The potential of genomic‐assisted breeding to improve Fusarium head blight resistance in winter durum wheat

Durum wheat is the most important tetraploid wheat mainly used for semolina and pasta production, but is notorious for its high susceptibility to Fusarium head blight (FHB). Our objectives were to identify and characterize quantitative trait loci (QTL) in winter durum and to evaluate the potential of genomic approaches for the improvement of FHB resistance. Here, we employed an international panel of 170 winter and 14 spring durum lines, phenotyped for Fusarium culmorum resistance at five environments. Heading date, plant height and mean FHB severity showed significant genotypic variation with high heritabilities and FHB resistance was negatively correlated with both heading date and plant height. The dwarfing gene Rht‐B1 significantly affected FHB resistance and the genome‐wide association scan identified eight additional QTL affecting FHB resistance, explaining between 1% and 14% of the genotypic variation. A genome‐wide prediction approach yielded only a slightly improved predictive ability compared to marker‐assisted selection based on the four strongest QTL. In conclusion, FHB resistance in durum wheat is a highly quantitative trait and in breeding programmes may best be tackled by classical high‐throughput recurrent phenotypic selection that can be assisted by genomic prediction if marker profiles are available.     

User‐friendly markers linked to Fusarium wilt race 1 resistance Fw gene for marker‐assisted selection in pea

Fusarium wilt is one of the most widespread diseases of pea. Resistance to Fusarium wilt race 1 was reported as a single gene, Fw, located on linkage group III. The previously reported AFLP and RAPD markers linked to Fw have limited usage in marker‐assisted selection due to their map distance and linkage phase. Using 80 F₈ recombinant inbred lines (RILs) derived from the cross of Green Arrow × PI 179449, we amplified 72 polymorphic markers between resistant and susceptible lines with the target region amplified polymorphism (TRAP) technique. Marker–trait association analysis revealed a significant association. Five candidate markers were identified and three were converted into user‐friendly dominant SCAR markers. Forty‐eight pea cultivars with known resistant or susceptible phenotypes to Fusarium wilt race 1 verified the marker–trait association. These three markers, Fw_Trap_480, Fw_Trap_340 and Fw_Trap_220, are tightly linked to and only 1.2 cM away from the Fw locus and are therefore ideal for marker‐assisted selection. These newly identified markers are useful to assist in the isolation of the Fusarium wilt race 1 resistance gene in pea.     

Effect of 3B, 5A and 3A QTL for Fusarium head blight resistance on agronomic and quality performance of Canadian winter wheat

The objectives of this study were to investigate (i) the correlations between Fusarium head blight (FHB) index, deoxynivalenol (DON) accumulation and percentage of Fusarium‐damaged kernels (FDK) with agronomic and quality traits and (ii) the effect associated with the presence of single QTLs for FHB resistance on agronomic and quality traits in winter wheat. The population was derived from the cross between ‘RCATL33' (FHB resistance derived from ‘Sumai 3’ and ‘Frontana’) and ‘RC Strategy’. Parental lines and recombinant inbred lines (RILs) were genotyped with SSR markers associated with the 3B, 5A and 3A QTLs. The population was planted in FHB‐inoculated nurseries and in agronomy trials. Lines in the 3B QTL class had the lowest FHB index, DON content and FDK level and did not have a significantly lower yield, thousand kernel weight or protein content compared with the lines grouped in other QTL classes (including no QTL class). Marker‐assisted selection of the 3B QTL for FHB resistance into high‐yielding FHB‐susceptible winter wheat is the recommended approach for the development of lines with increased FHB resistance without significant yield and quality penalties.     

Alleles on the two dwarfing loci on 4B and 4D are main drivers of FHB‐related traits in the Canadian winter wheat population “Vienna” × “25R47”

Fusarium head blight (FHB), leaf rust and stem rust are among the most destructive wheat diseases. High‐yielding, native disease resistance sources are available in North America. The objective of this study was to map loci associated with FHB traits, leaf rust, stem rust and plant height in a “Vienna”/”25R47” population. DArT markers were used to generate a genetic map, and quantitative trait loci (QTL) analysis was performed by evaluating 113 doubled haploid lines across three environments in Ontario, Canada. FHB resistance QTL were identified on chromosomes 4D, 4B, 2D and 7A, while a QTL for leaf and stem rust resistance was identified on chromosome 1B. The dwarfing alleles of both Rht‐B1 and Rht‐D1 were associated with increased FHB index and DON content.     

Identification of QTL controlling high levels of partial resistance to Fusarium solani f. sp. pisi in pea

Fusarium root rot is a common biotic restraint on pea yields, and genetic resistance is the most feasible method for improving pea production. This study was conducted to discover quantitative trait loci (QTL) controlling genetic partial resistance to Fusarium root rot caused by Fusarium solani (Mart.) Sacc. f.sp. pisi (F.R. Jones) W.C. Snyder & H.N. Hans (Fsp). A RIL population was screened in a Fusarium root rot field disease nursery for 3 years. Composite interval mapping was employed for QTL detection using the means of disease severity from three growing seasons. Five QTL were identified, including one QTL identified in all three years. The multiyear QTL Fsp‐Ps2.1 contributed to a significant portion of the phenotypic variance (22.1–72.2%), while a second QTL, Fsp‐Ps6.1, contributed 17.3% of the phenotypic variance. The other single growing season QTL are of additional interest as they colocate with previously reported pea–Fusarium root rot resistance QTL. QTL Fsp‐Ps2.1, Fsp‐Ps3.1, Fsp‐4.1 and Fsp‐Ps7.1 are flanked by codominant SSRs and may be useful in marker‐assisted breeding of pea for high levels of partial resistance to Fsp.     

Validation of a major QTL for scab resistance with SSR markers and use of marker-assisted selection in wheat

The objectives of this study were to validate the major quantitative trait locus (QTL) for scab resistance on the short arm of chromosome 3B in bread wheat and to isolate near‐isogenic lines for this QTL using marker‐assisted selection (MAS). Two resistant by susceptible populations, both using ‘Ning7840’ as the source of resistance, were developed to examine the effect of the 3BS QTL in different genetic backgrounds. Data for scab resistance and simple sequence repeat (SSR) markers linked to the resistance QTL were analyzed in the F_2:3 lines of one population and in the F_3:4 lines of the other. Markers linked to the major QTL on chromosome 3BS in the original mapping population (‘Ning7840’/‘Clark’) were closely associated with scab resistance in both validation populations. Marker‐assisted selection for the QTL with the SSR markers combined with phenotypic selection was more effective than selection based solely on phenotypic evaluation in early generations. Marker‐assisted selection of the major QTL during the seedling stage plus phenotypic selection after flowering effectively identified scab resistant lines in this experiment. Near‐isogenic lines for this 3BS QTL were isolated from the F₆ generation of the cross ‘Ning7840’/‘IL89‐7978’ based on two flanking SSR markers, Xgwm389 and Xbarc147. Based on these results, MAS for the major scab resistance QTL can improve selection efficiency and may facilitate stacking of scab resistance genes from different sources.     

QTL analysis of resistance to Fusarium head blight in wheat using a 'Wangshuibai'-derived population

Fusarium head blight (FHB) is a devastating disease that reduces the yield, quality and economic value of wheat. For quantitative trait loci (QTL) analysis of resistance to FHB, F₃ plants and F_3:5 lines, derived from a ‘Wangshuibai’ (resistant)/‘Seri82’(susceptible) cross, were spray inoculated during 2001 and 2002, respectively. Artificial inoculation was carried out under field conditions. Of 420 markers, 258 amplified fragment length polymorphism and 39 simple sequence repeat (SSR) markers were mapped and yielded 44 linkage groups covering a total genetic distance of 2554 cM. QTL analysis was based on the constructed linkage map and area under the disease progress curve. The analyses revealed a QTL in the map interval Xgwm533‐Xs18/m12 on chromosome 3BS accounting for up to 17% of the phenotypic variation. In addition, a QTL was detected in the map interval Xgwm539‐Xs15/m24 on chromosome 2DL explaining up to 11% of the phenotypic variation. The QTL alleles originated from ‘Wangshuibai’ and were tagged with SSR markers. Using these SSR markers would facilitate marker‐assisted selection to improve FHB resistance in wheat.     

Validation of two major quantitative trait loci for fusarium head blight resistance in Chinese wheat line W14

Identity of quantitative trait loci (QTL) governing resistance to fusarium head blight (FHB) initial infection (type I), spread (type II), kernel infection, and deoxynivalenol (DON) accumulation was characterized in Chinese wheat line W14. Ninety‐six double‐haploid lines derived from a cross of W14 × ’Pion2684’ were evaluated for FHB resistance in two greenhouse and one field experiment. Two known major QTL were validated on chromosomes 3BS and 5AS in W14 using the composite interval mapping method. The 3BS QTL had a larger effect on resistance than the 5AS QTL in the greenhouse experiments, whereas, the 5AS QTL had a larger effect in the field experiment. These two QTL together explained 33%, 35%, and 31% of the total phenotypic variation for disease spread, kernel infection, and DON concentration in the greenhouse experiments, respectively. In the field experiment, the two QTL explained 34% and 26% of the total phenotypic variation for FHB incidence and severity, respectively. W14 has both QTL, which confer reduced initial infection, disease spread, kernel infection, and DON accumulation. Therefore, marker‐assisted selection (MAS) for both QTL should be implemented in incorporating W14 resistance into adapted backgrounds. Flanking markers Xbarc133 and Xgwm493 on 3BS and Xbarc117 and Xbarc56 on 5AS are suggested for MAS.     

Identification of novel QTL for resistance to crown rot in the doubled haploid wheat population 'W21MMT70' × 'Mendos'

Crown rot (causal agent Fusarium pseudograminearum) is a fungal disease of major significance to wheat cultivation in Australia. A doubled haploid wheat population was produced from a cross between line ‘W21MMT70’, which displays partial seedling and adult plant (field) resistance to crown rot, and ‘Mendos’, which is moderately susceptible in seedling tests but partially resistant in field trials. Bulked segregant analysis (BSA) based on seedling trial data did not reveal markers for crown rot resistance. A framework map was produced consisting of 128 microsatellite markers, four phenotypic markers, and one sequence tagged site marker. To this map 331 previously screened AFLP markers were then added. Three quantitative trait loci (QTL) were identified with composite interval mapping across all of the three seedling trials conducted. These QTL are located on chromosomes 2B, 2D and 5D. The 2D and 5D QTL are inherited from the line ‘W21MMT70’, whereas the 2B QTL is inherited from ‘Mendos’. These loci are different from those associated with crown rot resistance in other wheat populations that have been examined, and may represent an opportunity for pyramiding QTL to provide more durable resistance to crown rot.     

Development of gene‐based markers for the Turnip mosaic virus resistance gene retr02 in Brassica rapa

Turnip mosaic virus (TuMV) is responsible for a serious disease that affects the production of Chinese cabbage. Previous studies have cloned a series of TuMV resistance genes and developed molecular markers. In this study, a derived cleaved amplified polymorphism sequence (dCAPS) marker and a Kompetitive Allele Specific PCR (KASP) marker were developed based on a single recessive gene, retr02, which confers broad‐spectrum TuMV resistance in Chinese cabbage by means of an additional G at the junction of exon 1 and intron 1. The two markers were able to detect the retr02 allele in Chinese cabbage accessions used in breeding programmes. Compared with the dCAPS marker, the KASP marker was flexible, cost‐effective and quick to process, which is likely to be beneficial in establishing high‐throughput assays for marker‐assisted selection.     

Quantitative trait loci for resistance to Fusarium head blight in recombinant inbred wheat lines from the cross Huapei 57-2 / Patterson

Fusarium head blight (FHB), primarily caused by Fusarium graminearum in North America, often results in significant losses in yield and grain quality of wheat (Triticum aestivum L.). Evaluation of FHB resistance is laborious and can be affected by environmental conditions. The development of DNA markers associated with FHB quantitative trait loci (QTL) and their use in breeding programs could greatly enhance selection. The objective of this study was to identify the location and effect of QTLs for FHB resistance using simple sequence repeat (SSR) markers. A population of wheat recombinant inbred lines derived from the cross ‘Huapei57-2’/‘Patterson’ was characterized for type II resistance in one field experiment and two tests under controlled conditions in the greenhouse. Bulked segregant analysis followed by QTL mapping was used to identify the major segregating QTLs. Results indicate that ‘Huapei 57-2’ may have the same resistance allele as ‘Sumai3’ at a QTL located on the short arm of chromosome 3B. Other QTLs of lower effect size were identified on the long arm of 3Band on chromosomes 3A and 5B. Our findings along with results from other studies demonstrate that the effect of the QTL on3BS is large and consistent across a wide range of genetic backgrounds and environments. Pyramiding this QTL with other FHB QTLs using marker-assisted selection should be effective in improving FHB resistance in a wheat breeding program. This revised version was published online in August 2006 with corrections to the Cover Date.     

Assessing diversity in Triticum durum cultivars and breeding lines for high versus low cadmium content in seeds using the CAPS marker usw47

Cadmium (Cd) is a toxic heavy metal that occurs naturally in soils. Durum wheat is known to accumulate generally more Cd than other cereal crops. The uptake of Cd in durum wheat is governed by the gene Cdu1, which co‐segregates with several DNA markers, such as the co‐dominant marker usw47 and the dominant marker ScOPC20. A panel of 314 durum wheat cultivars or lines originating from 16 countries or regions were assessed with usw47. The plant material was mainly comprised of cultivars and modern breeding lines. From this set, 165 durum wheat lines were classified as low‐Cd accumulators, 144 high‐Cd accumulators and five were heterogeneous. A smaller subset of 16 cultivars had previously been evaluated for Cd accumulation in replicated field trials. Lines with the high‐Cd allele showed a 2.4‐fold higher Cd content in the seeds than lines with the low‐Cd allele. We also compared the utility of markers usw47 and ScOPC20 as selection tools. Marker‐assisted selection appears as a robust and practicable tool for breeding durum cultivars with low‐Cd content.     

QTL mapping and marker-assisted selection for Fusarium head blight resistance in wheat: a review

During the past decade, numerous studies have been published on molecular mapping of Fusarium head blight (FHB) resistance in wheat. We summarize the relevant findings from 52 quantitative trait loci (QTL) mapping studies, nine research articles on marker-assisted selection and seven on marker-assisted germplasm evaluation. QTL for FHB resistance were found on all wheat chromosomes except chromosome 7D. Some QTL were found in several independent mapping studies indicating that such QTL are stable and therefore useful in breeding programmes. We summarize and update current knowledge on the genetics of FHB resistance in wheat resulting from QTL mapping investigations and review and suggest FHB breeding strategies based on the available information and DNA markers.     

QTLs for black-point resistance in wheat and the identification of potential markers for use in breeding programmes

Quantitative trait loci (QTLs) for black‐point resistance have been mapped in two doubled haploid‐derived wheat populations, each thought to contain unrelated sources of resistance. In the ‘Sunco’בTasman’‐derived population, QTLs were located on chromosomes 1D, 2B, 3D, 4A, 5A and 7A with each QTL explaining between 4 and 15% of the observed phenotypic variance. QTLs were contributed by both parents. In the ‘Cascades’בAUS1408’‐derived population, QTLs from ‘Cascades’ were identified on chromosomes 2A, 2D and 7A with each QTL explaining between 12 and 18% of the phenotypic variance. Several markers were identified which are promising candidates for use in marker‐assisted selection programmes. If one, two or three of these markers would have been used to select for black‐point resistance in the ‘Sunco’בTasman’ population, then with one marker 34 of 39 resistant lines, with two markers 23 of 32 and with three markers 17 of 32 would have been selected. At the same time, 67 false positives obtained by selecting with one marker are reduced to 24 by selection with two markers and to 11 by selection with three markers. Similarly, if one, two or three markers are used to select for black‐point resistance in the ‘Cascades’בAUS1408’ populations, then with one marker 25 of 31 resistant lines, with two markers 26 of 31 and with three markers 10 of 31 are selected. At the same time, 14 false positives are obtained with one marker are reduced to six by selection with two markers and no false positives are selected using three markers.     

Phenotypic selection for high resistance to Fusarium head blight after introgression of quantitative trait loci (QTL) from exotic spring wheat and verification by simple sequence repeat markers a posteriori

Fusarium head blight (FHB) has become an important disease of wheat. We introgressed three resistance quantitative trait loci (QTL) alleles on chromosomes 3B, 5A (from CM82036) and 3A (from ‘Frontana’) into European elite spring wheat and performed phenotypic selection among double‐cross (DC) derived progeny in generations DCF₂ and DCF₃. After recombination and selfing, we analysed 135 phenotypically selected progeny by simple sequence repeat (SSR) markers linked to the QTL. In a second experiment, we forwarded the best 20 progeny for a further two generations by pedigree selection. Progeny were inoculated at two to four locations with Fusarium culmorum and the percentage of infected spikelets per plot was estimated. Both experiments show that phenotypic selection was highly effective. One‐hundred out of 135 phenotypically selected DCF_1:3 progeny had the combination of donor‐QTL alleles (3B + 5A + 3A, 3B + 5A) with the highest effects on FHB resistance. In the subsequent generations, sufficient genotypic variance was detected. The best F_5:7 bulks had similar resistance to the donor CM82036. The FHB rating was reduced in total by 45% points compared to the parental mean. QTL with high effects can be detected solely by phenotypic selection after targeted introgression.     

Identification and marker‐assisted introgression of QTL conferring resistance to bacterial leaf blight in cowpea (Vigna unguiculata (L.) Walp.)

Bacterial leaf blight (BLB), caused by Xanthomonas axonopodis pv. vignicola (Xav), is widespread in major cowpea [Vigna unguiculata (L.) Walp.] growing regions of the world. Considering the resource poor nature of cowpea farmers, development and introduction of cultivars resistant to the disease is the best option. Identification of DNA markers and marker‐assisted selection will increase precision of breeding for resistance to diseases like bacterial leaf blight. Hence, an attempt was made to detect QTL for resistance to BLB using 194 F_2 : 3 progeny derived from the cross ‘C‐152’ (susceptible parent) × ‘V‐16’ (resistant parent). These progeny were screened for resistance to bacterial blight by the leaf inoculation method. Platykurtic distribution of per cent disease index scores indicated quantitative inheritance of resistance to bacterial leaf blight. A genetic map with 96 markers (79 SSR and 17 CISP) constructed from the 194 F₂ individuals was used to perform QTL analysis. Out of three major QTL identified, one was on LG 8 (qtlblb‐1) and two on LG 11 (qtlblb‐2 and qtlblb‐3). The PCR product generated by the primer VuMt337 encoded for RIN2‐like mRNA that positively regulate RPM1‐ and RPS2‐dependent hypersensitive response. The QTL qtlblb‐1 explained 30.58% phenotypic variation followed by qtlblb‐2 and qtlblb‐3 with 10.77% and 10.63%, respectively. The major QTL region on LG 8 was introgressed from cultivar V‐16 into the bacterial leaf blight susceptible variety C‐152 through marker‐assisted backcrossing (MABC).     

Use of non‐adapted quantitative trait loci for increasing Fusarium head blight resistance for breeding semi‐dwarf wheat

Semi‐dwarf wheat is an important prerequisite for releasing a successful commercial cultivar in high‐yielding environments. In Northern Europe, this aim is achieved by using one of the dwarfing genes Rht‐B1 (formerly known as Rht‐1) or Rht‐D1 (Rht‐2). Both genes, however, result in a higher susceptibility to Fusarium head blight (FHB). We analysed the possibility to use the two non‐adapted FHB resistance quantitative trait loci Fhb1 and Fhb5 (syn. QFhs.ifa‐5A) to counterbalance the negative effect of the dwarfing allele Rht‐D1b in a winter wheat population of 585 doubled‐haploid (DH) lines segregating for the three loci. All lines were inoculated with Fusarium culmorum at four locations and analysed for FHB severity, plant height, and heading date. The DH population showed a significant (p < 0.001) genotypic variation for FHB severity ranging from 3.6% to 65.9% with a very high entry‐mean heritability of 0.95. The dwarfing allele Rht‐D1b reduced plant height by 24 cm, but nearly doubled the FHB susceptibility (24.74% vs. 12.74%). The resistance alleles of Fhb1 and Fhb5 reduced FHB susceptibility by 6.5 and 11.3 percentage points, respectively. Taken all three loci together, Fhb5 alone was already able to reduce FHB susceptibility to the same extent as Rht‐D1b increased it. This opens new avenues for selecting semi‐dwarf wheat by marker‐assisted introgression of Fhb5 without the enhancement of FHB susceptibility.     

Identification and mapping of adult‐plant stripe rust resistance in soft red winter wheat cultivar ‘USG 3555’

Little is known about the extent or diversity of resistance in soft red winter wheat (Triticum aestivum L.) to stripe rust, caused by the fungal pathogen Puccinia striiformis f.sp. tritici. The soft red winter (SRW) wheat cultivar ‘USG 3555’ has effective adult‐plant resistance to stripe rust, which was characterized in a population derived from ‘USG 3555’/‘Neuse’. The mapping population consisted of 99 recombinant inbred lines, which were evaluated for stripe rust infection type (IT) and severity to race PST‐100 in field trials in North Carolina in 2010 and 2011. Genome‐wide molecular‐marker screenings with 119 simple sequence repeats and 560 Diversity Arrays Technology (DArT) markers were employed to identify quantitative trait loci (QTL) for stripe rust resistance. QTL on chromosomes 1AS, 4BL and 7D of ‘USG 3555’ explained 12.8, 73.0 and 13.6% of the variation in stripe rust IT, and 13.5, 72.3 and 10.5% of the variation in stripe rust severity, respectively. Use of these and additional diagnostic markers for these QTL will facilitate the introgression of this source of stripe rust resistance into SRW wheat lines via marker‐assisted selection.     

Development of molecular markers for crown rot resistance in wheat: mapping of QTLs for seedling resistance in a '2-49' × 'Janz' population

Crown rot, caused by Fusarium pseudograminearum, is an important disease of wheat in Australia and elsewhere. In order to identify molecular markers associated with partial seedling resistance to this disease, bulked segregant analysis and quantitative trait loci (QTL) mapping approaches were undertaken using a population of 145 doubled haploid lines constructed from ‘2‐49’ (partially resistant) × ‘Janz’ (susceptible) parents. Phenotypic data indicated that the trait is quantitatively inherited. The largest QTLs were located on chromosomes 1D and 1A, and explained 21% and 9% of the phenotypic variance, respectively. Using the best markers associated with five QTLs identified by composite interval mapping, the combined effect of the QTLs explained 40.6% of the phenotypic variance. All resistance alleles were inherited from ‘2‐49’ with the exception of a QTL on 2B, which was inherited from ‘Janz’. A minor QTL on 4B was loosely linked (19.8 cM) to the Rht1 locus in repulsion. None of the QTLs identified in this study were located in the same region as resistance QTLs identified in other populations segregating for Fusarium head blight, caused by Fusarium graminearum.     

The locus for resistance to Asian soybean rust in PI 587855

Asian soybean rust (ASR) caused by Phakopsora pachyrhizi is one of the most serious soybean (Glycine max) diseases in tropical and subtropical areas. A soybean line, PI 587855, showed a resistance phenotype against ASR pathogens in Japan and South America at high frequency; however, little is known of the genetic control of this resistance and chromosomal location of the corresponding locus. Therefore, the aim of this study was to study the inheritance of PI 587855 resistance and map the corresponding locus with SSR markers aiming to use the linked markers in marker‐assisted selection. In the segregating population, resistance to ASR appeared to be controlled by a single dominant gene. The ASR resistance locus was mapped near to the chromosomal region where the resistant loci, Rpp1 and Rpp1‐b, were previously mapped. Comparative genetic mapping and disease reaction profiles of other seven lines carrying Rpp1 or Rpp1‐b to four Brazilian ASR isolates revealed that the resistance reaction exhibited by PI 587855 was similar to that of Rpp1‐b‐carrying varieties which have useful resistance to South American ASR strains.