carbohydrate metabolism, oxidation-reduction reactions and stress response. Among the detected proteins, higher abundance of RuBisCO large and small subunits, RuBisCO activase, two Oxygen-evolving enhancer proteins, Ferredoxin-NADP reductase, Cytochrome P450-dependent fatty acid hydroxylase and 14-3-3 protein was associated with higher freezing tolerance level. Lower relative level of hypothetical ATP synthase beta subunit, uncharacterized mitochondrial protein AtMg00810 and ribosomal RNA small subunit methyltransferase G also seems to be important. The results of proteomic studies were complemented by the evaluation of photosynthetic apparatus acclimation, showing distinctive differences between the studied genotypes in the number of active PSII reaction centres (RC/CSm). Additionally, the analysis of antioxidative enzyme activities suggests the importance of H2O2 as a signalling molecule possibly involved in the initiation of cold-induced plant acclimation. However, in DH lines with high freezing tolerance, H2O2 generation during cold hardening treatment was accompanied by more stable activity of catalase, H2O2-decomposing enzyme.


SIGNIFICANCE
In the study, the changes in protein abundance induced by cold hardening treatment were analysed by two-dimensional gel electrophoresis in ten doubled haploid (DH) lines of winter barley. Harnessing DH technology resulted in distinctive widening of genetic variation with respect to freezing tolerance level. Both the cold-hardening effect on the protein pattern in an individual winter barley DH line as well as the variation among the selected DH lines were investigated, which resulted in the identification of 45 differentiated proteins classified as involved in 14 metabolic pathways and cellular processes. Among them, eight proteins: (1) the precursor of RuBisCO large subunit, (2) RuBisCO small subunit (partial), (3) RuBisCO activase small isoform, (4) the precursor of Oxygen-evolving enhancer protein 1-like (predicted protein), (5) Oxygen-evolving enhancer protein 2, (6) the leaf isozyme of Ferredoxin-NADP reductase, (7) hypothetical protein M569_12509 Cytochrome P450-dependent fatty acid hydroxylase-like and (8) hypothetical protein BRADI_1g11290 (14-3-3 protein A-like) were accumulated to a higher level in leaves of cold-hardened seedlings of freezing tolerant winter barley DH lines in comparison with susceptible ones. Three others: (9) hypothetical protein BRADI_5g05668 F1 ATP synthase beta subunit-like, (10) predicted protein uncharacterized mitochondrial protein AtMg00810-like and (11) BnaA02g08010D Ribosomal RNA small subunit methyltransferase G-like were detected at lower level in freezing tolerant seedlings in comparison with susceptible genotypes. The last two were for the first time linked to cold acclimation. The results of complementary analyses indicate that PSII activity and stability of antioxidative enzymes under low temperature are also very important for freezing tolerance acquisition.
The status of FAO and codex alimentarius developments on vegetable proteins
The FAO is interested in increasing world protein supplies both by advising and assisting developing countries to increase their production of staple foods and by encouraging the development of technology to produce protein concentrates both from vegetable and animal sources to supplement diets which are low in protein content. Recent revisions of the amount of protein required in the diet in relation to total per capita energy requirements and the difficulties encountered in introducing novel foods into established dietary patterns has led FAO to concentrate on increased agricultural productions in developing countries as the primary means of combatting malnutrition. In the meantime, in North America and Europe, advances in the technology of processing vegetable proteins has given highly acceptable products simulating meats which are being used as meat extenders and meat analogues. The consequent increase in vegetable protein production has revealed a need for international, agreement on standardization, and it has been agreed that this will be undertaken by the Joint FAO/WHO Food Standards’ Programme. The program operates chiefly through the Codex Alimentarius, Commission, an international governmental body, the purpose of which is to protect consumers against health hazard in food and against fraud. The Commission, which has 116 member governments, controls an intensive program of work covering the composition, labeling, additives, contaminants, pesticide residues, hygiene, sampling and analysis aspects of foods and has established a procedure which allows member governments ample opportunity to reach agreement on food standards and to accept them for incorporation into national legislation. The Codex Commission, at its 12th session, decided to establish a Committee on Vegetable Proteins with the following terms of references: “to elaborate definitions and world-wide standards for vegetable protein products deriving from soybeams, cottonseed, groundnuts, cereals and from other vegetable sources as they come into use for human consumption, and to elaborate guidelines on utilization of such vegetable protein products in the food supply system, on nutritional requirements and safety on labeling and on other aspects as may seem appropriate.” The question of which country will host the Committee is still under discussion.