A multiparental cross population for mapping QTL for relevant agronomic traits in durum wheat (Triticum durum Desf.)

Multiparental cross designs for mapping quantitative trait loci (QTL) in crops are efficient alternatives to conventional biparental experimental populations because they exploit a broader genetic basis and higher mapping resolution. We describe the development and deployment of a multiparental recombinant inbred line (RIL) population in durum wheat (Triticum durum Desf.) obtained by crossing four elite cultivars characterized by different traits of agronomic value. A linkage map spanning 2,663 cM and including 7,594 single nucleotide polymorphisms (SNPs) was produced by genotyping 338 RILs with a wheat-dedicated 90k SNP chip. A cluster file was developed for correct allele calling in the framework of the tetraploid durum wheat genome. Based on phenotypic data collected over four field experiments, a multi-trait quantitative trait loci (QTL) analysis was carried out for 18 traits of agronomic relevance (including yield, yield-components, morpho-physiological and seed quality traits). Across environments, a total of 63 QTL were identified and characterized in terms of the four founder haplotypes. We mapped two QTL for grain yield across environments and 23 QTL for grain yield components. A novel major QTL for number of grain per spikelet/ear was mapped on chr 2A and shown to control up to 39% of phenotypic variance in this cross. Functionally different QTL alleles, in terms of direction and size of genetic effect, were distributed among the four parents. Based on the occurrence of QTL-clusters, we characterized the breeding values (in terms of effects on yield) of most of QTL for heading and maturity as well as yield component and quality QTL. This multiparental RIL population provides the wheat community with a highly informative QTL mapping resource enabling the dissection of the genetic architecture of multiple agronomic relevant traits in durum wheat.

Selective Transport of Zinc, Manganese, Nickel, Cobalt and Cadmium in the Root System and Transfer to the Leaves in Young Wheat Plants

• Background and Aims The uptake, translocation and redistribution of the heavy metals zinc, manganese, nickel, cobalt and cadmium are relevant for plant nutrition as well as for the quality of harvested plant products. The long-distance transport of these heavy metals within the root system and the release to the shoot in young wheat (Triticum aestivum ‘Arina’) plants were investigated. • Methods After the application of 65Zn, 54Mn, 63Ni, 57Co and 109Cd for 24 h to one seminal root (the other seminal roots being excised) of 54-h-old wheat seedlings, the labelled plants were incubated for several days in hydroponic culture on a medium without radionuclides. • Key Results The content of 65Zn decreased quickly in the labelled part of the root. After the transfer of 65Zn from the roots to the shoot, a further redistribution in the phloem from older to younger leaves was observed. In contrast to 65Zn, 109Cd was released more slowly from the roots to the leaves and was subsequently redistributed in the phloem to the youngest leaves only at trace levels. The content of 63Ni decreased quickly in the labelled part of the root, moving to the newly formed parts of the root system and also accumulating transiently in the expanding leaves. The 54Mn content decreased quickly in the labelled part of the root and increased simultaneously in leaf 1. A strong retention in the labelled part of the root was observed after supplying 57Co. • Conclusions The dynamics of redistribution of 65Zn, 54Mn, 63Ni, 57Co and 109Cd differed considerably. The rapid redistribution of 63Ni from older to younger leaves throughout the experiment indicated a high mobility in the phloem, while 54Mn was mobile only in the xylem and 57Co was retained in the labelled root without being loaded into the xylem.

A systematic review of the evidence for use of herbal medicine for the treatment of acute diarrhea

Background. Acute diarrhea (AD) is an important cause of morbidity and mortality among both children and adults. An ideal antidiarrheal treatment should be safe, effective, compatible with Oral Rehydration Solution, and inexpensive. Herbal medicines, if effective, should fit these criteria as well or better than standard treatment. ^ Objective. The objective of the present study was to assess the effectiveness of plant preparations in patients with AD in reports of randomized and non-randomized controlled trials. ^ Aims. The aims of the present study were to identify effective antidiarrheal herbs and to identify potential antidiarrheal herbs for future studies of efficacy through well designed clinical trials in human populations. ^ Methods. Nineteen published studies of herbal management of AD were examined to identify effective plant preparations. Ten plant preparations including Berberine (Berberis aristata), tormentil root ( Potentialla tormentilla), baohauhau (from the baobaosan plant), carob (Ceratonia siliqua), pectin (Malus domestica), wood creosote (Creosote bush), guava (Psidium guajava L.), belladonna (Atropa belladonna), white bean (Phaseolis vulgaris), and wheat (Triticum aestivum) were identified. ^ Results. Qualitative data analysis of nineteen clinical trials indicated berberine’s potentially valuable antisecretory effects against diarrhea caused by Vibrio cholerae and enterotoxigenic Escherichia coli. Tormentil root showed significant efficacy against rotavirus-induced diarrhea; carob exhibited antidiarrheal properties not only by acting to detoxify and constipate but by providing a rich source of calories; guava and belladonna are antispasmodics and have been shown to relieve the symptoms of AD. Finally, white bean and wheat yielded favorable clinical and dietary outcomes in children with diarrhea. ^ Conclusion. The present study is the first to review the evidence for use of herbal compounds for treatment of AD. Future randomized controlled trials are needed to evaluate their efficacy and safety.^

Analysis of genetic variability in a sample of the durum wheat (Triticum durum Desf.) Spanish collection based on gliadin markers

In this work gliadin proteins were used to analyse the genetic variability in a sample of the durum wheat Spanish collection conserved at the CRF-INIA. In total 38 different alleles were identified at the loci Gli-A1, Gli-A3, Gli-B5, Gli-B1, Gli-A2 and Gli-B2. All the gliadin loci were polymorphic, possessed large genetic diversity and small and large differentiation within and between varieties, respectively. The Gli-A2 and Gli-B2 loci were the most polymorphic, the most fixed within varieties and the most useful to distinguish among varieties. Alternatively, Gli-B1 locus presented the least genetic variability out of the four main loci Gli-A1, Gli-B1, Gli-A2 and Gli-B2. The Gli-B1 alleles coding for the gliadin γ-45, associated with good quality, had an accumulated frequency of 69.7%, showing that the Spanish germplasm could be a good source for breeding quality. The Spanish landraces studied showed new gliadin alleles not catalogued so far. These new alleles might be associated with specific Spanish environment factors. The large number of new alleles identified also indicates that durum wheat Spanish germplasm is rather unique.

Creation and Validation of the Spanish Durum Wheat Core Collection.

Spanish wheat (Triticum spp.) landraces have a considerable polymorphism, containing many unique alleles, relative to other collections. The existence of a core collection is a favored approach for breeders to efficiently explore novel variation and enhance the use of germplasm. In this study, the Spanish durum wheat (Triticum turgidum L.) core collection (CC) was created using a population structure–based method, grouping accessions by subspecies and allocating the number of genotypes among populations according to the diversity of simple sequence repeat (SSR) markers. The CC of 94 genotypes was established, which accounted for 17% of the accessions in the entire collection. An alternative core collection (CH), with the same number of genotypes per subspecies and maximizing the coverage of SSR alleles, was assembled with the Core Hunter software. The quality of both core collections was compared with a random core collection and evaluated using geographic, agromorphological, and molecular marker data not previously used in the selection of genotypes. Both core collections had a high genetic representativeness, which validated their sampling strategies. Geographic and agromorphological variation, phenotypic correlations, and gliadin alleles of the original collection were more accurately depicted by the CC. Diversity arrays technology (DArT) markers revealed that the CC included genotypes less similar than the CH. Although more SSR alleles were retained by the CH (94%) than by the CC (91%), the results showed that the CC was better than CH for breeding purposes.

Calidad semolera y nuevas variantes al��licas prolaminas en variedades indígenas españolas d trigo duro (Triticum Turgidum L. Tell.)

Las prolaminas son las principales proteínas del endospermo de los trigos panaderos que están relacionadas con la calidad panadera. El objetivo de este trabajo es caracterizar gluteninas LMW según la nomenclatura de Liu et al y relacionarlas con la fuerza panadera.

Generalización de variables medioambientales mediante interpolación GIS

La investigación de esta tesis se centra en el estudio de técnicas geoestadísticas y su contribución a una mayor caracterización del binomio factores climáticos-rendimiento de un cultivo agrícola. El inexorable vínculo entre la variabilidad climática y la producción agrícola cobra especial relevancia en estudios sobre el cambio climático o en la modelización de cultivos para dar respuesta a escenarios futuros de producción mundial. Es información especialmente valiosa en sistemas operacionales de monitoreo y predicción de rendimientos de cultivos Los cuales son actualmente uno de los pilares operacionales en los que se sustenta la agricultura y seguridad alimentaria mundial; ya que su objetivo final es el de proporcionar información imparcial y fiable para la regularización de mercados. Es en este contexto, donde se quiso dar un enfoque alternativo a estudios, que con distintos planteamientos, analizan la relación inter-anual clima vs producción. Así, se sustituyó la dimensión tiempo por la espacio, re-orientando el análisis estadístico de correlación interanual entre rendimiento y factores climáticos, por el estudio de la correlación inter-regional entre ambas variables. Se utilizó para ello una técnica estadística relativamente nueva y no muy aplicada en investigaciones similares, llamada regresión ponderada geográficamente (GWR, siglas en ingl��s de “Geographically weighted regression”). Se obtuvieron superficies continuas de las variables climáticas acumuladas en determinados periodos fenol��gicos, que fueron seleccionados por ser factores clave en el desarrollo vegetativo de un cultivo. Por ello, la primera parte de la tesis, consistió en un análisis exploratorio sobre comparación de Métodos de Interpolación Espacial (MIE). Partiendo de la hipótesis de que existe la variabilidad espacial de la relación entre factores climáticos y rendimiento, el objetivo principal de esta tesis, fue el de establecer en qué medida los MIE y otros métodos geoestadísticos de regresión local, pueden ayudar por un lado, a alcanzar un mayor entendimiento del binomio clima-rendimiento del trigo blando (Triticum aestivum L.) al incorporar en dicha relación el componente espacial; y por otro, a caracterizar la variación de los principales factores climáticos limitantes en el crecimiento del trigo blando, acumulados éstos en cuatro periodos fenol��gicos. Para lleva a cabo esto, una gran carga operacional en la investigación de la tesis consistió en homogeneizar y hacer los datos fenol��gicos, climáticos y estadísticas agrícolas comparables tanto a escala espacial como a escala temporal. Para España y los Bálticos se recolectaron y calcularon datos diarios de precipitación, temperatura máxima y mínima, evapotranspiración y radiación solar en las estaciones meteorol��gicas disponibles. Se dispuso de una serie temporal que coincidía con los mismos años recolectados en las estadísticas agrícolas, es decir, 14 años contados desde 2000 a 2013 (hasta 2011 en los Bálticos). Se superpuso la malla de información fenol��gica de cuadrícula 25 km con la ubicación de las estaciones meteorol��gicas con el fin de conocer los valores fenol��gicos en cada una de las estaciones disponibles. Hecho esto, para cada año de la serie temporal disponible se calcularon los valores climáticos diarios acumulados en cada uno de los cuatro periodos fenol��gicos seleccionados P1 (ciclo completo), P2 (emergencia-madurez), P3 (floración) y P4 (floraciónmadurez). Se calcul�� la superficie interpolada por el conjunto de métodos seleccionados en la comparación: técnicas deterministas convencionales, kriging ordinario y cokriging ordinario ponderado por la altitud. Seleccionados los métodos más eficaces, se calcul�� a nivel de provincias las variables climatol��gicas interpoladas. Y se realizaron las regresiones locales GWR para cuantificar, explorar y modelar las relaciones espaciales entre el rendimiento del trigo y las variables climáticas acumuladas en los cuatro periodos fenol��gicos. Al comparar la eficiencia de los MIE no destaca una técnica por encima del resto como la que proporcione el menor error en su predicción. Ahora bien, considerando los tres indicadores de calidad de los MIE estudiados se han identificado los métodos más efectivos. En el caso de la precipitación, es la técnica geoestadística cokriging la más idónea en la mayoría de los casos. De manera unánime, la interpolación determinista en función radial (spline regularizado) fue la técnica que mejor describía la superficie de precipitación acumulada en los cuatro periodos fenol��gicos. Los resultados son más heterogéneos para la evapotranspiración y radiación. Los métodos idóneos para estas se reparten entre el Inverse Distance Weighting (IDW), IDW ponderado por la altitud y el Ordinary Kriging (OK). También, se identificó que para la mayoría de los casos en que el error del Ordinary CoKriging (COK) era mayor que el del OK su eficacia es comparable a la del OK en términos de error y el requerimiento computacional de este último es mucho menor. Se pudo confirmar que existe la variabilidad espacial inter-regional entre factores climáticos y el rendimiento del trigo blando tanto en España como en los Bálticos. La herramienta estadística GWR fue capaz de reproducir esta variabilidad con un rendimiento lo suficientemente significativo como para considerarla una herramienta válida en futuros estudios. No obstante, se identificaron ciertas limitaciones en la misma respecto a la información que devuelve el programa a nivel local y que no permite desgranar todo el detalle sobre la ejecución del mismo. Los indicadores y periodos fenol��gicos que mejor pudieron reproducir la variabilidad espacial del rendimiento en España y Bálticos, arrojaron aún, una mayor credibilidad a los resultados obtenidos y a la eficacia del GWR, ya que estaban en l��nea con el conocimiento agronómico sobre el cultivo del trigo blando en sistemas agrícolas mediterráneos y norteuropeos. Así, en España, el indicador más robusto fue el balance climático hídrico Climatic Water Balance) acumulado éste, durante el periodo de crecimiento (entre la emergencia y madurez). Aunque se identificó la etapa clave de la floración como el periodo en el que las variables climáticas acumuladas proporcionaban un mayor poder explicativo del modelo GWR. Sin embargo, en los Bálticos, países donde el principal factor limitante en su agricultura es el bajo número de días de crecimiento efectivo, el indicador más efectivo fue la radiación acumulada a lo largo de todo el ciclo de crecimiento (entre la emergencia y madurez). Para el trigo en regadío no existe ninguna combinación que pueda explicar más allá del 30% de la variación del rendimiento en España. Poder demostrar que existe un comportamiento heterogéneo en la relación inter-regional entre el rendimiento y principales variables climáticas, podría contribuir a uno de los mayores desafíos a los que se enfrentan, a día de hoy, los sistemas operacionales de monitoreo y predicción de rendimientos de cultivos, y éste es el de poder reducir la escala espacial de predicción, de un nivel nacional a otro regional. ABSTRACT This thesis explores geostatistical techniques and their contribution to a better characterization of the relationship between climate factors and agricultural crop yields. The crucial link between climate variability and crop production plays a key role in climate change research as well as in crops modelling towards the future global production scenarios. This information is particularly important for monitoring and forecasting operational crop systems. These geostatistical techniques are currently one of the most fundamental operational systems on which global agriculture and food security rely on; with the final aim of providing neutral and reliable information for food market controls, thus avoiding financial speculation of nourishments of primary necessity. Within this context the present thesis aims to provide an alternative approach to the existing body of research examining the relationship between inter-annual climate and production. Therefore, the temporal dimension was replaced for the spatial dimension, re-orienting the statistical analysis of the inter-annual relationship between crops yields and climate factors to an inter-regional correlation between these two variables. Geographically weighted regression, which is a relatively new statistical technique and which has rarely been used in previous research on this topic was used in the current study. Continuous surface values of the climate accumulated variables in specific phenological periods were obtained. These specific periods were selected because they are key factors in the development of vegetative crop. Therefore, the first part of this thesis presents an exploratory analysis regarding the comparability of spatial interpolation methods (SIM) among diverse SIMs and alternative geostatistical methodologies. Given the premise that spatial variability of the relationship between climate factors and crop production exists, the primary aim of this thesis was to examine the extent to which the SIM and other geostatistical methods of local regression (which are integrated tools of the GIS software) are useful in relating crop production and climate variables. The usefulness of these methods was examined in two ways; on one hand the way this information could help to achieve higher production of the white wheat binomial (Triticum aestivum L.) by incorporating the spatial component in the examination of the above-mentioned relationship. On the other hand, the way it helps with the characterization of the key limiting climate factors of soft wheat growth which were analysed in four phenological periods. To achieve this aim, an important operational workload of this thesis consisted in the homogenization and obtention of comparable phenological and climate data, as well as agricultural statistics, which made heavy operational demands. For Spain and the Baltic countries, data on precipitation, maximum and minimum temperature, evapotranspiration and solar radiation from the available meteorological stations were gathered and calculated. A temporal serial approach was taken. These temporal series aligned with the years that agriculture statistics had previously gathered, these being 14 years from 2000 to 2013 (until 2011 for the Baltic countries). This temporal series was mapped with a phenological 25 km grid that had the location of the meteorological stations with the objective of obtaining the phenological values in each of the available stations. Following this procedure, the daily accumulated climate values for each of the four selected phenological periods were calculated; namely P1 (complete cycle), P2 (emergency-maturity), P3 (flowering) and P4 (flowering- maturity). The interpolated surface was then calculated using the set of selected methodologies for the comparison: deterministic conventional techniques, ordinary kriging and ordinary cokriging weighted by height. Once the most effective methods had been selected, the level of the interpolated climate variables was calculated. Local GWR regressions were calculated to quantify, examine and model the spatial relationships between soft wheat production and the accumulated variables in each of the four selected phenological periods. Results from the comparison among the SIMs revealed that no particular technique seems more favourable in terms of accuracy of prediction. However, when the three quality indicators of the compared SIMs are considered, some methodologies appeared to be more efficient than others. Regarding precipitation results, cokriging was the most accurate geostatistical technique for the majority of the cases. Deterministic interpolation in its radial function (controlled spline) was the most accurate technique for describing the accumulated precipitation surface in all phenological periods. However, results are more heterogeneous for the evapotranspiration and radiation methodologies. The most appropriate technique for these forecasts are the Inverse Distance Weighting (IDW), weighted IDW by height and the Ordinary Kriging (OK). Furthermore, it was found that for the majority of the cases where the Ordinary CoKriging (COK) error was larger than that of the OK, its efficacy was comparable to that of the OK in terms of error while the computational demands of the latter was much lower. The existing spatial inter-regional variability between climate factors and soft wheat production was confirmed for both Spain and the Baltic countries. The GWR statistic tool reproduced this variability with an outcome significative enough as to be considered a valid tool for future studies. Nevertheless, this tool also had some limitations with regards to the information delivered by the programme because it did not allow for a detailed break-down of its procedure. The indicators and phenological periods that best reproduced the spatial variability of yields in Spain and the Baltic countries made the results and the efficiency of the GWR statistical tool even more reliable, despite the fact that these were already aligned with the agricultural knowledge about soft wheat crop under mediterranean and northeuropean agricultural systems. Thus, for Spain, the most robust indicator was the Climatic Water Balance outcome accumulated throughout the growing period (between emergency and maturity). Although the flowering period was the phase that best explained the accumulated climate variables in the GWR model. For the Baltic countries where the main limiting agricultural factor is the number of days of effective growth, the most effective indicator was the accumulated radiation throughout the entire growing cycle (between emergency and maturity). For the irrigated soft wheat there was no combination capable of explaining above the 30% of variation of the production in Spain. The fact that the pattern of the inter-regional relationship between the crop production and key climate variables is heterogeneous within a country could contribute to one is one of the greatest challenges that the monitoring and forecasting operational systems for crop production face nowadays. The present findings suggest that the solution may lay in downscaling the spatial target scale from a national to a regional level.

Temperature and precipitation effects on wheat yield across a European transect: a crop model ensemble analysis using impact response surfaces

This study explored the utility of the impact response surface (IRS) approach for investigating model ensemble crop yield responses under a large range of changes in climate. IRSs of spring and winter wheat Triticum aestivum yields were constructed from a 26-member ensemble of process-based crop simulation models for sites in Finland, Germany and Spain across a latitudinal transect. The sensitivity of modelled yield to systematic increments of changes in temperature (-2 to +9°C) and precipitation (-50 to +50%) was tested by modifying values of baseline (1981 to 2010) daily weather, with CO2 concentration fixed at 360 ppm. The IRS approach offers an effective method of portraying model behaviour under changing climate as well as advantages for analysing, comparing and presenting results from multi-model ensemble simulations. Though individual model behaviour occasionally departed markedly from the average, ensemble median responses across sites and crop varieties indicated that yields decline with higher temperatures and decreased precipitation and increase with higher precipitation. Across the uncertainty ranges defined for the IRSs, yields were more sensitive to temperature than precipitation changes at the Finnish site while sensitivities were mixed at the German and Spanish sites. Precipitation effects diminished under higher temperature changes. While the bivariate and multi-model characteristics of the analysis impose some limits to interpretation, the IRS approach nonetheless provides additional insights into sensitivities to inter-model and inter-annual variability. Taken together, these sensitivities may help to pinpoint processes such as heat stress, vernalisation or drought effects requiring refinement in future model development.

Plastid-localized acetyl-CoA carboxylase of bread wheat is encoded by a single gene on each of the three ancestral chromosome sets

5′-End fragments of two genes encoding plastid-localized acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) of wheat (Triticum aestivum) were cloned and sequenced. The sequences of the two genes, Acc-1,1 and Acc-1,2, are 89% identical. Their exon sequences are 98% identical. The amino acid sequence of the biotin carboxylase domain encoded by Acc-1,1 and Acc-1,2 is 93% identical with the maize plastid ACCase but only 80–84% identical with the cytosolic ACCases from other plants and from wheat. Four overlapping fragments of cDNA covering the entire coding region were cloned by PCR and sequenced. The wheat plastid ACCase ORF contains 2,311 amino acids with a predicted molecular mass of 255 kDa. A putative transit peptide is present at the N terminus. Comparison of the genomic and cDNA sequences revealed introns at conserved sites found in the genes of other plant multifunctional ACCases, including two introns absent from the wheat cytosolic ACCase genes. Transcription start sites of the plastid ACCase genes were estimated from the longest cDNA clones obtained by 5′-RACE (rapid amplification of cDNA ends). The untranslated leader sequence encoded by the Acc-1 genes is at least 130–170 nucleotides long and is interrupted by an intron. Southern analysis indicates the presence of only one copy of the gene in each ancestral chromosome set. The gene maps near the telomere on the short arm of chromosomes 2A, 2B, and 2D. Identification of three different cDNAs, two corresponding to genes Acc-1,1 and Acc-1,2, indicates that all three genes are transcriptionally active.

Patterns of Starchy Endosperm Acidification and Protease Gene Expression in Wheat Grains following Germination1

Cereal aleurone responses to gibberellic acid (GA3) include activation of synthesis of hydrolytic enzymes and acidification of the external medium. We have studied the effect of the pH of the incubation medium on the response of wheat (Triticum aestivum) aleurone cells to GA3. De-embryonated half grains show the capacity for GA3-activated medium acidification when incubation is carried out at pH 6.0 to 7.0 but not at lower pHs. In addition, the activating effect of GA3 on the expression of carboxypeptidase III and thiol protease genes is more efficient when the hormone treatment is carried out at neutral pH. In situ pH staining showed that starchy endosperm acidification takes place upon imbibition and advances from the embryo to the distal part of the grain. In situ hybridization experiments showed a similar pattern of expression of a carboxypeptidase III gene, which is up-regulated by GA3 in aleurone cells. However, aleurone gene expression precedes starchy endosperm acidification. These findings imply that in vivo GA perception by the aleurone layer takes place at neutral pH and suggest that the acidification of the starchy endosperm is regulated by GA3 in germinated wheat grains.

Circadian-Regulated Transcription of the psbD Light-Responsive Promoter in Wheat Chloroplasts1

The level of mRNAs derived from the plastid-encoded psbD light-responsive promoter (LRP) is controlled by a circadian clock(s) in wheat (Triticum aestivum). The circadian oscillations in the psbD LRP mRNA level persisted for at least three cycles in continuous light and for one cycle in continuous dark, with maxima in subjective morning and minima in subjective early night. In vitro transcription in chloroplast extracts revealed that the circadian cycles in the psbD LRP mRNA level were dominantly attributed to the circadian-regulated transcription of the psbD LRP. The effects of various mutations introduced into the promoter region on the psbD LRP activity in vitro suggest the existence of two positive elements located between −54 and −36, which generally enhance the transcription activity, and an anomalous core promoter structure lacking the functional “−35” element, which plays a crucial role in the circadian fluctuation and light dependency of psbD LRP transcription activity.

Detection of Ca2+-Dependent Transglutaminase Activity in Root and Leaf Tissue of Monocotyledonous and Dicotyledonous Plants

Protein extracted from root and leaf tissue of the dicotyledonous plants pea (Pisum sativum) and broad bean (Vicia faba) and the monocotyledonous plants wheat (Triticum aestivum) and barley (Hordeum vulgare) were shown to catalyze the incorporation of biotin-labeled cadaverine into microtiter-plate-bound N′,N′-dimethylcasein and the cross-linking of biotin-labeled casein to microtiter-plate-bound casein in a Ca2+-dependent manner. The cross-linking of biotinylated casein and the incorporation of biotin-labeled cadaverine into N′,N′-dimethylcasein were time-dependent reactions with a pH optimum of 7.9. Transglutaminase activity was shown to increase over a 2-week growth period in both the roots and leaves of pea. The product of transglutaminase's protein-cross-linking activity, ε-(γ-glutamyl)-lysine isodipeptide, was detected in root and shoot protein from pea, broad bean, wheat, and barley by cation-exchange chromatography. The presence of the isodipeptide was confirmed by reversed-phase chromatography. Hydrolysis of the isodipeptide after cation-exchange chromatography confirmed the presence of glutamate and lysine.

Expression and Localization of Phosphoenolpyruvate Carboxylase in Developing and Germinating Wheat Grains1

Phosphoenolpyruvate carboxylase (PEPC) activity and corresponding mRNA levels were investigated in developing and germinating wheat (Triticum aestivum) grains. During grain development PEPC activity increased to reach a maximum 15 d postanthesis. Western-blot experiments detected two main PEPC polypeptides with apparent molecular masses of 108 and 103 kD. The most abundant 103-kD PEPC subunit remained almost constant throughout the process of grain development and in the scutellum and aleurone layer of germinating grains. The less-abundant 108-kD polypeptide progressively disappeared during the second half of grain development and was newly synthesized in the scutellum and aleurone layer of germinating grains. PEPC mRNA was detected throughout the process of grain development; however, in germinating grains PEPC mRNA accumulated transiently in the scutellum and aleurone layer, showing a sharp maximum 24 h after imbibition. Immunolocalization studies revealed the presence of the enzyme in tissues with a high metabolic activity, as well as in the vascular tissue of the crease area of developing grains. A clear increase in PEPC was observed in the scutellar epithelium of grains 24 h after imbibition. The data suggest that the transiently formed PEPC mRNA in the scutellar epithelium encodes the 108-kD PEPC subunit.

Xyloglucan Octasaccharide XXLGol Derived from the Seeds of Hymenaea courbaril Acts as a Signaling Molecule1

Treatment of the xyloglucan isolated from the seeds of Hymenaea courbaril with Humicola insolens endo-1,4-β-d-glucanase I produced xyloglucan oligosaccharides, which were then isolated and characterized. The two most abundant compounds were the heptasaccharide (XXXG) and the octasaccharide (XXLG), which were examined by reference to the biological activity of other structurally related xyloglucan compounds. The reduced oligomer (XXLGol) was shown to promote growth of wheat (Triticum aestivum) coleoptiles independently of the presence of 2,4-dichlorophenoxyacetic acid (2,4-D). In the presence of 2,4-D, XXLGol at nanomolar concentrations increased the auxin-induced response. It was found that XXLGol is a signaling molecule, since it has the ability to induce, at nanomolar concentrations, a rapid increase in an α-l-fucosidase response in suspended cells or protoplasts of Rubus fruticosus L. and to modulate 2,4-D or gibberellic acid-induced α-l-fucosidase.

Genetic and economic analysis of a targeted marker-assisted wheat breeding strategy

The advent of molecular markers as a tool to aid selection has provided plant breeders with the opportunity to rapidly deliver superior genetic solutions to problems in agricultural production systems. However, a major constraint to the implementation of marker-assisted selection (MAS) in pragmatic breeding programs in the past has been the perceived high relative cost of MAS compared to conventional phenotypic selection. In this paper, computer simulation was used to design a genetically effective and economically efficient marker-assisted breeding strategy aimed at a specific outcome. Under investigation was a strategy involving the integration of both restricted backcrossing and doubled haploid (DH) technology. The point at which molecular markers are applied in a selection strategy can be critical to the effectiveness and cost efficiency of that strategy. The application of molecular markers was considered at three phases in the strategy: allele enrichment in the BC1F1 population, gene selection at the haploid stage and the selection for recurrent parent background of DHs prior to field testing. Overall, incorporating MAS at all three stages was the most effective, in terms of delivering a high frequency of desired outcomes and at combining the selected favourable rust resistance, end use quality and grain yield alleles. However, when costs were included in the model the combination of MAS at the BC1F1 and haploid stage was identified as the optimal strategy. A detailed economic analysis showed that incorporation of marker selection at these two stages not only increased genetic gain over the phenotypic alternative but actually reduced the over all cost by 40%.