Recombinant Substitution Lines Research Articles

A genetic system on chromosome arm 1BL of wild emmer causes distorted segregation in common wheat

Nonrandom segregation ratios of alleles 'segregation distortion' can have a striking impact on transmission genetics, and with widespread availability of genetic markers has been shown to be a frequent phenomenon. To investigate the possible effect of genetic interaction on segregation distortion and genetic map construction, the segregation and mapping of genetic markers locatedon wheat chromosomes 1A and 1B were followed in four recombinant substitution line (RSL) populations, produced using four chromosome-arm substitution lines (CASLs 1AS, 1AL, 1BS and 1BL) of wild emmer (Triticum turgidum var. dicoccoides, accession TTD140) in the background of the common wheat (T. aestivum) cultivar Bethlehem (BLH), each crossed to BLH itself. Using these four RSL populations, four genetic maps of chromosome 1 arms were constructed. A total of 22 genetic markers representing 19 loci were assigned to chromosome 1A, and 32 markers representing 30 loci were assigned to 1B. For chromosome 1B, two linkage maps were also constructed using RFLP data of an F2 population derived from the same cross combination as the RSLs. The RSL and F2 maps varied in genetic distances, but showed the same linear order of DNA markers. Segregation analysis revealed strong selection against BLH alleles on chromosome 1B, skewing the allelic frequency distribution in favour of TTD in both F2 and RSL populations at all marker loci. On the contrary, strong selection against TTD alleles on chromosome 1A was detected for some loci in the BLH × CASL1AL RSLs, and their distribution was significantly skewed to BLH. F2 populations always showed more segregation distortion than the corresponding RSLs. More markers near the region of chromosome 1B shared by both CASL1BS and 1BL (∼55 cM on chromosome 1B across the centromere) showed significantly distorted segregation in the BLH × CASL1BL population than in thecorresponding BLH × CASL1BS populations. Six markers located on chromosome 1A region shared by CASL1AS and 1AL showed significantly distorted segregation in 1AL-RSL, while no marker showed distorted segregation in 1AS-RSL. These results indicated that genetic factor(s) in the centromere region cause the distorted segregation of genetic markers on wheat chromosome 1B.

Chromosomal Location and Mapping of Quantitative Trait Locus Determining Technological Parameters of Grain and Flour in Strong-flour Bread Wheat Cultivar Saratovskaya 29

Bread wheat is the primary bread crop in the majority of countries in the world. The most important product that is manufactured from its grain and flour is yeast bread. In order to obtain an excellent bread, grain with high physical properties is needed for flour and dough. The Russian spring wheat cultivar Saratovskaya 29 is characterized by its exclusively high physical properties of flour and dough. The purpose of this work was to identify the chromosomes carrying the main loci for these traits in Saratovskaya 29 and to map them using recombinant substitution lines genotyped with molecular markers. A set of inter-varietal substitution lines Saratovskaya 29 (Yanetzkis Probat) was used to identify the “critical” chromosomes. The donor of individual chromosomes is a spring cultivar with average dough strength and tenacity. Substitution of 1D and 4D*7A chromosomes in the genetic background of Saratovskaya 29 resulted in a significant decrease in the physical properties of the dough. Such a deterioration in the case of 1D chromosome might be related to the variability of gluten protein composition. With the help of recombinant substitution double haploid lines obtained from a Saratovskaya 29 (Yanetzkis Probat 4D*7A) substitution line the region on the 4D chromosome was revealed in the strong-flour cultivar Saratovskaya 29, with the microsatellite locus Xgwm0165 to be associated with the unique physical properties of flour and dough. The detected locus is not related to the composition gluten proteins. These locus may be recommended to breeders for the selection of strong-flour cultivars. Additionally, a QTL associated with vitreousness of grain was mapped in the short arm of chromosome 7A.

Chromosome elimination of hexaploid common wheat mediated by interaction between Chinese Spring cytoplasm and a genetic factor(s) on chromosome arm 1BL of wild emmer

To create recombinant substitution lines from chromosome arm substitution lines (CASLs) of wild emmer, the common wheat cultivar ‘Bethlehem’ (BLH) was crossed with 4 CASLs of group 1 and the resulting F1s were backcrossed with corresponding BLH ditelosomic (DT) lines to produce monotelosomic plants (BC1F1, 2n = 6x = 41 + t). For each cross combination, about 100 BC1F2 plants derived from 47 to 49 BC1F1 plants were analyzed for their chromosome number and constitution. The frequencies of plants with 42 or 41 + t chromosomes in [DT1BS × (BLH × CASL1BL)] progenies were much lower than in crosses involving CASLs 1AS, 1AL and 1BS; while the frequency of [DT1BS × (BLH × CASL1BL)] progeny plants with 41 chromosomes was high. A total of 49 (BLH × CASL1BL) BC1F1 plants could be clearly classified into three types based on chromosome constitution of their 461 BC1F2 progenies. In type I (21 BC1F1 plants), 95 % of BC1F2 plants possessed 42 chromosomes (counting the telosomic chromosome as an entire chromosome). In type III (24 BC1F1 plants), most BC1F2 plants (75.6 %) had 41 chromosomes and no telosomic chromosome. In type II (4 BC1F1 plants), the expected number of ~48 % of BC1F2 plants had the telosomic chromosome, but about 70 % had fewer than the expected 42 chromosomes. All type III BC1F1 plants were confirmed to have experienced elimination of telosomic chromosome 1BS. Observation of BC1F1 seedling mitosis revealed that the elimination of 1BS occurred most probably during embryogenesis. However, type II BC1F1 plants experienced the elimination of different chromosomes, which was deduced to occur during meiosis. Reciprocal crosses indicated that both types of chromosome elimination occurred only when BLH DT1BS, which has Chinese Spring cytoplasm, was used as female parent. Collectively, these results reveal a genetic system on chromosome arm 1BL of wild emmer that interacts with the CS cytoplasm to cause chromosome elimination.

Genetic Mapping of Major‐Effect Seed Dormancy Quantitative Trait Loci on Chromosome 2B using Recombinant Substitution Lines in Tetraploid Wheat

ABSTRACTDurum wheat (Triticum turgidum L.) cultivars can benefit from having some level of seed dormancy to help reduce seed damage and lower grain quality caused by preharvest sprouting (PHS) occurring during wet harvesting conditions. Previously, a single chromosome substitution line carrying chromosome 2B of wild emmer [Triticum turgidum L. subsp. dicoccoides (Körn. ex Asch. & Graebn.) Thell.] in the durum cultivar Langdon background was found to have higher levels of seed dormancy and PHS tolerance. In this study, a population of recombinant substitution lines was developed and used to construct a single nucleotide polymorphism (SNP)‐based high‐density genetic linkage map. Seed germination tests were used to evaluate seed dormancy levels. Multiple interval mapping analysis revealed four quantitative trait loci (QTL) regions affecting seed dormancy. Two regions containing major‐effect QTL contributed by wild emmer were consistently expressed in four environments and explained 5.89 to 11.14% of the phenotypic variation. One QTL region was located near the centromere and the other on the long arm of chromosome 2B in Bayes credible intervals of 4 and 7 cM, respectively. Recombinant inbred lines (RILs) carrying both QTL had an average of 35% reduced rate of germination measured by weighted germination index compared with RILs carrying neither QTL. The two QTL regions identified in this study should be useful for improving PHS tolerance in wheat. Efforts to transfer the two QTL into elite durum cultivars are in progress to examine the effects of genetic background and environment on QTL expression and to evaluate the performance of other agronomic traits in the presence of the QTL.

Resistance to Oculimacula yallundae and Oculimacula acuformis is conferred by Pch2 in wheat

The recent report of a differential response of wheat lines containing the Pch2 gene to infection with the eyespot pathogens Oculimacula yallundae and O. acuformis has prompted this re‐examination of the response to these fungi by the recombinant lines used to map Pch2. Homozygous recombinant substitution lines (RSL) derived from the hybridization of Chinese Spring (CS) and the CS chromosome substitution line Cappelle Desprez 7A (CS/CD7A), previously evaluated for response to glucuronidase (GUS)‐transformed O. yallundae, were evaluated for response to infection with GUS‐transformed O. acuformis. Based on visual scores and on GUS expression level, which reflects fungal colonization of seedling plants, evidence of a quantitative trait locus (QTL) conferring resistance to O. acuformis was found in two separate growth chamber experiments (logarithm of the odds, LOD, = 2·7 and 6·7 at 305 and 289 cM, respectively) that was equivalent in location to that for resistance to O. yallundae (LOD = 13·2 and 11·4 at 289 and 304 cM, respectively). These results confirm that Pch2 confers some degree of resistance against both O. yallundae and O. acuformis under these conditions.

Development of consistently crossable wheat genotypes for alien wheat gene transfer through fine-mapping of the Kr1 locus

Breeders can force sexual hybridisation between wheat and related grass species to produce interspecific hybrids containing a dihaploid set of wheat and related chromosomes. This facilitates the introgression of desirable genes into wheat from the secondary gene pool. However, most elite European wheat varieties carry genes that suppress crossability, making the transfer of novel traits from exotic germplasm into elite wheat varieties difficult or impossible. Previous studies have identified at least five crossability loci in wheat. Here, the crossability locus with the largest effect, Kr1 on chromosome arm 5BL, was fine-mapped by developing a series of recombinant substitution lines in which the genome of the normally non-crossable wheat variety 'Hobbit sib' carries a recombinant 5BL chromosome arm containing segments from the crossable variety 'Chinese Spring'. These recombinant lines were scored for their ability to cross with rye over four seasons. Analysis revealed at least two regions on 5BL affecting crossability, including the Kr1 locus. However, the ability to set seed is highly dependent on prevailing environmental conditions. Typically, even crossable wheat lines exhibit little or no seed set when crossed with rye in winter, but show up to 90% seed set from similar crosses made in summer. By recombining different combinations of the two regions affecting crossability, wheat lines that consistently exhibit up to 50% seed set, whether crossed in the UK winter or summer conditions, were generated, thus creating a very important tool for increasing the efficiency of alien wheat transfer programmes.

Effects of chromosome 7B genes on grain protein concentration, yield and earliness in wheat (Triticum aestivumL.)

Grain protein concentration is a major factor in determining bread-making quality of wheat. We studied the effect of chromosome 7B of the high protein line F26–70 on protein content, yield and heading date, by testing 45 recombinant substitution lines derived from the cross Favorit//Favorit/F26-70(7B) in field trials, in 2005–2007. In 2006 and 2007 the lines were tested with and without nitrogen fertilization. Genes located on chromosome 7B of line F26–70 had significant effects on grain protein concentration and on heading date of recombinant substitution lines, but not on grain yield. In the conditions of our trials, protein content was not negatively associated with grain yield. Association of high protein content with earliness was significant in one year out of three. The interaction between RSLs and nitrogen fertilization was not significant, suggesting that the high protein gene(s) on chromosome 7B might be also useful at low nitrogen availability. Segregation of RSLs for grain protein concentration suggests a small number of genes, possibly one gene, responsible for the variation caused by chromosome 7B. We conclude that the high protein gene(s) on chromosome 7B might be potentially interesting for breeding.

Genetic mapping of a new flowering time gene on chromosome 3B of wheat

The single chromosome substitution lines of chromosome 3B of the Czech alternative wheat vari- ety Beska Plesivka (CP 3B) into two spring varieties Zlatka and Sandra, revealed clear diVerences in Xower- ing time compared to the recipient varieties. To map this gene(s), recombinant substitution lines for chromo- some 3B were produced from crosses of the substitu- tion lines with their recipient parents and genetic maps developed using SSR markers. Two populations were mapped, Sandra//Sandra 3B/Sandra (CP 3B) and Zlatka//Zlatka/Zlatka (CP 3B). Combining the geno- type data with phenotype data on Xowering time in Wve independent experiments under natural long day or controlled short day conditions revealed a single Xow- ering time QTL. This gene had an additive eVect of 1- 6 days, depending on environment and genetic back- ground, and was mapped in both populations to a position in the region of marker Xbarc164 near the centromere on the long arm of 3B. Comparisons of the genetic maps with other 3B maps developed by the authors indicated that the QTL may be homologous to a QTL segregating in UK germplasm.

Mapping quantitative trait loci for growth responses to exogenously applied stress induced hormones in wheat

Biotic and abiotic stresses are responsible for much of the variation in wheat production worldwide. The plant, in responding to stress events, increases the endogenous synthesis of hormones such as Abscisic Acid, Ethylene, Jasmonic Acid and Salicylic Acid. This paper was aimed at determining the location of plant-defence genes triggered by treatment with stress-induced hormones in a set of recombinant doubled haploid substitution lines involving variation for a single chromosome, 6A, previously shown to carry genes for aphid resistance. Eighty-three doubled haploid recombinant substitution (DHR) lines for chromosome 6A derived from the F1 of “Chinese Spring” (CS) × “Chinese Spring (Synthetic 6A)” (S6A) substitution line, were used as a mapping population. Plants of every genotype at the fully expanded 3rd leaf stage were subjected to four hormone treatments: ethylene (E), jasmonic acid (J), salicylic acid (S) and abscisic acid (A), together with an untreated control. The changes in a range of phenotypic traits were measured, aerial fresh weight (AFWC), aerial dry weight (ADWC), root fresh weight (RFWC), root dry weight (RDWC), foliar area (FAC). These were recorded twice: at the onset of the experiment before spraying with hormones and 72 h later. Nine QTLs were detected which were significantly associated with 6A molecular markers, explaining the variation for ADWC, RDWC and FAC. Most of the QTLs were associated with the interval between loci Xgwm459 and Xgwm334a, located in the telomeric region of the short arm of 6A. The dissection of complex agronomic traits such as tolerance to stress and QTLs related to exogenous hormone treatments could be used in marker-assisted selection for breeding wheat with tolerance to stresses.

Effect of rye Secale cereale L. chromosomes 1R and 3R on polyembryony expression in hybrid combinations between (Hordeum L.)-Triticum aestivum L. alloplasmic recombinant lines and wheat-rye substitution lines T. aestivum L.-S. cereale L.

The effect of rye chromosomes on polyembryony was studied for reciprocal hybrid combinations between (Hordeum vulgare L.)-Triticum aestivum L. alloplasmic recombinant lines and five wheat T. aestivum L. (cultivar Saratovskaya 29)-rye Secale cereale L. (cultivar Onokhosikaya) substitution lines: 1R(1D), 2R(2D), 3R(3B), 5R(5A), and 6R(6A), and for direct hybrid combinations between the [H. marinum ssp. gussoneanum (H. geniculatum All.)]-T. aestivum alloplasmic recombinant line and the wheat-rye substitution lines 1R(1A), 1R(1D), and 3R(3B). Chromosomes 1R and 3R of rye cultivar Onokhoiskaya proved to affect the expression of polyembryony in the hybrid combinations that involved the alloplasmic recombinant lines of common wheat as maternal genotypes. Based on this finding, polyembryony was regarded as a phenotypic expression of nuclear-cytoplasmic interactions where an important role is played by rye chromosomes 1R and 3R and the H. vulgare cytoplasm. Consideration is given to the association between the effect of rye chromosomes 1R and 3R on polyembryony in the [(Hordeum)-T. aestivum × wheat-rye substitution lines] hybrid combinations and their stimulating effect on the development on androgenic embryoids in isolated anther cultures of the wheat-rye substitution lines.

The high grain protein content gene Gpc-B1 accelerates senescence and has pleiotropic effects on protein content in wheat

High grain protein content (GPC) is a frequent target of wheat breeding programmes because of its positive effect on bread and pasta quality. A wild wheat allele at the Gpc-B1 locus with a significant impact on this trait was identified previously. The precise mapping of several senescence-related traits in a set of tetraploid recombinant substitution lines (RSLs) segregating for Gpc-B1 is reported here. Flag leaf chlorophyll degradation, change in peduncle colour, and spike water content were completely linked to the Gpc-B1 locus and to the differences in GPC within a 0.3 cM interval corresponding to a physical distance of only 250 kb. The effect of Gpc-B1 was also examined in different environments and genetic backgrounds using a set of tetraploid and hexaploid pairs of isogenic lines. The results were consistent with those observed in the RSLs. The high GPC allele conferred a shorter duration of grain fill due to earlier flag leaf senescence and increased GPC in all four genetic backgrounds. The effect on grain size was more variable, depending on the genotype-environment combinations. These results are consistent with a model in which the wild-type allele of Gpc-B1 accelerates senescence in flag leaves producing pleiotropic effects on nitrogen remobilization, total GPC, and grain size.

Fine Mapping of a Major Insect Resistance QTL in Soybean and its Interaction with Minor Resistance QTLs

ABSTRACTUtilization of native insect resistance genes can be an important component for managing insects in soybean [Glycine max (L.) Merr.]. A major quantitative trait locus (QTL‐M) for insect resistance from PI 229358, controlling antibiosis and antixenosis, was previously identified on linkage group (LG) M and was found to increase the effectiveness of a Bacillus thuringiensis (Bt) transgene in soybean. The objectives of this study were to fine‐map QTL‐M using recombinant substitution lines (RSLs) identified from a ‘Benning’ backcross population, and to evaluate the main effects and the epistatic interactions between QTL‐M and other resistance QTLs on LGs G and H using near‐isogenic lines (NILs) in a Benning genetic background. The effect of QTL‐M was still detectable in the Benning NILs when they were evaluated for resistance to corn earworm [CEW, Helicoverpa zea (Boddie)]. The two minor resistance QTLs only provided insect resistance when QTL‐M was also present in the Benning NILs. The QTL‐M was fine‐mapped to an approximately 0.52‐cM region after the first round of phenotyping the RSLs for resistance to CEW and soybean looper [SBL, Pseudoplusia includens (Walker)]. These results should increase the feasibility of cloning QTL‐M and help guide the development of insect resistant soybean cultivars.

Registration of Five Wheat Isogenic Lines for Leaf Rust and Stripe Rust Resistance Genes

Registration of Five Wheat Isogenic Lines for Leaf Rust and Stripe Rust Resistance Genes

High-temperature adult-plant (HTAP) stripe rust resistance gene Yr36 from Triticum turgidum ssp. dicoccoides is closely linked to the grain protein content locus Gpc-B1

Several new races of the stripe rust pathogen have become frequent throughout the wheat growing regions of the United States since 2000. These new races are virulent to most of the wheat seedling resistance genes limiting the resistance sources that can be used to combat this pathogen. High-temperature adult-plant (HTAP) stripe rust resistance has proven to be more durable than seedling resistance due to its non-race-specific nature, but its use is limited by the lack of mapping information. We report here the identification of a new HTAP resistance gene from Triticum turgidum ssp. dicoccoides (DIC) designated as Yr36. Lines carrying this gene were susceptible to almost all the stripe rust pathogen races tested at the seedling stage but showed adult-plant resistance to the prevalent races in California when tested at high diurnal temperatures. Isogenic lines for this gene were developed by six backcross generations. Field tests in two locations showed increased levels of field resistance to stripe rust and increased yields in isogenic lines carrying the Yr36 gene compared to those without the gene. Recombinant substitution lines of chromosome 6B from DIC in the isogenic background of durum cv. Langdon were used to map the Yr36 gene on the short arm of chromosome 6B completely linked to Xbarc101, and within a 2-cM interval defined by PCR-based markers Xucw71 and Xbarc136. Flanking locus Xucw71 is also closely linked to the grain protein content locus Gpc-B1 (0.3-cM). Marker-assisted selection strategies are presented to improve stripe rust resistance and simultaneously select for high or low Gpc-B1 alleles.

Nitrogen uptake and remobilization in tetraploid 'Langdon' durum wheat and a recombinant substitution line with the high grain protein gene Gpc-B1

Author(s): Kade, M; Barneix, AJ; Olmos, S; Dubcovsky, J | Abstract: Tetraploid wheat (Triticum turgidum L. var. durum) cv. 'Langdon' (LDN) and its near-isogenic recombinant substitution line no. 68 (RSL no. 68) carrying the high grain protein gene Gpc-B1 from emmer wheat, were compared in three greenhouse experiments to establish in which way Gpc-B1 increases grain protein concentration (GPC). At anthesis, RSL no. 68 had higher soluble protein and amino acids concentrations in the flag leaf than LDN. At maturity, both lines presented a similar above ground biomass and grain yield. However, RSL no. 68 showed a higher total N content in ears, grain and chaff than LDN; N harvest index (NHI) was also higher because of a lower straw N concentration and higher grain N concentration. When both lines were grown with a low N supply, and when N supply was interrupted before anthesis, similar trends were observed but the differences in GPC were smaller. It is concluded that RSL no. 68 accumulates a higher GPC than LDN mainly because of a more efficient N remobilization from the leaves to the ears during grain filling.

Mapping quantitative trait loci in wheat for resistance against greenbug and Russian wheat aphid

AbstractBreeding for genetic resistance against greenbug and Russian wheat aphid (RWA) is the most effective way of controlling these widespread pests in wheat. Earlier work had shown that chromosome 7D of a synthetic hexaploid wheat, ‘Synthetic’ (T. dicoccoides × Ae. squarrosa) (AABB × DD) gave resistance when transferred into the genetic background of an aphid‐susceptible cultivar, ‘Chinese Spring’, as the recipient. To map the genes involved, a set of 103 doubled haploid recombinant substitution lines was obtained from crossing the 7D substitution line with the recipient, and used to determine the number and chromosomal location of quantitative trait loci (QTL) controlling antixenosis and antibiosis types of resistance. Antixenosis to RWA was significantly associated with marker loci Xpsr687 on 7DS, and Xgwm437 on 7DL. Antibiosis to greenbug was associated with marker loci Xpsr490, Rc3 (on 7DS), Xgwm44, Xgwm111, Xgwm437, Xgwm121 and D67 (on 7DL). Similarly, antibiosis to RWA was linked to loci Xpsr490, Rc3, Xgwm44, Xgwm437 and Xgwm121. At least two QTL in repulsion phase, one close to the centromere either on the 7DS or 7DL arms, and a second distal on 7DL could explain antibiosis to RWA and, partially, this mechanism against greenbug.

New quantitative trait loci influencing grain texture and protein content in bread wheat

The object of this study was to discover new QTL controlling grain texture and protein quantity which could be of use for plant breeders for directed plant breeding by marker-assisted selection. A series of precise genetic stocks of wheat developed between the good bread-making quality parent, Avalon, and a biscuit wheat, Hobbit Sib, were used for this purpose. Analysis of a population of Hobbit Sib (Avalon 5A) recombinant substitution lines revealed two new year-on-year, consistent, putative (non-significant) QTL, one associated with grain protein content and one with grain texture. Analysis of Hobbit Sib (Avalon 5D) recombinant substitution lines revealed strong, statistically significant and consistent associations between the known hardness locus, Ha , and grain texture, and between a new QTL on the long arm, with grain protein content. Analysis of a population of Hobbit Sib x Avalon recombinant inbred lines revealed year-on-year consistent, statistically significant, new QTL for protein content on chromosomes 2B, 6B and 7A, and for texture on chromosome 1B.

New quantitative trait loci influencing grain texture and protein content in bread wheat

The object of this study was to discover new QTL controlling grain texture and protein quantity which could be of use for plant breeders for directed plant breeding by marker-assisted selection. A series of precise genetic stocks of wheat developed between the good bread-making quality parent, Avalon, and a biscuit wheat, Hobbit Sib, were used for this purpose. Analysis of a population of Hobbit Sib (Avalon 5A) recombinant substitution lines revealed two new year-on-year, consistent, putative (non-significant) QTL, one associated with grain protein content and one with grain texture. Analysis of Hobbit Sib (Avalon 5D) recombinant substitution lines revealed strong, statistically significant and consistent associations between the known hardness locus, Ha, and grain texture, and between a new QTL on the long arm, with grain protein content. Analysis of a population of Hobbit Sib x Avalon recombinant inbred lines revealed year-on-year consistent, statistically significant, new QTL for protein content on chromosomes 2B, 6B and 7A, and for texture on chromosome 1B.

Dwarfing effect associated with the threshability gene Q on wheat chromosome 5A

AbstractA dwarfing effect of the 44.1 cM chromosomal region between the threshability gene Q and Xfba068 on the long arm of hexaploid wheat chromosome 5A has been reported. To clarify whether Q or its adjacent region is responsible for regulating culm elongation, two precise genetic stocks of near‐isogenic lines (NIL), a single chromosome substitution line, ‘Chinese Spring’ (CS; ‘Cappelle‐Desprez’ 5A) (NIL‐Q) and a single chromosome recombinant substitution line (NIL‐q) were used. The target segment of NIL‐q included the q allele and QEet.ocs‐5A.1, an earlinessper se gene, from spelt wheat in the CS genetic background. They were grown under 16‐h day length with and/ or without vernalization treatment. Being independent of heading date, NIL‐Q showed shorter elongation in lower internodes and decreased internode differentiation in comparison with NIL‐q. The culm‐length reduction associated with Q was confirmed in the recombinant F5 population derived from the cross between NIL‐Q and NIL‐q. Vernalization promotion had a tendency to reduce this dwarfing effect.

Precise mapping of a locus affecting grain protein content in durum wheat.

Grain protein content (GPC) is an important factor in pasta and breadmaking quality, and in human nutrition. It is also an important trait for wheat growers because premium prices are frequently paid for wheat with high GPC. A promising source for alleles to increase GPC was detected on chromosome 6B of Triticum turgidum var. dicoccoides accession FA-15-3 (DIC). Two previous quantitative trait locus (QTL) studies found that the positive effect of DIC-6B was associated to a single locus located between the centromere and the Nor-B2 locus on the short arm of chromosome 6B. Microsatellite markers Xgwm508 and Xgwm193 flanking the QTL region were used in this study to develop 20 new homozygous recombinant substitution lines (RSLs) with crossovers between these markers. These 20 RSLs, plus nine RSLs developed in previous studies were characterized with four new RFLP markers located within this chromosome segment. Grain protein content was determined in three field experiments organized as randomized complete block designs with ten replications each. The QTL peaks for protein content were located in the central region of a 2.7-cM interval between RFLP markers Xcdo365 and Xucw67 in the three experiments. Statistical analyses showed that almost all lines could be classified unequivocally within low- and high- protein groups, facilitating the mapping of this trait as a single Mendelian locus designated Gpc-6B1. The Gpc-6B1 locus was mapped 1.5-cM proximal to Xcdo365 and 1.2-cM distal to Xucw67. These new markers can be used to reduce the size of the DIC chromosome segment selected in marker-assisted selection programs. Markers Nor-B2 and Xucw66 flanking the previous two markers can be used to select against the DIC segment and reduce the linkage drag during the transfer of Gpc-6B1 into commercial bread and pasta wheat varieties. The precise mapping of the high GPC gene, the high frequency of recombinants recovered in the targeted region, and the recent development of a tetraploid BAC library including the Gpc-6B1 DIC allele are the first steps towards the map-based cloning of this gene.