Differential accumulation of sulfur-rich and sulfur-poor wheat flour proteins is affected by temperature and mineral nutrition during grain development

Hard red spring wheat (Triticum aestivum cv Butte86) was grown under controlled environmental conditions and grain produced under 24/17 °C, 37/17 °C or 37/28 °C day/night regimens with or without post-anthesis N supplied as NPK. Flour proteins were analyzed and quantified by differential fractionation and RP-HPLC, and endosperm proteins were assessed by two-dimensional gel electrophoresis (2-DE). High temperature or NPK during grain fill increased protein percentage and altered the proportions of S-rich and S-poor proteins. Addition of NPK increased protein accumulation per grain under the 24/17 °C but not the 37/28 °C regimen. However, flour protein composition was similar for grain produced with NPK at 24/17 °C or 37/28 °C. 2-DE of gluten proteins during grain development revealed that NPK or high temperature increased the accumulation rate for S-poor proteins more than for S-rich proteins. Flour S content did not indicate S-deficiency, however, and addition of post-anthesis S had no effect on protein composition. Although, high-protein flour from grain produced under the 37/28 °C regimen with or without NPK had loaf volumes comparable to flour produced at 24/17 °C with NPK, mixing tolerance was decreased by the high temperature regimen.     

Anthocyanin content of grains in purple wheat is affected by grain position,assimilate availability and agronomic management

Purple wheat grains are a promising source of anthocyanins and have been proposed as raw material for baking. However, little information is available about the effect of physiological and management variables on grain anthocyanins. This research aimed to (i) characterize the anthocyanin dynamic of grains set at different positions within the spikelet, (ii) evaluate the effect of different source–sink ratios on anthocyanins, and (iii) assess the response of anthocyanins to magnesium fertilization and harvest timing. Four source–sink ratios (pre- and post-anthesis spike halving, spike-shading and control) and two agronomic practices (Mg fertilization and early harvesting) were evaluated in field trials. Grain position affected grain weight and anthocyanin content, decreasing at distal positions. Both variables showed positive response to higher source–sink ratio. Pre-anthesis halving increased anthocyanin content by 54 and 37% (Exps. 1 and 2), while post-anthesis halving increased this trait by 31 and 29%. Source reduction decreased anthocyanin content by 26% and 51%. Changes in anthocyanin content associated with source–sink treatments and with grain positions were the result of different maximum anthocyanin content reached by physiological maturity. Magnesium fertilization and early harvesting increased anthocyanin content and concentration by 65 and 39%, respectively, seeming promising to increase anthocyanins in purple wheat.     

Individual and combined effects of pre- and post-anthesis temperature on protein composition of two malting barley cultivars

The present study has investigated the individual and combined influence of pre- and post-anthesis temperatures and cultivars on the protein composition in barley grains. Two barley cultivars were grown in soil and hydroponic systems in daylight chambers with different pre- and post-anthesis temperatures. Size exclusion (SE)-HPLC was used to evaluate the protein composition in mature barley grains. The results showed that individual and interactive effects of pre- and post-anthesis temperatures and cultivar variations influenced protein composition in the barley grains. Pre-anthesis temperature greatly affected the amounts of total sodium dodecyl sulphate (SDS) extractable proteins (TOTE) and explained 30% of the variation in TOTE. The barley cultivars accounted for 20% of the variation in TOTE. Variation in malting barley cultivars was found to influence the SDS extractable small monomers (41% of the variation). Percentage of SDS un-extractable polymeric proteins in total amount of polymeric proteins (%UPP) was governed by post-anthesis temperature, accounting for 11% of the variation and cultivar differences accounted for 7% of the variation. Thus, the climatic conditions during the specific growing period and a choice of cultivars played a major role in determining the protein composition and ultimately the malting quality of spring barley.     

Grain weight,radiation interception and use efficiency as affected by sink-strength in Mediterranean wheats released from 1940 to 2005

Bread wheat has been frequently characterised as sink-limited during grain filling but the degree of sink-limitation could vary with the contribution of breeding in increasing the number of grains per unit land area. This hypothetical change in the level of sink-limitation due to breeding has been insufficiently documented. Two source–sink manipulation experiments under field conditions with three released cultivars and an advanced breeding line representing important steps in wheat breeding in the Mediterranean area of Spain were conducted in order to quantify whether genetic improvement of grain yield in Mediterranean wheat modified the source–sink balance during grain filling, and how it affected grain weight and post-anthesis photosynthetically active radiation intercepted by the crop (IPAR) and radiation use efficiency (RUE). Average grain weight of control and trimmed spikes during grain filling was not significantly affected by halving the number of growing grains in the two oldest cultivars, but it was significantly increased in the most modern line, and had an intermediate response in the intermediate cultivar. In those cases in which a certain degree of source-limitation during grain growth occurred the magnitude of the response reflected a co-rather than a source-limitation. Considering grains from different positions within the spikes the smaller (distal) grains responded more markedly than the larger (proximal) grains. No differences in post-anthesis IPAR were found between the trimmed and control sub-plots for any of the genotypes. However, trimming the spikes reduced post-anthesis RUE, a fact corroborated by a simultaneous reduction in leaf net photosynthetic rate at noon. It seems that bread wheat breeding has tended to reduce the strength of the sink-limitation during post-anthesis even under Mediterranean conditions, and consequently the most modern line presented a sort of co-limitation.     

Effect of short heat shocks applied during grain development on wheat (Triticum aestivum L.) grain proteome

In the field, developing cereal grains are often exposed to short periods of very high temperature (>35 °C) that may dramatically affect grain yield and flour quality. Here we report on the effect of 4 h of heat shock (HS) at 38 °C applied on four consecutive days during the linear phase of storage compound accumulation on grain proteome of the winter bread wheat. At maturity the average single grain dry mass and the total quantity of nitrogen per grain were 25% and 16%, respectively lower for the HS treatment than for the control, resulting in a higher (+1.6% dm) grain protein concentration in HS grains. Individual albumin–globulin, gliadin and glutenin protein fractions from grains collected just before the HS and 1, 8, and 26 (ripeness maturity) days after the HS were quantified then analysed by 2-dimensional electrophoresis followed by MALDI-TOF and MS/MS identification. The quantity per grain of 10 gliadins and 3 low molecular weight glutenin subunit proteins were significantly affected by HS. Thirty-eight HS responsive albumin and globulin proteins were identified, including several enzymes involved in carbohydrate, redox, and lipid metabolisms. One protein was transitorily induced in response to HS. Detailed discussion of the expression of these proteins is presented.     

Aspergillus flavus growth and aflatoxin production as influenced by total lipid content during growth and development of cottonseed

Aspergillus flavus infects several food and feed crops, such as corn (Zea mays L.), cotton (Gossypium hirsutum L.), peanuts (Arachis hypogaea L.), and tree nut crops and contaminates the seed with carcinogenic aflatoxins. These susceptible crops contain rich reserves of lipids and fatty acids. The nature of relationship between lipids and the ability of the fungus to infect and produce aflatoxins in mature cottonseed, a protein-rich animal feed, has been addressed previously. In this study, we tracked lipid accumulation in developing cottonseed (15–35 days post-anthesis [DPA]) and also the ability of an aflatoxigenic strain and an isogenic non-aflatoxigenic strain to grow and produce aflatoxins in planta. The aflatoxigenic strain Af-70 green fluorescent protein (GFP) and the isogenic, non-aflatoxigenic strain SRRC 1500 (fungal collections maintained at the Southern Regional Research Center) did not differ much in infection and colonization of cottonseed. The non-aflatoxigenic strain did not produce aflatoxin at all stages of cottonseed development, whereas the aflatoxigenic strain Af-70 GFP produced copious amounts of aflatoxin and it coincided with the increasing levels of lipids, especially in mature cottonseed (30–35 DPA). Fungal growth, as quantified by the GFP expression in the fungus, was highly correlated with toxin production.     

Cloning, expression and characterization of novel avenin-like genes in wheat and related species

B-Type avenin-like genes and proteins were characterized in 23 species of Triticeae. Southern blot analysis showed that the avenin-like genes belong to a multigene family. RT-PCR showed expression only in developing endosperms of wheat and related species, between 3 and 22 DPA (days post anthesis) with a peak between 11 and 15 DPA in wheat. The encoded proteins are cysteine-rich, containing 18–19 cysteine residues. An avenin-like protein from wheat was expressed in Escherichia coli, purified and used to raise polyclonal antibodies. These antibodies were used to detect b-type avenin-like proteins in endosperms of wheat and related species by western blotting.     

Effect of soil compaction in the sub-humid cropping environment in Pakistan on uptake of NPK and grain yield in wheat (Triticum aestivum): I. Compaction

Soil compaction is a major cause of decrease in crop yield. The most serious cause of soil compaction is continuous ploughing at the same depth which affects bulk density, porosity and root proliferation, consequently affecting concentration and uptake of nutrients by plants. The effects of soil compaction on concentration and uptake of Nitrogen, Phosphorus and Potassium (NPK) by wheat were studied at the Agriculture Research Institute, Mingora, Pakistan in two separate experiments, conducted during 2002–2003 and repeated in 2003–2004. The treatments in each experiment consisted of four compaction levels arranged in a randomized complete block design replicated four times. Subsoil compaction affected soil bulk density and total porosity. With increasing compaction, bulk density increased in the range of 15–26% while total porosity decreased in the range of 15–27%. Compaction treatments significantly and progressively decreased concentration and uptake of NPK in both years of the experiments. Higher nutrient concentration and uptake was recorded during the second year as compared to first year, probably as a result of higher seasonal rainfall. Concentration of NPK showed reductions of 5–20%, 10–53% and 9–21%, respectively, due to the compaction treatments over control. The uptake of NPK decreased due to the compaction treatments in the range of 7–26%, 11–54% and 11–28%, respectively, over control. Compaction treatments decreased the dry matter accumulation in the range of 2–9% whereas grain yield showed a reduction of 5–48%. Inverse relationships between bulk density, and concentration and uptake of NPK, dry matter accumulation and grain yield were recorded. The implications of these findings for intensive agricultural systems in Pakistan and similar environments are discussed.     

Effect of high temperature on albumin and globulin accumulation in the endosperm proteome of the developing wheat grain

The accumulation of KCl-soluble/methanol-insoluble albumins and globulins was investigated in the endosperm of developing wheat (Triticum aestivum, L. cv. Butte 86) grain produced under a moderate (24 °C/17 °C, day/night) or a high temperature regimen (37 °C/28 °C) imposed from 10 or 20 days post-anthesis (dpa) until maturity. Proteins were separated by 2-DE and developmental profiles for nearly 200 proteins were analyzed by hierarchical clustering. Comparison of protein profiles across physiologically equivalent stages of grain fill revealed that high temperature shortened, but did not substantially alter, the developmental program. Accumulation of proteins shifted from those active in biosynthesis and metabolism to those with roles in storage and protection against biotic and abiotic stresses. Few proteins responded transiently when plants were transferred to the high temperature regimens, but levels of a number of proteins were altered during late stages of grain development. Specific protein responses depended on whether the high temperature regimens were initiated early or mid development. Some of the heat responsive proteins have been implicated in gas bubble stabilization in bread dough and others are suspected food allergens.     

Interfacial Behaviour of Secalin and Rye Flour-milling Streams in Comparison with Gliadin

Secalin was extracted from rye flour and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The SDS-gel pattern showed that some proteins were composed of disulphide linked polypeptide chains. However, a gliadin sample contained more polymeric protein. The behaviour of gliadin and secalin at the air–water interface was compared using the surface balance technique. It was found that secalin was more surface active than gliadin, spreading faster and to a higher surface pressure. The influence of pH on the interfacial behaviour was also studied. The surface pressure after 45 min equilibrium of both gliadin and secalin decreased with decreasing pH. The effect was independent of the acid (hydrochloric acid, lactic acid or ascorbic acid) when compared at the same pH. The behaviour at the gas–liquid interface of five different rye flour-milling streams together with the whole (straight run) flour was also investigated. The fraction with highest protein content spread fastest and reached the highest surface pressure value. When spread on ascorbic acid at pH 3·7 the surface pressure of the flour stream with lowest protein content decreased to the greatest extent, whereas the fraction with the highest protein content was not affected. It was thus found that, although secalin showed an interfacial behaviour similar to gliadin, this behaviour was not necessarily shown by the total protein mixture in a rye flour.     

Water stress during grain development affects starch synthesis,composition and physicochemical properties in triticale

Triticale, a man-made cereal crop developed from a cross between wheat and rye, has excellent agronomic traits to produce starch for bioindustrial applications. The effects of different levels of water stress on expression of starch synthesis genes and starch composition and physicochemical properties were investigated in this study. Three triticale varieties from 5 days post-anthesis were treated with three levels of water stress: low water stress (LWS) at 55–60% of soil moisture, moderate water stress (MWS) at 30–35% soil moisture and severe water stress (SWS) at 10–15% soil moisture. Water stress led to a significant reduction in seed weight at SWS (35–45%). A decrease in starch content was noticeable from MWS onwards and the values were decreased by 42–55% at SWS across all varieties. Such decrease was associated with the reduced expression of starch synthesis genes at 19 days of water stress (DWS). MWS favoured an increase of amylose proportion in triticale starch and it was accompanied by a significant up-regulation of GBSSI expression throughout the grain development. Triticale starch synthesized under water stress showed a reduced population of small granules and an increase of A-type to B-type ratio. SWS caused pitting on starch granules but did not alter the biconcave disc shape of mature granules. An inverse relationship between water stress and a range of starch gelatinization temperature was observed and the MWS environment specifically decreased the peak temperature (Tp) and increased the enthalpy. Our results signify that starch morphology, composition and physicochemical properties in triticale grains could be altered if triticale is grown under drought conditions.     

Effects of gamma-irradiation of wheat on gluten proteins

The effects of ⁶⁰Co gamma-irradiation treatments (2·5, 5·0, 10·0 and 20·0 kGy) on the gluten proteins of two bread wheats and one durum wheat cultivar were investigated. Dough rheological properties of the flour processed from the irradiated wheat were also determined using a computerised micromixograph. Irradiation caused a significant deteriorating effect on all mixogram parameters. There was no observable effect of irradiation on gliadin proteins analysed by polyacrylamide gel electrophoresis. The 50% 1-propanol-insoluble (50 PI) glutenin fraction was highly affected by irradiation. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, reduced 50 PI glutenin showed a noticeable reduction in band intensities of both high (HMW) and low molecular weight (LMW) glutenin subunits (GS) with increasing irradiation dosage greater than 5 kGy. The irradiation effect on 50 PI glutenin was further studied and quantified by reversed-phase high-performance liquid chromatography of glutenin subunits; there was a progressive decrease in the quantity of subunits with increasing irradiation dose level. Compared to non-irradiated wheat, the relative decline in total insoluble glutenin at the 20 kGy dosage level ranged from 34–49% depending on cultivar. Increasing levels of irradiation also progressively reduced the ratio of HMW:LMW-GS up to 13–15% at 20 kGy indicating that irradiation had a greater effect on the largest polymers of glutenin. The observed weakening of dough mixing properties and concomitant decline in the quantity of 50 PI glutenin with increasing levels of gamma-irradiation are consistent with a degradation of glutenin to a lower average molecular size by depolymerisation and/or disaggregation.     

Histological development of the sunflower fruit pericarp as affected by pre- and early post-anthesis canopy shading

The dynamics of pericarp development compared to that of the embryo, as well as the effect of pre-anthesis and post-anthesis shading on pericarp histogenesis and dry weight dynamics of fruits from two sunflower (Helianthus annuus L.) hybrid cultivars along three capitulum positions, peripheral, mid and central were studied. During fruit formation, the cell division phase of the carpel takes place before anthesis. Eight days after anthesis the pericarp reached its final size, while its cell wall's sclerification was almost complete 13 days after anthesis. Pre-anthesis shading affected the carpel cell division period reducing (17–33%) the number of pericarp middle layer strata and increased the thickenings of the cell wall of the mid (19%) and central (33–63%) fruits. In central fruits, the dry weight accumulation period was reduced. In contrast, post-anthesis shading reduced both the cell wall thickness (16–64%) and the number (38–58%) of pericarp middle layer sclerified strata of fruits in the three positions of the capitulum. In the mid and central fruits, the dry weight accumulation period extended 11–16 and 3–4 days, respectively, over those of the control. Both shading treatments produced thinner and lighter pericarps, but with different anatomical features that were associated with differences in the efficiency of use of the fruit for industrial oil extraction.