Amylose content and starch properties generated by five variant <Emphasis Type="Italic">Wx</Emphasis> alleles for granule-bound starch synthase in common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

In common and durum wheats (Triticum aestivum L. and T. durum Desf.), variant waxy (Wx) alleles have been reported for three Wx proteins (Wx-A1, -B1 and -D1), responsible for amylose synthesis in flour starch. Five variant alleles, Wx-A1c, -A1e, -B1c, -B1d and -D1c, were examined to elucidate their effects on amylose content in flour starch. Common wheat lines carrying a Wx protein produced by one variant (e.g., Wx-A1c) and one control (e.g., Wx-A1a) allele were bred and their starches were compared. Results showed that Wx-A1e did not produce amylose (waxy phenotype), whereas three alleles (Wx-A1c, -B1c and -B1d) reduced amylose, and -D1c might have increased it slightly. Most data on blue value, swelling power and starch paste clarity in water and dimethyl sulphoxide also suggested the variant Wx alleles either reduced or increased amylose content.     

Amylose synthesis capacity of the three Wx genes of wheat cv. Chinese Spring

The Wx locus controls amylose synthesis in the cereal endosperm. Hexaploid wheat (Triticum aestivum L.) has the three Wx loci on chromosomes 7A ( Wx-A1), 4A (Wx-B1) and 7D (Wx-D1). To verify the effects of null alleles on reducing amylose content and determine the amylose synthesis capacity of each Wx gene independently and accurately, we produced eight possible types of recombinant lines carrying different null alleles at the Wx loci under the ‘Chinese Spring’ genetic background. Amylose content varied from 0% of the waxy ‘Chinese Spring’ to 25% of the ‘Chinese Spring’ normal type. The reducing effect of the single null alleles was the largest in Wx-B1b, and there was no significant difference between Wx-A1b and Wx-D1b. More than 3% reductions in amylose content were detected in the double null types. The results of the double null lines further demonstrated that for the capacity of amylose synthesis, Wx-B1a predominates and produces 21–22% amylose, followed by Wx-D1a (20–21%) and Wx-A1a (15–18%). These significant differences were partly correlated with variation in the amounts of the Wx proteins produced by different Wx genes. However, comparisons of the double null lines with the single null or normal lines indicated that amylose content was not linearly proportional to the number of the Wx genes, suggesting that the Wx genes act in an epistatic manner. This revised version was published online in July 2006 with corrections to the Cover Date.     

Polymorphism of two waxy proteins in the emmer group of tetraploid wheat, Triticum dicoccoides, T. dicoccum, and T. durum

Gel-electrophoretic analyses detected polymorphism of two waxy (Wx) proteins, Wx-Al and Wx-Bl, in 334 accessions of the emmer group of tetraploid wheat, Triticum dicoccoides, T. dicoccum and T. durum. The null allele for the Wx-Al protein (Wx-Alb) was found in one accession of T. dicoccoides and seven of T. dicoccum, but it was not present in those T. durum accessions analysed. The null allele for the Wx-Bl protein (Wx-Blb) did not occur in three emmer-wheat species. Wx-Al and Wx-Bl proteins showing alteration in mobility in SDS-PAGE gel or in isoelectric points were found in five accessions and considered to be the products of new alleles, Wx-Ald, Wx-Ale and/or Wx-Bld. Densitometric analysis of Wx protein bands revealed that Wx-Al was present in smaller amounts than Wx-B1 in almost all accessions.     

Polymorphism of waxy proteins in Iberian hexaploid wheats

A collection of 130 cultivars of bread wheat, 332 landraces of bread wheat and 144 spelt wheats was analysed for waxy proteins in the grain. The electrophoretic patterns showed very low polymorphism and most of the hexaploid wheats had the Wx-Ala, Wx-D1a and Wx-B1 alleles of ‘Chinese Spring’. Two alleles were detected at Wx-A1 (Wx-A1a, and Wx-A1b (null)), the latter was present in only 5.1% of the bread wheat landraces and 7.6% ofthe spelt wheats. No allelic variation was found at the Wx-D1 locus and all the hexaploid wheats had the Wx-D1a allele. Wx-B1 was the most polymorphic locus, with three alleles detected: Wx-B1a, Wx-B1b (null) and Wx-Blc coding for a Wx-B1 protein with a slightly different mobility from Wx-B1a. The null Wx-B1b allele was found in 10.8% of the bread wheat cultivars, 21.4% of the bread wheat landraces and 12.5% of the spelt wheats. Among the 604 hexaploid wheats analysed, only two bread wheat landraces (0.6%) and two spelt wheats (1.4%) had the null allele at both Wx-A1 and Wx-B1 loci.     

Microsatellite mapping of complementary genes for purple grain colour in bread wheat (Triticum aestivum) L.

Complementary genes for purple grain colour Pp1, Pp2, Pp3 (now designated Pp1, Pp3b, Pp3a, respectively) were mapped using crosses between purple-grained hexaploid wheats ‘Purple Feed’ – Pp1Pp1/Pp2Pp2 (Pp1Pp1/Pp3bPp3b), ‘Purple’ – Pp1Pp1/Pp3Pp3 (Pp1Pp1/Pp3aPp3a) with non-purple-grained cultivars ‘Novosibirskaya 67’ (‘N67’) and ‘Saratovskaya 29’ (‘S29’). The genes Pp2 (Pp3b) and Pp3 (Pp3a) were inherited as monofactorial dominant when purple-grained wheats were crossed to ‘N67’. Both were mapped in the centromeric region of the chromosome 2A. Therefore, they were suggested being different alleles at the same locus and designated Pp3a and Pp3b. In the crosses between purple-grained wheats and ‘S29’ a segregation ratio of 9 (purple) to 7 (non purple) was obtained suggesting a complementary interaction of two dominant genes, Pp1 and Pp3. To map Pp1 as a single gene, the influence of the other Pp gene was taken into consideration by determining the Pp3 genotype of the F₂ plants. The gene was mapped on chromosome 7BL, about 24 cM distal to the centromere. The Pp1gene was shown to be non allelic to the Rc-1 (red coleoptile) and Pc (purple culm) genes, contrary to what was previously suggested. The colouration caused by the Pp genes has no effect on pre-harvest sprouting.     

Genetic mapping of loci determining long glumes in the genus Triticum

Elongated glumes are present in thetetraploid wheat species T.polonicum, T. turanicum, T.durum convar. falcatum and in thehexaploid species T. petropavlovskyi.Inheritance of glume length was studiedwith the aim to map the respective lociusing wheat microsatellite markers. In T. polonicum and T. petropavlovskyiloci conferring long glume were mapped nearthe centromere on chromosome 7A. These twoloci are designated P-A ^pol 1 andP-A ^pet 1, respectively. It isshown that both are probably homoeoallelicto each other and to the P gene ofT. ispahanicum on chromosome 7B. The loci determining elongated glumes in T. turanicum and T. durum conv. falcatum are not homoeologous to the P loci in the centromeric region of thegroup 7 chromosomes.     

Genetic control of long glume phenotype in tetraploid wheat derived from Triticum petropavlovskyi Udacz. et Migusch.

The Chinese wheat landrace, Xinjiang rice wheat (T. petropavlovskyi Udacz. et Migusch., 2n = 42), known as ‘Daosuimai’ or rice-head wheat is characterized by long glumes, and was found in the agricultural areas in the west part of Talimu basin, Xinjiang, China in 1948. The gene for long glume from T. petropavlovskyi was introduced into a line of spring durum wheat, LD222. The gene for long glume is located approximately46.8 cm from the cn-A1 locus, which controls the chlorinatrait. Significant deviation from a 3:1 in the F₂ of LDN7D(7A)/ANW5C confirmed that the long glume of T. petropavlovskyi can be controlled by a gene located on chromosome 7A. The gene locates approximately 12.4 ± 0.5 cM from the centromere on the long arm of 7A. It is considered that the gene for long glume from T. petropavlovskyi is an allele on the P ₁ locus, and it should be designated as P _1a. It is suggested that T. petropavlovskyi originated from either the natural hybrid between T. aestivum that has an awn-like appendage on the glume and T. polonicum or a natural point mutation of T. aestivum. This revised version was published online in August 2006 with corrections to the Cover Date.     

Variation of high-molecular-weight glutenin subunits amongst cultivars of Triticum turgidum L. from Portugal

Summary Variation in high-molecular-weight (HMW) glutenin subunit composition amongst 63 varieties of Triticum turgidum L. from Portugal was investigated using SDS-PAGE. A total of thirteen Glu-A1 and Glu-B1 alleles were identified, and three of them were found to be different from those previously described in the literature. A number of the tetraploid wheats examined contained subunits known to have a beneficial effect of the bread-making properties of T.aestivum. Camara, a tetraploid wheat cultivar carrying a 1D/1B chromosomal substitution, is proposed as a bridge for the transfer of Glu-D1 alleles and of other protein fractions controlled by the 1D chromosomes (Gli-D1 locus) from hexaploid to tetraploid wheat.     

Resistance to stripe rust in Triticum turgidum,T. tauschii and their synthetic hexaploids

Resistance to stripe rust (caused by Puccinia striiformis Westend.) of 34 Triticum turgidum L. var.durum, 278 T. tauschii, and 267 synthetic hexaploid wheats (T. turgidum x T. tauschii) was evaluated at the seedling stage in the greenhouse and at the adult-plant stage at two field locations. Mexican pathotype 14E14 was used in all studies. Seedling resistance, expressed as low infection type, was present in all three species. One hundred and twenty-eight (46%) accessions of T. tauschii, 8 (23%) of T. turgidum and 31 (12%) of synthetic hexaploid wheats were highly resistant as seedlings. In the field tests, resistance was evaluated by estimating area under disease progress curve (AUDPC). Synthetic hexaploid wheats showed a wide range of variability for disease responses in both greenhouse and field tests, indicating the presence of a number of genes for resistance. In general, genotypes with seedling resistance were also found to be resistant as adult plants. Genotypes, which were susceptible or intermediate as seedlings but resistant as adult plants, were present in both T. turgidum and the synthetic hexaploids. Resistances from either T. turgidum or T. tauschii or both were identified in the synthetic hexaploids in this study. These new sources of resistance could be incorporated into cultivated hexaploid wheats to increase the existing gene pool of resistance to stripe rust.     

Waxy proteins and amylose content in diploid Triticeae species with genomes A, S and D

A collection of 89 accessions of diploid species of wheat was analysed for waxy protein in the grain: 39 accessions of Einkorn wheats, 41 accessions of Sitopsis section wheat and nine accessions of Triticum tauschii. The electrophoretic patterns showed low polymorphism. In each group of wheat, a single and different allele was detected. In accessions of Einkorn wheats that allele had a similar electrophoretical mobility to the Wx‐A1a allele of the bread wheat ‘Chinese Spring’, in accessions of the Sitopsis section it had a similar mobility to that of the Wx‐B1f allele of tetraploid wheat, and in the accessions of T. tauschii, it was similar to the Wx‐D1a allele of the bread wheat ‘Chinese Spring’. The accessions were also analysed for apparent amylose content. Results showed that amylose content ranged from 22 to 35% in Einkorn wheats, from 28 to 41% in the Sitopsis section and from 26 to 35% in accessions of T. tauschii.     

Chromosome location of genes for resistance to powdery mildew in common wheat (Triticum aestivum L.) 1. Mlk and other alleles at the Pm3 locus

Summary Several wheat cultivars/lines were inoculated with isolates of Erysiphe graminis tritici to identify new genes/alleles for resistance. The wheats were tested with 13 isolates that had been characterized from responses on differential lines with known resistance genes. Gene Mlk which occurs in cultivars Kolibri, Syros, Ralle and several other European common wheats was found to be an allele at the Pm3 locus and is now designated Pm3d. The mildew resistance in an old Australian wheat, W150, is conferred by a single gene also allelic to Pm3 and now designated Pm3e. The near-isogenic line Michigan Amber/8^*Cc possesses another allele now designated Pm3f. A Syrian land variety of common wheat shows mildew resistance that is conditioned by the combination of genes Pm1 and Pm3a. Finally, two accessions of Triticum aestivum ssp. sphaerococcum appeared to possess the Pm3c allele.     

Gene dosage effect of the wheat Wx alleles and their interaction on amylose synthesis in the endosperm

To find out gene dose effect of each of the three homoeologous Wx genes and their interaction on the production of granule-bound starch synthase (GBSS I) and amylose biosynthesis in the endosperm, Chinese Spring and its near-isogenic waxy types were crossed reciprocally and, obtained a plant population with varying doses of each Wx gene. The amount of GBSSI was increased linearly with increasing gene dose of either of Wxloci. In each of the three Wx loci, the change in amylose content was linear up to 3 doses, with a more potent capacity ofWx-B1a at any dose. Higher level of amylose production was observed in the reciprocal F₁ grains than the expected effect of dose/s of each gene or additive effect of different allelic combination by artificially blend starches which have amylose produced by equivalent number ofWx alleles to that of relevant F₁ cross. When Wx-B1a and Wx-A1a were combined, increase in amylase content was not in proportion to increase in gene dosage. The enhanced amylase synthesis was shown by 2-gene and 3-geneinteraction, indicating that not only type of the three Wx genes and its dose but the interaction among them have significant roles in determining the amylose content. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of near-isogenic lines of wheat carrying different null Wx alleles and their starch properties

The granule-bound starch synthase (GBSS I) encoded by the Wxgenes, is involved in amylose synthesis. For analyses of mechanisms of amylose synthesis and associated starch properties in hexaploid wheat, eight possible genotypes having different combinations of the three null alleles at the Wx loci with a common genetic background are a prerequisite. A near-isogenic population of doubled haploid (DH) lines was produced from Chinese Spring × waxy Chinese Spring F₁ plants using the wheat × maize method. The Wx protein phenotypes of the DH progeny were examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and found that the null alleles at each of the three Wx loci segregated in a Mendelian fashion. A field trial demonstrated no differences between the eight types for ear emergence time, plant height and grain yield traits. Amylose content in the endosperm starch was highest in the wild type while lowest in the waxy type having no Wx proteins. Comparison between single null types and double null types indicated that the amylose synthesis capacity of Wx-A1a allele is the lowest. Pasting properties of starch are the highest in the waxy type, followed by the double null types. Consequently, both peak viscosity and breakdown were negatively correlated with amylose content. The chain-length distribution analysis of amylopectin structure revealed no clear difference among the eight types,suggesting that the reduced GBSS I activity due to introgression of the null Wx alleles does not affect either the chain length or the degree of branching of amylopectin. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chromosomal regions associated with green plant regeneration in wheat (Triticum aestivum L.) anther culture

Genetic capacity for green plant regeneration in anther culture were mapped in a population comprising 50 doubled haploid lines from a cross between two wheat varieties ‘Ciano’ and ‘Walter’ with widely different capacity for green plant regeneration. Bulked segregant analysis with AFLP markers and composite interval mapping detected four QTLs for green plant percentage on chromosomes 2AL (QGpp.kvl-2A), 2BL (QGpp.kvl-2B.1 and QGpp.kvl-2B.2) and 5BL (QGpp.kvl-5B).The three QTLs detected on chromosome 2AL and 2BL all derived their alleles favouring green plant formation from the responsive parent ‘Ciano’.The remaining QTL on chromosome 5BL had the allele favouring green plants from the low responding parent ‘Walter’. In a multiple regression analysis the four QTLs could explain a total of 80% of the genotypic variation for green plant percentage. None of the chromosomal regions with QTLs for green plant percentage showed significant influence on either embryo formation or regeneration frequencies from the anther culture. The three major QTLs located on group two chromosomes were fixed in a second DH population derived from two parents ‘Ciano’ and ‘Benoist’,both with high capacity to produce green plants. A QTL explaining31.5% of the genetic variation for green plant formation were detected on chromosome 5BL in this cross as well. This revised version was published online in July 2006 with corrections to the Cover Date.     

Waxy proteins in diploid, tetraploid and hexaploid wheats

Electrophoretic analyses of waxy proteins, encoded by genes present at the Wx‐1 loci, present in several cultivars and accessions of hexaploid wheat, Triticum aestivum, have permitted the detection of null alleles at the Wx‐B1 and Wx‐D1 loci. Polymorphism at the Wx‐A1 and Wx‐B1 loci was also investigated in several accessions of tetraploid wheats, Triticum durum, Triticum dicoccoides and Triticum timopheevi, and in diploid species, Triticum urartu, Triticum boeoticum and Triticum monococcum. One null allele at the Wx‐A1 locus and three polymorphic alleles at Wx‐B1 locus were detected in T. durum; a new allele at one of the two waxy loci was identified in the tetraploid wheat T. timopheevi; no polymorphism was detected in diploid species. Polymerase chain reaction techniques made possible the detection of further polymorphism existing at the Wx‐1 loci and the reason for the lack of expression of the null genotypes to be investigated. The null forms detected at each locus have been used to produce complete sets of partial and total waxy lines in durum and bread wheat.     

Increased grain protein content and its association with agronomic and end-use quality in two hard red spring wheat populations derived from Triticum turgidum L. var. dicoccoides

Suitability of wheat (Triticum aestivum L.) for many food products depends on its unique protein. Elevated grain protein content (GPC) and its quality influences the bread making properties of wheat. The objective of this study was to examine the association of elevated GPC with agronomic and end-use quality in two hard red spring wheat recombinant inbred (RI) populations derived from wild emmer (Triticum turgidum L. var. dicoccoides). The two hard red spring populations (ND683/Bergen and Glupro/Bergen) were developed using a single-seed-descent method. ND683 and ‘Glupro’ are high in GPC (180 g kg^-1), presumably due to the introgression of gene(s) from Triticum turgidum L. var. dicoccoides and ‘Bergen’ is low in GPC (145 g kg^-1). From each of the two populations 12 high- and 12 low-GPC RI lines (F_5:7) were selected for replicated testing at two North Dakota (ND) locations in 1995. In both populations, the high-GPC lines had significantly (p < 0.05) higher values compared to the low-GPC lines for mean GPC and water absorption. Mean grain yield of the high-GPC lines was not significantly different from the low-GPC lines in both populations. In the ND683/Bergen population, the high-GPC lines had significantly (p < 0.05) higher values than the low-GPC lines for mean plant height, days to heading, and flour extraction. GPC was significantly (p < 0.05)and negatively associated with test weight and also significantly (p < 0.01) and positively associated with water absorption in the Glupro/Bergen population. In these populations, results suggested that it may be possible to select lines that combine higher GPC and acceptable yield level, but later in maturity and taller in plant height. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detecting and estimating segregation distortion and linkage between glufosinate tolerance and blackleg resistance in Brassica napus L.

Summary The segregation and linkage between glufosinate (transgenes ‘Rf3’ and ‘T177’) and blackleg resistance genes in canola (Brassica napus L.) were assessed using F₁ microspore-derived doubled haploid (DH) populations from four crosses including reciprocals, two involving the transgene ‘Rf3’ and the other two involving the transgene ‘T177’. To relax the assumption of no segregation distortion required for the conventional analysis of segregation and linkage, we employed Bailey's analysis that allows detecting segregation distortion at linked loci. The significant departures from the 1:1 segregation were detected in the crosses involving the transgene ‘T177’ but not in the crosses involving the transgene ‘Rf3’. The apparent deficit of the herbicide tolerant DH lines in the crosses with the transgene ‘T177’ is likely due to differential selection against the gametes carrying ‘T177’ during microspore culture. The linkage was strong between blackleg resistance and the transgene ‘Rf3’ but weak or absent between blackleg resistance and the transgene ‘T177’, suggesting that the two transgenes are probably inserted into distant regions of the genome. The observed linkage offers an opportunity to develop new canola cultivars with both glufosinate tolerance conferred by transgene ‘Rf3’ and blackleg resistance.     

Comparative Analysis of High-Molecular-Weight Glutenin Subunit Composition in Various Triticum Species

Variation in high-molecular-weight glutenin subunit composition amongst Triticum durum cvs. of different origins was investigated by SDS-PAGE and compared with that reported for T. dicoccum and T. aestivum. Tetraploid wheat collections (408 cvs.) were found to carry nearly twice as many Glu-A1 and Glu-B1 alleles as a hexaploid wheat sample of comparable size. In each of the taxa considered, allelic variation at the Glu-B1 locus was markedly greater than that observed for Glu-A1. However, because all the Glu-B1 subunits so far discovered exhibit a restricted and distinctive mobility range during. SDS-PAGE, it is suggested that they are derived from a single source, possibly from Aegilops searsii. Most durum cvs. carried a ‘null’Glu-A1 allele and therefore fewer subunits than dicoccums and common wheats. It is argued that differences in the frequency of occurrence of ‘null’Glu-1 alleles between taxa probably resulted from random samplings made by early agriculturalists and breeders, rather than from an inherent tendency of polyploid wheats to suppress the activity of “redundant” genes.     

Production and cytogenetics of intergeneric hybrids between Triticum durum-Dasypyrum villosum amphidiploid and Psathyrostachys huashanica

Summary Intergeneric crosses between Triticum durum-Dasypyrum villosum (2n=42, AABBVV), and Psathyrostachys huashanica (2n=14, N ^h N ^h ) were made, the seed set was 1.67%. Intergeneric hybrid were successfully obtained by means of embryo culture for first time. The average chromosome pairing in the hybrid (ABVN ^h ) was 26.61% univalents, and 0.69 bivalents. The chiasmata per cell was 0.69. The chiasmata was higher than that in Triticum durum dihaploid (AB), and lower than that in T. durum-Dasypyrum villosum trihaploid (ABV). The result indicated that the N ^h genome of Psathyrostachys huashanica has no homology with the V genome of Dasypyrum villosum, and the A and B genomes of Triticum durum. The coenocytism, micronuclei cell and variation in chromosome numbers were also observed. The F₁ hybrid was crossed with Triticum aestivum (AABBDD), and resulted in seed set. The hybrid of T. durum-D. villosum amphidiploid x P. huashanica showed partial fertility. It made the possibility for chromosome manipulation among Triticum aestivum, Dasypyrum villosum and Psathyrostachys huashanica.     

Wheat pollen dispersal under semiarid field conditions: potential outcrossing with Triticum aestivum and Triticum turgidum

Isolation distance is the main barrier to crop-to-crop gene-flow. A 3-year study assessed the maximum potential outcrossing under field conditions between two wheat cultivars (Triticum aestivum L.) and between wheat and durum wheat (Triticum turgidum L. var. durum). Outcrossing was measured by seed set on emasculated recipient plants placed at four sides with different distances from a 3 m × 3 m T. aestivum (cultivar Chinese Spring) pollen source. Frequencies of seed set at 0 m distance were 45% (37–56%) for T. aestivum cultivars and 18% (5–30%) with T. turgidum. These values agree with hybridization in non-limiting pollen conditions measured by manual crosses in greenhouse. The number of pollen grains and the outcrossing frequencies decreased at increasing distances influenced by the prevailing wind direction. Under semiarid conditions of this assay, viable pollen was found 14 m from the pollen source, with a maximum distance of 8 m at which cross-pollination decreases below 1%. Ambient conditions affect pollen viability, hybridization and pollen dispersal. Data presented in this paper emphasize the major role played by environmental conditions in outcrossing. Data obtained in one area may therefore not coincide with the prevailing situation in different locations and climates.