Diallelic analysis of quantitative traits in hexaploid wheat (Triticum aestivum L.)

Abstract

A complete diallel study of crosses between eight wheat varieties was carried out to determine the relative magnitude of components of genetic variation and heritability for important grain yield, quality and drought‐related traits. The data appeared adequate for the additive‐dominance model. The additive effects predominated for most traits, and consequently the narrow‐sense heritability was high to moderately high for flag leaf area, weight and venation, stomatal frequency and size, epidermal cell size, biomass, protein content, number of tillers, spike length, spike density, 1000‐grain weight and grain yield. These results appear promising for selecting better plants in the segregating populations with some degree of improvement for yield, quality and physiological efficiency.     

Suspension and protoplast culture of hexaploid wheat (Triticum aestivum L.)

Suspension cultures have been initiated from embryogenic callus of hexaploid wheat (Triticum aestivum L.). Most commonly, these suspensions are composed of callus-like clusters (up to 2 mm in diameter). Two rapidly-growing lines (MBE6 and C82d) have been obtained, which consist of smaller aggregates of cytoplasmic cells, and these have been maintained for more than 4 years. These lines show very limited morphogenetic capacity and only a single plantlet has been regenerated, from line MBE6, after 9 months in culture. Protoplasts isolated from line MBE6 are unable to divide, but protoplasts from line C82d consistently undergo sustained divisions to form callus or secondary cell suspensions.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog (1962) medium     

Mapping of quantitative trait loci determining agronomic important characters in hexaploid wheat ( Triticum aestivum L.)

A set of 114 recombinant inbred lines of the 'International Triticeae Mapping Initiative' mapping population was grown during the seasons 1997, 1998, 1999 and 2000 under several environments. Twenty morphological (glume colour, awn colour, waxiness, leaf erectness, peduncle length), agronomical (ear emergence time, flowering time, grain filling time, ear length, plant height, lodging, grain number, thousand-grain-weight, grain weight per ear, grain protein content, winter hardiness) and disease resistance (powdery mildew, yellow rust, leaf rust, fusarium) traits were studied. Not all traits were scored in each experiment. In total 210 QTLs with a LOD threshold of >2.0 (minor QTLs) were detected of which 64 reached a LOD score of >3.0 (major QTLs). Often QTLs were detected in comparable positions in different experiments. Homologous and homoeologous relationships of the detected QTLs, and already described major genes or QTLs determining the same traits in wheat or other Triticeae members, are discussed.     

Differential proteomic analysis of polyubiquitin-related proteins in chemical hybridization agent-induced wheat (Triticum aestivum L.) male sterility

Protein polyubiquitination is a significant regulator of diverse physiological functions, including sexual reproduction, in plants. Chemical hybridizing agents (CHA) SQ-1 has been shown to induce male sterility in wheat (Triticum aestivum L.) through inhibition of pollen development. This mechanism by which CHA induces male sterility in wheat is unclear. In this study, differential proteomic analysis of polyubiquitinated proteins associated with wheat male sterility was investigated. Wheat plants of the same genetic background were treated with or without CHA. Ubiquitinated proteins were then extracted and enriched for proteomic analysis. Differentially expressed polyubiquitinated proteins in trinuclear stage anther were identified by nanospray liquid chromatography/tandem mass spectrometry. A total of 127 and 131 differentially expressed polyubiquitinated proteins, including heat shock protein 70, ATPase subunit, glycosyltransferase, ubiquitin-related enzyme, and 20S proteasome subunit, were successfully identified by searching against wheat protein database and NCBInr database, respectively. Most of these proteins are related to photosynthesis, carbohydrate and energy metabolism, and multiple metabolic processes. These findings show that alteration of polyubiquitinated proteins is associated with male sterility in wheat.     

Transcript-specific, single-nucleotide polymorphism discovery and linkage analysis in hexaploid bread wheat (Triticum aestivum L.)

Food security is a global concern and substantial yield increases in cereal crops are required to feed the growing world population. Wheat is one of the three most important crops for human and livestock feed. However, the complexity of the genome coupled with a decline in genetic diversity within modern elite cultivars has hindered the application of marker‐assisted selection (MAS) in breeding programmes. A crucial step in the successful application of MAS in breeding programmes is the development of cheap and easy to use molecular markers, such as single‐nucleotide polymorphisms. To mine selected elite wheat germplasm for intervarietal single‐nucleotide polymorphisms, we have used expressed sequence tags derived from public sequencing programmes and next‐generation sequencing of normalized wheat complementary DNA libraries, in combination with a novel sequence alignment and assembly approach. Here, we describe the development and validation of a panel of 1114 single‐nucleotide polymorphisms in hexaploid bread wheat using competitive allele‐specific polymerase chain reaction genotyping technology. We report the genotyping results of these markers on 23 wheat varieties, selected to represent a broad cross‐section of wheat germplasm including a number of elite UK varieties. Finally, we show that, using relatively simple technology, it is possible to rapidly generate a linkage map containing several hundred single‐nucleotide polymorphism markers in the doubled haploid mapping population of Avalon × Cadenza.     

Identification of three Wx proteins in wheat (Triticum aestivum L.)

Nullisomic analysis of waxy (Wx) protein of hexaploid wheat (Triticum aestivum L.) cv. “Chinese Spring” using two-dimensional polyacrylamide gel electrophoresis revealed that threeWx loci,Wx-A1, Wx-B1, andWx-D1, located on chromosome arms 7AS, 4AL, and 7DS, produce three distinct Wx subunit groups, subunit group-A (SGA), SGB, and SGD, respectively. SGA has a higher molecular weight and a more basic isoelectric point (pI) than the other two. SGB and SGD have the same molecular weight but a slightly different pI range. Owing to the detection of these three subunit groups, we were able to identify the expression of three waxy genes in wheat endosperm and to find two types of mutants among Japanese wheat cultivars, one lacking SGA and the others SGB. These results suggest the possibility of breeding a waxy wheat.     

Mapping loci associated with flour colour in wheat (Triticum aestivum L.)

 An RFLP map constructed using 150 single seed descent (SSD) lines from a cross between two hexaploid wheat varieties (‘Schomburgk’בYarralinka’) was used to identify loci controlling flour colour. Flour colour data were obtained from field trials conducted over two seasons at different sites. The estimated heritability of this trait was calculated as 0.67. Two regions identified in the preliminary analysis on chromosomes 3A and 7A, accounted for 13% and 60% of the genetic variation respectively. A detailed analysis of the major locus on 7A was conducted through fine mapping of AFLP markers identified using bulked segregant analysis (BSA). Seven additional markers were identified by the BSA and mapped to the region of the 7A locus. The applicability of these markers to identify wheat lines with enhanced flour colour is discussed. Received: 30 September 1997 / Accepted: 4 February 1998     

Mapping loci associated with milling yield in wheat (Triticum aestivum L.)

A partial genetic linkage map constructed using 150 single seed descent (SSD) lines generated from a cross between the hexaploid wheat varieties Schomburgk and Yarralinka was used to identify loci controlling milling yield. Milling yield data were obtained using seed collected from field trials conducted at different sites over two seasons. The estimated broad-sense heritability of milling yield in this population was calculated as 0.48. In the preliminary analysis, two regions were identified on chromosomes 3A and 7D, which were significantly associated with milling yield and accounted for 22% and 19% of the genetic variation, respectively. Bulked segregant analysis in combination with AFLP identified other markers linked to these loci, as well as an additional region on chromosome 5A, which accounted for 19% of the genetic variation. The applicability of these markers as selection tools for breeding purposes is discussed.     

Linked sucrose synthase genes in group-7 chromosomes in hexaploid wheat (Triticum aestivum L.)

A cDNA library from developing wheat endosperm was screened for sucrose-synthase clones using a maize cDNA probe corresponding to the Sh1 locus under non-stringent conditions. Five positive clones were isolated and initially classified into two types on the basis of their relative ability to hybridize with the probe and of their partial restriction maps. Determination of the nucleotide sequences indicated homology between the two types of wheat clones, with type 1 showing higher homology to the maize Sh1 locus than to type-2 sequences. The inserts cloned in plasmids pST8 (type 1) and pST3 (type 2) were used as probes to determine the chromosomal locations of the two types of genes. DNAs from compensated nulli-tetrasomic and ditelosomic lines of wheat cultivar Chinese Spring were cleaved with EcoRI and analysed in Southern blots. DNA segments of the two types were thus identified in the short arms of chromosomes 7A, 7D, and, possibly, 7B. The two types of linked loci have been designated Ss1 and Ss2, respectively.     

Comparative mapping of genes for glume colouration and pubescence in hexaploid wheat (Triticum aestivum L.)

Microsatellite markers were used to map the major genes Bg (determining black glume colour), Rg1 and Rg3 (red glume), and a locus determining smokey-grey coloured glume to the distal ends of the short arms of the homoeologous group 1 chromosomes, proximally (or closely linked) to Xgwm1223 and distal to Xgwm0033. On this basis, we propose that these genes represent a set of homoeoloci, designated Rg-A1, Rg-B1, and Rg-D1. Rg3 and Bg appear to be variant alleles of Rg-A1. Both Rg3 and Bg are closely linked with the major glume pubescence gene Hg. Similarly, the hexaploid wheat smokey-grey glume gene and Rg2 represent alleles at Rg-D1. The microsatellite markers linked to the Rg genes were used to analyse a phenotypically and genotypically characterized set of Siberian spring wheats. A coincidence between the presence of the 264-bp allele of Xgwm0136 and Rg-A1b (Rg3) was observed; so Xgwm0136 can probably be used as a diagnostic marker for this gene.     

Embryo and endosperm development in wheat (Triticum aestivum L.) kernels subjected to drought stress

The aim of the present work was to reveal the histological alterations triggered in developing wheat kernels by soil drought stress during early seed development resulting in yield losses at harvest. For this purpose, observations were made on the effect of drought stress, applied in a controlled environment from the 5th to the 9th day after pollination, on the kernel morphology, starch content and grain yield of the drought-sensitive Cappelle Desprez and drought-tolerant Plainsman V winter wheat (Triticum aestivum L.) varieties. As a consequence of water withdrawal, there was a decrease in the size of the embryos and the number of A-type starch granules deposited in the endosperm, while the development of aleurone cells and the degradation of the cell layers surrounding the ovule were significantly accelerated in both genotypes. In addition, the number of B-type starch granules per cell was significantly reduced. Drought stress affected the rate of grain filling shortened the grain-filling and ripening period and severely reduced the yield. With respect to the recovery of vegetative tissues, seed set and yield, the drought-tolerant Plainsman V responded significantly better to drought stress than Cappelle Desprez. The reduction in the size of the mature embryos was significantly greater in the sensitive genotype. Compared to Plainsman V, the endosperm cells of Cappelle Desprez accumulated significantly fewer B-type starch granules. In stressed kernels of the tolerant genotype, the accumulation of protein bodies occurred significantly earlier than in the sensitive variety.     

Plant regeneration from long term suspension culture-derived protoplasts of hexaploid wheat (Triticum aestivum L.)

Highly regenerable callus cultures have been obtained from immature embryos of hexaploid wheat cv. Oderzo. Friable fast growing calli were induced at high frequency. Suspensions were initiated from the most friable callus lines: they became established in about two months. Suspensions consisted of cell aggregates of 30 to 1000 um in diameter. Upon plating on MS hormone-free medium, suspensions regenerated green plantlets, and their regenerative capability was maintained for at least 10 months. Protoplasts were isolated from 7–8 day old suspension cultures with a yield of 4–6×10⁶ protoplasts/g fresh weight cells. Protoplast culture was either in liquid medium or in a bead-type system with agarose beads. First divisions were detected at day 5. At day 14 visible colonies were detected and the plating efficiency was evaluated between 2 and 8% over the initial number of protoplasts plated. Protoplast-derived calli were cultured in the presence of 1 mg/l IAA and 0.5 mg/l zeatin and were used for reinitiating new suspension cultures. Upon plating onto MS hormone-free medium, with or without the addition of 0.1 mg/l GA3, calliclones were induced to differentiate. Regeneration of complete plantlets, with shoot and roots took about two months. Plantlets were grown in sterile conditions until 12–15 cm height, and were subsequently transplanted in soil.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - MS Murashige and Skoog (1962) medium - PCV packed cell volume - MES morpholinoethanesulfonic acid     

Chromosome localization of genes for soluble proteins in hexaploid wheat (triticum aestivum L.)

An electrophoretic spectra of proteins, extracted with tris-HCI buffer, pH 8.3 are studied. The ditelosomic lines of the Chinese Spring common wheat cultivar are analysed by the chromosomes of the B genome and of the ditelosomic lines of the same cultivar by first and third chromosomes of the D genome. It is found that structural genes for the synthesis of components Nos. 7, 8, 9 and 10 are localized in 1BL, 2BS, 4BS and 5B chromosomes respectively. The genetic control of the component No. 3 is realized by genes, localized in 1BL and 3D chromosomes, while for component No. 2, in the 3D chromosome.     

Mapping QTLs for grain hardness and puroindoline content in wheat ( Triticum aestivum L.)

Genes for puroindoline-a (Pin-a), puroindoline-b (Pin-b) and grain-softness proteins (GSP) have been shown to be linked to the dominant Ha locus responsible for the soft texture of the grain. Though linkage has been demonstrated of the puroindoline genes to the Ha locus, there is no clear evidence that puroindoline content is the product of the gene Ha. A segregating population of 115 recombinant inbred lines (RILs) originating from a cross between the hexaploid Synthetic wheat ( Triticum durum x Aegilops tauschii, W 7984) and the cultivar 'Opata' (M 85) was studied in two different experimental years to detect Quantitative Trait Loci (QTLs) for three traits: grain hardness (Hard), puroindoline-a (Pin-a) and puroindoline-b (Pin-b) contents. The detection of QTLs was performed using marker linear regression. Negative correlation coefficients (-0.86 and -0.80) were identified between grain hardness and puroindoline content (a and b, respectively) on data obtained in 1996. Results obtained in 1999 confirmed the negative correlation between Hard and Pin-a (-0.73); however a positive correlation coefficient was found with Pin-b content (0.41). Total phenotypic variation explained by each QTL was calculated (R2). For each of the Hard, Pin-a and Pin-b traits one major QTL was detected on the short arm of chromosome 5D, located close to the mta9 allele (puroindoline-a). For the first year (1996) the QTL in this region explained around 63% of the phenotypic variability in grain hardness, 77% in Pin-a and 45% in Pin-b contents. These values were confirmed in trials carried out in 1999 with a R2 value of 0.71, 0.72 and 0.25 for Hard, Pin-a and Pin-b, respectively. In 1996 and 1999 a second major QTL was detected for grain hardness on the long arm of the same chromosome. Present results indicate that it cannot be definitely concluded that puroindoline content represents a linear explanation for variations in grain hardness.     

Silicon absorption by wheat (Triticum aestivum L.)

Although silicon (Si) is a quantitatively major inorganic constituent of higher plants the element is not considered generally essential for them. Therefore it is not included in the formulation of any of the solution cultures widely used in plant physiological research. One consequence of this state of affairs is that the absorption and transport of Si have not been investigated nearly as much as those of the elements accorded 'essential' status. In this paper we report experiments showing that Si is rapidly absorbed by wheat (Triticum aestivum L.) plants from solution cultures initially containing Si at 0.5 mM, a concentration realistic in terms of the concentrations of the element in soil solutions. Nearly mature plants (headed out) 'preloaded' with Si absorbed it at virtually the same rate as did plants grown previously in solutions to which Si had not been added. The rate of Si absorption increased by more than an order of magnitude between the 2-leaf and the 7-8 leaf stage, with little change thereafter. This revised version was published online in June 2006 with corrections to the Cover Date.     

The wheat (Triticum aestivum L.) leaf proteome

The wheat leaf proteome was mapped and partially characterized to function as a comparative template for future wheat research. In total, 404 proteins were visualized, and 277 of these were selected for analysis based on reproducibility and relative quantity. Using a combination of protein and expressed sequence tag database searching, 142 proteins were putatively identified with an identification success rate of 51%. The identified proteins were grouped according to their functional annotations with the majority (40%) being involved in energy production, primary, or secondary metabolism. Only 8% of the protein identifications lacked ascertainable functional annotation. The 51% ratio of successful identification and the 8% unclear functional annotation rate are major improvements over most previous plant proteomic studies. This clearly indicates the advancement of the plant protein and nucleic acid sequence and annotation data available in the databases, and shows the enhanced feasibility of future wheat leaf proteome research.     

Expression analysis and physical mapping of low-molecular-weight glutenin loci in hexaploid wheat (Triticum aestivum L.).

Gliadins and glutenins are storage proteins important in determining the bread-, noodle-, and pasta-making quality of wheat. Glutenins consist of HMW and LMW subunits. The Glu-A3, Glu-B3, and Glu-D3 loci on the short arms of chromosomes 1A, 1B, and 1D, respectively, are the major loci for LMW glutenins. To construct physical maps of the Glu-3 loci, a set of 24 high-density filters representing a 3.1x genome coverage hexaploid wheat BAC library was screened by hybridization using a probe made of 3 LMW glutenin sequences. After 2 rounds of hybridization, a subset of 536 BAC clones were selected and fingerprinted. Three developing seed cDNA libraries were also constructed. A total of 5000-6000 ESTs were generated from each library, assembled into contigs and searched by homology for LMW glutenin sequences. In total, 90 full-length LMW glutenin sequences were found to cluster into 8 distinct groups representing at least 21 different LMW glutenin subunits. A set of 24 pairs of PCR primers was designed from these groups and used as markers on the BAC clones. The combined fingerprinting and marker data were used to build the physical maps using FPC software. A total of 91 contigs comprising 254 clones were obtained and 282 clones remained singletons.     

Association analysis of some drought related characters in hexaploid spring wheat (Triticum aestivum L.).

The association among yield components and their direct and indirect influence on the grain yield of wheat were investigated. 24 breeding lines were tested in a randomized complete block experiment design with three replications. According to the results the phenotypic correlation among the traits and their path coefficient were estimated. Positive significant correlation coefficients were obtained for association between survival rate treatment I, III, leaf venation, stomatal frequency, osmotic pressure, flag leaf area and number of tillers per plant with grain yield per plant at both phenotypic and genotypic levels. Negatively significant correlation between hygrophilic colloids and epidermal cell size with grain yield per plant was obtained at phenotypic and genotypic levels. Path coefficient was also computed to estimate the contribution of character to the yield. Path coefficient analysis revealed that flag leaf area, root/shoot ratio and survival rate II had the highest positive direct effects on grain yield, while hygrophilic colloids and osmotic pressure had negative direct effect on grain yield. These results thus obtained suggested that flag leaf area is an important component of yield and hence needs special attention in selection strategies.