Chromosomal location of wheat genes of the carotenoid biosynthetic pathway and evidence for a catalase gene on chromosome 7A functionally associated with flour b* colour variation

Knowledge of molecular and genetic mechanisms controlling wheat grain quality characteristics is significant for improving flour for end-product functionality. Flour b* colour is an important quality trait for breeding wheat varieties to produce grain for specific market requirements. The degree of flour yellowness is due to the accumulation of carotenoids in grain, particularly lutein. Flour b* is under polygenic control and quantitative trait loci (QTL) have frequently been reported on chromosome 7AL. Analysis of carotenoid genes showed that phytoene synthase (PSY) co-located to the QTL on 7AL but other genes at this locus are also thought to contribute flour b* colour variation. This study used the wheat genome survey sequence and identified the chromosomal location of all wheat carotenoid genes, but none other than PSY were located on 7AL and, therefore, other genes may control flour b* colour variation including oxidative genes that degrade carotenoids. An investigation of EST bin mapped to 7AL identified a gene encoding a catalase enzyme (Cat3-A1) that was phylogenetically related to other plant class III enzymes, co-located to the QTL for flour b* colour variation on 7AL in three mapping populations and expressed during seed development. Therefore, Cat3-A1 was functionally associated with flour b* colour variation. Catalase acts upon hydrogen peroxide as a substrate and it was postulated that Cat3-A1 alleles control varying degrees of bleaching action on lutein in developing wheat grain. Markers for Cat3-A1 developed in this study can be used in conjunction with other candidate gene markers including phytoene synthase and lycopene-ε-cylase to develop a molecular signature for selecting lines with specific flour b* colour values in wheat breeding.     

Characterization of puroindolines in the control of endosperm texture in common wheat lines with substitutions of homeologous group-5 chromosomes

The genetic control of grain hardness and its association with the specific friabilin content on starch granules of common wheat cultivars and lines with intervarietal substitutions of homeologous group-5 chromosomes were studied. A significant correlation was revealed between the technological parameters of grain hardness (mean size of flour particles) and the specific content of puroindolines on the starch surface estimated in terms of starch doses. The results obtained allowed the method of starch doses to be used to identify soft and hard wheat cultivars and lines based on an analysis of a single grain. The biochemical analysis confirmed the previously obtained estimates of flour-grinding properties of wheat cultivars and substitution lines and allowed specific genotypes to be characterized according to the composition of puroindolines. The influence of chromosomes 5D and 5A of donor wheat cultivars on the activity of the Ha loci of recipient cultivars was revealed and found to be associated with the composition of PIN products and with the expression of the Pina-D1 and Pinb-D1 genes.     

Biochemical Basis and Molecular Genetics of Processing and Nutritional Quality Traits of Wheat

Wheat is unique in the sense that large numbers of end-use products such as chapati, bread, biscuits, noodles and pasta products are made from it. This is possible because of visco-elastic property imparted to the dough by its gluten proteins. Grain texture, gluten constituents and starch composition in the endosperm are major determinants of end-product quality. Each end-use product has its own specific requirements. Therefore, to expedite the breeding programme for the development of product-specific varieties, understanding the grain components at biochemical/molecular level and their relationship with quality, is needed. Impressive advances have been made in understanding the subunit composition of glutenins, gliadins and starch at biochemical and molecular levels. Molecular markers for these components have been identified, and are being used in breeding for the improvement of wheat quality. Wheat is also deficient of certain micronutrients such as Fe and Zn and lysine content. There is increasing concern globally for the improvement of the nutritional quality of wheat for micronutrient content, and protein and starch quality. Transgenic approach will be helpful in manipulating metabolic pathways for enhancing micronutrient bioavailability and increased lysine content. In this review, information on recent developments that have taken place in understanding the biochemical basis and molecular genetics of processing and nutritional quality of wheat grain and its future implications, has been provided.     

Expression profiles of genes involved in starch synthesis in non-waxy and waxy wheat

Non-waxy and waxy types of wheat were used to study the expression profiles of genes involved in starch synthesis. During grain development, expression profiles and levels of AGPL and AGPS genes were similar to each other. SSI expression remained constant during the late grain development, while expression of SSII and SSIII was higher over the early to middle and middle grain development, and the GBSSI was actively expressed during the entire grain development. SBEIIa was higher expressed during early to middle stage, SBEIIb was active in middle and SBEI during middle to late grain development. During the entire grain development, expression levels of GBSSI and SSIII genes were higher in non-waxy type of wheat, while those of SBEI and SBEIIb were lower in the non-waxy type of wheat. Expression of all genes involved in starch synthesis was stage-specific and tissue-specific. In addition, the expression profiles of genes encoding starch synthase were in agreement with the activity changes of starch synthase during grain development.     

Swedish Spring Wheat Varieties with the Rare High Grain Protein Allele of NAM-B1 Differ in Leaf Senescence and Grain Mineral Content

Some Swedish spring wheat varieties have recently been shown to carry a rare wildtype (wt) allele of the gene NAM-B1, known to affect leaf senescence and nutrient retranslocation to the grain. The wt allele is believed to increase grain protein concentration and has attracted interest from breeders since it could contribute to higher grain quality and more nitrogen-efficient varieties. This study investigated whether Swedish varieties with the wt allele differ from varieties with one of the more common, non-functional alleles in order to examine the effect of the gene in a wide genetic background, and possibly explain why the allele has been retained in Swedish varieties. Forty varieties of spring wheat differing in NAM-B1 allele type were cultivated under controlled conditions. Senescence was monitored and grains were harvested and analyzed for mineral nutrient concentration. Varieties with the wt allele reached anthesis earlier and completed senescence faster than varieties with the non-functional allele. The wt varieties also had more ears, lighter grains and higher yields of P and K. Contrary to previous information on effects of the wt allele, our wt varieties did not have increased grain N concentration or grain N yield. In addition, temporal studies showed that straw length has decreased but grain N yield has remained unaffected over a century of Swedish spring wheat breeding. The faster development of wt varieties supports the hypothesis of NAM-B1 being preserved in Fennoscandia, with its short growing season, because of accelerated development conferred by the NAM-B1 wt allele. Although the possible effects of other gene actions were impossible to distinguish, the genetic resource of Fennoscandian spring wheats with the wt NAM-B1 allele is interesting to investigate further for breeding purposes.     

Kernel softness in wheat is determined by starch granule bound Puroindoline proteins

Wheat has a vital position in agriculture because it is a staple food for masses and variation in grain hardness governs its applications. Soft wheats have softer endosperm texture that mills easily, so needs less energy to mill, produces smaller particles, and small amount of starch is damaged after milling as compared to hard wheat. Soft texture results from higher level of friabilin whereas hard texture results from low level of friabilin on starch granule surface. Friabilin, a marker of kernel texture is primarily composed of Puroindolines (PINs) and its genes (Pins) are located on the Hardness (Ha) locus. The Pins are the molecular-genetic basis of kernel softness in wheat. When both Pins are in their ‘wild state’ (Pina-D1a and Pinb-D1a), wheat kernel is soft. Absence or mutation in one of the Pins results in hard grain texture with different effects on end use and milling qualities. Pina-D1b genotypes gave harder grain texture, higher protein content, water absorption of flour, damaged starch granules and greater flour yield than hard wheat. Recently, other Pins like genes, Pin b variant genes located on the long arm of chromosome 7A were reported in bread wheat with more than 70% similarity to Pinb (Pinb-D1a) at the DNA level. Other genes located on chromosomes 1A, 2A, 5A, 7A, 5B, 2D and 6D also affect kernel texture. However the main determinants are the variants in the allelic diversity of Puroindoline family genes. Contemporary studies show that Pins are multifunctional family of genes having a range of functions from grain hardness to natural defense against insects and pathogens such as viruses, bacteria and fungi.     

Genome‐wide association study of six quality traits reveals the association of the TaRPP13L1 gene with flour colour in Chinese bread wheat

Flour colour, kernel hardness, grain protein content and wet gluten content are important quality properties that determine end use in bread wheat. Here, a wheat 90K genotyping assay was used for a genome‐wide association study (GWAS) of the six quality‐related traits in Chinese wheat cultivars in eight environments over four years. A total of 846 significant single nucleotide polymorphisms (SNPs) were identified, explaining approximately 30% of the phenotypic variation on average, and 103 multienvironment‐significant SNPs were detected in more than four environments. Quantitative trait loci (QTL) mapping in the biparent population confirmed some important SNP loci. Moreover, it was determined that some important genes were associated with the six quality traits, including some known functional genes and annotated unknown functional genes. Of the annotated unknown functional genes, it was verified that TaRPP13L1 was associated with flour colour. Wheat cultivars or lines with TaRPP13L1‐B1a showed extremely significantly higher flour redness and lower yellowness than those with TaRPP13L1‐B1b in the Chinese wheat natural population and the doubled haploid (DH) population. Two tetraploid wheat lines with premature stop codons of the TaRPP13L1 gene mutagenized by ethyl methanesulfonate (EMS) showed extremely significantly higher flour redness and lower yellowness than wild type. Our data suggest that the TaRPP13L1 gene plays an important role in modulating wheat flour colour. This study provides useful information for further dissection of the genetic basis of flour colour and also provides valuable genes or genetic loci for marker‐assisted selection to improve the process of breeding quality wheat in China.     

Hydrolysis of wheat flour with amylase preparations and individual enzymes

Rheological properties of wheat flour were studied in the course of its processing (cooking and saccharification). The effects of commercial α-amylase preparations were compared during flour preparation. Test preparations were equally potent in decreasing the viscosity of an all-grain batch. Homogenous glucoamylases isolated from Aspergillus differed in the presence or absence of the starch-binding domain. The starch-binding domain provided for the high activity of glucoamylase on insoluble starch, but gave no advantages in saccharification of pretreated wheat flour.     

Sequence diversity, haplotype analysis, association mapping and functional marker development in the waxy and starch synthase IIa genes for grain-yield-related traits in hexaploid wheat (Triticum aestivum L.)

Development of high-yielding cereal crops could meet increasing global demands for food, feed and bio-fuels. Wheat is one of the world??s most important cereal crops. The biosynthesis of starch is the major determinant of yield in wheat. Two starch biosynthesis genes, the waxy (Wx) genes and the starch synthase IIa (SSIIa) genes, were amplified and sequenced in 92 diverse wheat genotypes using genome-specific primers. Nucleotide diversity, haplotype analysis and association mapping were performed. The first exon (5??-UTR) and the first intron of the three homoeologous Wx genes were isolated using expressed sequence tag sequences. The Wx genes contained 12 exons separated by 11 introns. SNP (single nucleotide polymorphism) frequency ranged from 1 SNP/3,648?bp for Wx-D1 to 1 SNP/135?bp for SSIIa-A1, with an average of 1 SNP/230?bp. The average SNP frequencies in exon and intron regions were 1 SNP/322?bp and 1 SNP/228?bp, respectively. Thirty, 23 and 5 SNPs were identified and formed five, six and five haplotypes for SSIIa-A1, SSIIa-B1 and SSIIa-D1, respectively. However, no association was found between these SNPs and seven yield-related traits. Twenty-two, 15 and 1 SNPs were detected and formed nine, five and two haplotypes for Wx-A1, Wx-B1 and Wx-D1, respectively. Three unique nucleotides C+A+T at SNP5, SNP6 and SNP12 formed Wx-B1-H3, which was significantly associated with increased grain weight, thousand kernel weight, and total starch content in three spring wheat genotypes and five winter wheat genotypes. Cost-effective and co-dominant SNP markers were developed using temperature-switch (TS)-PCR and are being used for marker-assisted selection of doubled haploid lines with enhanced grain yield and starch content in winter wheat breeding programs.