Advanced Backcross QTL Research Articles

Advanced backcross QTL analysis and comparative mapping with RIL QTL studies and GWAS provide an overview of QTL and marker haplotype diversity for resistance to Aphanomyces root rot in pea (Pisum sativum).

Aphanomyces euteiches is the most damaging soilborne pea pathogen in France. Breeding of pea resistant varieties combining a diversity of quantitative trait loci (QTL) is a promising strategy considering previous research achievements in dissecting polygenic resistance to A. euteiches. The objective of this study was to provide an overview of the diversity of QTL and marker haplotypes for resistance to A. euteiches, by integrating a novel QTL mapping study in advanced backcross (AB) populations with previous QTL analyses and genome-wide association study (GWAS) using common markers. QTL analysis was performed in two AB populations derived from the cross between the susceptible spring pea variety "Eden" and the two new sources of partial resistance "E11" and "LISA". The two AB populations were genotyped using 993 and 478 single nucleotide polymorphism (SNP) markers, respectively, and phenotyped for resistance to A. euteiches in controlled conditions and in infested fields at two locations. GWAS and QTL mapping previously reported in the pea-Aphanomyces collection and from four recombinant inbred line (RIL) populations, respectively, were updated using a total of 1,850 additional markers, including the markers used in the Eden x E11 and Eden x LISA populations analysis. A total of 29 resistance-associated SNPs and 171 resistance QTL were identified by GWAS and RIL or AB QTL analyses, respectively, which highlighted 10 consistent genetic regions confirming the previously reported QTL. No new consistent resistance QTL was detected from both Eden x E11 and Eden x LISA AB populations. However, a high diversity of resistance haplotypes was identified at 11 linkage disequilibrium (LD) blocks underlying consistent genetic regions, especially in 14 new sources of resistance from the pea-Aphanomyces collection. An accumulation of favorable haplotypes at these 11 blocks was confirmed in the most resistant pea lines of the collection. This study provides new SNP markers and rare haplotypes associated with the diversity of Aphanomyces root rot resistance QTL investigated, which will be useful for QTL pyramiding strategies to increase resistance levels in future pea varieties.

Genomic Insights Into Sclerotinia Basal Stalk Rot Resistance Introgressed From Wild Helianthus praecox Into Cultivated Sunflower (Helianthus annuus L.).

Crop wild relatives of the cultivated sunflower (Helianthus annuus L.) are a valuable resource for its sustainable production. Helianthus praecox ssp. runyonii is a wild sunflower known for its resistance against diseases caused by the fungus, Sclerotinia sclerotiorum (Lib.) de Bary, which infects over 400 broadleaf hosts including many important food crops. The objective of this research was to dissect the Sclerotinia basal stalk rot (BSR) resistance introgressed from H. praecox ssp. runyonii into cultivated sunflower. An advanced backcross quantitative trait loci (AB-QTL) mapping population was developed from the cross of a H. praecox accession with cultivated sunflower lines. The AB-QTL population was evaluated for BSR resistance in the field during the summers of 2017–2018 and in the greenhouse in the spring of 2018. Highly significant genetic variations (p < 0.001) were observed for the BSR disease in the field and greenhouse with a moderately high broad-sense heritability (H2) ranging from 0.66 to 0.73. Genotyping-by-sequencing approach was used to genotype the parents and the progeny lines of the AB-QTL population. A genetic linkage map spanning 1,802.95 cM was constructed using 1,755 single nucleotide polymorphism (SNP) markers mapped on 17 sunflower chromosomes. A total of 19 BSR resistance QTL were detected on nine sunflower chromosomes, each explaining 2.21%–16.99% of the phenotypic variance for resistance in the AB-QTL population. Sixteen of the 19 QTL had alleles conferring increased BSR resistance derived from the H. praecox parent. SNP markers flanking the identified QTL will facilitate marker-assisted breeding to combat the disease in sunflower.

NEW SELECTION TECHNIQUES TO DETECT SOURCES OF RESISTANCE AGAINST STRIPE RUST IN WHEAT

Stripe rust is the most important biotic constraint in wheat production. Deployment of resistant wheat cultivars is the most practical way to decrease yield losses attributed to stripe rust. Usage of wild germplasm of wheat is the best technique to detect new resistant genes for the evolution of new varieties and mainstream of these hold capacity for disease resistance. Traditional procedures of gene identification by using conventional screening through diversified races do not generate accurate data. In conventional breeding, transferring desired traits, for instance, stripe rust resistance, from a donor parent is offered into a genotype of interest, typically high yielding. Identification of the sources of resistance by conventional screening through diversified races and backcross breeding is time-consuming which could be avoided through the usage of DNA markers. Marker-assisted selection (MAS) is practiced for the improvement of a variety of traits in wheat around the world through introgression of disease resistance against diseases including stripe rust, which may in any case partially support in providing the anticipated result. MAS has been valuable for the development of quite a lot of significant traits such as resistance against diseases. Marker-assisted backcrossing, forward breeding, and MAS involving doubled haploid technology have been effectively used for this objective. Novel skills based on high throughput genotyping related with new marker systems (e.g., Diversity arrays technology DArT and as Single Nucleotide Polymorphism SNP), and new selection strategies such as (Advanced backcross QTL AB-QTL), mapping-as-you-go, marker-assisted recurrent selection, and genome-wide selection will have to be tried in future for upgrading of compound multigenic traits. The improvement made in all these features of marker-assisted wheat breeding, and the restrictions and future scenarios of this incipient technology have been reviewed in this article.

Fine-Mapping of a Wild Genomic Region Involved in Pod and Seed Size Reduction on Chromosome A07 in Peanut (Arachis hypogaea L.).

Fruit and seed size are important yield component traits that have been selected during crop domestication. In previous studies, Advanced Backcross Quantitative Trait Loci (AB-QTL) and Chromosome Segment Substitution Line (CSSL) populations were developed in peanut by crossing the cultivated variety Fleur11 and a synthetic wild allotetraploid (Arachis ipaensis × Arachis duranensis)4x. In the AB-QTL population, a major QTL for pod and seed size was detected in a ~5 Mb interval in the proximal region of chromosome A07. In the CSSL population, the line 12CS_091, which carries the QTL region and that produces smaller pods and seeds than Fleur11, was identified. In this study, we used a two-step strategy to fine-map the seed size QTL region on chromosome A07. We developed new SSR and SNP markers, as well as near-isogenic lines (NILs) in the target QTL region. We first located the QTL in ~1 Mb region between two SSR markers, thanks to the genotyping of a large F2 population of 2172 individuals and a single marker analysis approach. We then used nine new SNP markers evenly distributed in the refined QTL region to genotype 490 F3 plants derived from 88 F2, and we selected 10 NILs. The phenotyping of the NILs and marker/trait association allowed us to narrowing down the QTL region to a 168.37 kb chromosome segment, between the SNPs Aradu_A07_1148327 and Aradu_A07_1316694. This region contains 22 predicted genes. Among these genes, Aradu.DN3DB and Aradu.RLZ61, which encode a transcriptional regulator STERILE APETALA-like (SAP) and an F-box SNEEZY (SNE), respectively, were of particular interest. The function of these genes in regulating the variation of fruit and seed size is discussed. This study will contribute to a better knowledge of genes that have been targeted during peanut domestication.

Assessment of 16 Peanut (Arachis hypogaea L.) CSSLs Derived from an Interspecific Cross for Yield and Yield Component Traits: QTL Validation

Cultivated peanut is an allotetraploid (2n = 4× = 40) with narrow genetic diversity. In previous studies, we developed an advanced backcross quantitative trait loci (AB-QTL) population from the cross between the synthetic allotetraploid ((Arachis ipaensis × Arachis duranensis)4×) and the cultivated variety Fleur11, and mapped several quantitative trait loci (QTLs) involved in yield and yield components. We also developed a chromosome segment substitution line (CSSL) population as a way to mendelize the QTLs and analyzing their effects. In this study, 16 CSSLs were used for assessing the contribution of wild alleles in yield performance and stability across environments, as well as validating QTLs for pod and seed size. The CSSLs and the recurrent parent Fleur11, used as a check, were assessed using an alpha lattice design in three locations during two consecutive rainy seasons in Senegal, totaling six environments. Our results showed that the chromosome segments from the wild species, in general, have no yield disadvantage and contributed positive variation to yield-related traits. Most of the QTLs detected for pod and seed size in the AB-QTL on linkage groups A07, A08, A09, and B06 were also found in the CSSLs, showing that the CSSLs used in this study are accurate material for QTL validation. Several new QTLs have also been identified. Two CSSLs (12CS_031 and 12CS_069) showed consistently higher pod and seed size than Fleur11 in all environments, suggesting that the QTLs were consistent and stable. Our study opens the way for pyramiding these QTLs for peanut improvement.

Identification of Anther Length QTL and Construction of Chromosome Segment Substitution Lines of Oryza longistaminata.

Life histories and breeding systems strongly affect the genetic diversity of seed plants, but the genetic architectures that promote outcrossing in Oryza longistaminata, a perennial wild species in Africa, are not understood. We conducted a genetic analysis of the anther length of O. longistaminata accession W1508 using advanced backcross quantitative trait locus (QTL) analysis and chromosomal segment substitution lines (CSSLs) in the genetic background of O. sativa Taichung 65 (T65), with simple sequence repeat markers. QTL analysis of the BC3F1 population (n = 100) revealed that four main QTL regions on chromosomes 3, 5, and 6 were associated to anther length. We selected a minimum set of BC3F2 plants for the development of CSSLs to cover as much of the W1508 genome as possible. The additional minor QTLs were suggested in the regional QTL analysis, using 21 to 24 plants in each of the selected BC3F2 population. The main QTLs found on chromosomes 3, 5, and 6 were validated and designated qATL3, qATL5, qATL6.1, and qATL6.2, as novel QTLs identified in O. longistaminata in the mapping populations of 94, 88, 70, and 95 BC3F4 plants. qATL3, qATL5, and qATL6.1 likely contributed to anther length by cell elongation, whereas qATL6.2 likely contributed by cell multiplication. The QTLs were confirmed again in an evaluation of the W1508ILs. In several chromosome segment substitution lines without the four validated QTLs, the anthers were also longer than those of T65, suggesting that other QTLs also increase anther length in W1508. The cloning and diversity analyses of genes conferring anther length QTLs promotes utilization of the genetic resources of wild species, and the understanding of haplotype evolution on the differentiation of annuality and perenniality in the genus Oryza.

Genetic relationships between anther and stigma traits revealed by QTL analysis in two rice advanced-generation backcross populations

Anther and stigma size are critical floral traits that influence outcrossing in rice (Oryza sativa), a crop that is predominantly self-fertilizing. The efficiency of hybrid rice seed production depends on efficient outcrossing ability of parental lines, which is promoted by increased anther and stigma size. Phenotypic correlations between anther and stigma traits have been observed in many studies; however, evidence for this relationship is unclear and the genetic basis remains to be elucidated. To examine this relationship and to identify quantitative trait loci (QTLs) for increased anther and stigma size, we developed two advanced backcross QTL mapping populations derived from a cross between a Thai elite indica crop variety (SPR1) and an accession of common wild rice (O. rufipogon Griff.), which is predominantly outcrossing. One mapping population was selected for increased anther size while the other was selected for increased stigma size. We mapped QTLs for anther size and stigma size in both populations. Bulked segregant analysis was used to identify molecular markers associated with the selected traits. A total of 16 significant QTLs associated with anther and stigma traits were identified across the two populations, and these were located in five genomic regions on four chromosomes. Whereas three of these regions have been previously reported, two of them are newly identified and should be further explored for improving outcrossing ability in rice. The co-localization of QTL for anther and stigma traits strongly suggests some degree of shared developmental basis for these traits.

Advanced Backcross QTL Analysis of Fiber Strength and Fineness in a Cross between Gossypium hirsutum and G. mustelinum.

The molecular genetic basis of cotton fiber strength and fineness in crosses between Gossypium mustelinum and Gossypium hirsutum (Upland cotton) was dissected using 21 BC3F2 and 12 corresponding BC3F2:3 and BC3F2:4 families. The BC3F2 families were genotyped with simple sequence repeat markers from a G. hirsutum by G. mustelinum linkage map, and the three generations of BC3-derived families were phenotyped for fiber strength (STR) and fineness (Micronaire, MIC). A total of 42 quantitative trait loci (QTLs) were identified through one-way analysis of variance, including 15 QTLs for STR and 27 for MIC, with the percentage of variance explained by individual loci averaging 13.86 and 14.06%, respectively. Eighteen of the 42 QTLs were detected at least twice near the same markers in different generations/families or near linked markers in the same family, and 28 of the 42 QTLs were identified in both mixed model-based composite interval mapping and one-way variance analyses. Alleles from G. mustelinum increased STR for eight of 15 and reduced MIC for 15 of 27 QTLs. Significant among-family genotypic effects (P < 0.001) were detected in 13 and 10 loci for STR and MIC respectively, and five loci showed significant (P < 0.001) genotype × family interaction for MIC. These results support the hypothesis that fiber quality improvement for Upland cotton could be realized by introgressing G. mustelinum alleles although complexities due to the different effects of genetic background on introgressed chromatin might be faced. Building on prior work with G. barbadense, G. tomentosum, and G. darwinii, QTL mapping involving introgression of G. mustelinum alleles offers new allelic variation to Upland cotton germplasm.

A New Advanced Backcross Tomato Population Enables High Resolution Leaf QTL Mapping and Gene Identification.

Quantitative Trait Loci (QTL) mapping is a powerful technique for dissecting the genetic basis of traits and species differences. Established tomato mapping populations between domesticated tomato (Solanum lycopersicum) and its more distant interfertile relatives typically follow a near isogenic line (NIL) design, such as the S. pennellii Introgression Line (IL) population, with a single wild introgression per line in an otherwise domesticated genetic background. Here, we report on a new advanced backcross QTL mapping resource for tomato, derived from a cross between the M82 tomato cultivar and S. pennellii. This so-called Backcrossed Inbred Line (BIL) population is comprised of a mix of BC2 and BC3 lines, with domesticated tomato as the recurrent parent. The BIL population is complementary to the existing S. pennellii IL population, with which it shares parents. Using the BILs, we mapped traits for leaf complexity, leaflet shape, and flowering time. We demonstrate the utility of the BILs for fine-mapping QTL, particularly QTL initially mapped in the ILs, by fine-mapping several QTL to single or few candidate genes. Moreover, we confirm the value of a backcrossed population with multiple introgressions per line, such as the BILs, for epistatic QTL mapping. Our work was further enabled by the development of our own statistical inference and visualization tools, namely a heterogeneous hidden Markov model for genotyping the lines, and by using state-of-the-art sparse regression techniques for QTL mapping.

Advanced Backcross Quantitative Trait Locus Analysis of Fiber Elongation in a Cross between Gossypium hirsutum and G. mustelinum

In an effort to explore the secondary gene pool for the enhancement of upland cotton (Gossypium hirsutum L.) germplasm, we developed advanced‐generation backcross populations by crossing G. hirsutum (PD94042) and Gossypium mustelinum Miers ex Watt (AD4–8), then backcrossing to the G. hirsutum parent for three cycles. Genome‐wide mapping revealed introgressed alleles at an average of 13.8% of loci in each BC3F1 plant, collectively representing G. mustelinum introgression in 80.9% of the genome. Twenty‐one BC3F1 plants were selfed to generate BC3F2 families of 127 to 160 plants per family (totaling 3203 plants), which were field‐tested for fiber elongation and genetically mapped. One‐way ANOVA detected 15 nonoverlapping quantitative trail loci (QTLs) distributed over 12 chromosomes. Individual loci explained from 10.0 to 25.24% of phenotypic variance. Nine stringent QTLs were detected in one‐way ANOVAs and composite interval mapping; two of the nine QTLs explained more than 20% of variance and one was detected in four different families simultaneously with similar additive effects. Although the G. mustelinum parent does not produce spinnable fiber, G. mustelinum alleles contributed to increased fiber elongation for three of the nine stringent QTLs and five of the 15 total QTLs. Two‐way ANOVA detected significant (P &lt; 0.001) among‐family genotype effects and genotype × family interactions, suggesting that the phenotypic effects of some introgressed chromosomal segments are dependent on the existence of other chromosomal segments. Gossypium mustelinum alleles that contribute to increased fiber elongation are of great interest to be further exploited in cotton breeding.

Hybrid breeding boosted molecular genetics in rye

The history of rye (Secale cereale L.) breeding began from the first targeted selections made in Germany by the Probsteier Seed Cooperative around 1850, and over 150 years of breeding yielded a tremendous amount of results. Rye has also long been used as cytological subject due to its low number of chromosomes and their size. However, genetic findings in rye up to the early 1980s were rather scant. About 120 genes could be assigned to seven linkage groups. However, since then development of new methods such as C-banding procedures, in situ DNA hybridization, enzymology and molecular marker techniques achieved an enormous progress. Particularly, the latter of these was driven by the agronomic success of hybrid breeding in rye. The basic genetic knowledge resulting from intra- and interspecies genetic diversity assay and phylogenetic studies in the Secale genus used nuclear and cytoplasmic molecular markers to facilitate successful selection of parents for hybrid breeding and hence contribute to improvement of heterosis effect. The main achievements in rye hybrid breeding and genetic mechanisms exploited for different hybrid breeding strategies are discussed. The exploitation of landraces or wild rye species gene pools via advanced backcross QTL analysis or introgression libraries to increase the heterosis effect is a prospective strategy of marker-assisted hybrid breeding in rye. Prediction of hybrid performance can also be improved significantly by marker-assisted selection and genomic selection based on genome-wide marker data. The findings in rye genetics (including phylogenetic, mapping and population studies) are reviewed in their relation with the hybrid breeding purposes and demands. Overall, about 450 morphological and biochemical traits are mapped throughout the the genome plus about 5,000 DNA markers. They are not only associated with individual chromosome or segments but also efficiently used for comparative mapping (evolutionary studies), introgression monitoring, QTL tagging, and marker-assisted selection.

Genomics in agriculture.

Abstract Plant breeders mainly rely on phenotypic selection for the improvement of plant varieties. However, with the advent of molecular markers in the early 1980s, it became possible to select desirable traits more directly. Molecular markers, primarily DNA markers, are sections of the genome of the organisms that are used for recognition of a larger area of the genome. The genome represents the sum of all the genes in an organism and the study of the whole genome is known as genomics which comprises both structural as well as functional aspect of genomics. The first DNA-based genetic marker was restriction fragment length polymorphism. A molecular marker can either be located within the gene of interest or be linked to a gene determining a trait of interest. Easily detectable DNA markers can now be used in plant breeding through marker-assisted selection (MAS) for the development of improved varieties based on the genotype of plants rather than assessing the phenotype only. In addition, DNA markers are also used for germplasm evaluation, genetic diagnostics, phylogenetic analysis, study of genome organization and screening of transformants. The breeding strategies of MAS used most frequently for introgression of genes from breeding lines or wild relatives to cultivated species are: marker-assisted back-crossing, marker-assisted recurrent selection, advanced backcross-quantitative trait loci and pyramiding of genes. Current breeding objectives, for which MAS is regularly utilized in crops are mainly, disease and pest resistance and secondarily yield improvement, quality traits, and abiotic stress resistance.

Advanced backcross QTL analysis reveals complicated genetic control of rice grain shape in a &lt;i&gt;japonica&lt;/i&gt; × &lt;i&gt;indica&lt;/i&gt; cross

Grain shape is an important trait for improving rice yield. A number of quantitative trait loci (QTLs) for this trait have been identified by using primary F2 mapping populations and recombinant inbred lines, in which QTLs with a small effect are harder to detect than they would be in advanced generations. In this study, we developed two advanced mapping populations (chromosome segment substitution lines [CSSLs] and BC4F2 lines consisting of more than 2000 individuals) in the genetic backgrounds of two improved cultivars: a japonica cultivar (Koshihikari) with short, round grains, and an indica cultivar (IR64) with long, slender grains. We compared the ability of these materials to reveal QTLs for grain shape with that of an F2 population. Only 8 QTLs for grain length or grain width were detected in the F2 population, versus 47 in the CSSL population and 65 in the BC4F2 population. These results strongly suggest that advanced mapping populations can reveal QTLs for agronomic traits under complicated genetic control, and that DNA markers linked with the QTLs are useful for choosing superior allelic combinations to enhance grain shape in the Koshihikari and IR64 genetic backgrounds.

Mapping and introgression of QTL for yield and related traits in two backcross populations derived from Oryza sativa cv. Swarna and two accessions of O. nivara.

Advanced backcross QTL (AB-QTL) analysis was carried out in two Oryza nivara-derived BC2F2 populations. For nine traits, we identified 28 QTL in population 1 and 26 QTL in population 2. The two most significant yield-enhancing QTL, yldp9.1 and yldp2.1 showed an additive effect of 16 and 7 g per plant in population 1, while yld2.1 and yld11.1 showed an additive effect of 11 and 10 g per plant in population 2. At least one O. nivara-derived QTL with a phenotypic variance of >15% was detected for seven traits in population 1 and three traits in population 2. The O. nivara-derived QTL ph1.1, nt12.1, nsp1.1, nfg1.1, bm11.1, yld2.1 and yld11.1 were conserved at the same chromosomal locations in both populations. Two major QTL clusters were detected at the marker intervals RM488-RM431 and RM6-RM535 on chromosomes 1 and 2, respectively. The colocation of O. nivara-derived yield QTL with yield meta-QTL on chromosomes 1, 2 and 9 indicates their accuracy and consistency. The major-effect QTL reported in this study are useful for marker-assisted breeding and are also suitable for further fine mapping and candidate gene identification.

Advanced Backcross QTL Analysis for the Whole Plant Growth Duration Salt Tolerance in Rice (Oryza sativa L.)

Salinity is a major factor limiting rice yield in coastal areas of Asia. To facilitate breeding salt tolerant rice varieties, the whole-plant growth duration salt tolerance (ST) was genetically dissected by phenotyping two sets of BC2F5 introgression lines (ILs) for four yield traits under severe natural salt stress and non-stress filed conditions using SSR markers and the methods of advanced backcross QTL (AB-QTL) analysis and selective introgression. Many QTLs affecting four yield traits under salt stress and non-stress conditions were identified, most (>90%) of which were clustered in 13 genomic regions of the rice genome and involved in complex epistasis. Most QTLs affecting yield traits were differentially expressed under salt stress and non-stress conditions. Our results suggested that genetics complementarily provides an adequate explanation for the hidden genetic diversity for ST observed in both IL populations. Some promising Huanghuazhan (HHZ) ILs with favorable donor alleles at multiple QTLs and significantly improved yield traits under salt stress and non-stress conditions were identified, providing excellent materials and relevant genetic information for improving rice ST by marker-assisted selection (MAS) or genome selection.

Quantitative trait loci for quality and agronomic traits in two advanced backcross populations in oat (Avena sativa L.)

AbstractUsing the advanced backcross quantitative trait loci (AB‐QTL) strategy, we successfully transferred and mapped valuable allelic variants from the high β‐glucan (BG) accession IAH611 (PI 502955), into the genome of cultivar ‘Iltis’. By backcrossing one BC1F1 plant to ‘Iltis’, we developed two BC2F2‐6 populations A and B, comprising 98 and 72 F2‐individuals, respectively. Genotyping of BC2F2 individuals with predominantly AFLP markers resulted in 12 linkage groups with a map size of 455.4 cM for Population A and 11 linkage groups with a map size of 313.5 cM for Population B. Both populations were grown at three sites in Germany over a three‐year period. Individuals were then phenotyped for 13 traits including grain yield (YD) and β‐glucan content (BG). QTL analysis via stepwise regression detected a total of 33 QTLs, most of which were clustered in three linkage groups. Two dense linkage groups A1 and B13 were found to be putatively homologous to groups KO_6 and KO_11 of the ‘Kanota’/‘Ogle’ map, respectively.

Use of Major Quantitative Trait Loci to Improve Grain Yield of Rice

Further improvement of rice productivity remains a challenge. Breeding is perceived as an important option to increase rice yield. However, the genetic progress of grain yield in most rice breeding programs was slow in the last decades. Although great progress in rice genomics and molecular biology has been achieved, the effect of such technological innovations on rice breeding is far small. Marker-assisted selection (MAS) for a few target quantitative trait loci (QTLs) has significant effects in improving qualitative traits, such as disease resistance. The success of MAS has therefore motivated breeders to identify and use major QTLs for yield and yield component traits. In this review, we summarized the recent methods in QTL identification, including novel statistical methods for linkage and association mapping, special population types, and whole-genome sequencing. We reviewed the successful application of marker-assisted gene introgression and gene pyramiding to improve grain yield and discussed the design of efficient MAS schemes to further increase the success rate of breeding programs. The use of well-characterized major QTLs through introgression and gene pyramiding is proven effective in improving grain yield, particularly yield under abiotic stress. Major QTLs that are stable across genetic background and growing environments are often found in less adapted germplasms, such as landraces and wild relatives. Advanced backcross QTL analysis and introgression lines, which integrate QTL discovery and utilization, are important methods for exploiting major QTLs contained in such germplasms. Nextgeneration sequencing substantially increases mapping resolution and accelerates the identification of casual genes underlying major QTLs. Practical guidelines derived from theoretical and empirical studies are given to guide the design of efficient marker-assisted gene introgression and pyramiding schemes.

Identification of quantitative trait loci contributing to yield and seed quality parameters under terminal drought in barley advanced backcross lines

Drought is a major limiting factor for barley production, especially in the primary areas of its cultivation. Wild barley represents a major source of favourable alleles for increasing the genetic variation for multiple traits including resistance to both biotic and abiotic stresses. We used advanced backcross quantitative trait locus (AB-QTL) analysis of a BC3-doubled haploid population developed between the cultivated parent Brenda (Hordeum vulgare ssp. vulgare) and the wild accession HS584 (H. vulgare ssp. spontaneum) to study the contribution of wild barley in improving various agronomic and seed quality traits under post-anthesis drought. The experiment was carried out at two different locations (IPK, Gatersleben and Nordsaat, Bohnshausen) and terminal drought was imposed by withholding water or spraying with potassium iodide at 10 days after flowering under greenhouse or field conditions, respectively. QTL analysis indicated that wild barley contributed favourably to most of the traits studied under both control and drought conditions. A total of seven hot-spot QTL regions with co-localizing QTL for various traits harboured more than 80 % of the stable QTL detected in the present study. For yield and thousand-grain weight and their respective drought tolerance indices, most of the QTL were derived from Brenda. On the other hand, for traits like seed length and seed nitrogen content, all the QTL were contributed by HS584, the parent having higher trait value. A significantly reduced carbon/nitrogen (C/N) ratio in the selected contrasting inferior lines compared to superior ones suggests that C/N ratio could be a potential parameter for screening not just seed quality parameters but also grain weight performance under terminal drought.

AB-QTL analysis reveals new alleles associated to proline accumulation and leaf wilting under drought stress conditions in barley (Hordeum vulgare L.).

BackgroundLand plants have evolved several measures to maintain their life against abiotic stresses. The accumulation of proline is the most generalized response of plants under drought, heat or salt stress conditions. It is known as an osmoprotectant which also acts as an instant source of energy during drought recovery process. But, both its role and genetic inheritance are poorly understood in agriculture crops. In the present work, advanced backcross quantitative trait locus (AB-QTL) analysis was performed to elucidate genetic mechanisms controlling proline accumulation and leaf wilting in barley under drought stress conditions.ResultsThe analysis revealed eight QTL associated to proline content (PC) and leaf wilting (WS). QTL for PC were localized on chromosome 3H, 4H, 5H and 6H. The strongest QTL effect QPC.S42.5H was detected on chromosome 5H where drought inducible exotic allele was associated to increase PC by 54%. QTL effects QPC.S42.3H, QPC.S42.4H and QPC.S42.6H were responsible to heighten PC due to the preeminence of elite alleles over the exotic alleles which ranged from 26% to 43%. For WS, QTL have been localized on chromosome 1H, 2H, 3H and 4H. Among these, QWS.S42.1H and QWS.S42.4H were associated to decrease in WS due to the introgression of exotic alleles. In addition, two digenic epistatic interaction effects were detected for WS where the additive effect of exotic alleles imparted a favorable increase in the trait value.ConclusionsThe present data represents a first report on whole-genome mapping of proline accumulation and leaf wilting in barley. The detected QTL are linked to new alleles from both cultivated and wild accessions which bring out an initial insight on the genetic inheritance of PC and WS. These QTL alleles are fixed in the isogenic background of Scarlett, which will allow for positional cloning of underlying genes and to develop drought resilient barley cultivars.

Use of the advanced backcross-QTL method to transfer seed mineral accumulation nutrition traits from wild to Andean cultivated common beans

Iron deficiency anemia and zinc deficiency are major health concerns across the world and can be addressed by biofortification breeding of higher mineral concentration in staple crops, such as common bean. Wild common beans have for the most part had higher average seed mineral concentration than cultivars of this species but have small un-commercial seeds. A logical approach for the transfer of the seed mineral trait from wild beans to cultivated beans is through the advanced backcross breeding approach. The goal of this study was to analyze a population of 138 BC(2)F(3:5) introgression lines derived from the very high iron wild genotype G10022 backcrossed into the genetic background of the commercial-type variety 'Cerinza', a large-red seeded bush bean cultivar of the Andean genepool. In addition to measuring seed mineral accumulation traits and the quantitative trait loci (QTL) controlling these traits we were interested in simultaneously testing the adaptation of the introgression lines in two replicated yield trials. We found the cross to have high polymorphism and constructed an anchored microsatellite map for the population that was 1,554-cM long and covered all 11 linkage groups of the common bean genome. Through composite interval mapping (CIM) and single point analysis (SPA), we identified associations of markers and mineral traits on b01, b06, b07, b08, b10 and b11 for seed iron concentration, and markers on b01, b04 and b10 for seed zinc concentration. The b07 and b08 QTL aligned with previous QTL for iron concentration. A large number of QTL were found for seed weight (9 with CIM and 36 with SPA analysis) and correlations between seed size and mineral content affected the identification of iron and zinc contents' QTL on many linkage groups. Segregation distortion around domestication genes made some areas difficult to introgress. However, in conclusion, the advanced backcross program produced some introgression lines with high mineral accumulation traits using a wild donor parent.