Multi-parent Advanced Generation Inter-cross Population Research Articles

Loci discovery, network-guided approach, and genomic prediction for drought tolerance index in a multi-parent advanced generation intercross (MAGIC) cowpea population

Cowpea is a nutrient-dense legume that significantly contributes to the population’s diet in sub-Saharan Africa and other regions of the world. Improving cowpea cultivars to be more resilient to abiotic stress such as drought would be of great importance. The use of a multi-parent advanced generation intercross (MAGIC) population has been shown to be efficient in increasing the frequency of rare alleles that could be associated with important agricultural traits. In addition, drought tolerance index has been reported to be a reliable parameter for assessing crop tolerance to water-deficit conditions. Therefore, the objectives of this study were to evaluate the drought tolerance index for plant growth habit, plant maturity, flowering time, 100-seed weight, and grain yield in a MAGIC cowpea population, to conduct genome-wide association study (GWAS) and identify single nucleotide polymorphism (SNP) markers associated with the drought tolerance indices, to investigate the potential relationship existing between the significant loci associated with the drought tolerance indices, and to conduct genomic selection (GS). These analyses were performed using the existing phenotypic and genotypic data published for the MAGIC population which consisted of 305 F8 recombinant inbred lines (RILs) developed at University of California, Riverside. The results indicated that: (1) large variation in drought tolerance indices existed among the cowpea genotypes, (2) a total of 14, 18, 5, 5, and 35 SNPs were associated with plant growth habit change due to drought stress, and drought tolerance indices for maturity, flowering time, 100-seed weight, and grain yield, respectively, (3) the network-guided approach revealed clear interactions between the loci associated with the drought tolerance traits, and (4) the GS accuracy varied from low to moderate. These results could be applied to improve drought tolerance in cowpea through marker-assisted selection (MAS) and genomic selection (GS). To the best of our knowledge, this is the first report on marker loci associated with drought tolerance indices in cowpea.

Genome-Wide Association Study Using a Multiparent Advanced Generation Intercross (MAGIC) Population Identified QTLs and Candidate Genes to Predict Shoot and Grain Zinc Contents in Rice

Zinc (Zn) is an essential trace element for the growth and development of both humans and plants. Increasing the accumulation of Zn in rice grains is important for the world’s nutrition and health. In this study, we used a multiparent advanced generation intercross (MAGIC) population constructed using four parental lines and genotyped using a 55 K rice SNP array to identify QTLs related to Zn2+ concentrations in shoots at the seedling stage and grains at the mature stage. Five QTLs were detected as being associated with shoot Zn2+ concentration at the seedling stage, which explained 3.7–5.7% of the phenotypic variation. Six QTLs were detected as associated with grain Zn2+ concentration at the mature stage, which explained 5.5–8.9% of the phenotypic variation. Among the QTLs, qSZn2-1/qGZn2 and qSZn3/qGZn3 were identified as being associated with both the shoot and grain contents. Based on gene annotation and literature information, 16 candidate genes were chosen in the regions of qSZn1, qSZn2-1/qGZn2, qSZn3/qGZn3, qGZn7, and qGZn8. Analysis of candidate genes through qRT-PCR, complementation assay using the yeast Zn-uptake-deficient double-mutant ZHY3, and sequencing of the four parental lines suggested that LOC_Os02g06010 may play an important role in Zn2+ accumulation in indica rice.

QTL identification for nine seed-related traits in Brassica juncea using a multiparent advanced generation intercross (MAGIC) population

Agronomic traits are usually determined by multiple quantitative trait loci (QTLs) that can have pleiotropic effects. A multiparent advanced generation intercross (MAGIC) population is well suited for genetically analysing the effects of multiple QTLs on the traits of interest because it contains more QTL alleles than a biparental population and can overcome the problem of confounding the population structure of the natural germplasm population. We previously developed the B. juncea MAGIC population, derived from eight B. juncea lines with great diversity in agronomic and quality traits. In this study, we show that the B. juncea MAGIC population is also effective for the evaluation of multiple QTLs for complex agronomic traits in B. juncea. A total of twenty-two QTLs for nine seed-related traits were identified, including one QTL for each oil content, seed number per silique and thousand-seed weight; two QTLs for each acid detergent lignin and neutral detergent fibre; three QTLs for each acid detergent fibre and protein content; four QTLs for the seed maturity time; and five QTLs for the white index. Some of these QTLs overlapped. These results should be helpful for further fine mapping, gene cloning, plant breeding and marker-assisted selection (MAS) in B. juncea.

Genetic Dissection for Maize Forage Digestibility Traits in a Multi-Parent Advanced Generation Intercross (MAGIC) Population

Forage feedstock is the greatest source of energy for livestock. Unfortunately, less than 50% of their fiber content is actually digested and assimilated by the ruminant animals. This recalcitrance is mainly due to the high concentration of plant cell wall material and to the limited digestion of the fiber by the microorganisms. A Genome-Wide Association Study (GWAS) was carried out in order to identify Single Nucleotide Polymorphisms (SNPs) associated with forage digestibility traits in a maize Multi-Parent Advanced Generation Intercross (MAGIC) population. We identified seven SNPs, corresponding to five Quantitative Trait Loci (QTL), associated to digestibility of the organic matter, 11 SNPs, clustered in eight QTLs, associated to Neutral Detergent Fiber (NDF) content and eight SNPs corresponding with four QTL associated with Acid Detergent Fiber (ADF). Candidate genes under the QTL for digestibility of the organic matter could be the ones involved in pectin degradation or phenylpropanoid pathway. Transcription factor genes were also proposed for the fiber QTL identified, in addition to genes induced by oxidative stress, or a gene involved in lignin modifications. Nevertheless, for the improvement of the traits under study, and based on the moderate heritability value and low percentage of the phenotypic variability explained by each QTL, a genomic selection strategy using markers evenly distributed across the whole genome is proposed.

Insights into the Intraspecific Variability of the above and Belowground Emissions of Volatile Organic Compounds in Tomato.

The in-vivo monitoring of volatile organic compound (VOC) emissions is a potential non-invasive tool in plant protection, especially in greenhouse cultivation. We studied VOC production from above and belowground organs of the eight parents of the Multi-Parent Advanced Generation Intercross population (MAGIC) tomato population, which exhibits a high genetic variability, in order to obtain more insight into the variability of constitutive VOC emissions from tomato plants under stress-free conditions. Foliage emissions were composed of terpenes, the majority of which were also stored in the leaves. Foliage emissions were very low, partly light-dependent, and differed significantly among genotypes, both in quantity and quality. Soil with roots emitted VOCs at similar, though more variable, rates than foliage. Soil emissions were characterized by terpenes, oxygenated alkanes, and alkenes and phenolic compounds, only a few of which were found in root extracts at low concentrations. Correlation analyses revealed that several VOCs emitted from foliage or soil are jointly regulated and that above and belowground sources are partially interconnected. With respect to VOC monitoring in tomato crops, our results underline that genetic variability, light-dependent de-novo synthesis, and belowground sources are factors to be considered for successful use in crop monitoring.

Evaluation of the Suitability of Some Cowpea Genotype for Maize-Cowpea Intercrop in Northern Ghana

Council for Scientific and Industrial Research-Savanna Agricultural Research Institute (CSIR-SARI) in collaboration with University of California, Riverside phenotyped 300 Recombinant Inbred Lines (RILs) of Multi-parent Advanced Generation Inter-Cross (MAGIC) cowpea population from eight elite cowpea cultivars in Northern Ghana. Among the traits targeted in the phenotyping is extra early maturity suitable for Sudan Savanna agro ecological zone of Ghana. Ten selected extra early genotypes from the MAGIC population were intercropped with maize to identify genotype(s) that can maintain agronomic performances and grain yield. A field experiment was carried out at the Manga Station of SARI, Ghana during the 2018 and 2019 growing season to evaluate the ten extra-early cowpea genotypes in maize/cowpea intercrop. The experimental design used was split plot with three replications. The cropping patterns (row, strip and sole cropping) were assigned to the main plot. Ten cowpea genotypes (MAGIC 008, MAGIC 043, MAGIC 048, MAGIC 055, MAGIC 076, MAGIC 118, MAGIC 154, MAGIC 176, CB27, and SARC 1-57-2) were assigned to sub-plots. The results indicated that the number of seed per pod of the cowpea was not affected by cowpea genotype and intercrop pattern interaction; however, the interaction influenced grain yield, pod per plant, plant height, 50% flowering and 100 seed weight of cowpea. MAGIC genotypes, M008, M048, M055, M154, recorded higher grain yield under both strip intercropping and sole cropping. SARC1-57-2 also recorded the highest grain yield under row intercropping. M048, M055, M076, M176 and SARI’s collection SARC1-57-2 were the top five genotypes in fodder production. Intercropping advantage was compared with sole cropping. Land equivalent ratio greater than 1 was observed for all the genotypes with MAGIC 048 recording the highest LER at strip intercrop. Benefit Cost Ratio also showed that there is advantage of intercropping than sole cropping.

The genetic control of leaf and petal allometric variations in Arabidopsis thaliana

BackgroundOrgan shape and size covariation (allometry) factors are essential concepts for the study of evolution and development. Although ample research has been conducted on organ shape and size, little research has considered the correlated variation of these two traits and quantitatively measured the variation in a common framework. The genetic basis of allometry variation in a single organ or among different organs is also relatively unknown.ResultsA principal component analysis (PCA) of organ landmarks and outlines was conducted and used to quantitatively capture shape and size variation in leaves and petals of multiparent advanced generation intercross (MAGIC) populations of Arabidopsis thaliana. The PCA indicated that size variation was a major component of allometry variation and revealed negatively correlated changes in leaf and petal size. After quantitative trait loci (QTL) mapping, five QTLs for the fourth leaf, 11 QTLs for the seventh leaf, and 12 QTLs for petal size and shape were identified. These QTLs were not identical to those previously identified, with the exception of the ER locus. The allometry model was also used to measure the leaf and petal allometry covariation to investigate the evolution and genetic coordination between homologous organs. In total, 12 QTLs were identified in association with the fourth leaf and petal allometry covariation, and eight QTLs were identified to be associated with the seventh leaf and petal allometry covariation. In these QTL confidence regions, there were important genes associated with cell proliferation and expansion with alleles unique to the maximal effects accession. In addition, the QTLs associated with life-history traits, such as days to bolting, stem length, and rosette leaf number, which were highly coordinated with climate change and local adaption, were QTL mapped and showed an overlap with leaf and petal allometry, which explained the genetic basis for their correlation.ConclusionsThis study explored the genetic basis for leaf and petal allometry and their interaction, which may provide important information for investigating the correlated variation and evolution of organ shape and size in Arabidopsis.

Stability and transferability assessment of the cotton fiber strength QTL qFS-c7-1 on chromosome A07

Previously we identified a major cotton fiber strength QTL (qFS-c7-1) on chromosome A07 using a multi-parent advanced generation intercross (MAGIC) population. To assess the stability and transferability of this QTL and its utility in cotton breeding, we made ten new populations. These populations were developed from crosses between MAGIC recombinant inbred lines, or between cotton cultivars that are different from the MAGIC parents. A total of 2801 F2 plants were grown and their fiber quality traits were measured. We also selected a subset of F3 seeds from two populations, and grew F3 progeny plots to further evaluate the stability of this QTL. Our results showed that the peak of qFS-c7-1 is at 70–72 Mb region. This QTL had a major effect on fiber strength explaining 21.9% phenotypic variance. Its effect on other fiber quality attributes such as micronaire, short fiber content, length and uniformity varied between populations, and no effect on fiber elongation was observed. The QTL effects were stable in the populations analyzed, and in different generations of the same population. The SSR and SNP markers near and within the QTL peak reported herein will assist selecting superior fiber quality traits in breeding, with a recommendation that the parental cotton lines should be analyzed using the seven DNA markers within the QTL peak before fully implementing marker assisted selection in a cotton breeding program.

Genetic mapping for agronomic traits in a MAGIC population of common bean (Phaseolus vulgaris L.) under drought conditions

BackgroundCommon bean is an important staple crop in the tropics of Africa, Asia and the Americas. Particularly smallholder farmers rely on bean as a source for calories, protein and micronutrients. Drought is a major production constraint for common bean, a situation that will be aggravated with current climate change scenarios. In this context, new tools designed to understand the genetic basis governing the phenotypic responses to abiotic stress are required to improve transfer of desirable traits into cultivated beans.ResultsA multiparent advanced generation intercross (MAGIC) population of common bean was generated from eight Mesoamerican breeding lines representing the phenotypic and genotypic diversity of the CIAT Mesoamerican breeding program. This population was assessed under drought conditions in two field trials for yield, 100 seed weight, iron and zinc accumulation, phenology and pod harvest index.Transgressive segregation was observed for most of these traits. Yield was positively correlated with yield components and pod harvest index (PHI), and negative correlations were found with phenology traits and micromineral contents. Founder haplotypes in the population were identified using Genotyping by Sequencing (GBS). No major population structure was observed in the population. Whole Genome Sequencing (WGS) data from the founder lines was used to impute genotyping data for GWAS. Genetic mapping was carried out with two methods, using association mapping with GWAS, and linkage mapping with haplotype-based interval screening. Thirteen high confidence QTL were identified using both methods and several QTL hotspots were found controlling multiple traits. A major QTL hotspot located on chromosome Pv01 for phenology traits and yield was identified. Further hotspots affecting several traits were observed on chromosomes Pv03 and Pv08. A major QTL for seed Fe content was contributed by MIB778, the founder line with highest micromineral accumulation. Based on imputed WGS data, candidate genes are reported for the identified major QTL, and sequence changes were identified that could cause the phenotypic variation.ConclusionsThis work demonstrates the importance of this common bean MAGIC population for genetic mapping of agronomic traits, to identify trait associations for molecular breeding tool design and as a new genetic resource for the bean research community.

Morphoagronomic characterization and whole-genome resequencing of eight highly diverse wild and weedy S. pimpinellifolium and S. lycopersicum var. cerasiforme accessions used for the first interspecific tomato MAGIC population

The wild Solanum pimpinellifolium (SP) and the weedy S. lycopersicum var. cerasiforme (SLC) are largely unexploited genetic reservoirs easily accessible to breeders, as they are fully cross-compatible with cultivated tomato (S. lycopersicum var. lycopersicum). We performed a comprehensive morphological and genomic characterization of four wild SP and four weedy SLC accessions, selected to maximize the range of variation of both taxa. These eight accessions are the founders of the first tomato interspecific multi-parent advanced generation inter-cross (MAGIC) population. The morphoagronomic characterization was carried out with 39 descriptors to assess plant, inflorescence, fruit and agronomic traits, revealing the broad range of diversity captured. Part of the morphological variation observed in SP was likely associated to the adaptation of the accessions to different environments, while in the case of SLC to both human activity and adaptation to the environment. Whole-genome resequencing of the eight accessions revealed over 12 million variants, ranging from 1.2 to 1.9 million variants in SLC and from 3.1 to 4.8 million in SP, being 46.3% of them (4,897,803) private variants. The genetic principal component analysis also confirmed the high diversity of SP and the complex evolutionary history of SLC. This was also reflected in the analysis of the potential footprint of common ancestors or old introgressions identified within and between the two taxa. The functional characterization of the variants revealed a significative enrichment of GO terms related to changes in cell walls that would have been negatively selected during domestication and breeding. The comprehensive morphoagronomic and genetic characterization of these accessions will be of great relevance for the genetic analysis of the first interspecific MAGIC population of tomato and provides valuable knowledge and tools to the tomato community for genetic and genomic studies and for breeding purposes.

MAGIC - Multi-parent Advanced Generation Inter-Cross

The development of Multi-parent Advanced Generation Inter-Cross populations (MAGIC) in rice and discusses potential applications for mapping quantitative trait loci (QTLs) and for rice varietal development. The 4 multi-parent populations: indica MAGIC (8 indica parents); MAGIC plus (8 indica parents with two additional rounds of 8-way F1 inter-crossing); japonica MAGIC (8 japonica parents); and Global MAGIC (16 parents – 8 indica and 8 japonica). The parents used in creating these populations are improved varieties with desirable traits for biotic and abiotic stress tolerance, yield, and grain quality. The purpose is to fine map QTLs for multiple traits and to directly and indirectly use the highly recombined lines in breeding programs. These MAGIC populations provide a useful germplasm resource with diverse allelic combinations to be exploited by the rice community.

GWAS reveals consistent QTL for drought and salt tolerance in a MAGIC population of 550 lines derived from intermating of 11 Upland cotton (Gossypium hirsutum) parents.

Cotton is grown in arid and semi-arid regions where abiotic stresses such as drought and salt are prevalent. There is a lack of studies that simultaneously address the genetic and genomic basis of tolerance to drought and salt stress. In this study, a multi-parent advanced generation inter-cross (MAGIC) population of 550 recombinant inbred lines (RILs) together with their 11 Upland cotton parents with a total of 473,516 polymorphic SNP markers was used to identify quantitative trait loci (QTL) for drought tolerance (DT) and salt tolerance (ST) at the seedling stage based on two replicated greenhouse tests. Transgressive segregation occurred in the MAGIC-RILs, indicating that tolerant and sensitive alleles recombined for tolerance to the abiotic stress during the intermating process for the population development. A total of 20 QTL were detected for DT including 13 and 7 QTL based on plant height (PH) and dry shoot weight (DSW), respectively; and 23 QTL were detected for ST including 12 and 11 QTL for PH and DSW, respectively. There were several chromosomes with QTL clusters for abiotic stress tolerance including four QTL on chromosome A13 and three QTL on A01 for DT, and four QTL on D08 and three QTL on A11 for ST. Nine QTL (21% of the 43 QTL) detected were in common between DT and ST, indicating a common genetic basis for DT and ST. The narrow chromosomal regions for most of the QTL detected in this study allowed identification of 53 candidate genes associated with responses to salt and drought stress and abiotic stimulus. The QTL identified for both DT and ST have significantly augmented the repertoire of QTL for abiotic stress tolerance that can be used for marker-assisted selection to develop cultivars with resilience to drought and/or salt and further genomic studies towards the identification of drought and salt tolerance genes in cotton.

Identification and cross-validation of genetic loci conferring resistance to Septoria nodorum blotch using a German multi-founder winter wheat population

Key messageWe identified allelic variation at two major loci, QSnb.nmbu-2A.1 and QSnb.nmbu-5A.1, showing consistent and additive effects on SNB field resistance. Validation of QSnb.nmbu-2A.1 across genetic backgrounds further highlights its usefulness for marker-assisted selection.Septoria nodorum blotch (SNB) is a disease of wheat (Triticum aestivum and T. durum) caused by the necrotrophic fungal pathogen Parastagonospora nodorum. SNB resistance is a typical quantitative trait, controlled by multiple quantitative trait loci (QTL) of minor effect. To achieve increased plant resistance, selection for resistance alleles and/or selection against susceptibility alleles must be undertaken. Here, we performed genetic analysis of SNB resistance using an eight-founder German Multiparent Advanced Generation Inter-Cross (MAGIC) population, termed BMWpop. Field trials and greenhouse testing were conducted over three seasons in Norway, with genetic analysis identifying ten SNB resistance QTL. Of these, two QTL were identified over two seasons: QSnb.nmbu-2A.1 on chromosome 2A and QSnb.nmbu-5A.1 on chromosome 5A. The chromosome 2A BMWpop QTL co-located with a robust SNB resistance QTL recently identified in an independent eight-founder MAGIC population constructed using varieties released in the United Kingdom (UK). The validation of this SNB resistance QTL in two independent multi-founder mapping populations, regardless of the differences in genetic background and agricultural environment, highlights the value of this locus in SNB resistance breeding. The second robust QTL identified in the BMWpop, QSnb.nmbu-5A.1, was not identified in the UK MAGIC population. Combining resistance alleles at both loci resulted in additive effects on SNB resistance. Therefore, using marker assisted selection to combine resistance alleles is a promising strategy for improving SNB resistance in wheat breeding. Indeed, the multi-locus haplotypes determined in this study provide markers for efficient tracking of these beneficial alleles in future wheat genetics and breeding activities.

Application of the Cottonscope for determining fiber maturity and fineness of an upland cotton MAGIC population

AbstractFineness and maturity are two cotton (Gossypium spp.) fiber properties that respectively estimate the fiber diameter and degree of internal fiber thickening. Cotton researchers have been searching for accurate and efficient ways of measuring them due to their important roles in lint yield, fiber quality, and downstream textile performance. The recent development of the Cottonscope has offered an improved way of quantitatively assessing fineness and maturity based on the tests with reference cotton materials composed of clean and homogenous fibers. However, cotton geneticists and breeders have not widely adopted the new technology, since it has not been tested with a genetic population consisting of a large number of fiber samples with high levels of variations within a sample and among replicates. Thus, we measured a multiparent advanced generation intercross (MAGIC) population using the Cottonscope in comparison with a conventional Advanced Fiber Information System (AFIS). Comparative analyses showed a correlation of the fineness values between AFIS and Cottonscope, but a discrepancy in the maturity ratio values between them. Extensive phenotypic analyses suggested that Cottonscope technology using snippets and a normalization with a fiber width might help improve the accuracy of maturity from various fiber samples of the MAGIC population. The information may help cotton scientists measuring and interpreting fiber maturity from experimental materials for genetic and genomic analyses.

QTL mapping of adult plant and seedling resistance to leaf rust (Puccinia triticina Eriks.) in a multiparent advanced generation intercross (MAGIC) wheat population

Key messageThe Bavarian MAGIC Wheat population, comprising 394 F6:8 recombinant inbred lines was phenotyped for Puccinia triticina resistance in multi-years’ field trials at three locations and in a controlled environment seedling test. Simple intervall mapping revealed 19 QTL, corresponding to 11 distinct chromosomal regions.The biotrophic rust fungus Puccinia triticina is one of the most important wheat pathogens with the potential to cause yield losses up to 70%. Growing resistant cultivars is the most cost-effective and environmentally friendly way to encounter this problem. The emergence of leaf rust races being virulent against common resistance genes increases the demand for wheat varieties with novel resistances. In the past decade, the use of complex experimental populations, like multiparent advanced generation intercross (MAGIC) populations, has risen and offers great advantages for mapping resistances. The genetic diversity of multiple parents, which has been recombined over several generations, leads to a broad phenotypic diversity, suitable for high-resolution mapping of quantitative traits. In this study, interval mapping was performed to map quantitative trait loci (QTL) for leaf rust resistance in the Bavarian MAGIC Wheat population, comprising 394 F6:8 recombinant inbred lines (RILs). Phenotypic evaluation of the RILs for adult plant resistance was carried out in field trials at three locations and two years, as well as in a controlled-environment seedling inoculation test. In total, interval mapping revealed 19 QTL, which corresponded to 11 distinct chromosomal regions controlling leaf rust resistance. Six of these regions may represent putative new QTL. Due to the elite parental material, RILs identified to be resistant to leaf rust can be easily introduced in breeding programs.

The Dawn of the Age of Multi-Parent MAGIC Populations in Plant Breeding: Novel Powerful Next-Generation Resources for Genetic Analysis and Selection of Recombinant Elite Material.

The compelling need to increase global agricultural production requires new breeding approaches that facilitate exploiting the diversity available in the plant genetic resources. Multi-parent advanced generation inter-cross (MAGIC) populations are large sets of recombinant inbred lines (RILs) that are a genetic mosaic of multiple founder parents. MAGIC populations display emerging features over experimental bi-parental and germplasm populations in combining significant levels of genetic recombination, a lack of genetic structure, and high genetic and phenotypic diversity. The development of MAGIC populations can be performed using “funnel” or “diallel” cross-designs, which are of great relevance choosing appropriate parents and defining optimal population sizes. Significant advances in specific software development are facilitating the genetic analysis of the complex genetic constitutions of MAGIC populations. Despite the complexity and the resources required in their development, due to their potential and interest for breeding, the number of MAGIC populations available and under development is continuously growing, with 45 MAGIC populations in different crops being reported here. Though cereals are by far the crop group where more MAGIC populations have been developed, MAGIC populations have also started to become available in other crop groups. The results obtained so far demonstrate that MAGIC populations are a very powerful tool for the dissection of complex traits, as well as a resource for the selection of recombinant elite breeding material and cultivars. In addition, some new MAGIC approaches that can make significant contributions to breeding, such as the development of inter-specific MAGIC populations, the development of MAGIC-like populations in crops where pure lines are not available, and the establishment of strategies for the straightforward incorporation of MAGIC materials in breeding pipelines, have barely been explored. The evidence that is already available indicates that MAGIC populations will play a major role in the coming years in allowing for impressive gains in plant breeding for developing new generations of dramatically improved cultivars.

Assessing diversity in canopy architecture, photosynthesis, and water-use efficiency in a cowpea magic population.

Optimizing crops to improve light absorption and CO2 assimilation throughout the canopy is a proposed strategy to increase yield and meet the needs of a growing population by 2050. Globally, the greatest population increase is expected to occur in Sub‐Saharan Africa where large yield gaps currently persist; therefore, it is crucial to develop high‐yielding crops adapted to this region. In this study, we screened 50 cowpea (Vigna unguiculata (L.) Walp) genotypes from the multi‐parent advanced generation inter‐cross (MAGIC) population for canopy architectural traits, canopy photosynthesis, and water‐use efficiency using a canopy gas exchange chamber in order to improve our understanding of the relationships among those traits. Canopy architecture contributed to 38.6% of the variance observed in canopy photosynthesis. The results suggest that the light environment within the canopy was a limiting factor for canopy CO2 assimilation. Traits favoring greater exposure of leaf area to light such as the width of the canopy relative to the total leaf area were associated with greater canopy photosynthesis, especially in canopies with high biomass. Canopy water‐use efficiency was highly determined by canopy photosynthetic activity and therefore canopy architecture, which indicates that optimizing the canopy will also contribute to improving canopy water‐use efficiency. We discuss different breeding strategies for future programs aimed at the improvement of cowpea yield for the Sub‐Saharan African region. We show that breeding for high biomass will not optimize canopy CO2 assimilation and suggest that selection should include multiple canopy traits to improve light penetration.

Identification of novel genetic sources for agronomic and quality traits in soybean using multi-trait allele specific genic marker assays

Natural allelic variation among the genes governing various agronomic and quality traits of plant provides opportunity to develop desirable plant type in crops. In soybean [Glycine max (L.) Merr.], natural allelic variation is known for many functionally characterized genes conferring superior phenotype for their respective governing trait. Exploring this natural allelic variation in germplasm pool through gene based allele specific multiplex marker genotyping is one way to identify and combine the alleles of multiple traits in desired genetic background. Using known alleles of 15 genes governing 10 different agronomic and quality traits, we have developed two multi-trait allele specific genic marker assays of 22-plex and 7-plex alleles. These two multi-trait allele specific genic marker assays were used to genotype a large set of diverse soybean germplasm accessions, which identified several novel genetic sources of rare alleles conferring superior or desirable phenotype. Selected genetic sources of rare alleles for days to flowering and maturity, hard-seededness, pod shattering and oleic acid content were validated phenotypically. Genotyping and phenotyping in a multi-parent advanced generation intercross population validated the utility of multi-trait allele specific genic marker assays in marker assisted selection of multiple traits from multiple parents. The novel genetic sources identified in this study will help in the selection of parents with multiple desirable traits for hybridization. The multi-trait allele specific genic marker assays developed will speed up selection of multiple traits in routine soybean breeding programmes for cultivar development and improvement.

QTL identification for salt tolerance related traits at the seedling stage in indica rice using a multi-parent advanced generation intercross (MAGIC) population

Salinity is one of the most important abiotic stresses, which seriously affects rice production. In this study, a multiparent advanced generation intercross population, DC1, derived from intercrossing four elite indica varieties, was used to identify QTLs conferring salt tolerance. The whole population and four parents were genotyped with a 55K rice SNP array. A total of 7 QTLs delineated from 186 significant marker-trait associations were detected on chromosomes 1, 2, 5 and 9, which accounted for 7.42–9.38% of the total phenotypic variations. Among these QTLs, one novel QTL (qRRL2) on chromosome 2 for relative root length and one multi-trait QTL (qSLST1/qRDSW1/qRB1) on chromosome 1 affecting shoot length, root dry weight and root biomass under salt treatment were detected. Gene expression analysis revealed that a transcription factor gene (LOC_Os01g66280) within the multi-trait QTL is the potential candidate gene for salt tolerance. Interestingly, we identified one significant nonsynonymous SNP in the coding region of this candidate gene. These results will facilitate fine mapping of the candidate gene and QTL pyramiding to genetically improve salt tolerance in rice.

Resistance and susceptibility QTL identified in a rice MAGIC population by screening with a minor-effect virulence factor from Xanthomonas oryzae pv. oryzae.

SummaryEffective and durable disease resistance for bacterial blight (BB) of rice is a continuous challenge due to the evolution and adaptation of the pathogen, Xanthomonas oryzae pv. oryzae (Xoo), on cultivated rice varieties. Fundamental to this pathogens’ virulence is transcription activator‐like (TAL) effectors that activate transcription of host genes and contribute differently to pathogen virulence, fitness or both. Host plant resistance is predicted to be more durable if directed at strategic virulence factors that impact both pathogen virulence and fitness. We characterized Tal7b, a minor‐effect virulence factor that contributes incrementally to pathogen virulence in rice, is a fitness factor to the pathogen and is widely present in geographically diverse strains of Xoo. To identify sources of resistance to this conserved effector, we used a highly virulent strain carrying a plasmid borne copy of Tal7b to screen an indica multi‐parent advanced generation inter‐cross (MAGIC) population. Of 18 QTL revealed by genome‐wide association studies and interval mapping analysis, six were specific to Tal7b (qBB‐tal7b). Overall, 150 predicted Tal7b gene targets overlapped with qBB‐tal7b QTL. Of these, 21 showed polymorphisms in the predicted effector binding element (EBE) site and 23 lost the EBE sequence altogether. Inoculation and bioinformatics studies suggest that the Tal7b target in one of the Tal7b‐specific QTL, qBB‐tal7b‐8, is a disease susceptibility gene and that the resistance mechanism for this locus may be through loss of susceptibility. Our work demonstrates that minor‐effect virulence factors significantly contribute to disease and provide a potential new approach to identify effective disease resistance.