Mapping QTLs with main and epistatic effects underlying grain yield and heading time in soft winter wheat

There is increasing awareness that epistasis plays a role for the determination of complex traits. This study employed an association mapping approach in a large panel of 455 diverse European elite soft winter wheat lines. The genotypes were evaluated in multi-environment trials and fingerprinted with SSR markers to dissect the underlying genetic architecture of grain yield and heading time. A linear mixed model was applied to assess marker-trait associations incorporating information of covariance among relatives. Our findings indicate that main effects dominate the control of grain yield in wheat. In contrast, the genetic architecture underlying heading time is controlled by main and epistatic effects. Consequently, for heading time it is important to consider epistatic effects towards an increased selection gain in marker-assisted breeding.     

Characterisation of grain yield and nitrogen level in relation to flag leaf senescence of wheat varieties

Three wheat cultivars ( Triticum aestivum L.), Splendeur, Hobbit and Maris Huntsman grown in pots were compared. Especially when compared to Splendeur, the flag leaf senesced most rapidly in Maris Huntsman, which presented the most rapid loss of moisture, chlorophyll and nitrogen. The uptake of exogenous nitrogen during the post-anthesis period was lower in the rapidly than in the slowly senescing variety. A higher concentration of free amino nitrogen in the flag leaf at a given sampling date was associated with a lower percentage decrease of soluble proteins at the following date. Acid proteinase activity in the flag leaf was inversely related to moisture percentage and free amino nitrogen level, but unrelated to the nitrogen loss of the flag leaf. Acid proteinase activity in the flag leaf was directly related to grain nitrogen percentage, but inversely related to grain yield. Grain yield was also directly related to the mean soluble protein content of the flag leaf through senescence.     

Heritability of grain yield,plant height and test weight of a population of winter wheat adapted to Southwestern Ontario

Summary Grain yield, plant height and test weight were studied in a population of winter wheat (Triticum aestivum L.). The population consisted of F₂ bulk populations of 42 crosses among 11 genotypes adapted to S. W. Ontario. Heritabilities were: 0.30±0.32 for yield, 0.77±0.15 for height and 0.98±0.08 for test weight. Predicted genetic gain with 10% selection intensity was 0.15 t/ha for yield, 10.1 cm for height and 3.00 kg/hl for test weight. The low heritability for yield indicates that effective selection would require pedigree information and progeny tests, while the high heritabilities for height and test weight indicate that selection for these traits using single plots would be appropriate.     

Mapping QTL for grain yield, yield components, and spike features in a doubled haploid population of bread wheat

A doubled haploid (DH) population derived from a cross between the Japanese cultivar 'Fukuho-kumogi' and the Israeli wheat line 'Oligoculm' was used to map genome regions involved in the expression of grain yield, yield components, and spike features in wheat (Triticum aestivum L). A total of 371 markers (RAPD, SSR, RFLP, AFLP, and two morphological traits) were used to construct the linkage map that covered 4190 cM of wheat genome including 28 linkage groups. The results of composite interval mapping for all studied traits showed that some of the quantitative trait loci (QTL) were stable over experiments conducted in 2004 and 2005. The major QTL located in the Hair-Xpsp2999 interval on chromosome 1A controlled the expression of grains/spike (R(2) = 12.9% in 2004 and 22.4% in 2005), grain weight/spike (R(2) = 21.4% in 2004 and 15.8% in 2005), and spike number (R(2) = 15.6% in 2004 and 5.4% in 2005). The QTL for grain yield located on chromosomes 6A, 6B, and 6D totally accounted for 27.2% and 31.7% of total variation in this trait in 2004 and 2005, respectively. Alleles inherited from 'Oligoculm' increased the length of spikes and had decreasing effects on spike number. According to the data obtained in 2005, locus Xgwm261 was associated with a highly significant spike length QTL (R(2) = 42.33%) and also the major QTL for spikelet compactness (R(2) = 26.1%).     

Heritability of and correlations among genotype-by-environment stability statistics for grain yield in bread wheat

Several genotype-by-environment stability measures are in use, but little information exists about their inheritance or genetic inter-relationships. Among those measures in common use are the linear regression coefficient (b), deviations from regression (s_b), coefficient of determination (R²), coefficient of phenotypic variation (CPV) and, more recently, interaction principal components (IPCA) of the additive-main-effect-and-multiplicative-interaction (AMMI) model. Because of the factorial structure of the data, the diallel cross is well suited to study these parameters and their relationship to quantitative traits. For this study a complete diallel cross, derived by mating eight lines from a broad based bread wheat breeding population, was grown for several growing seasons at two Ugandan locations, one of which was prone to yellow rust. Stability parameters and grain yield were measured for each cross. CPV had the highest narrow-sense heritability (h²=0.522) followed by IPCA1 of the AMMI (h²=0.461). Lowest narrow-sense heritabilities were calculated for b and R² (h²=0.150 and 0.100 respectively). There were high additive genetic correlations (r_A) between grain yield and CPV (r_A=−0.933), grain yield and IPCA1 (r_A=0.707), and grain yield and IPCA2 (r_A=0.751). The genetic association between CPV and IPCA1 was also high and negative (r_A= −0.934). These results suggest that it may be possible to select simultaneously for high and stable grain yield in this broad-based bread wheat breeding pool by selecting outyielders that exhibit a low CPV. Received: 25 July 2000 / Accepted: 7 December 2000     

QTLs for cell membrane stability and flag leaf area under drought stress in a wheat RIL population

A population of 206 recombinant inbred lines (RILs F₉–F₁₀) derived from wheat cross WL711/C306 was phenotyped for morpho-physiological traits such as flag leaf area (FLA), flag leaf length (FLL), flag leaf width (FLW), and cell membrane stability (CMS) under water deficit stress (WDS) environment. High yielding cultivar, WL711 had higher FLA than the medium yielding cultivar C306 across trials under both environments. Parent cultivar C306 maintained membrane integrity while WL711 showed higher membrane damage under WDS. The RIL population showed considerable variation, normal distribution and transgressive segregation for FLA, FLL, FLW and CMS under WDS. The genetic linkage map of WL711/C306 RIL population was constructed comprising of 346 markers. The total map distance was 4526.8 cM with an averaged interval of 12.9 cM between adjacent markers. Major consistent QTL for FLA, FLL, FLW, and CMS were identified on chromosomes 2DS and 3BS respectively in the WL711/C306 RIL population under WDS. The major QTL for FLA, qFLAWD.2D.1 which expressed in multiple environments and for CMS, qCMSWD.3B.3 and qCMSWD.3B.4, accounted for a large proportion of phenotypic variance (PV) with positive allele being contributed by C306, a drought resistant (DR) parent. QTL qFLAWD.2D.1 for FLA co-located with QTL for grain number (GN) and days to flowering (DTF) while QTL qCMSWD.3B.3 and qCMS.3B.4 co-located with QTL for grain yield and its components, days to flowering, canopy temperature and coleoptiles length as reported in our previous publications on the WL711/C306 population (Shukla et al. in Euphytica 203:449–467, 2015; Singh et al. in J Plant Biochem Biotechnol 24:324–330, 2015). Two candidate genes Ghd7 for grain yield and heading date and OsCDK4 for calcium dependent protein kinases were identified in the 2DS and 3BS QTL regions respectively on comparison with gene content of rice chromosomes 7 and 1 respectively. Hence, QTLs qFLAWD.2D.1 and qCMSWD.3B.3 are potential target regions for fine mapping and marker assisted selection for FLA and CMS respectively in wheat under water deficit environments.

     

Detection of QTLs for heading time and photoperiod response in wheat using a doubled-haploid population.

The genetic basis of heading time in wheat (Triticum aestivum L.) was investigated through the study of flowering under normal autumn sown field conditions as well as photoperiod responses under a controlled environment. Quantitative trait loci (QTLs) for these traits were mapped in a doubled-haploid (DH) population derived from a cross between the wheat cultivars 'Courtot' and 'Chinese Spring'. A molecular marker linkage map of this cross that was previously constructed based on 187 DH lines and 380 markers was used for QTL mapping. The genome was well covered (85%) except for chromosomes 1D and 4D, and a set of anchor loci regularly spaced over the genome (one marker each 15.5 cM) was chosen for marker regression analysis. The presence of a QTL was declared at a significance threshold of alpha = 0.005. The population was grown under field conditions in Clermont-Ferrand, France during two years (1994-1995), in Norwich, U.K. over one year (1998), and also under controlled environments in Norwich. For each trait, between 2 and 4 QTLs were identified with individual effects ranging between 6.3% and 44.4% of the total phenotypic variation. Two QTLs were detected that simultaneously affected heading time and photoperiod response. For heading time, these two QTLs were detected in more than one year. One QTL located on chromosome arm 2BS near the locus Xfbb121-2B, co-segregated with the gene Ppd-B1 known to be involved in photoperiod response. This chromosome region explained a large part of the variation (23.4-44.4% depending on the years or the traits). Another region located on chromosome arm 7BS between the loci Xfbb324-7B and Xfbb53-7B also had a strong effect (7.3-15.3%). This region may correspond to a QTL for earliness per se.     

Relationships between grain yield and morphological characters in the winter wheat genotypes of the Belgian national list

Yield per shoot and to a much lesser extend yield per unit area were related to morphological characters. The flag sheath was better related to shoot yield than were any of the three uppermost leaf laminae. Among these the areas of the two lower leaves showed a better relationship to the yield than did the flag leaf lamina. Variation in main shoot yield was associated mainly with variation in grain number. More attention should be given to morphological character related to spike development before anthesis.     

Identification of novel QTLs for seedling and adult plant leaf rust resistance in a wheat doubled haploid population

Pyramiding of genes that confer partial resistance is a method for developing wheat (Triticum aestivum L.) cultivars with durable resistance to leaf rust caused by Puccinia triticina. In this research, a doubled haploid population derived from the cross between the synthetic hexaploid wheat (SHW) (×Aegilotriticum spp.) line TA4152-60 and the North Dakota breeding line ND495 was used for identifying genes conferring partial resistance to leaf rust in both the adult plant and seedling stages. Five QTLs located on chromosome arms 3AL, 3BL, 4DL, 5BL and 6BL were associated with adult plant resistance with the latter four representing novel leaf rust resistance QTLs. Resistance effects of the 4DL QTL were contributed by ND495 and the effects of the other QTLs were contributed by the SHW line. The QTL on chromosome arm 3AL had large effects and also conferred seedling resistance to leaf rust races MJBJ, TDBG and MFPS. The other major QTL, which was on chromosome arm 3BL, conferred seedling resistance to race MFPS and was involved in a significant interaction with a locus on chromosome arm 5DS. The QTLs and the associated molecular markers identified in this research can be used to develop wheat cultivars with potentially durable leaf rust resistance.     

Mapping of main and epistatic effect QTLs associated to grain protein and gluten strength using a RIL population of durum wheat

Quality, specifically protein content and gluten strength are among the main objectives of a durum wheat breeding program. The aim of this work was to validate quantitative trait loci (QTLs) associated with grain protein content (GPC) and gluten strength measured by SDS sedimentation volume (SV) and to find additional QTLs expressed in Argentinean environments. Also, epistatic QTL and QTL x environmental interactions were analyzed. A mapping population of 93 RILs derived from the cross UC1113 x Kofa showing extreme values in gluten quality was used. Phenotypic data were collected along six environments (three locations, two years). Main effect QTLs associated with GPC were found in equivalent positions in two environments on chromosomes 3BS (R² = 21.0-21.6%) and 7BL (R² = 12.1-13%), and in one environment on chromosomes 1BS, 2AL, 2BS, 3BL, 4AL, 5AS, 5BL and 7AS. The most important and stable QTL affecting SV was located on chromosome 1BL (Glu-B1) consistently detected over the six environments (R² = 20.9- 54.2%). Additional QTLs were found in three environments on chromosomes 6AL (R² = 6.4-12.5%), and in two environments on chromosomes 6BL (R² = 11.5-12.1%), 7AS (R² = 8.2-10.2%) and 4BS (R² = 11–16.4%). In addition, pleiotropic effects were found affecting grain yield, test weight, thousand-kernel- weight and days to heading in some of these QTLs. Epistatic QTLs and QTL x environment interactions were found for both quality traits, mostly for GPC. The flanking markers of the QTLs detected in this work could be efficient tools to select superior genotypes for the mentioned traits.     

Relationships between grain protein content and grain yield components through quantitative trait locus analyses in a recombinant inbred line population derived from two elite durum wheat cultivars

Grain protein content (GPC) in durum wheat (Triticum turgidum var. durum) is negatively correlated with grain yield. To evaluate possible genetic interrelationships between GPC and grain yield per spike, thousand-kernel weight and kernel number per spike, quantitative trait loci (QTL) for GPC were mapped using GPC-adjusted data in a covariance analysis on yield components. Phenotypic data were evaluated in a segregating population of 120 recombinant inbred lines derived from crossing the elite cultivars Svevo and Ciccio. The material was tested at five environments in southern Italy. QTL were determined by composite interval mapping based on the Svevo?×?Ciccio linkage map described in Gadaleta et al. (2009) and integrated with DArT markers. The close relationship between GPC and yield components was reflected in the negative correlation between the traits and in the reduction of variance when GPC values were adjusted to yield components. Ten independent genomic regions involved in the expression of GPC were detected, six of which were associated with QTL for one or more grain yield components. QTL alleles with increased GPC effects were associated with QTL alleles with decreased effects on one or more yield component traits, or vice versa (i.e. the allelic effects were in opposite direction). Four QTL for GPC showed always significant effects, and these QTL should represent genes that influence GPC independently from variation in the yield components. Such genes are of special interest in wheat breeding since they would allow an increase in GPC without a concomitant decrease in grain yield.     

The novel ethylene antagonist, 3-cyclopropyl-1-enyl-propanoic acid sodium salt (CPAS), increases grain yield in wheat by delaying leaf senescence

Orchid is a major floral crop around the world and Dendrobium hybrids are considered to be one of the most popular orchids. In vitro germination of hybrid seeds is a common practice among orchid growers, however, in many cross pollinations the embryos may not develop to maturity, leading to poor seed germination. The effect of seed maturity and sucrose concentration were investigated via asymbiotic germination of nobile Dendrobium hybrids. Capsules were harvested from two hybrids (Den. Lucky Girl × Den. Second Love ‘Kirameki’ and Den. Lucky Girl × Den. Hamana Lake ‘Kumi’) and one selfing of Den. Second Love ‘Kirameki’ at 2, 3, 4, and 5 months after pollination and immature seeds were taken. Immature seeds from 3- to 5-month old capsules could be successfully germinated on Hyponex based medium. Immature seeds from 4-month old capsules showed greatest germination rate of tested treatments, whereas 3-month old immature seeds showed the least germination. After 6 weeks of in vitro culture, protocorms derived from embryos developed on every concentration of sucrose, but germination was greater at lower concentrations. Greater concentration of sucrose decreased normal-developed protocorms.     

Rate and duration of grain filling in relation to flag leaf senescence and grain yield in spring wheat (Triticum aestivum) exposed to different concentrations of ozone

Effects of different concentrations of ozone on grain filling, flag leaf senescence and final grain yield in field‐grown spring wheat (Triticum aestivum L. cv. Dragon) were studied using open‐top chambers. The hypothesis tested was that an ozone‐induced reduction in grain yield is mainly related to an enhanced senescence and a shortening of the grain‐filling period. The plants were exposed to filtered air (F), non‐filtered air without extra ozone (NF) or non‐filtered air with 3 different levels of ozone added (NF1+, NF2+ and NF3+). The mean daytime (08.00–20.00 h) ozone concentrations during the exposure period (31 days) were 7, 20, 34, 48 and 62 nmol mol?1 in F, NF, NF1+, NF2+ and NF3+, respectively. The corresponding ozone doses, expressed as the accumulated exposure over a concentration threshold of 40 nmol mol?1 (AOT40), were 0, 12, 1 989, 5 881 and 10 375 nmol mol?1 h, respectively, and 884, 2 594, 4 557, 6 188 and 7 900 μmol m?2, respectively, expressed as the calculated cumulative flag leaf ozone flux (CFO30). The flag leaves senesced earlier and the grain‐filling duration was significantly shorter at higher ozone exposure compared to F (?5, ?13 and ?18% in NF1+, NF2+ and NF3+, respectively). The relative grain‐filling rate did not differ between the treatments. The 1000‐grain weights were 10, 28 and 37% lower, and the grain yields were 15, 29 and 46% lower than F in NF1+, NF2+ and NF3+, respectively. Ozone exposure had no significant effect on the number of grains per unit ground area or on straw yield, but significantly reduced the harvest index and increased the grain protein concentration in NF2+ and NF3+ compared to F. The grain yield was negatively correlated with the ozone dose, expressed either as AOT40 or as CFO3 with or without an ozone flux threshold. The 1000‐grain weight was positively correlated with the grain‐filling duration (R2=0.998), which in turn was positively correlated with the leaf area duration (R2=0.989).     

Genetic control of grain yield and grain physical characteristics in a bread wheat population grown under a range of environmental conditions

Key message

Genetic analysis of the yield and physical quality of wheat revealed complex genetic control, including strong effects of photoperiod-sensitivity loci.

Abstract

Environmental conditions such as moisture deficit and high temperatures during the growing period affect the grain yield and grain characteristics of bread wheat (Triticum aestivum L.). The aim of this study was to map quantitative trait loci (QTL) for grain yield and grain quality traits using a Drysdale/Gladius bread wheat mapping population grown under a range of environmental conditions in Australia and Mexico. In general, yield and grain quality were reduced in environments exposed to drought and/or heat stress. Despite large effects of known photoperiod-sensitivity loci (Ppd-B1 and Ppd-D1) on crop development, grain yield and grain quality traits, it was possible to detect QTL elsewhere in the genome. Some of these QTL were detected consistently across environments. A locus on chromosome 6A (TaGW2) that is known to be associated with grain development was associated with grain width, thickness and roundness. The grain hardness (Ha) locus on chromosome 5D was associated with particle size index and flour extraction and a region on chromosome 3B was associated with grain width, thickness, thousand grain weight and yield. The genetic control of grain length appeared to be largely independent of the genetic control of the other grain dimensions. As expected, effects on grain yield were detected at loci that also affected yield components. Some QTL displayed QTL-by-environment interactions, with some having effects only in environments subject to water limitation and/or heat stress.     

Analysis of QTLs for yield, yield components, and malting quality in a BC3-DH population of spring barley

Advanced backcross (AB)-quantitative trait locus (QTL) analysis has been successfully applied for detecting and transferring QTLs from unadapted germplasm into elite breeding lines in various plant species. Here, we describe the application of a modified AB breeding scheme to spring barley. A BC₃-doubled haploid (DH) population consisting of 181 lines derived from the German spring barley cultivar Brenda (Hordeum vulgare subsp. vulgare) as the recurrent parent and the wild species line HS213 (H. vulgare subsp. spontaneum) as the donor line was evaluated for yield and its components as well as malting quality traits. A set of 60 microsatellite markers was used to genotype the population, and phenotypic data were collected at two locations in Germany in continuous years. Altogether, 25 significant QTLs were detected by single-marker regression analysis and interval mapping. Most positive QTLs originated from the recurrent parent Brenda. A QTL, Qhd2.1, on chromosome 2HS from Brenda explained 18.3% and 20.7% of the phenotypic variation for yield and heading date, respectively. Due to the small percentage of donor-parent genome of 6.25%, the BC₃-DH lines could be directly used for the extraction of near-isogenic lines (NILs) for Qhd2.1. Consequently, it was possible to determine the precise location of the locus hd2.1 within a region of 6.5 cM, using an F₂ population consisting of 234 individuals developed from a cross between an NIL containing a defined donor segment at this locus and Brenda. The location of this QTL was consistent with the presence of a major photoperiod response gene, Ppd-H1, previously reported in this region, which is associated with pleiotropic effects on yield components. In summary, the analysis of a BC₃-DH population in barley provides a compromise between the analysis of QTLs by means of an AB scheme and the generation of defined substitution lines. Several lines carrying defined different donor segments for only one single chromosome or trait in the genetic background of Brenda could be selected for further genetic studies.     

Mapping of QTLs associated with resistance to common bunt,tan spot,leaf rust,and stripe rust in a spring wheat population

Spring wheat (Triticum aestivum L.) breeding goals in western Canada include good agronomic characteristics and good end-use quality, and also moderate to elevated resistance to diseases of economic importance. In this study, we aimed to identify quantitative trait loci (QTL) associated with resistance to common bunt (Tilletia tritici and Tilletia laevis), tan spot (Pyrenophora tritici-repentis), leaf rust (Puccinia triticina), and stripe rust (Puccinia striiformis f. sp. tritici). A total of 167 recombinant inbred lines (RILs) derived from a cross between two spring wheat cultivars, ‘Attila’ and ‘CDC Go’, were evaluated for reactions to the four diseases in nurseries from three to eight environments, and genotyped with the Wheat 90K SNP array and three gene-specific markers (Ppd-D1, Vrn-A1, and Rht-B1). The RILs exhibited transgressive segregation for all four diseases, and we observed several lines either superior or inferior to the parents. Broad-sense heritability varied from 0.25 for leaf rust to 0.48 for common bunt. Using a subset of 1203 informative markers (1200 SNPs and 3 gene-specific markers) and average disease scores across all environments, we identified two QTLs (QCbt.dms-1B.2 and QCbt.dms-3A) for common bunt, and three QTLs each for tan spot (QTs.dms-2B, QTs.dms-2D, and QTs.dms-6B), leaf rust (QLr.dms-2D.1, QLr.dms-2D.2, and QLr.dms-3A), and stripe rust (QYr.dms-3A, QYr.dms-4A, and QYr.dms-5B). Each QTL individually explained between 5.9 and 18.7% of the phenotypic variation, and altogether explained from 21.5 to 26.5% of phenotypic and from 52.2 to 86.0% of the genetic variation. The resistance alleles for all QTLs except one for stripe rust (QYr.dms-5B) were from CDC Go. Some of the QTLs are novel, while others mapped close to QTLs and/or genes reported in other studies.     

The relationships between seedling root screens,root growth in the field and grain yield for wheat

Plant and Soil - The development of fertile patches within an infertile matrix is a common phenomenon in drylands. Shrub-centered expansion of fertile islands is generally attributed to processes...     

Insect infestations, incidence of viral plant diseases, and yield of winter wheat in relation to planting date in the northern Great Plains

Planting date effects on arthropod infestation and viral plant disease are undocumented for winter wheat, Triticum aestivum L., in South Dakota and the northern Great Plains. Winter wheat was planted over three dates (early, middle, and late; generally from late August to late September) to determine the effect on abundance of insect pests, incidence of plant damage, incidence of viral plant disease, and grain yield. The study was conducted simultaneously at two sites in South Dakota over three consecutive cropping seasons for a total of six site yr. Cereal aphids (Homoptera: Aphididae) were abundant in three site yr. Rhopalosiphum padi (L.), bird cherry-oat aphid, was the most abundant cereal aphid at the Brookings site, whereas Schizaphis graminum (Rondani), greenbug, predominated at Highmore. Aphid-days were greater in early versus late plantings. Aphid abundance in middle plantings depended on aphid species and site, but it usually did not differ from that in early plantings. Incidence of Barley yellow dwarf virus (family Luteoviridae, genus Luteovirus, BYDV) declined with later planting and was correlated with autumnal abundance of cereal aphids. Incidence of BYDV ranged from 24 to 81% among 1999 plantings and was < 8% in other years. Damage to seedling wheat by chewing insects varied for two site-years, with greater incidence in early and middle plantings. Wheat streak mosaic virus, spring infestations of cereal aphids, wheat stem maggot, and grasshoppers were insignificant. Yield at Brookings was negatively correlated with BYDV incidence but not cereal aphid abundance, whereas yield at Highmore was negatively correlated with aphid abundance but not BYDV incidence. Planting on 20 September or later reduced damage from chewing insects and reduced cereal aphid infestations and resulting BYDV incidence.     

Relationships between early vigour, grain yield, leaf structure and stable isotope composition in field grown barley

Fast growth and early development in barley are used in breeding programmes to improve the water use efficiency and transpiration efficiency of this crop in Mediterranean conditions. Here, we examine the use of several simple traits based on the structure and stable isotope composition of seedling leaves to assess differences in early vigour, phenology and grain yield, and also the interaction with low temperatures in barley. A set of 260 F₈ lines of two-row barley (Hordeum vulgare L.) derived from the cross of Tadmor and WI 2291 were cultivated in two locations in northwest Syria. Total chlorophyll content on an area basis (SPAD) and specific leaf dry weight (SLDW) were measured in recently fully expanded intact leaves of seedlings. Total leaf area and total dry weight per seedling were evaluated in the same seedlings. The stable isotope compositions of carbon and nitrogen (δ¹³C and δ¹⁵N, respectively) were analyzed in the same leaves on a subset of 75 genotypes. Number of days from planting to heading and grain yield were recorded at both sites. The grain yield measured at both locations was positively correlated with the SPAD value of seedlings, but showed no relationship with SLDW. Days to heading was negatively correlated with SPAD values. Regarding early vigour, a negative relationship between the SLDW and the total leaf area of seedlings was observed. However, no relationship between the δ¹³C of seedlings and early vigour was observed, except when only the genotypes most resistant to low temperatures (i.e. showing the highest SPAD values) were considered. This subset of genotypes showed negative relationships between δ¹³C and either total leaf area or total dry weight. In addition, δ¹⁵N was negatively correlated with SPAD only within the high-SPAD genotypes. This suggests that within the genotypes resistant to low temperatures, those with higher chlorophyll content assimilate more nitrogen from nitrate.