Puroindoline genotype,starch granule size distribution and milling quality of wheat

Genetically-diverse wheat samples from the Australian Winter Cereals Collection propagated in two environments were sequenced to identify puroindoline genotypes then the relationships between flour yield, genotype, starch granule size distribution and starch-bound puroindoline protein content were investigated. The Pina-D1a, Pinb-D1b genotype resulted in a higher average flour yield than either the Pina-D1b, Pinb-D1a or the Pina-D1a, Pinb-D1a but the ranges of flour yields for the three genotypes showed considerable overlap. For both hard wheat genotypes (Pina-D1a, Pinb-D1b or Pina-D1b, Pinb-D1a), a higher proportion of type A to type C starch granules was associated with higher flour yield and this relationship accounted for between 31% and 33% of the variation in flour yield. This result is consistent with previously reported findings for soft wheat. For the Pina-D1a, Pinb-D1b genotype, increased flour yield was also associated with a decrease in starch granule-bound puroindoline protein, which accounted for 31–35% of the variation in flour yield across the two environments. The combined effect of starch granule type and associated puroindoline content accounted for 68% of the variation in flour yield within the Pina-D1a, Pinb-D1b genotype.     

Effect of tolerance to Septoria tritici blotch on grain yield,yield components and grain quality in Argentinean wheat cultivars

Septoria tritici blotch (STB) caused by Zymoseptoria tritici (Mycosphaerella graminicola) is a major disease of wheat worldwide due to significant losses in grain yield and quality. Disease tolerance is the ability to maintain yield performance in the presence of disease symptoms. Therefore, it could be a useful tool in the management of the disease. Although it is known, that there is disease tolerance to STB in some wheat cultivars, this aspect has not been studied among Argentinean cultivars. The aims of this study were to evaluate genotypic differences in tolerance to STB among Argentinean cultivars, considering the relationship between the area under disease progress curve or the green leaf area or the non-green leaf area duration with the grain yield. In addition the effect of the disease on yield, yield components, test weight, grain protein concentration, wet and dry gluten concentration and the influence of tolerance on these traits was investigated. Field experiments were carried out with ten cultivars in a split-split-plot design during 2010 and 2011. Inoculation treatments were the main plots and cultivars, the subplots. STB significantly reduced grain yield, their components, test weight and increase grain protein and gluten concentration. Cultivar Baguette 10 showed major tolerance to STB, indicated by a consistent low regression slope between the green area duration and yield, while Klein Chaja was non-tolerant due to a high regression slope. However, many cultivars such as Buck Brasil, Buck 75 Aniversario, Klein Escorpion and Klein Flecha had considerably similar regression slopes to Baguette 10, provided good levels of tolerance. Other cultivars presented no significant differences. The correlation coefficient between tolerance and grain yield potential was not significant, suggesting that tolerant high-yielding cultivars can be obtained. No relationship was found between quality group or tolerance with the increase in protein and gluten concentration due to STB either.     

Effects of drought and the presence of the 1BL/1RS translocation on grain vitreosity, hardness and protein content in winter wheat

Grain texture is an important component of end-use quality in wheat. The effects of water availability on the components of texture; vitreosity, determined using a Light Transflectance meter (LTm), grain hardness measured using the single-kernel characterisation system (SKCS), and protein content, were studied in field experiments of winter wheat in the UK in 2001/2002 and 2002/2003. Experiments were grown on a drought prone soil and employed a mapping population of 46 doubled haploid (DH) lines and their parents, Beaver (+1BL/1RS, soft wheat) and Soissons (1B, hard wheat). The results showed that drought increased hardness in both seasons, but the effect was never sufficient to move a line from the soft class into the hard class. Puroindoline (PIN)-a:b peak height ratio explained ca. 78% of the variation in hardness, and drought also appeared to increase the amounts of PINs in the grain. Minor quantitative trait loci (QTLs) were found for hardness on chromosomes 2A, 2D, 3A and 6D, also associated with QTLs for PINs. Vitreosity also increased in response to drought in both seasons. Variation in vitreosity explained 7–11% of the overall variation in texture within a hardness class, with hardness increasing on average by 2.2 SKCS units for each 10% increase in the proportion of vitreous grains. The relationship between vitreosity and protein content was poor, despite the fact that protein content also increased in response to drought. Minor QTLs associated with both protein content and vitreosity were found on chromosomes 1B, 4D and 5D. A minor QTL for vitreosity was also found on chromosome 2D. However, there appeared to be no direct relationship between alleles at the Ha locus, the gene which controls the difference between hard and soft wheats, and vitreosity. A positive relationship between the presence of the 1BL/1RS translocation and the proportion of vitreous grains was identified, suggesting that vitreosity was strongly linked to changes in protein quality.     

Modeling plant carbon flow and grain starch accumulation in wheat

The process of starch accumulation in grain directly influences the yield and quality formation in wheat. Since few studies have been aimed at modeling the grain starch accumulation, this study was undertaken to develop a simplified process model for predicting the rate of starch accumulation in wheat grain by focusing on the variation of plant carbon dynamics post-anthesis. Five different experiments involving genotypes, nitrogen rates and water regimes were conducted to support model development and model evaluation. The model proposed that the starch accumulation rate (STR) in individual grain was determined by the availability of carbon source in plant (GCA_i) and the ability of starch synthesis in grain (f(A_i)), as influenced by the factors of temperature, water and nitrogen conditions within plants. The f(A_i) could be described in a two-section curve with post-anthesis growing degree days (GDD), first exponential increase and then linear decrease. The GCA_i was determined by post-anthesis carbon assimilation and carbon remobilization from vegetative organs to grains. A genotypic parameter was incorporated into the model algorithm, i.e. the maximum rate of individual grain starch accumulation, to differentiate the ability of starch accumulation among cultivars. The overall performance of the model was validated with different data sets from three field experiments spanning 3 years and comprising various genotypes, nitrogen and water levels. The RMSE values for all treatments were averaged as 12.51%, indicating a good fit between the simulated and observed data. It appears that the model can give a reliable prediction for grain starch accumulation of different wheat cultivars under various growing conditions.     

Optimal numbers of environments to assess slopes of joint regression for grain yield,grain protein yield and grain protein concentration under nitrogen constraint in winter wheat

Plant breeders are interested in rationally reducing the number of testing environments for breeding new genotypes adapted to diverse conditions. One way to characterize the adaptation of a genotype is to use the joint regression model. Our objectives were to estimate the stability for grain yield (GY), grain protein yield (GPY) and grain protein content (GPC) of a set of wheat genotypes grown under varying nitrogen conditions and then to determine optimal numbers of environments for assessing the slopes of joint regression.     

Combined nitrogen and sulphur fertilisation and its effect on wheat quality and protein composition measured by SE-FPLC and proteomics

Soil sulphur deficiency, which is increasingly prevalent in Western Europe, lowers wheat yields, and also affects the gluten quality of the flour. Differences in S availability may change the proportion of S-poor to S-rich gliadins and glutenin subunits. This may cause unpredictable and unwanted variations in wheat quality. The combined effects of nitrogen (N) and sulphur (S) fertilisers and split application of S and N on wheat gluten quality and composition were investigated. The results revealed effects of S fertilisation on dough quality. At high N fertilisation levels significant responses to S fertilisation were found which emphasised the need for precision application of S in intensive wheat production systems. Protein fractionation by SE-FPLC showed that quality differences were associated with changing proportions of high M_r polymeric proteins. Changes in protein composition of salt soluble proteins were also confirmed by proteomics. Glyceraldehyde-3-phosphate dehydrogenase and one of the serpin protein spots increased at high N, combined with the lower S level. The enzymes also increased in samples with increased S fertilisation combined with low N, but was not changed at higher N levels. Furthermore, at high N the serpin protein spot, and also a 27 K protein and one unidentified protein spot decreased with increasing S.     

Effect of endosperm starch granule size distribution on milling yield in hard wheat

Increased flour yield in hard wheat is associated with increased endosperm rheology index, calculated from strength and stiffness as measured by the SKCS. A study of the fractured endosperm of hard wheat varieties grouped according to similar rheology index values was performed using environmental scanning electron microscopy (ESEM). Differing microstructures and fracture patterns were observed between each group. Specifically, the group representing high rheology index had a greater concentration of small starch granules in prismatic cells. Samples of diverse wheat germplasm were grown at two sites and subjected to laboratory milling. Starch granule size distribution (SGSD) analysis using a laser diffraction method was undertaken on a subset of samples in triplicate representing a range in flour yield. The results supported an hypothesis for a significant influence of SGSD on flour yield of hard wheat varieties. In addition, a significant part (R²>0.40 (p<0.05) at two sites) of the association appeared to be under genetic control. Results indicate a more even gradation of distributions involving an increase in the sample volume % of small granule (types B and C) and decrease in type A granules. This was associated with increased rheology index values and higher flour yield. The ratio of type A:C starch granules accounted for up to 58% (p<0.05) of the variation in flour yield in the samples studied. Thus, rheological parameters measured using a rapid SKCS screening method can be linked to the genetic regulation of SGSD with implications for the improvement of commercial processing performance of hard wheat.     

The effect of gluten on the retrogradation of wheat starch

The retrogradation of amylopectin in a wheat starch and a wheat starch/gluten (10:1) blend prepared by extrusion and containing 34% water (wet weight basis) was studied using X-ray diffraction, differential scanning calorimetry and NMR relaxometry during storage at constant water content and temperature (25 °C). For both samples, amylopectin ‘fully’ retrograded after 2–3 days storage, i.e. the different parameters monitored with time to follow the retrogradation had reached their maximum value, and crystallised predominantly into the A polymorph. Under the experimental conditions used, there was no evidence of any significant effects of the presence of gluten on the kinetics, extent or polymorphism of amylopectin retrogradation.     

The influence of temperature extremes on some quality and starch characteristics in bread,biscuit and durum wheat

Environmental conditions during grain-fill can affect the duration of protein accumulation and starch deposition, and thus play an important role in grain yield and flour quality of wheat. Two bread-, one durum- and one biscuit wheat were exposed to extreme low (−5.5 °C for 3 h) and high (32 °C/15 °C day/night for three days) temperatures during grain filling under controlled conditions for two consecutive seasons. Flour protein content was increased significantly in one bread wheat, Kariega, under heat stress. Cold stress significantly reduced SDS sedimentation in both bread wheats. Kernel weight and diameter were significantly decreased at both stress treatments for the two bread wheats. Kernel characteristics of the biscuit wheat were thermo stable. Kernel hardness was reduced in the durum wheat for the heat treatment. Durum wheat had consistently low SDS sedimentation values and the bread wheat high values. Across the two seasons, the starch content in one bread wheat was significantly reduced by both high and low temperatures, as is reflected in the reduction of weight and diameter of these kernels. In the durum wheat, only heat caused a significant reduction in starch content, which is again reflected in the reduction of kernel weight and diameter.     

Pre-anthesis high temperature acclimation alleviates the negative effects of post-anthesis heat stress on stem stored carbohydrates remobilization and grain starch accumulation in wheat

The potential role of pre-anthesis high temperature acclimation in alleviating the negative effects of post-anthesis heat stress on stem stored carbohydrate remobilization and grain starch accumulation in wheat was investigated. The treatments included no heat-stress (CC), heat stress at pre-anthesis only (HC), heat at post-anthesis only (CH), and heat stress at both stages (HH). Post-anthesis heat stress decreased grain starch content, reduced the content of fructans and depressed activities of related synthesis enzymes of sucrose:sucrose fructosyltransferase and fructan:fructan fructosyltransferase. Interestingly, HH plants had significantly higher grain yield than the CH plants. In addition, post-anthesis high temperature lowered grain starch content and increased percentages of volume, number and surface area of B-type starch granules in CH and HH than in CC treatment. However, HH plants had much higher starch content, and caused less modified B-type starch granule size indicators than the CH plants. Our results indicated that, compared with the non-acclimated plants, the pre-anthesis high temperature acclimation effectively enhanced carbohydrate remobilization from stems to grains, led to less changed starch content and starch granule size distribution in grains of wheat under post-anthesis heat stress.     

Effects of species and breeding on wheat protein composition

Wheat proteins are characterized by their excellent contribution to technological and baking properties. However, wheat proteins, especially gluten and amylase-trypsin inhibitors (ATIs), are also known to be responsible for a broad range of intolerances and allergies. In order to evaluate the impact of genetic variability on the composition of these functional but immunogenic protein types, a set of different Triticum species, including common wheat, durum, spelt, emmer and einkorn, was examined regarding ATI and gluten concentrations by RP-HPLC. Additionally, inhibition towards trypsin was determined by an enzymatic assay. Based on the results, none of the investigated wheat species can be considered to be less ‘immunogenic’. Nevertheless, due to the large variability of ATI and gluten amounts among different genotypes, the selection of less-immunoreactive wheat varieties for individuals that suffer from wheat related diseases (WRDs) might be possible. The impact of breeding was assessed for different varieties of common wheat from different breeding periods. Results revealed significant improvement of technologically valuable parameters such as the amount of high molecular weight (HMW) glutenins and the gliadin to glutenin ratio, but no increase in immunogenic proteins.     

Fungicide treatments affect yield and moisture content of grain and straw in winter wheat

Straw and grain yield and water content in both grain and straw were measured during 3 years in 17 varieties of winter wheat following treatments with different fungicides. The water content of the straw varied significantly dependent on year, variety and fungicide treatment. In 1998, the water content in straw was significantly higher after the use of strobilurins compared with untreated and EBI fungicides (ergosterol biosyntesis inhibitors). On average water content in 10 varieties increased from 15% in untreated to 31% after applying two full dosages of azoxystrobin. In 1999 and 2000, lower dosages of azoxystrobin were used and water content increased only by 2–4 percentage units. The results indicate that in some years and with high doses precaution has to be taken following the use of strobilurins regarding handling and removing of the straw, which likely will include postponement of baling. Water content in grain was also increased significantly from fungicide treatments, but the increase was minor and at the maximum increase found to be 1.1% percentage unit. The effect of fungicide input on water content in grain was reduced much faster during the ripening period than for water content in straw. Fungicide treatments and varieties significantly influenced straw yields. The yield increases in straw varied between 0 and 1.0 tonnes/ha depending on year and variety, and was on average 0.42 tonnes/ha for two fungicide treatments. Two fungicide applications carried out at GS 31 and 45–55 gave only slightly higher increases compared with one application around ear emergence. Varieties showed variable increases following the use of fungicides. The variation was not influenced by the straw length (crop height). The increases in straw yield from fungicide treatments were relatively low compared to the increases in grain yield and no clear correlation was found between grain yield increases and straw yield increases.     

The influence of different nitrogen treatments on the size distribution of protein fractions in hard and soft wheat

The relative quantity of specific proteins, protein subunits, as well as amount and size-distribution of polymeric proteins in wheat kernels may vary due to environmental conditions. In this study, the effect of different nitrogen treatments on polymeric and monomeric proteins in wheat was determined. Two soft white biscuit wheat cultivars, a cracker wheat and a hard red bread wheat were planted under irrigation in a randomized complete block design with three replications in two successive years at six different nitrogen treatments. SE-HPLC was used to determine the amount of monomeric and polymeric proteins, and various quality characteristics were measured after harvesting. The large and small SDS-extractable polymeric proteins were not influenced by different N levels. There was a strong cultivar influence, where some cultivars had a larger reaction to N treatments than others. The later application (at flag leaf stage) of nitrogen did not increase the protein fractions. The total amount of N given seemed to have a greater influence on the protein fractions than the timing of the fertilizer application. The lowest N treatment consistently gave the lowest flour protein content value. There was a strong correlation between flour protein content and large monomeric proteins.     

Improving the protein content and composition of cereal grain

Cereals are important sources of protein for human nutrition but have low quality due to limitations in the amounts of essential amino acids, notably lysine. These deficiencies result from the low levels of these amino acids in the prolamin storage proteins and hence are exacerbated when high levels of nitrogen fertiliser are used to increase yield and total protein content.Genetic and genetic engineering strategies to increase both total protein content and the composition of essential amino acids have been employed. These include the exploitation of mutant high lysine genes and the use of transformation to either express additional proteins which are rich in lysine and/or methionine or to increase the free pools of these amino acids.     

Effects of genetic improvements on grain yield and agronomic traits of winter wheat in the Yangtze River Basin of China

Wheat is an important food crop worldwide. Genetic improvements have contributed much to wheat production since the 1960s. Verifying the evolution of agronomic traits and the physiological basis of grain yield will facilitate breeders and agronomists in developing new wheat cultivars, with the aim of stable and high yields. Thirty-five wheat cultivars, bred or widely planted in the Yangtze River Basin from 1950 to 2005, were grown in field experiments under three N rates (0, 112.5, and 225 kg N ha⁻¹) from 2006 to 2009 in Nanjing, China. Wheat grain yield, kernels per spike, 1000-kernel weight (TKW), and harvest index (HI) increased linearly with cultivar development from the 1950s to the 2000s, whereas spikes per unit land decreased significantly with cultivar development during the same period, and stabilized with further genetic improvements in cultivars. Grain yield, kernels per spike, and TKW differed with N rate and with cultivar. Grain yield, spikes per unit land, and kernels per spike increased significantly with increasing N fertilizer, but TKW and HI decreased. Cultivar height decreased with cultivar development from the 1950s to the 1980s, and remained relatively stable in subsequent cultivars. The proportion of the length of the top internode to total plant height increased with cultivar development from the 1950s to the 1980s and thereafter fell, while the length of the basal internode (BI) maintained a shortening trend. Leaf area per culm, leaf area index (LAI), net photosynthetic rate (Pn), and photosynthetic activity duration (PAD) of the flag leaf increased with cultivar development. Leaf area, LAI, and Pn increased significantly with increasing N fertilizer, while PAD did not. Single spike yield increased linearly with genetic development in cultivars, and these increases mainly resulted from increasing kernel number and weight, which were closely related to source size and cultivar. Grain yield was positively correlated to leaf area, LAI, Pn_Max, PAD, and single spike yield; single spike yield was positively correlated to leaf area, LAI, Pn_Max, and PAD, suggesting that grain yield improvements were mainly associated with improvements in the source (leaf area, LAI, Pn, PAD, etc.) and sink (single spike yield). Sink-source ratios increased with genetic development of cultivars, suggesting that productivity per leaf improved and that sink-source relationships have reached close to optimum in the Yangtze River Basin. Furthermore, breeding for high yield should be related to improvement in kernels per spike and TKW per unit land and increased sink-source ratios with a feasible LAI, and N fertilizer management should be considered during breeding for higher yields.     

Influence of variety and year of wheat cultivation on the chemical composition of starch and properties of glucose hydrolysates

The effect of chemical composition of starch samples derived from selected wheat cultivars harvested in different years (lipid content, amylose–lipid complex (AML) content, amylose concentration and the degree of starch damage) on physicochemical properties of glucose syrups produced from these starches was determined. It was found that glucose syrups obtained from the same wheat cultivar but harvested in different years displayed different physicochemical attributes such as concentration of reducing compounds, rate of filtration and coefficients of colour and transparency. Besides, the analyses showed that AML changed their form from amorphous to crystalline during enzymatic starch liquefaction by α-amylase. This in turn rendered starch less susceptible to enzymatic digestion and affected physicochemical features of glucose syrups. Presented results provide evidence that not only the total concentration of lipids and AML but also the AML polymorph present in starch hydrolysates (on completion of starch liquefaction) have an impact on quality of glucose syrups.     

Root growth,soil water variation,and grain yield response of winter wheat to supplemental irrigation

Water shortage threatens agricultural sustainability in the Huang-Huai-Hai Plain of China. Thus, we investigated the effect of supplemental irrigation (SI) on the root growth, soil water variation, and grain yield of winter wheat in this region by measuring the moisture content in different soil layers. Prior to SI, the soil water content (SWC) at given soil depths was monitored to calculate amount of irritation water that can rehydrate the soil to target SWC. The SWC before SI was monitored to depths of 20, 40, and 60 cm in treatments of W20, W40, and W60, respectively. Rainfed treatment with no irrigation as the control (W0). The mean root weight density (RWD), triphenyl tetrazolium chloride reduction activity (TTC reduction activity), soluble protein (SP) concentrations as well as catalase (CAT), and superoxide dismutase (SOD) activities in W40 and W60 treatments were significantly higher than those in W20. The RWD in 60–100 cm soil layers and the root activity, SP concentrations, CAT and SOD activities in 40–60 cm soil layers in W40 treatment were significantly higher than those in W20 and W60. W40 treatment is characterized by higher SWC in the upper soil layers but lower SWC in the 60–100-cm soil layers during grain filling. The soil water consumption (SWU) in the 60–100 cm soil layers from anthesis after SI to maturity was the highest in W40. The grain yield, water use efficiency (WUE), and irrigation water productivity were the highest in W40, with corresponding mean values of 9169 kg ha^?1, 20.8 kg ha^?1 mm^?1, and 35.5 kg ha^?1 mm^?1. The RWD, root activities, SP concentrations, CAT and SOD activities, and SWU were strongly positively correlated with grain yield and WUE. Therefore, the optimum soil layer for SI of winter wheat after jointing is 0–40 cm.     

Genotypic variation in linear rate of grain growth and contribution of stem reserves to grain yield in wheat

Grain growth in wheat depends on current photosynthesis and stem water-soluble carbohydrates (WSC). In semiarid regions with terminal drought, grain filling in wheat crops may depend more on stem WSC content than on current assimilates. Reduction in grain yield under drought is attributed to shorter duration of linear grain growth despite increased contribution of stem reserves to grain yield. The amount of stem reserves is measured either by changes in stem dry weight (indirect method) or by stem WSC content (direct method). Genotypic variation in the rate and duration of linear grain growth and in percent contribution of stem reserves to grain yield has not been evaluated in wheat. The objectives of this study were: (i) to quantify the relationship between the direct and indirect measurement of stem reserves during and across the grain-filling period and (ii) to measure the extent of genotypic variation in rate and duration of linear grain growth and in percent contribution of stem reserves to grain yield. Dry weight, WSC content and grain yield of the main stem were measured at 10-day intervals in 11 diverse wheat genotypes under well-watered and droughted-field conditions across 2 years. Drought reduced stem WSC content from 413 to 281 mg and grain yield from 4.6 to 2.5 t ha⁻¹. Stem WSC content and dry weight were positively correlated. Genotypic differences in linear rate of grain growth were significant in well-watered (ranging from 48.9 to 72.4 mg spike⁻¹ day⁻¹) and in droughted-field (ranging from 33.2 to 59.9 mg spike⁻¹ day⁻¹) conditions. Drought, on average, reduced the linear rate and duration of grain growth by 20 and 50%, respectively. Reduction in linear rate ranged from 13 to 43%. The amount of current assimilates and stem reserves contributed to grain yield was reduced, respectively, by 54 and 11% under drought. Genotypic differences in percent contribution of stem reserves to grain yield were significant in well-watered (ranging from 19.1 to 53.6%) and in droughted-field (ranging from 36.6 to 65.4%) conditions. The wheat genotypes responded differently to drought. Main spike grain yield was reduced by 43% under drought due to 26 and 11% reduction in grain weight and number of grains, respectively. Grain yield was correlated with linear grain growth under well-watered (r = 0.96) and droughted (r = 0.83) conditions. The genotypic variation observed indicates that breeding for a higher rate of linear grain growth and greater contribution of stem reserves to grain yield should be possible in wheat to stabilize grain yield in stressful environments.     

Influence of wheat variety and dough preparation on FODMAP content in yeast-leavened wheat breads

FODMAPs are fermentable oligo-, di, and monosaccharides and polyols, which can have adverse effects on human health. Especially in individuals with gastrointestinal disorders such as irritable bowel syndrome, ingestion of FODMAPs may trigger or aggravate symptoms like abdominal pain, bloating, or diarrhea. Our objectives were to investigate the FODMAP content in the flour of 21 wheat varieties and in their 42 yeast-leavened breads prepared with a short (110 min) or a long dough fermentation (24 h), the latter according to a typical baker's recipe. Fructan, glucose, fructose and excess fructose were measured using enzymatic kits. On average, the breads of both dough fermentations had a FODMAP content reduced by > 65% compared to that of the whole grain wheat flour. Average FODMAP content of both dough fermentations was 0.22 g per 100 g fresh bread. FODMAP content tended to be lower in the long compared to the short dough fermentation, however, the difference was not statistically significant. We also found that the 21 wheat varieties differed up to five times in their potential to form FODMAPs in bread. In conclusion, the choice of a low fructan wheat variety coupled with long dough fermentation with sourdough and yeast is best to minimize FODMAPs in breads.     

Influence of competition,irrigation levels and nitrogen fertilization on protein content and protein yield of three spring wheat (Triticum aestivum L.) cultivars

Protein content and protein yield of three spring wheat cultivars differing in morphological and physiological growth characters were found to be influenced by intercultivar competition, irrigation levels and nitrogen fertilization. The protein content of the tall cultivar C 306 and the protein yield of the dwarf cultivar HD 2160 were more than the other cultivars. Binary mixed stands were not superior to the better component cultivar. Intercultivar competition increased the protein content of dwarf and semi-dwarf cultivars, but decreased the protein content of tall cultivar. On the other hand, protein yield of the dwarf cultivar decreased and that of tall cultivar increased when grown in mixed stands. Protein yield of semi-dwarf cultivar increased when grown with dwarf cultivar, but decreased when grown with tall cultivar. Two or three irrigations increased the protein content and protein yield of all the three cultivars and their mixed stands over one irrigation. Protein content and protein yield of the cultivars and their mixed stands were higher when 150 kg N/ha was applied than when 80 kg N/ha was applied.