The response of four winter wheat (Triticum aestivum L.) varieties to field water deficit: leaf anti-oxidative protection and proteolytic activity

Introduction: Drought is one of the most significant factors that limit plant productivity. Oxidative stress is a secondary event in many unfavorable environmental conditions. Intracellular proteases have a role in the metabolism reorganisation and nutrient remobilization under stress. In order to under stand the relative significance of oxidative stress and proteolysis in the yield reduction under drought, four varieties of Triticum aestivum L. with different field drought resistance were examined. Methods: A two-year field experiment was conducted. Analyses were performed on the upper most leaf of control plants and plants under water deficitat the stages most critical for yield reduction under drought (from jointing till milk ripeness). Leaf water deficit and electrolyte leakage, malondyaldehyde level, activities and isoenzymes of superoxide dismutase, catalase and peroxidase, leaf protein content and proteolytic activity were studied. Yield components were analyzed. Results: A general trend of increasing the membrane in stability and accumulation of lipid hydroperoxides was observed with some differences among varieties, especially under drought. The anti-oxidative enzyme activities were progressively enhanced, as well as the azocaseinolytic activities. The leaf protein content decreased under drought at the last phase. Differences among varieties were observed in the parameters under study. They were compared to yield components` reduction under water deprivation.

Die Auffindung des wilden Emmers (Triticum Dicoccum) in Nordpalästina

Aaron Aaronsohn

Die Auffindung des wilden Emmers (Triticum Dicoccum) in Nordpalästina

G. Schweinfurth

Light intensity selectively influences the distribution and further redistribution of macro- and micronutrients in hydroponically grown wheat ( Triticum aestivum L.)

Investigations were focused on light effects on allocation of root-borne macronutrients (calcium, magnesium and potassium) and micronutrients (iron, manganese, zinc and copper) in roots, shoots and harvested grains of wheat (Triticum aestivum L.). Plants were exposed to low (100 μmol photons m−2 s−1) or high light (380 μmol photons m−2 s−1). High light stimulated both root and shoot growth. While the total contents per plant of some nutrients were markedly higher (calcium and potassium) or lower (copper) under high light, no major differences were observed for other nutrients. The distribution of nutrients and the further redistribution within the shoot were influenced by the light intensity in an element-specific manner. Nutrients were selectively directed to the leaves of the main shoot (low light) or to the tillers (high light). The quality of the harvested grains was also affected by the light intensity.

Respuesta productiva a diferentes programaciones de riego en trigo (Triticum aestivum L.) : región semiárida de Córdoba, Argentina

En la evaluación regional de riego con equipos presurizados, se observa que la mayoría de los establecimientos no hacen uso de programas de riego basados en necesidades hídricas, dando lugar a un uso inadecuado de los recursos hídricos. Por ello, en el presente trabajo se evalúa la producción de trigo (Triticum aestivum L.) bajo riego programado considerando las variables clima-suelo-cultivo. La experiencia se desarrolló en el campo experimental de la Universidad Nacional de Río Cuarto durante los ciclos 1999, 2000 y 2001. Se contó con la información climática de la estación meteorológica instalada en el lugar. En los ensayos se consideró un diseño estadístico en parcelas totalmente al azar con seis repeticiones. Los tratamientos fueron los siguientes: 1. sin riego; 2. reponiendo desde un umbral de marchitez incipiente hasta capacidad de campo; 3. reponiendo desde umbral de marchitez incipiente hasta un nivel que permita el aprovechamiento del agua de lluvia con un 75 % de probabilidad de ocurrencia; 4. riego sólo en el período comprendido entre las etapas fenológicas de macollaje y grano lechoso y 5. considerando el modelo de programación YACU. Los resultados de esta experiencia en los tres años considerados establecen una respuesta significativa al riego complementario, con una producción promedio de los tratamientos bajo riego de 4980 kg.ha-1 en 1999, 5895 kg.ha-1 en el 2000 y 6271 kg.ha-1 en el 2001, contra 3296 kg.ha-1, 4054 kg.ha-1 y 3197 kg.ha-1, respectivamente, para los tratamientos sin riego. Dentro de los tratamientos bajo riego, en los dos primeros años se obtuvieron valores significativamente menores en las parcelas regadas sólo entre macollaje y grano lechoso: 4704 kg.ha-1 en 1999 y 4973 kg.ha-1 en el 2000.

Caracterización del patrón de gluteninas de bajo peso molecular de trigo pan (Triticum aestivum L.) y su efecto sobre el potencial uso final de la harina

El trigo pan [Tritivum aestivum L.]tiene diversos usos en la industria alimenticia debido a las características particulares de la harina y es el ingrediente principal en la elaboración de pan, galletitas, productos de repostería, pastas frescas, etc. La calidad intrínseca de las harinas está influenciada por la cantidad y calidad de proteínas del gluten: Gluteninas de alto [HMW-Gs]y bajo peso molecular [LMW-Gs]y Gliadinas [Gli]. A su vez, la calidad reológica está afectada por factores ambientales y la relación Genotipo x Ambiente. Por lo mencionado, resulta importante conocer los efectos de los patrones proteicos sobre la calidad industrial de trigo pan y comprender la incidencia de la disponibilidad de Nitrógeno y temperaturas elevadas durante el llenado de los granos sobre las fracciones proteicas que componen el gluten y la reolgía de las masas. Se utilizaron proteinogramas [SDS-PAGE, unidimensionales]para la identificación proteica de 74 cultivares argentinos de trigo pan y el test de sedimentación [SDSS]para estimar la fuerza de gluten, como indicador de calidad intrínseca y para ver el efecto del ambiente se utilizaron 26 cultivares de trigo pan sembrados en el mismo sitio y con las mismas condiciones de manejo, en dos años [2008 y 2009]con temperaturas contrastantes durante el llenado de los granos y dos niveles de fertilización nitrogenada. La combinación total de las proteínas del gluten [HMW-Gs+LMWGs+Gli]tuvo más influencia sobre la calidad del gluten que cada fracción proteica en particular. En general, las altas temperaturas aumentaron la cantidad del Gli y la extensibilidad de las masas [L], en tanto que la disponibilidad de N incrementó todos los grupos proteicos, la fuerza [W]y la extensibilidad de la masa. Igualmente, se observó un efecto diferencial de dichos factores sobre los patrones proteicos y la calidad final, según la composición del gluten.

Effects of N fertilisation, year and prolamin alleles on guten quality in durum wheat (Triticum turgidum L. ssp. turgidum) landraces from Spain

A subset of durum wheat Spanish landraces, previously evaluated for yield at low and high nitrogen (N) levels, was analysed for quality, protein content (P) and sodium dodecyl sulphate sedimentation (SDSS) test. The evaluation was carried out at the two N rates and in two years. The influence of prolamin alleles at the Glu-1, Glu-3, Glu-B2 and Gli-1 loci on quality parameters was also studied. The non significant Variety-by-Year or Variety-by-N interactions suggested that year and N affected all the varieties in a similar manner. Year and N effects were larger than variety effect for P, which increased with N. In contrast, variety genotype exhibited a stronger influence on SDSS test, which was not affected by year and fertilizer. Variety effects on P did not reflect the variety differences for SDSS test. A high positive influence of some prolamin alleles on quality parameters was detected, mainly for SDSS values. No correlation between yield and P was detected in the landraces adapted to low N. Based on the results of yield and quality evaluations, four landraces with high yield and high gluten strength were pre-selected for low N production.

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.

Comparison of several models for calculating ozone stomatal fluxes on a Mediterranean wheat cultivar (Triticum durum Desf. cv. Camacho)

Ozone stomatal fluxes were modeled for a 3-year period following different approaches for a commercial variety of durum wheat (Triticum durum Desf. cv. Camacho) at the phenological stage of anthesis. All models performed in the same range, although not all of them afforded equally significant results. Nevertheless, all of them suggest that stomatal conductance would account for the main percentage of ozone deposition fluxes. A new modeling approach was tested, based on a 3-D architectural model of the wheat canopy, and fairly accurate results were obtained. Plant species-specific measurements, as well as measurements of stomatal conductance and environmental parameters, were required. The method proposed for calculating ozone stomatal fluxes (FO(3_3-D)) from experimental gs data and modeling them as a function of certain environmental parameters in conjunction with the use of the YPLANT model seems to be adequate, providing realistic estimates of the canopy FO(3_3-D), integrating and not neglecting the contribution of the lower leaves with respect to the flag leaf, although a further development of this model is needed.

Soil water availability modulation over estimated relative yield losses in wheat (Triticum aestivum L.) due to ozone exposure

The approach developed by Fuhrer in 1995 to estimate wheat yield losses induced by ozone and modulated by the soil water content (SWC) was applied to the data on Catalonian wheat yields. The aim of our work was to apply this approach and adjust it to Mediterranean environmental conditions by means of the necessary corrections. The main objective pursued was to prove the importance of soil water availability in the estimation of relative wheat yield losses as a factor that modifies the effects of tropospheric ozone on wheat, and to develop the algorithms required for the estimation of relative yield losses, adapted to the Mediterranean environmental conditions. The results show that this is an easy way to estimate relative yield losses just using meteorological data, without using ozone fluxes, which are much more difficult to calculate. Soil water availability is very important as a modulating factor of the effects of ozone on wheat; when soil water availability decreases, almost twice the amount of accumulated exposure to ozone is required to induce the same percentage of yield loss as in years when soil water availability is high.

A PCR-based method for discriminating between high molecular weight glutenin subunits Bx7 and Bx7* in Triticum aestivum L

The correct assignment of high molecular weight glutenin subunit variants is a key task in wheat breeding. However, the traditional analysis by protein electrophoresis is sometimes difficult and not very precise. This work describes a novel DNA marker for the accurate discrimination between the Glu-B1 locus subunits Bx7 and Bx7*. The analysis of one hundred and forty two bread wheat cultivars from different countries has highlighted a great number of misclassifications in the literature that could lead to wrong conclusions in studies of the relationship between glutenin composition and wheat quality.

Diversity and Genetic structure of the Spanish collection of durum wheat (Triticum turgidum L) landraces

The objectives of this study were to assess diversity and genetic structure of a collection of Spanish durum wheat (Triticum turgidum L) landraces, using SSRs, DArTs and gliadin-markers, and to correlate the distribution of diversity with geographic and climatic features, as well as agro-morphological traits. A high level of diversity was detected in the genotypes analyzed, which were separated into nine populations with a moderate to great genetic divergence among them. The three subspecies taxa, dicoccon, turgidum and durum, present in the collection, largely determined the clustering of the populations. Genotype variation was lower in dicoccon (one major population) and turgidum (two major populations) than in durum (five major populations). Genetic differentiation by the agro-ecological zone of origin was greater in dicoccon and turgidum than in durum. DArT markers revealed two geographic substructures, east-west for dicoccon and northeast-southwest for turgidum. The ssp. durum had a more complex structure, consisting of seven populations with high intra-population variation. DArT markers allowed the detection of subgroups within some populations, with agro-morphological and gliadin differences, and distinct agro-ecological zones of origin. Two different phylogenetic groups were detected; revealing that some durum populations were more related to ssp. turgidum from northern Spain, while others seem to be more related to durum wheats from North Africa

Estudio molecular de gluteninas de alto y bajo peso molecular en Triticum aestivum ssp. vulgare L. y su relación con la calidad panadera

El trigo blando (Triticum aestivum ssp vulgare L., AABBDD, 2n=6x=42) presenta propiedades viscoélasticas únicas debidas a la presencia en la harina de las prolaminas: gluteninas y gliadinas. Ambos tipos de proteínas forman parte de la red de gluten. Basándose en la movilidad en SDS-PAGE, las gluteninas se clasifican en dos grupos: gluteninas de alto peso molecular (HMW-GS) y gluteninas de bajo peso molecular (LMW-GS). Los genes que codifican para las HMW-GS se encuentran en tres loci del grupo 1 de cromosomas: Glu-A1, Glu-B1 y Glu-D1. Cada locus codifica para uno o dos polipéptidos o subunidades. La variación alélica de las HMW-GS es el principal determinante de de la calidad harino-panadera y ha sido ampliamente estudiado tanto a nivel de proteína como de ADN. El conocimiento de estas proteínas ha contribuido sustancialmente al progreso de los programas de mejora para la calidad del trigo. Comparadas con las HMW-GS, las LMW-GS forman una familia proteica mucho más compleja. La mayoría de los genes LMW se localizan en el grupo 1 de cromosomas en tres loci: Glu-A3, Glu-B3 y Glu-D3 que se encuentran estrechamente ligados a los loci que codifican para gliadinas. El número de copias de estos genes ha sido estimado entre 10-40 en trigo hexaploide, pero el número exacto aún se desconoce debido a la ausencia de un método eficiente para diferenciar los miembros de esta familia multigénica. La nomenclatura de los alelos LMW-GS por electroforesis convencional es complicada, y diferentes autores asignan distintos alelos a la misma variedad lo que dificulta aún más el estudio de esta compleja familia. El uso de marcadores moleculares para la discriminación de genes LMW, aunque es una tarea dificil, puede ser muy útil para los programas de mejora. El objetivo de este trabajo ha sido profundizar en la relación entre las gluteninas y la calidad panadera y desarrollar marcadores moleculares que permitan ayudar en la correcta clasificación de HMW-GS y LMW-GS. Se han obtenido dos poblaciones de líneas avanzadas F4:6 a partir de los cruzamientos entre las variedades ‘Tigre’ x ‘Gazul’ y ‘Fiel’ x ‘Taber’, seleccionándose para los análisis de calidad las líneas homogéneas para HMW-GS, LMW-GS y gliadinas. La determinación alélica de HMW-GS se llevó a cabo por SDS-PAGE, y se complementó con análisis moleculares, desarrollándose un nuevo marcador de PCR para diferenciar entre las subunidades Bx7 y Bx7*del locus Glu-B1. Resumen 2 La determinación alélica para LMW-GS se llevó a cabo mediante SDS-PAGE siguiendo distintas nomenclaturas y utilizando variedades testigo para cada alelo. El resultado no fue concluyente para el locus Glu-B3, así que se recurrió a marcadores moleculares. El ADN de los parentales y de los testigos se amplificó usando cebadores diseñados en regiones conservadas de los genes LMW y fue posteriormente analizado mediante electroforesis capilar. Los patrones de amplificación obtenidos fueron comparados entre las distintas muestras y permitieron establecer una relación con los alelos de LMW-GS. Con este método se pudo aclarar la determinación alélica de este locus para los cuatro parentales La calidad de la harina fue testada mediante porcentaje de contenido en proteína, prueba de sedimentación (SDSS) y alveógrafo de Chopin (parámetros P, L, P/L y W). Los valores fueron analizados en relación a la composición en gluteninas. Las líneas del cruzamiento ‘Fiel’ x ‘Taber’ mostraron una clara influencia del locus Glu-A3 en la variación de los valores de SDSS. Las líneas que llevaban el nuevo alelo Glu-A3b’ presentaron valores significativamente mayores que los de las líneas con el alelo Glu-A3f. En las líneas procedentes del cruzamiento ‘Tigre ’x ‘Gazul’, los loci Glu-B1 y Glu-B3 loci mostraron ambos influencia en los parámetros de calidad. Los resultados indicaron que: para los valores de SDSS y P, las líneas con las HMW-GS Bx7OE+By8 fueron significativamente mejores que las líneas con Bx17+By18; y las líneas que llevaban el alelo Glu-B3ac presentaban valores de P significativamente superiores que las líneas con el alelo Glu-B3ad y significativamente menores para los valores de L . El análisis de los valores de calidad en relación a los fragmentos LMW amplificados, reveló un efecto significativo entre dos fragmentos (2-616 y 2-636) con los valores de P. La presencia del fragmento 2-636 estaba asociada a valores de P mayores. Estos fragmentos fueron clonados y secuenciados, confirmándose que correspondían a genes del locus Glu-B3. El estudio de la secuencia reveló que la diferencia entre ambos se hallaba en algunos SNPs y en una deleción de 21 nucleótidos que en la proteína correspondería a un InDel de un heptapéptido en la región repetida de la proteína. En este trabajo, la utilización de líneas que difieren en el locus Glu-B3 ha permitido el análisis de la influencia de este locus (el peor caracterizado hasta la fecha) en la calidad panadera. Además, se ha validado el uso de marcadores moleculares en la determinación alélica de las LMW-GS y su relación con la calidad panadera. Summary 3 Bread wheat (Triticum aestivum ssp vulgare L., AABBDD, 2n=6x=42) flour has unique dough viscoelastic properties conferred by prolamins: glutenins and gliadins. Both types of proteins are cross-linked to form gluten polymers. On the basis of their mobility in SDS-PAGE, glutenins can be classified in two groups: high molecular weight glutenins (HMW-GS) and low molecular weight glutenins (LMW-GS). Genes encoding HMW-GS are located on group 1 chromosomes in three loci: Glu-A1, Glu-B1 and Glu-D1, each one encoding two polypeptides, named subunits. Allelic variation of HMW-GS is the most important determinant for bread making quality, and has been exhaustively studied at protein and DNA level. The knowledge of these proteins has substantially contributed to genetic improvement of bread quality in breeding programs. Compared to HMW-GS, LMW-GS are a much more complex family. Most genes encoded LMW-GS are located on group 1 chromosomes. Glu-A3, Glu-B3 and Glu-D3 loci are closely linked to the gliadin loci. The total gene copy number has been estimated to vary from 10–40 in hexaploid wheat. However, the exact copy number of LMW-GS genes is still unknown, mostly due to lack of efficient methods to distinguish members of this multigene family. Nomenclature of LMW-GS alleles is also unclear, and different authors can assign different alleles to the same variety increasing confusion in the study of this complex family. The use of molecular markers for the discrimination of LMW-GS genes might be very useful in breeding programs, but their wide application is not easy. The objective of this work is to gain insight into the relationship between glutenins and bread quality, and the developing of molecular markers that help in the allele classification of HMW-GS and LMW-GS. Two populations of advanced lines F4:6 were obtained from the cross ‘Tigre’ x ‘Gazul’ and ‘Fiel’ x ‘Taber’. Lines homogeneous for HMW-GS, LMW-GS and gliadins pattern were selected for quality analysis. The allele classification of HMW-GS was performed by SDS-PAGE, and then complemented by PCR analysis. A new PCR marker was developed to undoubtedly differentiate between two similar subunits from Glu-B1 locus, Bx7 and Bx7*. The allele classification of LMW-GS was initially performed by SDS-PAGE following different established nomenclatures and using standard varieties. The results were not completely concluding for Glu-B3 locus, so a molecular marker system was applied. DNA from parental lines and standard varieties was amplified using primers designed in conserved domains of LMW genes and analyzed by capillary electrophoresis. The pattern of amplification products obtained was compared among samples and related to the protein allele classification. It was possible to establish a correspondence between specific amplification products and almost all LMW alleles analyzed. With this method, the allele classification of the four parental lines was clarified. Flour quality of F4:6 advanced lines were tested by protein content, sedimentation test (SDSS) and alveograph (P, L, P/L and W). The values were analyzed in relation to the lines prolamin composition. In the ‘Fiel’ x ‘Taber’ population, Glu-A3 locus showed an influence in SDSS values. Lines carrying new allele Glu-A3b’, presented a significantly higher SDSS value than lines with Glu-A3f allele. In the ‘Tigre ’x ‘Gazul’ population, the Glu-B1 and Glu-B3 loci also showed an effect in quality parameters, in SDSS, and P and L values. Results indicated that: for SDSS and P, lines with Bx7OE+By8 were significantly better than lines with Bx17+By18; lines carrying Glu-B3ac allele had a significantly higher P values than Glu-B3ad allele values. lines with and lower L The analysis of quality parameters and amplified LMW fragments revealed a significant influence of two peaks (2-616 y 2-636) in P values. The presence of 2-636 peak gave higher P values than 2-616. These fragments had been cloned and sequenced and identified as Glu-B3 genes. The sequence analysis revealed that the molecular difference between them was some SNPs and a small deletion of 21 nucleotides that in the protein would produce an InDel of a heptapeptide in the repetitive region. In this work, the analysis of two crosses with differences in Glu-3 composition has made possible to study the influence of LMG-GS in quality parameters. Specifically, the influence of Glu-B3, the most interesting and less studied loci has been possible. The results have shown that Glu-B3 allele composition influences the alveograph parameter P (tenacity). The existence of different molecular variants of Glu-B3 alleles have been assessed by using a molecular marker method. This work supports the use of molecular approaches in the study of the very complex LMW-GS family, and validates their application in the analysis of advanced recombinant lines for quality studies.

Calidad semolera y nuevas variantes alélicas 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.