Cereal cyst nematode resistance gene <Emphasis Type="Italic">CreV</Emphasis> effective against <Emphasis Type="Italic">Heterodera filipjevi</Emphasis> transferred from chromosome 6VL of <Emphasis Type="Italic">Dasypyrum villosum</Emphasis> to bread wheat

Cereal cyst nematodes (CCN) are a global economic problem for cereal production. Heterodera filipjevi is one of the most commonly identified and widespread CCN species found in many wheat production regions of the world. Transferring novel genes for resistance to H. filipjevi from wild relatives of wheat is a promising strategy for protection of wheat crops. A set of wheat–Dasypyrum villosum chromosome addition lines, T6V#4S·6AL translocation lines and their donor parental lines were tested for their response to the nematode. D. villosum and wheat–D. villosum disomic addition line DA6V#4 were resistant. As T6V#4S·6AL translocation lines were susceptible, resistance was presumed to be located on chromosome 6V#4L. The objective of this study was to produce and characterize wheat–6V#4L translocations and confirm the chromosome location of the resistance. Introgression lines T6V#4L·6AS, T6V#4L-4BL·4BS and DT6V#4L were developed and subjected to molecular cytogenetic analysis. These and four additional wheat–6V#4 introgression lines were tested for response to H. filipjevi in the greenhouse. The results indicated that introgression lines DA6V#4, T6V#4L·6AS, T6V#4L-4BL·4BS, T6V#4L·6V#4S-7BS and DT6VL#4 had higher levels of H. filipjevi resistance than their recurrent parent. However, Del6V#4L-1 and translocation line T6V#4S·6AL were equally susceptible to wheat cv. Chinese Spring. The CCN resistance gene, temporarily named CreV, was therefore physically mapped to chromosome arm 6V#4L FL 0.80–1.00. Translocation chromosomes T6V#4L·6AS transferred to a modern wheat cv. Aikang 58 with its co-dominant molecular markers could be utilized as a novel germplasm for CCN resistance breeding in wheat.     

QTL mapping of Fusarium head blight resistance in three related durum wheat populations

Key message

The QTL Fhb1 was successfully introgressed and validated in three durum wheat populations. The novel germplasm and the QTL detected will support improvement of Fusarium resistance in durum wheat.

Abstract

Durum wheat (Triticum durum Desf.) is particularly susceptible to Fusarium head blight (FHB) and breeding for resistance is hampered by limited genetic variation within this species. To date, resistant sources are mainly available in a few wild relative tetraploid wheat accessions. In this study, the effect of the well-known hexaploid wheat (Triticum aestivum L.) quantitative trait locus (QTL) Fhb1 was assessed for the first time in durum wheat. Three F7-RIL mapping populations of about 100 lines were developed from crosses between the durum wheat experimental line DBC-480, which carries an Fhb1 introgression from Sumai-3, and the European T. durum cultivars Karur, Durobonus and SZD1029K. The RILs were evaluated in field experiments for FHB resistance in three seasons using spray inoculation and genotyped with SSR as well as genotyping-by-sequencing markers. QTL associated with FHB resistance were identified on chromosome arms 2BL, 3BS, 4AL, 4BS, 5AL and 6AS at which the resistant parent DBC-480 contributed the positive alleles. The QTL on 3BS was detected in all three populations centered at the Fhb1 interval. The Rht-B1 locus governing plant height was found to have a strong effect in modulating FHB severity in all populations. The negative effect of the semi-dwarf allele Rht-B1b on FHB resistance was compensated by combining with Fhb1 and additional resistance QTL. The successful deployment of Fhb1 in T. durum was further substantiated by assessing type 2 resistance in one population. The efficient introgression of Fhb1 represents a significant step forward for enhancing FHB resistance in durum wheat.

     

Novel sources of resistance to Septoria nodorum blotch in the Vavilov wheat collection identified by genome-wide association studies

Key message

The fungus Parastagonospora nodorum causes Septoria nodorum blotch (SNB) of wheat. A genetically diverse wheat panel was used to dissect the complexity of SNB and identify novel sources of resistance.

Abstract

The fungus Parastagonospora nodorum is the causal agent of Septoria nodorum blotch (SNB) of wheat. The pathosystem is mediated by multiple fungal necrotrophic effector–host sensitivity gene interactions that include SnToxA–Tsn1, SnTox1–Snn1, and SnTox3–Snn3. A P. nodorum strain lacking SnToxA, SnTox1, and SnTox3 (toxa13) retained wild-type-like ability to infect some modern wheat cultivars, suggesting evidence of other effector-mediated susceptibility gene interactions or the lack of host resistance genes. To identify genomic regions harbouring such loci, we examined a panel of 295 historic wheat accessions from the N. I. Vavilov Institute of Plant Genetic Resources in Russia, which is comprised of genetically diverse landraces and breeding lines registered from 1920 to 1990. The wheat panel was subjected to effector bioassays, infection with P. nodorum wild type (SN15) and toxa13. In general, SN15 was more virulent than toxa13. Insensitivity to all three effectors contributed significantly to resistance against SN15, but not toxa13. Genome-wide association studies using phenotypes from SN15 infection detected quantitative trait loci (QTL) on chromosomes 1BS (Snn1), 2DS, 5AS, 5BS (Snn3), 3AL, 4AL, 4BS, and 7AS. For toxa13 infection, a QTL was detected on 5AS (similar to SN15), plus two additional QTL on 2DL and 7DL. Analysis of resistance phenotypes indicated that plant breeders may have inadvertently selected for effector insensitivity from 1940 onwards. We identify accessions that can be used to develop bi-parental mapping populations to characterise resistance-associated alleles for subsequent introgression into modern bread wheat to minimise the impact of SNB.

     

Development of V chromosome alterations and physical mapping of molecular markers specific to <Emphasis Type="Italic">Dasypyrum villosum</Emphasis>

Wheat-Dasypyrum villosum translocations were induced in the progeny of the amphiploid Triticum durum-D. villosum (AABBVV) by pollen irradiation. The rearranged V genome chromosomes were characterized by genomic/fluorescence in situ hybridization (GISH/FISH) and molecular markers. Twenty wheat-D. villosum translocation chromosomes were selected, including four centric, seven large segments, and nine small segments in a Chinese Spring (CS) background. The four centric translocations were subsequently identified by GISH/FISH and by molecular markers specific to chromosome arms of the Triticeae linkage groups. They were T5DL.4VL, T4BL.7VS, and T4BS.7VL as well as the compensating translocation T7AL.7VS. Using a combination of previously developed V chromosome alterations, 52 translocations or deletions that divided V chromosomes into 42 bins were employed for deletion mapping of molecular markers specific to D. villosum in a wheat background. Ninety-five expressed sequence tag (EST)-sequence-tagged site (STS) and seven SSR markers that were previously reported, as well as 72 STS markers screened in the present study, were physically allocated into 37 of 42 chromosome bins of D. villosum. Multiple loci of EST-STS markers were also mapped using CS nullisomic tetrasomic (NT) and ditelosomic (DT) genetic stocks. Most EST-STS homoeoloci were located on homoeologous chromosomes, suggesting a high degree of homology between the genomes of D. villosum and wheat. Four 4VL-specific markers detected homoeoloci on group 7 chromosomes of wheat, indicating that chromosome 4V of D. villosum shows some affinity to both wheat homoeologous groups 4 and 7. This is the first physical map of D. villosum, which will provide insight into the V genome for molecular breeding.     

Structural chromosome rearrangements and polymorphisms identified in Chinese wheat cultivars by high-resolution multiplex oligonucleotide FISH

Key message

High-resolution multiplex oligonucleotide FISH revealed the frequent occurrence of structural chromosomal rearrangements and polymorphisms in widely grown wheat cultivars and their founders.

Abstract

Over 2000 wheat cultivars including 19 founders were released and grown in China from 1949 to 2000. To understand the impact of breeding selection on chromosome structural variations, high-resolution karyotypes of Chinese Spring (CS) and 373 Chinese cultivars were developed and compared by FISH (fluorescence in situ hybridization) using an oligonucleotide multiplex probe based on repeat sequences. Among them, 148 (39.7%) accessions carried 14 structural rearrangements including three single translocations (designated as T), eight reciprocal translocations (RT), one pericentric inversion (perInv), and two combined variations having both the deletion and single translocations. Five rearrangements were traced to eight founders, including perInv 6B detected in 57 cultivars originating from Funo, Abbondanza, and Fan 6, T 1RS?1BL in 47 cultivars derived from the Lovrin series, RT 4AS?4AL-1DS/1DL?1DS-4AL in 31 varieties from Mazhamai and Bima 4, RT 1RS?7DL/7DS?1BL in three cultivars was from Aimengniu, and RT 5BS?5BL-5DL/5DS?5DL-5BL was only detected in Youzimai. In addition to structural rearrangements, 167 polymorphic chromosome blocks (defined as unique signal patterns of oligonucleotide repeat probes distributed within chromosomes) were identified, and 59 were present in one or more founders. Some specific types were present at high frequencies indicating selective blocks in Chinese wheat varieties. All cultivars and CS were clustered into four groups and 15 subgroups at chromosome level. Common block patterns occurred in the same subgroup. Origin, geographic distribution, probable adaptation to specific environments, and potential use of these chromosomal rearrangements and blocks are discussed.

     

The genetic architecture of seedling resistance to Septoria tritici blotch in the winter wheat doubled-haploid population Solitär × Mazurka

Breeding for resistance to Septoria tritici blotch (STB), caused by Mycosphaerella graminicola (anamorph: Septoria tritici), is an essential component in controlling this important foliar disease of wheat. Inheritance of seedling resistance to seven worldwide pathogen isolates has been studied in a doubled-haploid (DH) population derived from a cross between the field resistant cultivar Solitär and the susceptible cultivar Mazurka. Multiple quantitative trait locus (QTL) mapping revealed major and minor genetic effects on resistance as well as several epistatic relationships in the seedling stage. Solitär conferred resistance to isolate IPO323, governed by Stb6 on chromosome 3A, as well as to IPO99015, IPO92034, Hu1 and Hu2 controlled by a QTL on chromosome arm 1BS, possibly corresponding to Stb11 and minor QTL on chromosomes 1B, 3D, 6B and 7D. Resistance of Mazurka to IPO90015 and BBA22 was caused by a QTL located in a region on 4AL which harbours Stb7 or Stb12. QTL specific to pycnidial coverage on 3B and specific to necrosis on 1A could be discovered for isolate IPO92034. Pairwise epistatic interactions were reliably detected with five isolates. Although their contributions to the total variance are generally low, the genotypic effect of the QTL by QTL interaction of 4AL (Stb7 or Stb12) and 3AS (Stb6) made up almost 15% of disease expression. Altogether, the results suggest a complex inheritance of resistance to STB in the seedling stage in terms of isolate-specificity and resistance mechanisms, which have implications for marker-assisted breeding in an attempt to pyramid STB resistance genes.     

Characterization and genome-wide association mapping of resistance to leaf rust,stem rust and stripe rust in a geographically diverse collection of spring wheat landraces

The challenge posed by rapidly changing wheat rust pathogens, both in virulence and in environmental adaptation, calls for the development and application of new techniques to accelerate the process of breeding for durable resistance. To expand the resistance gene pool available for germplasm improvement, a panel of 159 landraces plus old cultivars was evaluated for seedling and adult plant resistance (APR) to over 35 Australian pathotypes of Puccinia triticina, Puccinia graminis f. sp. tritici, and Puccinia striiformis f. sp. tritici. Known seedling resistance (SR) genes for leaf rust (Lr2a, Lr3a, Lr13, Lr23, Lr16, and Lr20), stem rust (Sr12, Sr13, Sr23, Sr30, and Sr36), and stripe rust (Yr3, Yr4, Yr5, Yr9, Yr10, Yr17, and Yr27) were postulated. The APR genes identified via field assessments and marker analyses included the pleiotropic genes (Lr34/Yr18/Sr57, Lr46/Yr29/Sr58, Lr67/Yr46/Sr55, and Sr2/Lr27/Yr30), Lr68, Lr74, and uncharacterized APR. A genome-wide association analysis using linear mixed models detected 79 single nucleotide polymorphism (SNP) markers significantly associated with rust resistance, which were mapped on chromosomes 1A, 1B, 1D, 2A, 2B, 3A, 3B, 3D, 4A, 5A, 5B, 6A, 6B, 6D, 7A, 7B and 7D. SNPs associated with multiple rust resistances probably indicate the presence of new pleiotropic or closely linked genes. SNPs were mapped on chromosome positions (1AL, 1DS, 2AL, 4AS, 5BS, 6DL, and 7AL) that have not been known to carry APR genes. This study revealed the presence of a range of possibly unidentified effective seedling and APRs among the landraces, which might represent new sources of rust resistance for the ongoing effort to develop improved wheat cultivars.     

Identification,mapping, and marker development of stem rust resistance genes in durum wheat ‘Lebsock’

Wheat production in many wheat-growing regions is vulnerable to stem rust, caused by Puccinia graminis f. sp. tritici (Pgt). Several previous studies showed that most of the durum cultivars adapted to the upper Great Plains in the USA have good resistance to the major Pgt pathotypes, including the Ug99 race group. To identify the stem rust resistance (Sr) genes in the durum cultivar ‘Lebsock’, a tetraploid doubled haploid (DH) population derived from a cross between Lebsock and Triticum turgidum ssp. carthlicum PI 94749 was screened with the Pgt races TTKSK, TRTTF, and TTTTF. The stem rust data and the genotypic data previously developed were used to identify quantitative trait loci (QTL) associated with resistance. We identified one QTL each on chromosome arms 4AL, 6AS, 6AL, and 2BL. Based on marker and race-specification analysis, we postulated that the QTL on 4AL, 6AS, 6AL, and 2BL correspond to Sr7a, Sr8155B1, Sr13, and likely Sr9e, respectively. The results indicated that most of the US durum germplasm adapted to the upper Great Plains likely harbors the four major Sr genes characterized in this study. Among these genes, Sr8155B1 was recently identified and shown to be unique in that it conferred susceptibility to TTKSK but resistance to variant race TTKST. Two, three, and one thermal asymmetric reverse PCR (STARP) markers were developed for Sr7a, Sr8155-B1, and Sr13, respectively. Knowledge of the Sr genes in durum germplasm and the new STARP markers will be useful to pyramid and deploy multiple Sr genes in future durum and wheat cultivars.     

<Emphasis Type="Italic">Pm62</Emphasis>, an adult-plant powdery mildew resistance gene introgressed from <Emphasis Type="Italic">Dasypyrum villosum</Emphasis> chromosome arm 2VL into wheat

Key message

Pm62, a novel adult-plant resistance (APR) gene against powdery mildew, was transferred from D. villosum into common wheat in the form of Robertsonian translocation T2BS.2VL#5.

Abstract

Powdery mildew, which is caused by the fungus Blumeria graminis f. sp. tritici, is a major disease of wheat resulting in substantial yield and quality losses in many wheat production regions of the world. Introgression of resistance from wild species into common wheat has application for controlling this disease. A Triticum durum-Dasypyrum villosum chromosome 2V#5 disomic addition line, N59B-1 (2n?=?30), improved resistance to powdery mildew at the adult-plant stage, which was attributable to chromosome 2V#5. To transfer this resistance into bread wheat, a total of 298 BC₁F₁ plants derived from the crossing between N59B-1 and Chinese Spring were screened by combined genomic in situ hybridization and fluorescent in situ hybridization, 2V-specific marker analysis, and reaction to powdery mildew to confirm that a dominant adult-plant resistance gene, designated as Pm62, was located on chromosome 2VL#5. Subsequently, the 2VL#5 (2D) disomic substitution line (NAU1825) and the homozygous T2BS.2VL#5 Robertsonian translocation line (NAU1823), with normal plant vigor and full fertility, were identified by molecular and cytogenetic analyses of the BC₁F₂ generation. The effects of the T2BS.2VL#5 recombinant chromosome on agronomic traits were also evaluated in the F₂ segregation population. The results suggest that the translocated chromosome may have no distinct effect on plant height, 1000-kernel weight or flowering period, but a slight effect on spike length and seeds per spike. The translocation line NAU1823 has being utilized as a novel germplasm in breeding for powdery mildew resistance, and the effects of the T2BS.2VL#5 recombinant chromosome on yield-related and flour quality characters will be further assessed.

     

Genome-wide association mapping for resistance to leaf rust,stripe rust and tan spot in wheat reveals potential candidate genes

Key message

Genome-wide association mapping in conjunction with population sequencing map and Ensembl plants was used to identify markers/candidate genes linked to leaf rust, stripe rust and tan spot resistance in wheat.

Abstract

Leaf rust (LR), stripe rust (YR) and tan spot (TS) are some of the important foliar diseases in wheat (Triticum aestivum L.). To identify candidate resistance genes for these diseases in CIMMYT’s (International Maize and Wheat Improvement Center) International bread wheat screening nurseries, we used genome-wide association studies (GWAS) in conjunction with information from the population sequencing map and Ensembl plants. Wheat entries were genotyped using genotyping-by-sequencing and phenotyped in replicated trials. Using a mixed linear model, we observed that seedling resistance to LR was associated with 12 markers on chromosomes 1DS, 2AS, 2BL, 3B, 4AL, 6AS and 6AL, and seedling resistance to TS was associated with 14 markers on chromosomes 1AS, 2AL, 2BL, 3AS, 3AL, 3B, 6AS and 6AL. Seedling and adult plant resistance (APR) to YR were associated with several markers at the distal end of chromosome 2AS. In addition, YR APR was also associated with markers on chromosomes 2DL, 3B and 7DS. The potential candidate genes for these diseases included several resistance genes, receptor-like serine/threonine-protein kinases and defense-related enzymes. However, extensive LD in wheat that decays at about 5?×?10⁷ bps, poses a huge challenge for delineating candidate gene intervals and candidates should be further mapped, functionally characterized and validated. We also explored a segment on chromosome 2AS associated with multiple disease resistance and identified seventeen disease resistance linked genes. We conclude that identifying candidate genes linked to significant markers in GWAS is feasible in wheat, thus creating opportunities for accelerating molecular breeding.

     

An intercalary translocation from <Emphasis Type="Italic">Agropyron</Emphasis><Emphasis Type="Italic">cristatum</Emphasis> 6P chromosome into common wheat confers enhanced kernel number per spike

Main conclusion

This study explored 6P chromosomal translocations in wheat, and determined the effects of 6P intercalary chromosome segments on kernel number per wheat spike. Exploiting and utilising gene(s) from wild relative species has become an essential strategy for wheat crop improvement. In the translocation line Pubing2978, the intercalary 6P chromosome segment from Agropyron cristatum (L.) Gaertn. (2n = 4x = 28, PPPP) carried valuable multi-kernel gene(s) and was selected from the offspring of the common wheat plant Fukuho and the irradiated wheat-A. cristatum 6P disomic substitution line 4844-8. Genomic in situ hybridisation (GISH), dual-colour fluorescence in situ hybridisation (FISH), and molecular markers were used to detect the small segmental 6P chromosome in the wheat background and its translocation breakpoint. Cytological studies demonstrated that Pubing2978 was a T1AS-6PL-1AS·1AL intercalary translocation with 42 chromosomes. The breakpoint was located near the centromeric region on the wheat chromosome 1AS and was flanked by the markers SSR12 and SSR283 based on an F₂ linkage map. The genotypic data, combined with the phenotypic information, implied that A. cristatum 6P chromosomal segment plays an important role in regulating the kernel number per spike (KPS). By comparison, the mean value of KPS in plants with translocations was approximately 10 higher than that in plants without translocations in three segregated populations. Moreover, the improvement in KPS was likely achieved by increasing both the spikelet number per spike (SNS) and the kernel number per spikelet. These excellent agronomic traits laid the foundation for further investigation of valuable genes and make the Pubing2978 line a promising germplasm for wheat breeding.

     

<Emphasis Type="Italic">Pm61</Emphasis>: a recessive gene for resistance to powdery mildew in wheat landrace Xuxusanyuehuang identified by comparative genomics analysis

Key message

A single recessive powdery mildew resistance gene Pm61 from wheat landrace Xuxusanyuehuang was mapped within a 0.46-cM genetic interval spanning a 1.3-Mb interval of the genomic region of chromosome arm 4AL.

Abstract

Epidemics of powdery mildew incited by the biotrophic fungus Blumeria graminis f. sp. tritici (Bgt) have caused significant yield reductions in many wheat (Triticum aestivum)-producing regions. Identification of powdery mildew resistance genes is required for sustainable improvement of wheat for disease resistance. Chinese wheat landrace Xuxusanyuehuang was resistant to several Bgt isolates at the seedling stage. Genetic analysis based on the inoculation of Bgt isolate E09 on the F₁, F₂, and F_2:3 populations produced by crossing Xuxusanyuehuang to susceptible cultivar Mingxian 169 revealed that the resistance of Xuxusanyuehuang was controlled by a single recessive gene. Bulked segregant analysis and simple sequence repeat (SSR) mapping placed the gene on chromosome bin 4AL-4-0.80-1.00. Comparative genomics analysis was performed to detect the collinear genomic regions of Brachypodium distachyon, rice, sorghum, Aegilops tauschii, T. urartu, and T. turgidum ssp. dicoccoides. Based on the use of 454 contig sequences and the International Wheat Genome Sequence Consortium survey sequence of Chinese Spring wheat, four EST-SSR and seven SSR markers were linked to the gene. An F₅ recombinant inbred line population derived from Xuxusanyuehuang?×?Mingxian 169 cross was used to develop the genetic linkage map. The gene was localized in a 0.46-cM genetic interval between Xgwm160 and Xicsx79 corresponding to 1.3-Mb interval of the genomic region in wheat genome. This is a new locus for powdery mildew resistance on chromosome arm 4AL and is designated Pm61.

     

Combination of all-stage and high-temperature adult-plant resistance QTL confers high-level,durable resistance to stripe rust in winter wheat cultivar Madsen

Key message

Wheat cultivar Madsen has a new gene on the short arm of chromosome 1A and two QTL for all-stage resistance and three QTL for high-temperature adult-plant resistance that in combination confer high-level, durable resistance to stripe rust.

Abstract

Wheat cultivar Madsen has maintained a high-level resistance to stripe rust over 30 years. To map quantitative trait loci (QTL) underlying the high-level, durable resistance, 156 recombinant inbred lines (RILs) developed from cross Avocet S?×?Madsen were phenotyped with selected races of Puccinia striiformis f. sp. tritici in the greenhouse seedling tests, and in naturally infected fields during 2015–2017. The RILs were genotyped by SSR and SNP markers from genotyping by sequencing and the 90 K wheat SNP chip. Three QTL for all-stage resistance were mapped on chromosomes 1AS, 1BS and 2AS, and two QTL for high-temperature adult-plant (HTAP) resistance were mapped on 3BS and 6BS. The most effective QTL on 2AS, explaining 8.97–23.10% of the phenotypic variation in seedling tests and 8.60–71.23% in field tests, contained Yr17 for all-stage resistance and an additional gene for HTAP resistance. The 6BS QTL, detected in all field tests, was identified as Yr78. The 1AS QTL, conferring all-stage resistance, was identified as a new gene, which explained 20.45 and 30.23% of variation in resistance to races PSTv-37 and PSTv-40, respectively, and contributed significantly to field resistance at Pullman in 2015-2017, but was not detected at Mount Vernon. The interactions among QTL were mostly additive, and RILs with all five QTL had the highest level of resistance in the field, similar to Madsen. Genotyping 148 US Pacific Northwest wheat cultivars with markers for the 1AS, 2AS and 6BS QTL validated the genes and markers, and indicated their usefulness for marker-assisted selection.

     

A cytogenetic ladder-map of the wheat homoeologous group-4 chromosomes

We report the results of chromosome maps of wheat homoeologous chromosomes 4A, 4B, and 4D using 40 RFLP markers and 39 homozygous deletion lines. Deletion breakpoints divide the chromosomes into 45 subarm intervals with 32 intervals distinguished by molecular markers. The chromosome maps confirm the homoeology of arms 4AS to 4BL and 4DL, and 4AL to 4BS and 4DS. The chromosome map of 4A reveals novel information concerning the 4AL-5AL-7BS cyclical translocation. The presence of homoeologous group-4 long-arm markers, Xksu G10 and Xpsr 1051, intervening between the translocated 5AL and 7BS chromosome segments in 4AL suggests that the translocation events are more complex than was earlier believed. Chromosome maps confirm a pericentric inversion in Chinese Spring chromosome 4B. The consensus chromosome map is compared to the genetic map of wheat to construct a cytogenetic ladder-map (CLM). The CLM reveals an unequal distribution of recombination along the length of the chromosome arms. Recombination is highest in the distal half, and low in the proximal half, of the chromosome arms.     

QTL mapping of pre-harvest sprouting resistance in a white wheat cultivar Danby

Key message

One major and three minor QTLs for resistance to pre-harvest sprouting (PHS) were identified from a white wheat variety “Danby.” The major QTL on chromosome 3A is TaPHS1, and the sequence variation in its promoter region was responsible for the PHS resistance. Additive?×?additive effects were detected between two minor QTLs on chromosomes 3B and 5A, which can greatly enhance the PHS resistance.

Abstract

Pre-harvest sprouting (PHS) causes significant losses in yield and quality in wheat. White wheat is usually more susceptible to PHS than red wheat. Therefore, the use of none grain color-related PHS resistance quantitative trait loci (QTLs) is essential for the improvement in PHS resistance in white wheat. To identify PHS resistance QTLs in the white wheat cultivar “Danby” and determine their effects, a doubled haploid population derived from a cross of Danby?×?“Tiger” was genotyped using genotyping-by-sequencing markers and phenotyped for PHS resistance in two greenhouse and one field experiments. One major QTL corresponding to a previously cloned gene, TaPHS1, was consistently detected on the chromosome arm 3AS in all three experiments and explained 21.6–41.0% of the phenotypic variations. A SNP (SNP?222) in the promoter of TaPHS1 co-segregated with PHS in this mapping population and was also significantly associated with PHS in an association panel. Gene sequence comparison and gene expression analysis further confirmed that SNP?222 is most likely the causal mutation in TaPHS1 for PHS resistance in Danby in this study. In addition, two stable minor QTLs on chromosome arms 3BS and 5AL were detected in two experiments with allele effects consistently contributed by Danby, while one minor QTL on 2AS was detected in two environments with contradicted allelic effects. The two stable minor QTLs showed significant additive?×?additive effects. The results demonstrated that pyramiding those three QTLs using breeder-friendly KASP markers developed in this study could greatly improve PHS resistance in white wheat.

     

Cloning of <Emphasis Type="Italic">TaTPP-6AL1</Emphasis> associated with grain weight in bread wheat and development of functional marker

Trehalose 6-phosphate phosphatase (TPP) dephosphorylates trehalose 6-phosphate to trehalose, an important growth regulator, and is involved in starch accumulation and grain yield. In this study, wheat TPP homologs were isolated from chromosomes 6AL, 6BL, and 6DL, designated as TaTPP-6AL1, TaTPP-6BL1, and TaTPP-6DL1, respectively. Sequence alignment showed a single-nucleotide polymorphism (SNP) at TaTPP-6AL1 locus between cultivars with contrasting thousand grain weight (TGW), forming alleles TaTPP-6AL1a and TaTPP-6AL1b, respectively. A cleaved amplified polymorphic sequence (CAPS) marker, TaTPP-6AL1-CAPS, was developed to differentiate the two alleles. TaTPP-6AL1 was mapped within the interval of IWB65749 and IWB60449 in a recombinant inbred line (RIL) population derived from Zhou8425B/Chinese Spring using the wheat 90K SNP assay. A QTL for TGW identified in the interval explained 12.1–19.1% of the phenotypic variance across five environments. Association analysis on 141 Chinese wheat cultivars also indicated a significant correlation of TaTPP-6AL1 with TGW. In conclusion, TaTPP-6AL1 and its functional marker are valuable to improve grain yield in wheat breeding.     

Identification and mapping of resistance to stem rust in the European winter wheat cultivars Spark and Rialto

A mapping population of 126 doubled haploid (DH) lines derived from a cross between the English winter wheat cultivars Spark and Rialto was evaluated for response to Puccinia graminis f. sp. tritici in the greenhouse and in artificially inoculated field plots at two locations over 3 years (2011, 2012 and 2013). Genetic analysis indicated the involvement of two seedling genes (Sr5 and Sr31, contributed by Rialto) and three adult plant resistance genes. QTL analyses of field data showed the involvement of three consistent effects QTL on chromosome arms 1BS (contributed by Rialto), and 3BS and chromosome 5A (contributed by Spark) in the observed resistance to stem rust. These QTLs explained average phenotypic variation of 78.5, 9.0 and 5.9 %, respectively. With the presence of virulence for Sr5 and absence of Sr31 virulence in the field, the QTL detected on 1BS (QSr.sun-1BS) was attributed to the major seedling resistance gene Sr31. The QTL located on chromosome arm 3BS (QSr.sun-3BS) was closely associated with SSR marker gwm1034, and the QTL detected on 5A (QSr.sun-5A) was closely linked with SSR marker gwm443. DH lines carrying the combination of QSr.sun-3BS and QSr.sun-5A exhibited lower stem rust responses indicating the additive effects of the two APR genes in reducing disease severity. The markers identified in this study can be useful in pyramiding these QTLs with other major or minor genes and marker assisted selection for stem rust resistance in wheat.     

Structure and expression of the <Emphasis Type="Italic">TaGW7</Emphasis> in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

OsGW7 (also known as OsGL7) is homologous to the Arabidopsis thaliana gene that encodes LONGIFOLIA protein, which regulates cell elongation, and is involved in regulating grain length in rice. However, our knowledge on its ortholog in wheat, TaGW7, is limited. In this study, we identified and mapped TaGW7 in wheat, characterized its nucleotide and protein structures, predicted the cis-elements of its promoter, and analysed its expression patterns. The GW7 orthologs in barley (HvGW7), rice (OsGW7), and Brachypodium distachyon (BdGW7) were also identified for comparative analyses. TaGW7 mapped onto the short arms of group 2 chromosomes (2AS, 2BS, and 2DS). Multiple alignments indicated GW7 possesses five exons and four introns in all but two of the species analysed. An exon–intron junction composed of introns 3–4 and exons 4–5 was highly conserved. GW7 has a conserved domain (DUF 4378) and two neighbouring low complexity regions. GW7 was mainly expressed in wheat spikes and stems, in barley seedling crowns, and in rice anthers and embryo-sacs during early development. Drought and heat significantly increased and decreased GW7 expression in wheat, respectively. In barley, GW7 was significantly down-regulated in paleae and awns but up-regulated in seeds under drought treatment and down-regulated under Fusarium and stem rust inoculation. In rice, OsGW7 expression differed significantly under drought treatments. Collectively, these results provide insights into GW7 structure and expression in wheat, barley and rice. The GW7 sequence structure and expression data are the foundation for manipulating GW7 and uncovering its roles in plants.     

Intraspecific divergence in wheats of the Timopheevi group as revealed by in situ hybridization with tandem repeats of the Spelt1 and Spelt52 families

Fluorescent in situ hybridization (FISH) was used to study the distribution of the Spelt1 and Spelt52 repetitive DNA sequences on chromosomes of ten accessions representing three polyploid wheat species of the Timopheevi group: Triticum araraticum (7), T. timopheevii (2), and T. kiharae (1). Sequences of both families were found mostly in the subtelomeric chromosome regions of the G genome. The total number of Spelt1 sites varied from 8 to 14 in the karyotypes of the species under study; their number, location, and size differed among the seven T. araraticum accessions and were the same in the two T. timopheevii accessions and T. kiharae, an amphidiploid T. timopheevii-Aegilops tauschii hybrid. The Spelt52 tandem repeat was detected in the subtelomeric regions of chromosomes 1-4; its sites did not coincide with the Spelt1 sites. The chromosome distribution and signal intensity of the Spelt52 repeats varied in T. araraticum and were the same in T. timopheevii and T. kiharae. The chromosome distributions of the Spelt1 and Spelt52 repeats were compared for the polyploid wheats of the Timopheevi group and diploid Ae. speltoides, a putative donor of the G genome. The comparison revealed a decrease in hybridization level: both the number of sites per genome and the size of sites were lower. The decrease was assumed to result from repeat elimination during polyploidization and subsequent evolution of wheat and from the founder effect, since the origin of Timopheevi wheats might involve the genotype of Ae. speltoides, which is highly polymorphic for the distribution of Spelt1 and Spelt52 sequences and is similar in the chromosome location of the repeats to modern wheat.