Impact of puroindoline b alleles on the genetic variation for hardness in soft × hard wheat crosses

Wheat grain hardness is controlled by one major genetic factor, the puroindoline hardness (ha) locus on the short arm of chromosome 5D, but there is also evidence for other minor genetic factors modifying the effect of puroindoline alleles. In this study, the progeny of nine soft × hard wheat crosses was evaluated for kernel texture, and the allelic state of puroindoline b (pinB) was assessed by polymerase chain reaction. The F₂ populations of all nine crosses showed the expected 1:2:1 segregation ratio of homozygous soft, heterozygous medium and homozygous hard offspring. A model of variance components was constructed to separate the effects of pinB‐D1 allelic variation from other genetic factors affecting endosperm hardness. This model showed that pinB‐D1 allelic variation could explain 75‐93% of the genetic variation for hardness in the F₂, but there were also significant contributions from other genetic factors in all the crosses. The feasibility of pinB‐D1 alleles as a molecular marker for hardness is demonstrated, and the results also indicate the possibility of breeding wheat varieties with true medium hardness.     

Quantitative trait locus analysis of wheat quality traits

Milling and baking quality traits in wheat (Triticum aestivum L.) were studied by QTL analysis in the ITMI population, a set of 114 recombinant inbred lines (RILs) generated from a synthetic-hexaploid (W7985) × bread-wheat (Opata 85) cross. Grain from RILs grown in U.S., French, and Mexican wheat-growing regions was assayed for kernel-texture traits, protein concentration and quality, and dough strength and mixing traits. Only kernel-texture traits showed similar genetic control in all environments, with Opata ha alleles at the hardness locus Ha on chromosome arm 5DS increasing grain hardness, alkaline water retention capacity, and flour yield. Dough strength was most strongly influenced by Opata alleles at 5DS loci near or identical to Ha. Grain protein concentration was associated not with high-molecular-weight glutenin loci but most consistently with the Gli-D2 gliadin locus on chromosome arm 6DS. In Mexican-grown material, a 2DS locus near photoperiod-sensitivity gene Ppd1 accounted for 25% of variation in protein, with the ppd1-coupled allele associated with higher (1.1%) protein concentration. Mixogram traits showed most influence from chromosomal regions containing gliadin or low-molecular-weight glutenin loci on chromosome arms 1AS, 1BS, and 6DS, with the synthetic hexaploid contributing favorable alleles.Some RI lines showed quality values consistently superior to those of the parental material, suggesting the potential of further evaluating new combinations of alleles from diploid and tetraploid relatives, especially alleles of known storage proteins, for improvement of quality traits in wheat cultivars.Contribution number 06-77J from the Kansas Agricultural Experimental Station.     

Genetic control of kernel modification found in South African quality protein maize inbred lines

Summary The genetic control of endosperm modification in 12 opaque-2 maize (Zea mays L.) inbred lines was investigated by means of a diallel cross experiment conducted across two environments. Kernel vitreousness and kernel hardness were determined by partially dominant genes. Additive gene action was largely responsible for kernel modification. A favourable general combining ability for kernel vitreousness and kernel hardness was positively correlated with an accumulation of dominant kernel modifying genes. South African sources of endosperm modifiers have been found to be similar to those used in other quality protein maize breeding programmes. Certain inbred lines displayed sufficient genetic potential for use in a quality protein maize hybrid breeding programme.     

Molecular mapping of cereal cyst nematode resistance in <Emphasis Type="Italic">Triticum monococcum</Emphasis> L. and its transfer to the genetic background of cultivated wheat

Triticum monococcum, the diploid A genome species, harbours enormous variability for resistance to biotic stresses. A spring type T. monococcum acc. 14087 was found to be resistant to Heterodera avenae (cereal cyst nematode, CCN). A recombinant inbred line population (RIL) developed by crossing this accession with a CCN susceptible T. boeoticum acc. 5088 was used for studying the inheritance and map location of the CCN resistance. Based on composite interval mapping two QTL, one each on chromosome 1AS and 2AS, were detected. The QTL on 1A, designated as Qcre.pau-1A, appeared to be a major gene with 26% contribution to the overall phenotypic variance whereas the QTL on 2A designated as Qcre.pau-2A contributed 13% to total phenotypic variation. Qcre.pau-1A is novel, being the only CCN resistance gene mapped in any ‘A’ genome species and none of the other known genes have been mapped on chromosome 1A. The QTL Qcre.pau-2A might be allelic to Cre5, a CCN resistance gene transferred from Ae. ventricosa and mapped on 2AS. The Qcre.pau-1A was transferred to cultivated wheat using T. durum cv. PBW114 as the bridging species. Selected CCN resistant F₈ lines showed introgression for the molecular markers identified to be linked with CCN resistance locus Qcre.pau-1A. Thus, this gene alone could impart complete resistance against CCN. These introgression lines can be used for marker-assisted transfer of Qcre.pau-1A to elite wheat cultivars.     

A polymorphic microsatellite in the 3'end of 'waxy' genes of wheat, Triticum aestivum

Potential polymorphism of an (AT)_N microsatellite at the 3’end of waxy genes in bread wheat was examined. Primers were designed from a published cDNA sequence of a wheat waxy gene. Polymerase chain reaction (PCR) amplification of genomic DNA from 135 mainly Australian cultivars revealed eight alleles on chromosome 7A. This polymorphic microsatellite is a potential codominant marker for the Wx-A1 locus in breeding programmes. A distinguishable fragment was also amplified from chromosome 7D. This fragment was absent where a plant was null for the waxy gene on chromosome 7D, being a dominant marker for the Wx-D1 locus. The primers were also useful for amplifying genomic DNA from barley, rye and triticale and can be used to detect potential polymorphism in these species.     

The effect of introgressions of wheat D-genome chromosomes into 'Presto' triticale

Hexaploid triticale (X Triticosecale Wittmack) (2n= 6x= 42, AABBRR) and wheat (Triticum aestivum L.) (2n= 6x= 42, AABBDD) differ in their R and D-genomes. This produces differences in both agronomic and end-use quality characteristics. Our objective was to determine how introgressions of individual chromosomes from the D-genome of wheat affect these characteristics of a winter triticale 'Presto'. We studied the effects of 18 D-genome chromosome substitution lines, 15 sib-lines as controls, and five check cultivars at Lincoln, NE in 1996, using a randomized complete block design with two replications. The experiment was repeated at Lincoln and Mead, NE in 1997 and 1998 with 15 substitution lines that survived the first winter in Lincoln, along with their 12 control sibs and five check cultivars. Few D-genome chromosomes had positive effects. Chromosomes 2D, 4D, and 6D significantly reduced plant height when substituted for 2R, 4B, and 6R, respectively. No grain yield increases were associated with any of the D-genome chromosomes tested, but three substitutions decreased the grain yield. Depending on the allele of the hardness gene present, chromosome 5D increased or decreased kernel hardness when substituted for 5R or 5A, respectively. Introgressions of chromosomes 1D and 6D improved end-use quality characteristics of Presto. These results suggest that apart from beneficial effects of individual loci located on the D-genome chromosomes, no major benefit can be expected from D-genome chromosome substitutions.     

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.     

Mapping the <Emphasis Type="Italic">compactum</Emphasis> locus in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) and its relationship to other spike morphology genes of the Triticeae

The spikes of club wheat are significantly more compact than spikes of common wheat due to the action of the dominant allele of the compactum (C) locus. Little is known about the location of C on chromosome 2D and the relationship between C and to other spike-compacting genes. Thus, a study was undertaken to place C on linkage maps and a chromosome deletion bin, and to assess its relatedness to the spike compacting genes zeocriton (Zeo) from barley and soft glume (Sog) from T. monococcum. Genetic mapping was based on recombinant inbred lines (RILs) from a cross between the cultivars Coda (club) and Brundage (common) and F₂ progeny from a cross between the club wheat Corrigin and a chromosome 2D substitution line [Chinese Spring (Ae. tauschii 2D)]. The C locus was flanked by Xwmc144 and Xwmc18 in the RIL population and it was completely linked to Xcfd116, Xgwm358 and Xcfd17 in the F₂ population. C could not be unambiguously placed to a chromosome bin because markers that were completely linked to C or flanked this locus were localized to chromosome bins on either side of the centromere (C-2DS1 and C-2DL3). Since C has been cytogenetically mapped to the long arm of chromosome 2D, we suspect C is located in bin C-2DL3. Comparative mapping suggested that C and Sog were present in homoeologous regions on chromosomes 2D and 2A^m, respectively. On the other hand, C and Zeo, on chromosome 2H, did not appear to be orthologous.     

Identification and genetic mapping of a powdery mildew resistance gene in wild emmer (<Emphasis Type="Italic">Triticum dicoccoides</Emphasis>) accession IW72 from Israel

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a devastating disease of wheat (Triticum aestivum) in China and worldwide, causing severe yield losses annually. Wild emmer (T. dicoccoides) accession IW72 collected from Israel is resistant to powdery mildew at the seedling and adult stages. Genetic analysis indicated that the resistance was controlled by a single dominant gene, temporarily designated MlIW72. The F₂ population and F₃ families derived from a hybrid between IW72 and susceptible durum wheat line Mo75 were used for molecular mapping of the resistance gene. MlIW72 was linked with SSR loci Xgwm344, Xcfa2040, Xcfa2240, Xcfa2257 and Xwmc525 on the long arm of chromosome 7A. In addition, two STS markers, MAG2185 (derived from RFLP marker PSR680) and MAG1759 (developed from EST CD452874), were mapped close to MlIW72. All these markers were physically located in the terminal bin 0.86–1.00 of 7AL. The chromosome location and genetic mapping results suggested that the powdery mildew resistance gene identified in wild emmer accession IW72 might be a new allele at the Pm1 locus or a new locus closely linked to Pm1.     

Mapping of the leaf rust resistance gene <Emphasis Type="Italic">Lr38</Emphasis> on wheat chromosome arm 6DL using SSR markers

Leaf rust caused by the fungus Puccinia triticina is one of the most important diseases of wheat (Triticum aestivum) worldwide. The use of resistant wheat cultivars is considered the most economical and environment-friendly approach in controlling the disease. The Lr38 gene, introgressed from Agropyron intermedium, confers a stable seedling and adult plant resistance against multiple isolates tested in Europe. In the present study, 94 F₂ plants resulting from a cross made between the resistant Thatcher-derived near-isogenic line (NIL) RL6097, and the susceptible Ethiopian wheat cultivar Kubsa were used to map the Thatcher Lr38 locus in wheat using simple sequence repeat (SSR) markers. Out of 54 markers tested, 15 SSRs were polymorphic between the two parents and subsequently genotyped in the population. The P. triticina isolate DZ7-24 (race FGJTJ), discriminating Lr38 resistant and susceptible plants, was used to inoculate seedlings of the two parents and the segregating population. The SSR markers Xwmc773 and Xbarc273 flanked the Lr38 locus at a distance of 6.1 and 7.9 cM, respectively, to the proximal end of wheat chromosome arm 6DL. The SSR markers Xcfd5 and Xcfd60 both flanked the locus at a distance of 22.1 cM to the distal end of 6DL. In future, these SSR markers can be used by wheat breeders and pathologists for marker assisted selection (MAS) of Lr38-mediated leaf rust resistance in wheat.     

Development of a kompetitive allele‐specific PCR marker for selection of the mutated Wx‐D1d allele in wheat breeding

Waxy (Wx) protein is a key enzyme for synthesis of amylose in endosperm. Amylose content in wheat grain influences the quality of end‐use products. Seven alleles have been described at the Wx‐D1 locus, but only two of them (Wx‐D1b, Wx‐D1e) were genotyped with codominant markers. The waxy wheat line K107Wx1 developed by treating ‘Kanto 107’ seeds with ethyl methanesulphonate carries the Wx‐D1d allele. However, no molecular basis supports this nomenclature. In the present study, DNA sequence analysis confirmed that a single nucleotide polymorphism in the sixth exon of Wx‐D1 changed tryptophan at position 301 into a termination codon. Based on this sequence variation, a PCR‐based KASP marker was developed to detect this point mutation using 68 BC₈F₁ plants and 297 BC₈F₂ lines derived from the cross ‘Ningmai 14’*9/K107Wx1. Combined with codominant markers for the Wx‐A1 and Wx‐B1 alleles, waxy and non‐waxy near‐isogenic lines were distinguished. The KASP marker was efficient in identifying the mutant allele and can be used to transfer waxiness to elite lines.     

The use of wheat/goatgrass introgression lines for the detection of gene(s) determining resistance to septoria tritici blotch (Mycosphaerella graminicola)

At the IPK Gatersleben a series of 85 bread wheat (T. aestivum)/goatgrass (Aegilops tauschii) introgression lines was developed recently. Based on the knowledge that chromosome 7D of this particular Ae. tauschii is a donor of resistance to septoria tritici blotch (Mycosphaerella graminicola), a sub-set of thirteen chromosome 7D introgression lines was investigated along with the susceptible recipient variety ‘Chinese Spring’ (CS) and the resistant donor line ‘CS (Syn 7D)’. The material was inoculated with two Argentinian isolates of the pathogen (IPO 92067 and IPO 93014) at both the seedlings (two leaf) and adult (tillering) stages at two locations over 2 years (2003, 2004). The resistance was effective against both isolates and at both developmental stages, and the resistance locus maps to the centromeric region of chromosome arm 7DS. On the basis of its relationship with the microsatellite marker Xgwm44, it is likely that the gene involved is Stb5. Stb5 is therefore apparently effective against M. graminicola isolates originating from both Europe and South America.     

Location of a Gene for Resistance to Eyespot (Pseudocercosparella herpotrichoides) on Chromosome 7D of Bread Wheat

Chromosome 7D of the wheat line VPM1 derived from a cross of Aegilops ventricosa with wheat confers resistance to the facultative fungal parasite Pseudocercosporella herpotrichoides. To determine the number of genes responsible fur this resistance, homozygous recombinant lines were developed from an F₁ between the wheat variety ‘Hobbit sib’ and a substitution line carrying chromosome 7D of VPM1 in a ‘Hobbit sib’ background. Resistance to Pseudocercosporella herpotrichoides is shown to be determined by a single gene located distally on the long arm of chromosome 7D. EpD1b, a unique allele of a gene encoding the readily detectable isoenzyme — endopeptidase, maps without recombination to Pch1 suggesting for two separate genes a maximum recombination value of 0.03 (P 0.05). Resistance to Pherpotrichoides could alter-natively be a product of Ep-D1b. Pch1 is also mapped against a gene for adult plant resistance to brown rust (Puccinia recondita), to Rc3 which confers coleoptile colour, and to α-Amy-D2, an isozyme that encodes α-amylase production.     

Preferential Transmission in Bread Wheat of a Chromosome Segment Derived from Thinopyrum distichum (Thunb.) Löve

A Thinopyrum distichum chromosome segment translocated on chromosome arm 7DL. of the line ‘Indis’, was shown to be preferentially transmitted in crosses with other bread wheats. The translocated segment carries a gene for leaf rust resistance and produces a null condition for the endopeptidase product, EP-Dla. These characters were used to follow the transmission of the translocated chromosome in segregating and testcross progenies derived by crossing ‘Indis’ to four bread wheat cultivars. The severity of the gametocidal response in the heterozygotes ranged from a virtually exclusive transfer of the translocation to an almost normal transmission of the homologues. In some genetic backgrounds an intermediate level of transmission occurred. In the F₁ with a gametocidal response, the transmission of the normal chromosome 7D was reduced in both sexes, but the reduction may be more severe in the male germline.     

Cytological and microsatellite mapping of mutant genes for spherical grain and compact spikes in durum wheat

Two mutants for sphaerococcoid seed (MA 16219) and compact spike (MA 17648) were isolated from M₃ progeny of durum wheat cultivar, Altaiskaya Niva, mutagenized with chemical mutagens. The chromosomal locations of the genes involved were determined by the use of a complete set of D-genome disomic substitutions in durum cultivar, Langdon. The gene for sphaerococcoid grain, s ¹⁶²¹⁹ , was allelic to S2, located in the centromeric region of chromosome 3B in hexaploid wheat. The gene for compact spike, C ¹⁷⁶⁴⁸ , was located on chromosome 5AL distal to the Q locus. Using microsatellite markers, C ¹⁷⁶⁴⁸ and awn inhibitor B1 were located in the F₂ of LD222 × MA17648. The gene order was Xbarc319—C ¹⁷⁶⁴⁸ —Xgwm179—Xgwm126—Xgwm291—B1.     

Molecular characterization of endosperm and amino acids modifications among quality protein maize inbreds

Modifier loci in QPM play a vital role in achieving acceptable degree of kernel hardness and accumulation of lysine and tryptophan. This study was undertaken to characterize a set of diverse QPM inbreds using SSRs linked to endosperm and amino acids modifier loci for their effective utilization in the breeding programme. Significant variation was observed for endosperm modification (25–100% opaqueness), tryptophan (0.056–0.111%) and lysine (0.223–0.444%). Generally, inbreds with soft endosperm possessed more tryptophan and lysine than inbreds with higher vitreousness. SSRs generated 341 alleles with two to seven alleles per locus. The frequency of unique and rare alleles was more for amino acid modifications, compared to endosperm modifications. Phylogenetic analyses grouped the inbreds into three major clusters, and the study identified suitable crosses for accumulation of endosperm and amino acids modifiers. QPM inbreds with desirable modifications identified here would serve as suitable donor for both opaque2 and modifier loci in the marker‐assisted backcross breeding. Further, contrasting inbreds can be used for generating mapping populations to identify new modifier loci underlying both endosperm and amino acids modifications.     

Characterization of a set of common wheat–Hordeum chilense chromosome 7Hch introgression lines and its potential use in research on grain quality traits

Chromosome 7H^ch from Hordeum chilense has potential for improving seed carotenoid content in wheat as it carries a Phytoene synthase 1 (Psy1) gene, which has a major role in this trait. Structural changes in chromosome 7H^ch were obtained in common wheat background by crossing the wheat disomic substitution line 7H^ch(7D) with a disomic addition line carrying chromosome 2C^c from Aegilops cylindrica in common wheat cv. ‘Chinese Spring’. Rearranged 7H^ch chromosomes were cytologically characterized by FISH. A set of 24 molecular markers and the Psy1 gene were used to identify the H. chilense chromosome segments involved in the introgressions. Six structural rearrangements of chromosome 7H^ch were identified. They included three homozygous wheat–H. chilense centromeric translocations, one involving the 7H^chS arm (T‐7H^chS·A/B) and two involving the 7H^chL arm (T1‐7H^chL·A/B and T2‐7H^chL·A/B). In addition, one 7H^chS arm deletion, one 7H^chL·7H^chL isochromosome and one 7H^chS telosome were obtained in hemizygous condition. These genetic stocks will be useful for studying the effect of chromosome 7H^ch on wheat flour colour.     

An approach to DNA polymorphism screening in <Emphasis Type="Italic">SBEIIa</Emphasis> homeologous genes of polyploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> L.)

The non-transgenic manipulation of starch properties in common wheat (Triticum aestivum L.) generally implies combining mutant alleles of the particular gene copies in all three subgenomes (A, B and D). The redundancy of the hexaploid wheat chromosome set substantially complicates the identification of recessive mutations and breeding. Nevertheless, naturally occurring or induced genetic polymorphism has already been successfully exploited for the production of waxy (GBSSI-deficient) and elevated amylose (SSIIa-deficient) wheats. However, in order to achieve the amylose content above 50% of wheat endosperm starch, it may be necessary to inactivate the starch branching enzyme (SBEIIa) isoforms, as the RNAi repression results and gene expression data strongly suggest. The identification of null SBEIIa alleles and their combination in a single genotype is therefore a promising approach to the production of non-transgenic high-amylose wheat; however, wheat SBEIIa polymorphism has not been characterized as of yet. In order to develop an approach to SBEIIa mutation screening, we sequenced the SBEIIa central region (exons 9–12) from the three subgenomes of common wheat cv. Chinese Spring and the A genome of diploid einkorn T. monococcum. The genome-specific primers were developed that amplify the exons downstream from intron 11 selectively from each homeologous gene. Using a single-stranded DNA conformation polymorphism (SSCP) approach, we screened 60 wheat cultivars, landraces, and rare species for naturally occurring SNPs in exons 12, 13 and 14 of the three SBEIIa homeologs. In total, 13 SNPs were discovered in the A and B wheat genomes. Two of these SNPs affect the amino acid sequences of SBEIIa isoforms and may change the enzyme functional properties. The presence of restriction site polymorphism at SNP positions enables their easy genotyping with CAPS assays. Our results indicate that the mining for naturally occurring sequence polymorphism in starch biosynthesis genes of wheat can be successfully performed at the DNA level, providing the starting point for a search for SBEIIa mutants at a larger scale.     

Transfer of the Glu-D1 locus encoding high molecular weight glutenin subunits 5+10 from breadwheat to diploid rye

A fragment of chromosome 1DL of breadwheat (Triticum aestivum L.) with the locus Glu-D1 encoding high molecular weight glutenin subunits 5+10 was translocated in hexaploid triticale (× Triticosecale Wittmack) to chromosome 1RL of rye (Secale cereale L.) where it replaced a corresponding fragment containing locus Sec-3 encoding rye secalins. The translocated chromosome 1R was transferred to diploid rye through backcrosses. During the transfer, at least two crossover events must have taken place that reduced the lengths of the 1DL inserts to about 5–8% of 1RL. These short inserts were selected on the basis of normal male transmission from heterozygotes and by low pairing with chromosome 1D in the F₁ hybrids with wheat, and tested by the in situ hybridization with total genomic DNA. While the wheat introgression in rye did not affect plant morphology or fertility, preliminary observations of the first population of homozygotes suggested that grain yield was lower, probably as a result of about 15% reduction of the 1000 kernel weight. The presence of a single wheat glutenin locus was insufficient to create rye with wheat-like breadmaking properties. However, relative to controls, the SDS-sedimentation value increased by about 75% and loaf volume was greater in test bakes using the procedure adapted for wheat-rye blends. Loaf volume for bread baked using the procedure for rye flour was not affected. Ryes with various glutenin subunits could be used in wheat-rye blends. This revised version was published online in July 2006 with corrections to the Cover Date.