Contribution of autochthonous maize populations for adaptation to European conditions

Early vigor, earliness and cold tolerance are the main potential contributions of European maize (Zea mays L.) for breeding programs for adaptation to areas with short growing seasons and cold springs. The objective of this research was to determine the potential contributions of populations from different European regions to breeding for adaptation. Six Spanish and six French maize populations differing on variability for earliness, vigor and cold tolerance were crossed in a complete diallel without reciprocals. The populations and their crosses were evaluated in the field and in a cold chamber. Minimum temperatures were the main environmental trait affecting genotype × environment interaction, probably due to the cold sensitivity of the genotypes with the best performance in the field. The best population cross, based on specific heterosis for adaptation-related traits in the field, was Viana × Rastrojero, but this cross was cold sensitive. Tuy × Lazcano should be the best choice for a breeding program for adaptation, based on performance in the field and cold tolerance. As conclusions, there was variability for earliness, vigor and cold tolerance among the populations and crosses involved in this study, being tolerant to cold conditions the populations with medium growing cycle originated in areas with short growing seasons. The highest yielding crosses were cold sensitive.     

Drought stress‐induced changes in starch yield and physiological traits in potato

Drought stress is an important limitation for potato (Solanum tuberosum L.) production as potato depends on appropriate water availability for high yields of good quality. Therefore, especially in the background of climate change, it is an important goal in potato breeding to improve drought stress tolerance. In this study, 34 European starch potato cultivars were evaluated for drought stress tolerance by growing under well‐watered and long‐term drought stress conditions in rainout shelters in 2 years’ pot trials. Besides yield, six physiological traits, that is free proline content, osmolality, total soluble sugar content, chlorophyll content (SPAD), cell membrane stability and crude protein content, were determined in leaves sampled during vegetative growth and during flowering to investigate their association with drought tolerance. ANOVA revealed significant treatment effects for all physiological traits and increased genotypic effects at flowering. The sensitivity of physiological traits to drought was significantly higher during flowering than during vegetative growth. Drought stress decreased starch yield significantly (p < .001), on average by 55%. Starch yield was significantly influenced by genotype and genotype × treatment interactions. Stress tolerance index (STI) calculated from starch yield ranged from 0.26 (sensitive) to 0.76 (tolerant) with significant genotype effects (p ≤ .001). STI correlated positively with cell membrane stability (r = .59) and crude protein content (r = .38) and negatively with osmolality (r = ?.57) and total soluble sugar content (r = ?.71). These contrary correlations suggest a dual adaptation strategy in potato under long‐term drought stress conditions including increased membrane stability combined with an increased osmolality due to an increased soluble sugar content.     

The nabicol: A horticultural crop in northwestern Spain

Nabicol (B. napus L. var. pabularia) is a traditional crop in the Northwest of Iberian Peninsula (South of Galicia and North of Portugal) where it is grown during the winter season on small farms and gardens using traditional varieties. A collection of 36 populations of nabicol from Galicia (northwestern Spain) was evaluated during 2002 and 2003 in two locations and two growing seasons (spring/summer and autumn/spring) for 28 agronomic and morphological traits. The objectives of this study were to: (i) evaluate a collection of nabicol landraces from northwestern Spain, (ii) determine the suitability of this germplasm as a summer crop and (iii) study the genetic diversity among local populations. Significant differences were observed among populations for most traits. Genotype × environment interaction was significant for most of them. Spring/summer growing season could be recommended for growing nabicol but resistance to Lepidoptera pests attacking Brassica crops should be improved. Most populations had an agronomic value similar to the commercial variety. The most promising variety for horticultural use was MBG-BRS0063, which showed the highest yield. Morphological and agronomic data were subjected to cluster analysis and four groups were defined with a group clustering most populations. The low genetic diversity could be explained because populations were collected in close geographical areas and the selection made by farmers was always for a horticultural use. These results give information about the diversity and breeding value of the nabicol Spanish germplasm, which could be useful in breeding programs.     

Variation within and among Ladino white clover ecotypes for agronomic traits

Summary One set of 285 genotypes from three landraces and one natural population and a second set of 165 genotypes from two landraces and one natural population were evaluated in separate trials. Variation among populations was mostly accounted for by the difference between landrace and natural population genotype groups, the former showing features related to higher seed yield and short-term aerial dry matter (DM) yield and to lower persistence, cold tolerance and protein content. Compared to variance among ecotypes, that within ecotypes was never lower and it was at least four-fold higher for important traits such as aerial and total (aerial plus stolon) DM yield, stolon density, production of seed heads and cold tolerance. Landraces and natural populations showed similar levels of intra-population variation for all traits except cold tolerance, for which landraces tended to somewhat greater variation. Trait associations were less definite at the genotype level than what previously found in terms of mean values of the ecotypes. This finding and the relatively high broad sense heritability which was observed for all components of DM and seed yield can support the breeding activity on this clover. The level of variation and of heritability found for protein content do not suggest the routine evaluation of this trait in breeding programmes.     

Variation for dry matter yield,seed yield and other agronomic traits in Ladino white clover landraces and natural populations

Summary A set of Ladino white clover ecotypes including the last available landraces and some natural populations collected from old permanent meadows, together with some white clover varieties of various origins, were evaluated as monocultures for dry matter (DM) and seed yield assessed in separate field experiments. Other agronomic traits were also recorded. The best performing variety, Espanso, was significantly outyielded by some ecotypes for DM and seed production. Compared to landraces, natural populations yielded on average far less seed but as much DM and tended towards higher persistence indicating that domestication of the native Ladino at the farm level only improved seed yield of the crop. Natural populations showed, besides lower values of most seed yield components, a distinct canopy architecture characterized by smaller leaves, longer internodes and denser stolons. Seed weight proved the best character discriminating between the two ecotype groups. Significant variation for most traits was found both among landraces, in which it was mostly relatable to differences among provenances, and among natural populations. The relations between the main multivariate patterns of phenotypic variation and a set of environment collecting variables suggested that evolutive adaptation to different agronomic practices was a major determinant of the overall variation found in landraces. Evidence is provided that a relevant part of the variation observed among ecotypes for some important traits could be genetic.Abbreviations DM dry matter - DMY dry matter yield - NRH number of ripe heads - NFH number of florets per head - PL petiole length - LS leaf size - IL internode length - NS number of stolons - PRO protein content - SY seed yield - TSW thousand seed weight - NSF number of seeds per floret - PCA principal components analysis - PC principal component     

Estimates of combining ability for resistance of winter rye to Fusarium culmorum head blight

Summary Head blight caused by Fusarium culmorum and F. graminearum is damaging in all winter rye (Secale cereale L.) growing areas. For hybrid breeding, the relative magnitude of general (GCA) and specific combining ability (SCA) is a crucial parameter for developing appropriate selection procedures. Forty single-cross hybrids were produced by crossing six and seven inbred lines of the Petkus and Carsten gene pool, respectively, in a factorial design. Hybrids were evaluated in two years with artificial F. culmorum inoculation. Resistance traits were head blight rating and grain weight relative to the non-inoculated control. Both resistance traits were closely correlated across both years (r-0.8, P=0.01). Significant genotypic variation was found for both traits with medium to high estimates of heritability (h²=0.6-0.8). Components of variance for GCA were, across years, 10 and 6 times larger than those for SCA for head blight rating and relative grain weight, respectively. Significant SCA effects were found for 15 to 20% of all cross combinations across both traits in each year. SCA effects were, however, inconsistent over years leading to a high SCA-year interaction. In conclusion, resistance to Fusarium head blight among the interpool hybrids tested was conditioned mainly by additive gene action that could be utilized by recurrent selection in multi-environment trials.Abbreviations GCA general combining ability - SCA specific combining ability     

Tolerance to paraquat is correlated with the traits associated with water stress tolerance in segregating F2 populations of barley and wheat

To identify scorable marker traits that can be used in cereal breeding programs for selecting drought tolerant individuals, we investigated the correlation among the drought-associated traits in two F₂ populations derived from the crosses made between drought tolerant and sensitive barley and wheat parental genotypes. The parental genotypes of these crosses also differed by at least three other traits – paraquat tolerance, leaf size, and the relative water content. These three traits were scored in two F₂populations of 80 individuals for each barley and wheat cross. Analysis of results indicated that the enhanced tolerance to paraquat was correlated with reduced leaf size and increased relative water content, two traits associated with water stress phenotypes of the drought tolerant barley and wheat parents. Our results suggested that the selection based on paraquat tolerance istechnically less demanding and thus useful for rapid screening of individuals for enhanced drought tolerance in segregating populations. This revised version was published online in August 2006 with corrections to the Cover Date.     

Mass selection for improved cold and density tolerance of two maize populations

Summary Data of planting and plant density are two cultural practices influencing grain yield of maize (Zea mays L.). Our study was designed to evaluate the usefulness of a mass selection scheme to improve cold and density tolerance of the BS2 and BS3 maize populations. Populations were planted at an early planting date and a high plant density, and three cycles of mass selection for well-filled ears on erect plants were conducted at earch of three Corn Belt locations (i.e., Waseca, MN; Ames, IA; Portageville, MO).Results showed that selection improved cold tolerance traits of BS3, but not of BS2. Mass selection did not increase density tolerance of either population at any location. Selection did not improve response to planting dates, although the early planting date did improve agronomic performance and grain yield of all entries. We concluded that mass selection at high plant densities and early planting dates at diverse geographical locations did not produce cycles adapted to specific environmental conditions.     

Genetic relationships between attributes in sugarcane clones closely related to Saccharum spontaneum

Summary Saccharum spontaneum is being used in sugarcane breeding programs in attempt to improve characteristics such as ratooning ability and stress tolerance. A population of F₁ (Saccharum officinarum or commercial variety x S. spontaneum) and F₁ x F₁ sugarcane clones was evaluated for sugar yield and a range of yield components in a plant and two ratoon crops. The aim was to determine genetic correlations between attributes in clones with a large component of S. spontaneum, that could be used to help derive appropriate selection indices in such populations.There were close associations between the same attributes measured in different crop-years and the associations between different attributes were generally similar across crop-years. Stalk number and fibre content were positively correlated, as were stalk weight and CCS. The latter two attributes (which are low in S. spontaneum but high in S. officinarum) were negatively correlated with the former two (high in S. spontaneum, low in S. officinarum). Sugar yield was more closely associated with stalk weight and CCS than with stalk number but became more closely associated with stalk number with successive ratoon crops.CCS was positively correlated (r_g=0.55) with cane yield in the plant crop but showed a small negative correlation with cane yield (r_g=–0.20) in the second ratoon crop. CCS (measured in any crop) also had a negative correlation with cane yield in the ratoon crops expressed as a percentage of plant cane yield. This suggests that CCS is negatively correlated with levels of traits contributing to ratooning ability. Intensive selection among such populations for CCS without consideration of ratooning performance may reduce the frequency of favourable specific ratooning characteristics.Abbreviations CCS commercial cane sugar     

外源调节物质对棉花幼苗耐寒生理特性的效应

以棉花幼苗为试材,叶面分别喷施外源调节物质水杨酸(SA)、壳聚糖(CTS)、水杨酸+壳聚糖(SCM)、水杨酸+壳聚糖+硝酸钙(SCCa M)后,在5℃低温下进行处理,分别于0、24 h、48 h及恢复处理24 h时测定相对电导率、MDA含量、叶绿素含量,SOD、POD、CAT的活性和可溶性蛋白、可溶性糖、脯氨酸的含量。结果表明:低温胁迫前用0.7 mmol·L-1水杨酸、0.6 mmol·L-1壳聚糖及复配制剂处理棉花幼苗,其叶片的SOD、POD、CAT活性和叶绿素、可溶性蛋白、可溶性糖、脯氨酸含量与对照相比均有不同程度的提高,相对电导率和MDA含量有所降低;复配制剂处理效果优于水杨酸和壳聚糖单独施用的效果,而水杨酸、壳聚糖、硝酸钙三元复配的效果比水杨酸和壳聚糖两元复配的效果好。表明水杨酸、壳聚糖和硝酸钙在诱导棉花幼苗耐寒性中具有协同效应。     

Breeding tomato for pollen tolerance to low temperatures by gametophytic selection

Haploid selection for traits related to pollen cold tolerance in tomato was performed in segregating populations derived from a Lycopersicon esculentum × L. pennellii hybrid. BC₁ populations were obtained by combining normal and low temperature treatments on two stages of pollen development: pollen formation, and germination and pollen tube growth. F₁ hybrids were cultivated under low and normal temperatures and their pollen was used to pollinate L. esculentum plants at low and normal temperatures. The four BC₁ populations obtained were tested for the quality and quantity of pollen produced at low temperatures. The population obtained by cold treatment at both stages had a significantly improved pollen germination ability at low temperatures. The two other coldselected BC₁ populations showed no differences compared with the unselected BC population. A second cycle of pollen selection, corresponding to BC₂, was applied in order to test its persistence in the subsequent generations and the possibility to further improve the character. This second cycle showed no improvement although some plants retained the high pollen germination ability at low temperatures that was observed in the first cycle. Hence, gametophytic selection of some characters related with tomato pollen performance may be feasible, at least for the first cycle of selection.     

Phenotypic and QTL analyses of herbage production-related traits in perennial ryegrass (<Emphasis Type="Italic">Lolium perenne</Emphasis> L.)

Phenotypic and genetic evaluation of morphological traits associated with herbage biomass production was undertaken in a perennial ryegrass (Lolium perenne L.) biparental F₁ mapping population (n = 200) with parent plants from cultivars ‘Grasslands Impact’ and ‘Grasslands Samson’. Morphological traits measured on three clonal replicates of the parental genotypes and 200 F₁ progeny in a glasshouse in two separate trials (autumn and spring) included: dry weight (DW), leaf elongation rate (LER), initial tiller number (TNs), final tiller number (TNe), site filling (Fs), tiller weight (TW), leaf lamina length, leaf tip and ligule appearance rates (ALf, ALg) and leaf elongation duration (LED). Principal component analysis of patterns of trait association identified negative correlation between TNs or TNe, and TW as the primary basis for morphological difference and indicated that either high LER or long LED could reduce TN. Plants with higher LER tended to have increased DW. Quantitative trait loci (QTL) were detected on all seven linkage groups (LG) of a perennial ryegrass linkage map for all but three traits. A total of 61 QTL were identified, many of which clustered at 15 shared genome locations. Significant genotype by environment effects were encountered, evidenced both by variation between experiments in genotype rankings and by a general lack of commonality for QTL for the same traits in the different experiments. Only five QTL, for ALf, ALg and TN, were conserved between autumn and spring trials. A QTL for TN and DW on LG6 is a strong candidate for application of MAS in future plant improvement work and was found to be co-linear with QTL for equivalent traits reported on chromosome 2 in rice.     

Alien species as sources of physiological traits for wheat improvement

Summary Enhanced levels of drought, cold and salinity tolerance plus leaf photosynthesis and high protein content have been identified in species of Agropyron (Elytrigia) and Triticum (Aegilops). Because these characters, is most cases, are products of numerous metabolic pathways, they likely will have complex inheritance patterns and therefore may be difficult to transfer to bread wheat. However, all of the alien species in question are crossable with wheat. Most of the hybrids have been made and in many cases the material has been advanced to the state of addition lines which are available for screening. Where adequate screening tests are available, breeding techniques such as recurrent selection should be effective in transferring the traits.Contribution No. 781 Ottawa Research Station.     

Genetic variation in a Norwegian germplasm collection of white clover (Trifolium repens L.)

Phenotypic (r_p), genotypic (r_g), genotype × location (r_gl) and error (r_e)correlations for important agronomic characters were estimated for eleven Norwegian populations of white clover (Trifolium repens L.), originating from between 58^°52′N and 69^°30′N latitude, and from altitudes of 10 to 450 m a.s.l. The populations were evaluated in monoculture clonal field plots for two years at two locations in Norway. All pairwise correlations between foliage height, leaflet length, dry matter yield (DMY) and general performance were positive and significant. Winter survival was positively correlated with general performance, spring growth and DMY, but negatively associated with internode length, while seed yield showed a small positive genotypic correlation with foliage height. Considerable differences were, however, revealed among populations with regard to sign and magnitude of the estimated correlations. Within-population estimates of r_g were consistently positive between DMY, general performance and most other characters; between foliage height on the one hand and leaflet length, winter survival and seed yield on the other hand, and between internode length and leaflet length, indicating pleiotropy. Correlations involving other character-combinations varied substantially among populations, and imply presence of linkage and/or coadaptation. Path coefficient analysis was used to partition the genotypic correlations between some of the characters into direct and indirect effects. Path coefficients revealed that foliage height had the highest positive direct effect on DMY, followed by winter survival and internode length. Leaflet length, which showed a positive, significant simple correlation with DMY, exhibited a negligible direct influence, counterbalanced nearly completely by the positive indirect effect on DMY via foliage height. Positive indirect effects on DMY via foliage height were also found for winter survival and internode length. The coastal populations from North and Middle Norway, and a southern highland population showed the largest expected responses to phenotypic selection for most of the characters. The results demonstrate that there is enough genetic variation in adapted local populations to develop new improved cultivars adapted to high-latitude environments that combine upright growth habit with sufficient winter-hardiness and persistency. This revised version was published online in July 2006 with corrections to the Cover Date.     

Variation among Andean races of maize for cold tolerance during heterotrophic and early autotrophic growth

Cold in the initial growth stages is an important stressfactor for maize grown in regions with a temperate climate,particularly in case of early sowing. Sources of tolerancehave been identified in adapted genotypes, but promisinggenes for cold tolerance should also be found in materialdeveloped under the lower-temperature margins of the cropdistribution. This research was conducted in order to testAndean maize accessions for cold tolerance expressed duringboth the heterotrophic and early autotrophic growth stages.Experiments were conducted in controlled environments tostudy cold tolerance traits (germination %, germinationindex and plant growth rate) at continuous 10^°C (heterotrophic growth) and at varying 10–16^°C (autotrophic growth). An experiment was also performed inthe field with early sowing (both heterotrophic and autotrophic growth). In each experiment, a control trialwas conducted in more favourable conditions (i.e. continuous25^°C in a controlled environment or late planting inthe field) so that cold tolerance traits could also beexamined as the ratio between the stress and the controltrial. None of the accessions was superior for all coldtolerance traits. However, several Andean maize accessionsoutperformed the US Corn-belt hybrid checks for one or moretraits, both in heterotrophic and autotrophic growth. Overall, BOZM 855, PMS 636, Poblacion D, Poblacion E andBOZM 696 were the best accessions, suggesting that they canbe a promising source of genes for improving cold toleranceof adapted maize genotypes.     

Genetic variability and interrelationships of seed yield and yield components in switchgrass

Information on the genetic variability of seed yield and yield components is limited and no information is available on correlations among seed yield and yield components and direct and indirect effects of the yield components on seed yield in switchgrass, Panicum virgatum L. Accordingly, we conducted replicated experiments at Chickasha and Perkins, OK, in 1998 involving 11 lowland type switchgrass populations to assess genetic variation for seed yield and yield components, quantify interrelationships among them, and determine direct and indirect effects of yield components on seed yield through path coefficient analysis to identify traits for indirect selection of seed yield. Significant (P ≤ 0.01) variation existed among the 11 populations over locations for percent seed set and 100-seed weight. Seed yield/plant and the seed yield components panicle number/plant, spikelet number/panicle, and seed number/panicle had significant (P ≤ 0.05) population × location interactions, indicating substantial environmental influence on these traits for these populations. Accordingly, data for these traits were analyzed separately for each location revealing significant (P ≤ 0.01) differences among populations at both locations for each of these traits. Phenotypic correlation between seed yield/plant and seed number/panicle was positive (r = 0.76** at Chickasha and r = 0.72** at Perkins). Path coefficient analyses revealed that seed number/panicle had the highest positive direct effect on seed yield at both locations. Ample genetic variability was present among the switchgrass populations studied to allow breeding improvement of seed yield. Selection for increased seed number/panicle would be the most effective means of indirectly selecting for higher seed yield within this germplasm. Correlation and path coefficient analyses among biomass yield, seed yield, and harvest index indicated that, at least within the switchgrass germplasm studied, it would be possible to breed switchgrass cultivars with enhanced biomass yielding ability and sufficient seed production capability for their commercial propagation.     

Cold acclimation of winter and spring peas: carbon partitioning as affected by light intensity

Like most plants, pea (Pisum sativum L.) becomes tolerant to frost if it is first exposed to low non-freezing temperatures, a process known as cold acclimation. Cold acclimation is a complex process involving many physiological and metabolic changes. Two spring dry peas, two winter dry peas and one winter forage line were exposed to cold temperature in a controlled environment in two experiments, one using low light intensity and the other regular light intensity. Plants were harvested throughout the experiment and dry matter accumulation, water content, soluble and insoluble sugar concentrations were determined from shoot and root samples. Cold acclimation did not occur when temperatures were low if light intensity was low, even in winter peas. In contrast, with regular light intensity, the winter peas acquired more freezing tolerance than spring peas and a close relationship was found between the soluble sugar concentration of leaves just before the frost and the degree of freezing tolerance obtained by the different genotypes. Relationships between freezing tolerance and carbon partitioning between shoot and roots are discussed.     

Genetic variability and interrelationship of traits in the industrial oil crop Vernonia galamensis

Vernonia galamensis is a wild plant from the family Asteraceae which is endemic to East Africa and has the potential to become a new oil crop for industrial uses. Its seed oil is rich in vernolic acid, a fatty acid of high interest for oleochemical applications. However, a breeding program for Vernonia galamensis cultivars with high seed and oil yields requires knowledge about the genetic variability of traits that influence seed and oil production. This study was undertaken to examine phenotypic and genotypic variability, broad-sense heritability, genetic advance under selection and interrelationships of agronomic and seed quality traits. A total of 122 Vernonia accessions, 115 collected from different regions of Ethiopia and seven introduced, were grown at two locations in Ethiopia (Alemaya and Babile), in 2001/2002 and were analyzed for 20 traits including phenology, yield, yield components, and seed quality with special emphasis on fatty acid composition. The collections exhibited significant variation for all traits except for days to emergence. Genotypes and locations interacted significantly (P 0.01) for all traits. Broad-sense heritability estimates ranged from 11% (for days to emergence) up to 79% (for days to maturity). Expected genetic advance was between 1.3% (for days to emergence) and 44.8% (for seed oil yield). Genetic correlation analysis revealed that seed yield per plant is highly and positively correlated with seed weight and head number; highly significant and negative correlations (r = –0.59, –0.82, –0.85, and –0.89) were found between vernolic acid and palmitic, stearic, oleic, and linoleic acid, respectively. Highly significant positive correlations (r = 0.55, 0.44, and 0.36) were observed between vernolic acid and oil content, meal protein content and seed oil yield, respectively. Path-coefficient analysis indicated seed weight and secondary head number to be the most important components of seed yield per plant. Vernolic acid, oleic acid and linoleic acid had positive direct effects and stearic acid had a negative direct effect on oil content. The direct positive effect of oleic acid on oil content was, however, compensated by the negative indirect effects of stearic and vernolic acid resulting in a negative correlation (r = –0.60) between oleic acid and oil content. These observations will support the selection of accessions with high seed and oil yield, high meal protein contents, and high vernolic acid content.     

Nutritional and culinary quality of bush bean populations intercropped with maize

Summary The intereropping of common bean (Phaseolus vulgaris L.) and maize (Zea mays L.) is a common practice in the Northwest of the Iberian Peninsula. In this experiment, 25 populations of bush bean were evaluated for nutritional and culinary traits of dry seed and immature pod in sole crop and in intercrop in two years and in two locations, in order to determine whether selection of bush bean in sole crop can result in genetic progress for intercrop.There were no differences in the population means between cropping systems. Significant differences were detected among populations for all traits tudied except starch content, crude ash, water absorption and coat proportion. Population x environment interaction was significant for pod thickness. Bean populations did not interact with cropping systems for any trait, although there were differences between cropping systems for crude protein. These results suggest that a sole crop system provides sufficient information to select bean populations efficiently for the bush bean-maize intercropping system, although the advanced generations of the breeding program should be tested for quantitative traits such as protein content in the appropriate cropping system (intercropping) to know the competitive ability with the associated species. This would permit to choose bush bean populations which complement more efficiently and would be less competitive with the maize population.     

Improved frost tolerance and winter hardiness in proline overaccumulating winter wheat mutants obtained by in vitro-selection is associated with increased carbohydrate,soluble protein and abscisic acid (ABA) levels

In previous studies in vitro-selection of proline overaccumulating lines of winter wheat (Triticum sativum L. cv. Jo 3063) with increased frost tolerance was reported. These traits were found to be genetically stable. In the present study the improvement of frost tolerance (winter hardiness) under field conditions is confirmed for F₇ progenies of the mutants. Moreover, the mutants accumulated higher levels of glucose and fructose, soluble protein and abscisic acid (ABA) in addition to proline than the wild type under cold hardening conditions in a growth chamber as well as under cold hardening field conditions. ABA and proline levels peaked when the temperature decreased, whereas carbohydrate levels increased more slowly at decreasing temperature. Soluble protein levels also increased during cold hardening, but in addition showed sharp declines during frost periods. Increased carbohydrate levels of the mutants were associated with lower osmotic potential values. The differences in carbohydrate, protein and ABA levels between the mutants and the wild type are probably due to pleiotropic effects of the mutation.