J. M. Lasa

The Spanish barley core collection

Spanish barleys constitute a germplasm group of particular interest for breeding purposes, as Spain has been proposed as a possible centre of origin of the crop. The Spanish National Germplasm Bank (Banco Nacional de Germoplasma, BNG), holds a collection of about 2000 barley accessions, mostly landraces collected in Spain prior to extensive introduction of modern varieties. The objective of this work is to create a core collection of barleys representative of old barley genotypes grown in Spain. The core collection will be constituted by three groups of germplasm: successful old varieties (15); entries in common with previously existing barley core collections (15); and 2-row (8) and 6-row (122) entries from the BNG, for a total of 160 entries. Entries were allocated by stratified sampling in agro-ecological uniform zones of barley cultivation in Spain. Classification of agro-ecological regions for barley was based on historical yield records for Spanish provinces. The number of entries for each region was determined in proportion to the logarithm of historical barley acreage. Final choice of accessions within provinces tried to maximize the diversity and avoid duplications by looking at passport data, and to agronomic evaluation data available for a group of about 900 accessions.

Patterns of genetic and eco-geographical diversity in Spanish barleys

The pool of Western Mediterranean landraces has been under-utilised for barley breeding so far. The objectives of this study were to assess genetic diversity in a core collection of inbred lines derived from Spanish barley landraces to establish its relationship to barleys from other origins, and to correlate the distribution of diversity with geographical and climatic factors. To this end, 64 SSR were used to evaluate the polymorphism among 225 barley (Hordeum vulgare ssp. vulgare) genotypes, comprising two-row and six-row types. These included 159 landraces from the Spanish barley core collection (SBCC) plus 66 cultivars, mainly from European countries, as a reference set. Out of the 669 alleles generated, a large proportion of them were unique to the six-row Spanish barleys. An analysis of molecular variance revealed a clear genetic divergence between the six-row Spanish barleys and the reference cultivars, whereas this was not evident for the two-row barleys. A model-based clustering analysis identified an underlying population structure, consisting of four main populations for the whole genotype set, and suggested further possible subdivision within two of these populations. Most of the six-row Spanish landraces clustered into two groups that corresponded to geographic regions with contrasting environmental conditions. The existence of wide genetic diversity in Spanish germplasm, possibly related to adaptation to a broad range of environmental conditions, and its divergence from current European cultivars confirm its potential as a new resource for barley breeders, and make the SBCC a valuable tool for the study of adaptation in barley.

Heading date QTL in a spring × winter barley cross evaluated in Mediterranean environments

Heading date is a key trait for the adaptation of barley to Mediterranean environments. We studied the genetic control of flowering time under Northern Spanish (Mediterranean) conditions using a new population derived from the spring/winter cross Beka/Mogador. A set of 120 doubled haploid lines was evaluated in the field, and under controlled temperature and photoperiod conditions. Genotyping was carried out with 215 markers (RFLP, STS, RAPD, AFLP, SSR), including markers for vernalization candidate genes, HvBM5 (Vrn-H1), HvZCCT (Vrn-H2), and HvT SNP22 (Ppd-H1). Four major QTL, and the interactions between them, accounted for most of the variation in both field (71–92%) and greenhouse trials (55–86%). These were coincident with the location of the major genes for response to vernalization and short photoperiod (Ppd-H2 on chromosome 1H). A major QTL, near the centromere of chromosome 2H was the most important under autumn sowing conditions. Although it is detected under all conditions, its action seems not independent from environmental cues. An epistatic interaction involving the two vernalization genes was detected when the plants were grown without vernalization and under long photoperiod. The simultaneous presence of the winter Mogador allele at the two loci produced a marked delay in heading date, beyond a mere additive effect. This interaction, combined with the effect of the gene responsive to short photoperiod, Ppd-H2, was found responsible of the phenomenon known as short-day vernalization, present in some of the lines of the population.

Field responses of grain sorghum to a salinity gradient

Abstract Grain sorghum is a potential crop for moderately saline areas, having been identified as fairly tolerant to salinity, and shown to contain intraspecific variability for that trait. The aim of this work was to describe the responses of grain sorghum to saline irrigation, assess the responses of a set of genotypes to salinity, and to analyze the relationships between several agronomic and physiological traits and salinity tolerance. In an experiment during three years, eleven public inbred lines and one cultivar were exposed to a salinity gradient (NaCl and CaCl2, 1:1 w/w) created with a triple line source sprinkler system. The traits most affected by salinity were grain yield, number of grains per head, shoot dry weight (both grain and stover), harvest index, and leaf chloride, sodium, calcium, and potassium concentrations. Plant height, head length, and head number per plot were moderately affected by salinity, whereas flowering time, and total number of leaves per plant were unaffected. Two sets of three genotypes were identified with consistently contrasting responses to salinity across the three years. The differences in tolerance between these two groups were not associated with differences in total shoot biomass, but rather with different patterns of biomass partitioning under the most saline conditions. There were significant differences between the tolerant and susceptible genotypes in leaf chloride and potassium concentrations. The possible implications of the latter in the determination of the contrasting genotypic responses to salinity are discussed.

Expression analysis of vernalization and day-length response genes in barley (Hordeum vulgare L.) indicates that VRNH2 is a repressor of PPDH2 (HvFT3) under long days

The response to vernalization and the expression of genes associated with responses to vernalization (VRNH1, VRNH2, and VRNH3) and photoperiod (PPDH1 and PPDH2) were analysed in four barley (Hordeum vulgare L.) lines: ‘Alexis’ (spring), ‘Plaisant’ (winter), SBCC058, and SBCC106 (Spanish inbred lines), grown under conditions of vernalization and short days (VSD) or no vernalization and long days (NVLD). The four genotypes differ in VRNH1. Their growth habits and responses to vernalization correlated with the level of expression of VRNH1 and the length of intron 1. ‘Alexis’ and ‘Plaisant’ behaved as expected. SBCC058 and SBCC106 showed an intermediate growth habit and flowered relatively late in the absence of vernalization. VRNH1 expression was induced by cold for all genotypes. Under VSD, VRNH1 expression was detected in the SBCC genotypes later than in ‘Alexis’ but earlier than in ‘Plaisant’. VRNH2 was repressed under short days while VRNH1 expression increased in parallel. VRNH3 was detected only in ‘Alexis’ under NVLD, whereas it was not expressed in plants with the active allele of VRNH2 (SBCC058 and ‘Plaisant’). Under VSD, PPDH2 was expressed in ‘Alexis’, SBCC058, and SBCC106, but it was only expressed weakly in ‘Alexis’ under NVLD. Further analysis of PPDH2 expression in two barley doubled haploid populations revealed that, under long days, HvFT3 and VRNH2 expression levels were related inversely. The timing of VRNH2 expression under a long photoperiod suggests that this gene might be involved in repression of PPDH2 and, indirectly, in the regulation of flowering time through an interaction with the day-length pathway.

CHARACTERIZATION AND GENETIC CONTROL OF GERMINATION-EMERGENCE RESPONSES OF GRAIN SORGHUM TO SALINITY

SummaryWhen grain sorghum is grown in saline soils, one cause of low yield is poor crop establishment. The objectives of this study were to assess the response of grain sorghum to salinity in the germination-emergence stages, study the inheritance of salt tolerance at this stage, and determine the relative contribution to final emergence of salt effects during imbibition, and after onset of germination. Twelve inbred lines and 18 F1 hybrids, resulting from an incomplete 6×6 factorial mating design, were tested for germination and emergence in folded paper at 10 salt concentrations, from 1.8 to 36 dSm-1. The mean EC50 (the electrical conductivity at which the variable score declines by 50%) for emerged seedlings production was 21.2 dSm-1. Large genotypic differences were observed for salt tolerance at germination and emergence stages, which were not related to the viability of seeds, and poorly related to seed weight (considered as an estimate of intrinsic seed vigor). In the hybrids, these differences were due to SCA and female GCA for emergence, and female GCA for germination, though the male GCA was also significant for both characters. Line per se performance was significantly correlated to individual GCA estimates for emergence, but not for germination. Heterosis was only detected in three crosses for final emergence and in one cross for germination. The genetic differences in final emergence were mainly due to effects occurring after the onset of germination rather than a consequence of effects during imbibition.

Joint analysis for heading date QTL in small interconnected barley populations

The purpose of the present work is to validate the effect of the main QTL determining heading date in a set of 281 doubled haploid lines of barley, derived from 17 small interconnected populations, whose parents are cultivars commonly used in the Spanish barley breeding program. We used 72 molecular markers distributed across the seven chromosomes, particularly in regions known to contain flowering time genes or QTL. A combined linkage map over the 17 populations was constructed. The lines were evaluated in four field trials: two autumn sowings and two winter sowings, and in two treatments at a greenhouse trial, under controlled conditions of photoperiod and temperature. We have found that it is possible to carry out QTL detection in a complex germplasm set, representative of the materials used in an active breeding programme. In most cases two alleles per QTL were detected, though polymorphism of flanking markers was notably higher. The results revealed that there is a set of QTL that accounts for an important percentage of the phenotypic variation, suitable for marker assisted selection. Also, the role of the regions carrying the photoperiod response genes Ppd-H1 and Ppd-H2, the vernalization response genes Vrn-H1 and Vrn-H2, and the earliness per se locus Eam6, of which allele-specific or closely linked markers were available, was confirmed. These results support the use of this kind of approach for the validation of QTL found in single cross population studies, or to survey allelic diversity in plant breeding sets of materials.

Introgression of an intermediate VRNH1 allele in barley (Hordeum vulgare L.) leads to reduced vernalization requirement without affecting freezing tolerance

The process of vernalization is mainly controlled by two genes in winter barley (Hordeum vulgare L.), VRNH1 and VRNH2. A recessive allele at VRNH1 and a dominant allele at VRNH2 must be present to induce a vernalization requirement. In addition, this process is usually associated with greater low-temperature tolerance. Spanish barleys originated in areas with mild winters and display a reduced vernalization requirement compared with standard winter cultivars. The objective of this study was to investigate the genetic origin of this reduced vernalization requirement and its effect on frost tolerance. We introgressed the regions of a typical Spanish barley line that carry VRNH1 and VRNH2 into a winter cultivar, Plaisant, using marker-assisted backcrossing. We present the results of a set of 12 lines introgressed with all four possible combinations of VRNH1 and VRNH2, which were evaluated for vernalization requirement and frost tolerance. The reduced vernalization requirement of the Spanish parent was confirmed, and was found to be due completely to the effect of the VRNH1 region. The backcross lines showed no decline in frost tolerance compared with that of the recurrent parent unless they carried an extra segment of chromosome 5H. This extra segment, a carryover of the backcross process, apparently contained the well-known frost tolerance quantitative trait locus Fr-H2. We demonstrate that it is possible to manipulate the vernalization requirement with only minor effects on frost tolerance. This finding opens the path to creating new types of barley cultivars that are better suited to specific environments, especially in a climate-change scenario.

Olive oil quality and ripening in super-high-density Arbequina orchard

BACKGROUND The aim of this work was to evaluate the evolution of the quality of extra virgin olive oil obtained from a super-high-density Arbequina orchard, under a drip irrigation system, throughout the ripening process. For this objective, physicochemical, nutritional and sensory parameters were studied. In addition, the oxidative stability, pigment content and colour evolution of olive oil were analysed during the ripening process. RESULTS Free acidity increased slightly throughout the ripening process, while peroxide value and extinction coefficient decreased. Total phenol content and oxidative stability showed a similar trend, increasing at the beginning of ripening up to a maximum and thereafter decreasing. α-Tocopherol and pigment contents decreased with ripening, leading to changes in colour coordinates. Sensory parameters were correlated with total phenol content, following a similar trend throughout the maturation process. CONCLUSION By sampling and monitoring the ripeness index weekly, it would be possible to determine an optimal harvesting time for olives according to the industrial yield and the physicochemical, nutritional and sensory properties of the olive oil.

Identification of quantitative trait loci for resistance to powdery mildew in a Spanish barley landrace

The Spanish landrace-derived inbred line SBCC97, together with other lines from the Spanish Barley Core Collection, displays high resistance to powdery mildew, caused by the fungus Blumeria graminis f. sp. hordei. The objective of this study was to map quantitative trait loci (QTLs) for resistance to powdery mildew in a recombinant inbred line population derived from a cross between SBCC97 and the susceptible cultivar ‘Plaisant’. Phenotypic analysis was performed using four B. graminis isolates, and genetic maps were constructed with mainly simple sequence repeat (SSR) markers, following a sequential genotyping strategy. Two major QTLs with large effects were identified on chromosome 7H, and they accounted for up to 45% of the total phenotypic variance. The alleles for resistance at each QTL were contributed by the Spanish parent SBCC97. One locus was mapped to the short arm of chromosome 7HS, and was flanked by the resistance gene analogue (RGA) marker S9202 and the SSR GBM1060. This corresponded to the same chromosomal region in which a major race-specific resistance gene from Hordeum vulgare ssp. spontaneum, designated as mlt, had been identified previously. The second QTL was linked tightly to marker EBmac0755, and it shared its chromosomal location with the qualitative resistance gene Mlf, which has only been described previously in the wild ancestor H. spontaneum. This is the first report of these two QTLs occurring together in cultivated barley, and it paves the way for their use in barley breeding programs that are designed to transfer resistance alleles into elite cultivars.