The contribution of ear photosynthesis to grain filling in bread wheat (Triticum aestivum L.)

The contribution of ear photosynthesis to grain filling in wheat (Triticum aestivum L.) is not well known. The main objective of this work was to evaluate this contribution through three different experimental approaches: (1) ear photosynthesis was reduced by removing awns or shading the ears (in combination with a defoliation treatment), (2) grain weight per ear was compared in an ‘all shaded’ crop versus plants where only the vegetative parts were shaded (‘ear emerging’), and (3) ear photosynthesis was reduced with DCMU (3-(3,4-dichlorophenyl)-1,1-dimethylurea), a specific inhibitor of photosystem II.     

Waxy endosperm accompanies increased fat and saccharide contents in bread wheat (Triticum aestivum L.) grain

The contents of fat, starch, pentosan, fructan, β-glucan and several mono- and oligosaccharides in grain were evaluated to find out the possible effects of the Wx-D1 gene of bread wheat using two sets of near-isogenic waxy and non-waxy lines and two low-amylose mutant lines with a common genetic background of Kanto 107. These materials have two non-functional Wx-A1b and Wx-B1b alleles in common. Waxy near-isogenic lines with a non-functional Wx-D1d allele showed consistently increased contents of fat, total fructan, β-glucan, glucose, fructose, sucrose, 1-kestose, 6-kestose, neokestose, nystose and bifurcose compared with non-waxy lines with a functional Wx-D1a allele throughout three growing/harvest seasons. Starch and total pentosan contents were inconsistently influenced by the allelic status of the Wx-D1 locus, while water-soluble pentosan and raffinose contents were not affected. The compositional changes of a low-amylose mutant line with an almost non-functional Wx-D1f allele were closely similar to those of waxy near-isogenic lines, while significantly different changes were barely observed in another low-amylose mutant line with a partly functional Wx-D1g allele in two seasons. These results showed that the Wx-D1 gene has pleiotropic effects on the fat and saccharide contents of bread wheat grain.     

Development of simple and co-dominant PCR markers to genotype puroindoline a and b alleles for grain hardness in bread wheat (Triticum aestivum L.)

Grain hardness is one of the most important quality characteristics of cultivated bread wheat (Triticum aestivum L.). A large deletion in the puroindoline a (Pina) gene or single nucleotide polymorphisms (SNPs) in the puroindoline b (Pinb) gene results in hard grain texture. So far, nine Pina alleles (Pina-D1a–Pina-D1b, Pina-D1k–Pina-D1q) and seventeen Pinb alleles (Pinb-D1a–Pinb-D1g, Pinb-D1p–Pinb-D1ab) have been identified in bread wheat. The major Pina and Pinb alleles identified in hard wheat cultivars are Pina-D1b, Pinb-D1b, Pinb-D1c and Pinb-D1d. In this study, a three-primer PCR system was employed to develop nine co-dominant STS markers for genotyping Pina-D1a and Pina-D1b, whereas temperature-switch (TS) PCR was used to develop six co-dominant SNP markers for genotyping the Pinb-D1a, Pinb-D1b, Pinb-D1c and Pinb-D1d alleles. These STS and TS-PCR markers were used to verify the grain hardness genotype of 100 wheat cultivars. The reliability and genotyping accuracy of TS-PCR markers were confirmed through sequencing of PCR products and a comparison with previously published results. Therefore, STS and TS-PCR markers offer a simple, cost-effective and reliable method for high-throughput genotyping Pina and Pinb alleles to select grain hardness in wheat quality breeding programs and for wheat market classification.     

Development of STS markers and establishment of multiplex PCR for Glu-A3 alleles in common wheat (Triticum aestivum L.)

Low-molecular-weight glutenin subunits (LMW-GS) play a key role in determining the processing quality of the end-use products of common wheat. The objectives of this study were to identify genes at Glu-A3 locus, develop the STS markers, and establish multiplex PCR with the STS markers for Glu-A3 alleles. Gene-specific PCR primers were designed to amplify six near-isogenic lines (NILs) and Glenlea with different Glu-A3 alleles (a, b, c, d, e, f and g) defined by the protein electrophoretic mobility. Three Glu-A3 genes with complete coding sequence were cloned, designated as GluA3-1, GluA3-2 and GluA3-3, respectively. Seven dominant allele-specific STS (sequence tagged sites) markers were designed based on the SNPs (single nucleotide polymorphisms) among different allelic variants for the discrimination of the Glu-A3 protein alleles a, b, c, d, e, f and g. Four multiplex PCRs were established including Glu-A3b + Glu-A3f, Glu-A3d + Glu-A3f, Glu-A3d + Glu-A3g, and Glu-A3b + Glu-A3e. These markers and multiplex-PCR systems were validated on 141 CIMMYT wheat varieties and advanced lines with different Glu-A3 alleles, confirming that they can be efficiently used in marker-assisted breeding.     

Analysis of starch swelling power in Australian breeding lines of hexaploid wheat (Triticum aestivum L.)

Starch is the major component of wheat (Triticum aestivum L.) grain and is composed of two large glucan molecules, amylose and amylopectin. The ratio between the two polymers types influences the water absorbing properties of starch upon heating, and thus affects the end-use of grain and purified starch. In this study, we evaluated the starch swelling power (SSP) values in seven wheat populations developed from crosses involving low-SSP lines. Analysis of starch produced by the F₂ generation plants showed that the largest SSP variation (11.4–16.2) and lowest SSP mean (13.9) was obtained for a population derived from doubled haploid lines SM1028 (SSP = 14.5) and VK306 (SSP = 13.6). The population of 360 lines was advanced by single seed descent to the following generations for further studies. Starch analysis of grain produced by F₄ generation lines in two field locations during 2006 and in a greenhouse environment during 2005 showed that SSP values were relatively stably inherited. The average broad-sense heritability was 73% and significant (P < 0.001) genotype × genotype and genotype × environment interactions were seen. Starches with the highest and lowest SSP values were inversely related to amylose concentration determined by high pressure liquid chromatography (HPLC)–size exclusion chromatography (SEC) of debranched starch. Developed lines with the lowest SSP values surpassed 40% in apparent amylose concentration. The study illustrates that screening for SSP in early generations can be used to develop wheat lines with desired starch swelling characteristics.     

Partial purification and characterization of polyphenoloxidase from durum wheat (Triticum durum L.)

A bright yellow color of pasta is an important qualitative trait for the durum wheat industry. Final color is the result of the balance between yellow and brown components in semolina. Polyphenoloxidase (PPO) is implicated as playing a significant role in darkening. This study aimed to characterize PPO activity of durum wheats. PPO was extracted and partially purified by ion-exchange chromatography on a column packed with diethyaminoethyl cellulose (DEAE). This procedure led to 26.33-fold purification with 24.7% recovery. The optimum temperature and pH of PPO were found to be 40 °C and 6.5, respectively. Heat stability of durum wheat PPO decreased as the temperatures increased from 30 to 80 °C. The z-value was calculated as 23.4 °C. It increased to 26.3 and 48.4 °C in the presence of 40% sucrose and 1 M NaCl, respectively. Durum wheat PPO was shown to use several phenolic compounds as substrate. Among the substrates used, the greatest substrate specificity was observed with catechol. Durum wheat PPO was sensitive to inhibitors such as ascorbic acid, cysteine, oxalic acid and citric acid. Ascorbic acid was the most effective inhibitor.     

Zinc nutrition influences the protein composition of flour in bread wheat (Triticum aestivum L.)

Zinc (Zn) deficiency and heat stress during grain filling occur in a number of important wheat growing regions around the world. The changes in grain protein composition due to high temperature are well documented, but little is known about the effect of grain Zn and its interaction with heat stress. Six field experiments were conducted at sites differing in grain filling temperatures to examine these effects. Two varieties of bread wheat were grown at six rates of Zn, including foliar sprays of Zn. The relative amounts of gliadin and polymeric protein were measured by size exclusion HPLC. Applying Zn increased grain yield at three sites and altered protein quality at two of these. Foliar Zn applications doubled grain Zn concentration, reduced the proportion of gliadin and SDS-unextractable polymeric protein and increased the proportion of SDS-extractable polymeric protein. Heat stress during grain filling was associated with a high proportion of gliadin and low proportions of the polymeric protein in the grain. However, the proportions of gliadin and of SDS-extractable polymeric protein were less affected by high temperatures in grain with high Zn concentrations. The results demonstrate that Zn nutrition can alter protein composition and the effects of Zn may interact with grain filling temperatures.     

Association of Puroindoline b-B2 variants with grain traits,yield components and flag leaf size in bread wheat (Triticum aestivum L.) varieties of the Yellow and Huai Valleys of China

A total of 169 wheat (Triticum aestivum L.) varieties (landraces and cultivars) were used to asses the relationship between Puroindoline D1 alleles and Puroindoline b-B2 variants and grain hardness, other grain traits, yield components, and flag leaf size. Results indicated that the average SKCS hardness of Pinb-B2v3 varieties was significantly greater than that of Pinb-B2v2 varieties within the soft Puroindoline D1 haplotype sub-group. Conversely, no statistically significant difference was obtained for SKCS hardness between varieties with the Pinb-B2v3 vs. Pinb-B2v2 alleles within the two hard Puroindoline D1 haplotypes (Pinb-D1b and Pinb-D1p sub-groups). Therefore, the Puroindoline b-B2 gene may have a bigger impact on soft wheat varieties than hard. Across all varieties, thousand-kernel weight, grain weight per spike, grain diameter, grain number per spike, flag leaf width and area of Pinb-B2v3 varieties were significantly greater than those possessing Pinb-B2v2. These results indicated that the Pinb-B2v3 allele was associated with preferable grain yield traits compared to the Pinb-B2v2 allele in bread wheat. This study provides evocative information for better understanding the molecular and genetic basis of wheat grain yield.     

Development of somaclonal variants of wheat (Triticum aestivum L.) for yield traits and disease resistance suitable for heat stressed and zero-till conditions

In a substantial rice–wheat cropping system area of South Asia, wheat sowing often gets too delayed and exposed to terminal heat stress. Therefore, farmers prefer varieties that are able to perform well under a short growing period. Tissue culturally regenerated plants of wheat variety cv. HUW 234, the most widely cultivated variety of North Eastern Plain Zone (NEPZ) of India were screened using immature embryo as explant. Days to heading and maturity, yield and other yield components and resistance to leaf rust and spot blotch were evaluated. A few somaclones in R₃ and R₄ generations displayed significant earliness for days to heading and maturity, improved yield traits and resistance to leaf rust and spot blotch diseases. The superior performance of two of the variants was confirmed in the R₅ generation in 3 years of testing under two dates of conventional and zero-till sowing. Stability analysis also suggested superiority of the two somaclones across 12 environments. This appeared to confirm the possibility of obtaining useful somaclonal variants of wheat for very late sown as well as zero-till managed agriculture. The superior performing somaclones can be used as parents in the ongoing breeding programmes targeting late sown wheat in South Asia exposed to terminal heat stress.     

In vitro pullulanase activity of wheat (Triticum aestivum L.) limit-dextrinase type starch debranching enzyme is modulated by redox conditions

Expression of a limit-dextrinase (LD) type starch debranching enzyme (EC 3.2.1.41) was observed in developing wheat (Triticum aestivum L.) endosperm and germinating grains, indicating a role for the enzyme in both biosynthesis and degradation of starch. A full-length cDNA, TaLD1, encoding LD in wheat developing kernels was isolated and predicted to encode a 98.6 kDa mature protein active in amyloplasts. Isolated cDNA was expressed in Escherichia coli to produce a recombinant His-tagged LD, which mainly accumulated in inclusion bodies as an inactive enzyme. Extraction of His-tagged LD from the inclusion bodies followed by dialysis under thiol/disulphide redox conditions allowed partial refolding of the protein and detection of pullulanase specific activities by zymogram analysis and enzyme assays. Several active conformations were demonstrated by the recombinant TaLD1 and pullulanase activity could be modulated by redox conditions in vitro. The results suggest that cellular redox conditions may regulate the level of LD activity in wheat tissues.     

Seasonal conditions influence dormancy and preharvest sprouting tolerance of wheat (Triticum aestivum L.) in the field

Preharvest sprouting occurs following rainfall after maturity and reduces grain quality and value. Dormancy at maturity is a trait frequently used by wheat breeders to improve tolerance to sprouting. To determine the environmental influence on the predicative relationship between dormancy at maturity and improved preharvest sprouting tolerance, dormancy (germination index) at maturity and grain quality (falling number) at maturity and then, after rainfall, was measured over three seasons. Based on the results it was possible to draw three main conclusions. Firstly, genotypes with strong dormancy (germination index <0.20) which have the embryo and seed coat component of dormancy maintained a falling number >300 s at all sites and seasons for the 2-month period after maturity despite receiving up to 122 mm of rain. Adequate preharvest sprouting tolerance also occurred in dormant genotypes, with just the embryo component; in all but the most severe conditions. Secondly, though the effect of environment and interaction of genotype and environment was significant, the G by E interaction did not account for a large proportion of the variation (<6%) in sprouting tolerance (measured by falling number after rainfall) or change the relative rankings of preharvest sprouting tolerance. Finally, other defects associated with rainfall during grain filling such as black point and fungal staining may slightly reduce dormancy estimates and preharvest sprouting tolerance. For this reason grain defects need to be improved in parallel with preharvest sprouting tolerance. In conclusion, strong dormancy provides a reliable source of preharvest sprouting tolerance in the field.     

Apigenin di-C-glycosides (ACG) content and composition in grains of bread wheat (Triticum aestivum) and related species

Apigenin di-C-glycosides (ACGs) are present in the grain of bread wheat and other related cereals primarily as one or two sets of Wesseley-Moser isomers containing either arabinose and glucose (ACG1) or arabinose and galactose (ACG2) on the A ring of apigenin. These compounds may contribute to the yellow colour of wheat-based products made under alkaline conditions and in addition, have possible roles in a number of plant physiology processes and human health. The aims of this investigation were to survey genetic variation for ACG content and composition in hexaploid bread wheat (Triticum aestivum L.) and to examine ACGs in the putative progenitors of hexaploid wheat and available genetic stocks as a first step towards understanding the mechanisms involved in their biosynthesis and genetic control. Substantial variation in both grain ACG content and the ratio, ACG1/ACG2, were identified within bread wheat cultivars and related species. Genotype controlled the major portion of the variation. ACG content appeared to be a multigenic trait whereas variation in ACG1/ACG2 was associated with a limited number of chromosomes, in particular chromosomes 1B, 7B and 7D. The results suggest that it should be possible to manipulate both ACG content and composition traits through breeding.     

Isolation and characterization of EMS-induced Dy10 and Ax1 high molecular weight glutenin subunit deficient mutant lines of elite hexaploid wheat (Triticum aestivum L.) cv. Summit

The mixing properties of the dough are critical in the production of bread and other food products derived from wheat. The high molecular weight glutenin subunits (HMW-GS) are major determinants of wheat dough processing qualities. The different alleles of the HMW-GS genes in hexaploid wheat vary in their effect on dough quality. To determine the contribution of the individual HMW-GS alleles, lines deficient in HMW-GS proteins were generated by chemical mutagenesis in the elite bread wheat Triticum aestivum cv. Summit. In this report we describe the identification and characterization of Dy10 and Ax1 deficient lines. Examination of the effect of Dy10 and Ax1 deficiency on dough rheological properties by mixography showed shorter mixing time to reach peak resistance, and weaker and less extensible doughs relative to the wild type control. This is the first time that the role of Dy10 in vivo has been examined apart from the Dx5 + Dy10 allelic pair combination.     

Influence of high temperature and terminal moisture stress on dormancy in wheat (Triticum aestivum L.)

Preharvest sprouting is common in cereals that lack grain dormancy if maturing grain is exposed to rain. Over three successive seasons wheat genotypes with a range of dormancy levels, were exposed to moisture stress and periods of high temperature stress (>30 °C) in controlled field trials. Dormancy assessments were based on a germination index of hand threshed grain throughout grain filling. There were three main results. First, moisture stress combined with consistently high temperature during grain filling was associated with induced dormancy in Cunderdin, (germination index of 0.41) in a normally non-dormant genotype (germination index normally >0.80), but no additional dormancy in DM 2001, a dormant genotype (germination index normally <0.10). In contrast sudden heat shocks (>30 °C max. for >12 days) at 30–50 days post-anthesis reduced dormancy, germination index increase of 0.42 on average across five genotypes. Secondly, whilst dormancy was affected by moisture and heat stress, genotypes maintained their relative rankings across environments and genotype had the most effect on dormancy (70–92% of the variation in germination index) with DM 2001 and DH 22 more dormant than DH 56, DH 45 and Cunderdin. Finally, the effect of environment was different for different genotypes; those with partial dormancy (germination index usually 0.20–0.50, DH 56 and DH 45) were most influenced by the environmental conditions with germination indexes ranging from 0.06 to 0.85 depending on environment. Consequently avoidance of high temperatures, moisture stress, and maturity × stress interactions, are important prerequisites in screening for genotypes with genetic differences in dormancy.     

In winter wheat (Triticum aestivum L.), post-anthesis nitrogen uptake and remobilisation to the grain correlates with agronomic traits and nitrogen physiological markers

In wheat, nitrogen (N) uptake and remobilisation after flowering contributes largely, in Northern countries, to grain yield and grain protein content. The aim of our study was first to estimate the proportion of N taken up and remobilised to the grain as well as their relative efficiency using ¹⁵NO₃⁻-labelling at flowering. The validity of the technique was assessed in comparison to the N budget calculation method on five winter wheat cultivars grown for 2 years at low and high fertilization input. We estimated that on average 71.2% of grain N originates from remobilisation with significant genotypic differences. Among the five genotypes, significant differences were also found for both N remobilisation efficiency (from 69.8 to 88.8%) and N translocation efficiency (from 89.7 to 93.4%). In parallel, during 1 year, we monitored physiological markers representative of N assimilation and recycling at two sampling dates during the grain filling period. We then examined if there was any relationship between these physiological markers, N absorption and remobilisation estimates and agronomic traits related to yield and grain N content. Nitrate reductase (NR) activity was highly correlated to N absorbed post-flowering and to grain protein content. Glutamine synthetase (GS) activity was even more highly correlated than NR activity to the amount of N remobilised and grain yield. The use of physiological traits such as NR and GS activities as markers of the wheat N status is discussed.     

Effect of genotype,environment and genotype-by-environment interaction on metabolite profiling in durum wheat (Triticum durum Desf.) grain

The aim of this study was to assess the relative roles of genotype, environment and genotype-by-environment interactions in determining the metabolite profile of durum wheat grain. Four durum wheat cultivars were grown under conventional and organic farming systems over three consecutive years. The use of a high-throughput gas chromatography–mass spectrometry platform allowed the analysis of sets of different polar and non-polar compounds, including amino acids, sugars, organic acids, fatty acids (saturated and unsaturated), and sterols. Statistical analysis of the data showed a small impact of genotype and large effects of both year and genotype-by-environment interaction on the metabolite composition and quality of the wheat grain. Overall, the data from this study highlight the potential role of metabolic profiling in the analysis of durum wheat quality and production.     

Reduced height alleles (Rht) and Hagberg falling number of wheat

Near-isogenic lines varying for alleles for reduced height (Rht) and photoperiod insensitivity (Ppd-D1) in cv. Mercia (2005/6–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht8c+Ppd-D1a, Rht-D1c, Rht12) and cvs Maris Huntsman and Maris Widgeon (2007/8–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht-B1b+Rht-D1b, Rht-D1b+Rht-B1c) were compared at one field site, but within different systems (‘organic’, O, 2005/6–2007/8 v. ‘intensive’, I, 2005/6–2010/11). Further experiments at the site (2006/7–2008/9) compared 64 lines of a doubled-haploid (DH) population [Savannah (Rht-D1b) × Renesansa (Rht-8c+Ppd-D1a)]. Gibberellin (GA) insensitive dwarfing alleles (Rht-B1b; Rht-B1c; Rht-D1b; Rht-D1c) could reduce α-amylase activity and/or increase Hagberg falling number (HFN) but effects depended greatly on system, background and season. Only Rht-B1c increased grain dormancy despite producing plants taller than Rht-D1c. The GA-sensitive Rht8c+Ppd-D1a in Mercia was associated with reduced HFN but analysis of the DH population suggested this was more closely linked with Ppd-D1a, rather than Rht8c. The GA-sensitive severe-dwarfing allele Rht12 was associated with reduced HFN. Instability in HFN over season tended to increase with degree of dwarfing. There was a negative association between mean grain weight and HFN that was in addition to effects of Rht and Ppd-D1 allele.