Root‐lodging resistance in maize as an example for high‐throughput genetic mapping via single nucleotide polymorphism‐based selective genotyping

Large‐scale selective genotyping and high‐throughput analysis are two important strategies for low‐cost and high‐effective genetic mapping. In this study, selective genotyping was applied to four maize F₂ populations. Thirty plants were selected from each of the two tails of the original F₂ populations to represent extreme resistant and susceptible plants to root lodging, and genotyped individually with 1536 single nucleotide polymorphisms (SNPs). A quantitative trait locus (QTL) was declared when at least three closely linked SNPs showed significant allele frequency difference between the two tails. Nine QTL were identified for root lodging across the four populations, which were located on chromosomes 2, 4, 5, 7, 8 and 10 and one of them was shared between two populations. A total of 20 segregation distortion regions (SDRs) were identified across the four populations, one of which was co‐localized with a QTL on chromosome 4. The tightly linked SNPs identified in this study can be used for marker‐assisted selection for root lodging. Selective genotyping, when combined with pooled DNA analysis, can be used to develop strategies for high‐throughput genetic mapping for all crops.     

Marker‐assisted backcross breeding to combine multiple rust resistance in wheat

A widely grown but rust susceptible Indian wheat variety HD2932 was improved for multiple rust resistance by marker‐assisted transfer of genes Lr19, Sr26 and Yr10. Foreground and background selection processes were practised to transfer targeted genes with the recovery of the genome of HD2932. The near‐isogenic lines (NILs) of HD2932 carrying Lr19, Sr26 and Yr10 were individually produced from two backcrosses with recurrent parent HD2932. Marker‐assisted background selection of NILs with 94.38–98.46% of the HD2932 genome facilitated rapid recovery of NILs carrying Lr19, Sr26 and Yr10. In the BC₂F₂ generation, NILs were intercrossed and two gene combinations of Lr19+Yr10, Sr26 + Yr10 and Lr19+Sr26 were produced. A total of 16 progeny of two gene combinations of homozygous NILs of HD2932 have been produced, which are under seed increase for facilitating the replacement of the susceptible HD2932 with three of the sixteen improved backcross lines with resistance to multiple rusts.     

Genetic-diversity assessed by microsatellites in tropical maize populations submitted to a high-intensity reciprocal recurrent selection

BR-105 and BR-106 are important tropical maize populations, which were submitted to a high-intensity reciprocal recurrent selection, generating the IG-3 and IG-4synthetics. Using 30-microsatellite loci,we measured and compared the genetic diversity of these populations and their synthetics. The populations did not differ significantly regarding the amount of genetic diversity. As a consequence of selection, genetic variability losses, in terms of mean number of alleles per locus,proportion of polymorphic loci, and gene diversity did occur and were greater in the synthetic IG-3 than IG-4. In the synthetics, the number of loci in adherence to Hardy-Weinberg proportions was superior to that observed in the populations. The Wright's mean fixation index was higher than the mean value expected for outcrossing species (5%) indicating as light excess of homozygotic individuals in both populations. The genetic distances confirmed the favourable effects of one cycle of recurrent selection, as the synthetics became more isolated in comparison to the original populations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Breeding maize lines for complete and partial resistance to maize streak virus (MSV)

Summary S₁ to S₅ inbred lines, derived from a maize population bred for its overall resistance to three tropical viruses, were screened for resistance to maize streak virus (MSV) by artificial plant infection using viruliferous leafhoppers. Symptoms were rated and intra-line frequency distributions studied for all pedigree inbred lines. Mortality due to MSV was very low among these inbreds. Symptoms appeared later, developed slower and were less severe than in the susceptible control hybrid. Results of a study of 500 S₁ and 93 S₂ lines suggested that resistance is under genetic control via a system involving loci with major genes (with dominance for resistance) controlling high to complete resistance, associated with a genetic system involving loci with minor genes controlling partial resistance. Lines expressing complete resistance to MSV were developed from 5 cycles of inbreeding and selection. The relevance of such complete and partial resistance is discussed.Abbreviations MRPS Mean Rating for Plants exhibiting Symptoms     

A genetic linkage map in southern‐by‐spring oat identifies multiple quantitative trait loci for adaptation and rust resistance

We developed 178 recombinant inbred lines from a southern‐by‐spring oat population designated as “TxH.” These lines were genotyped to generate a high‐quality linkage map that resolved 6,902 markers into 21 linkage groups that matched closely with the latest hexaploid oat consensus map. Three major quantitative trait loci (QTLs) affecting heading date were found in locations that are consistent with known QTLs and candidate genes, and two other QTLs affecting heading date were found in novel locations. Five QTLs affecting plant height were found. Both sets of QTLs are responsible for transgressive segregation observed for these two traits. Four QTLs affecting resistance to crown rust, caused by the pathogen Puccinia coronata f. sp. avenae, were identified. Two of these QTLs are consistent with known clusters of rust resistance genes, while two may represent new locations of novel rust resistance genes. A complete set of SNP sequences suitable for generating markers for molecular selection is provided.     

Identification of potential gene‐associated major traits using GBS‐GWAS for Korean apple germplasm collections

Genomewide association study (GWAS), which queries the association between loci and a particular trait by examining single nucleotide polymorphisms (SNPs) of the entire genome, is used in many fields of study. The development of next‐generation sequencing techniques has facilitated GWASs by decreasing the sequencing costs and time. In particular, genotyping by sequencing (GBS) is useful for sequencing many samples simultaneously and at a moderate price. Herein, we describe a potential GWAS using GBS, focused on the apple germplasm, with the goal of developing an effective apple breeding strategy through the identification of useful markers. From 308 Korean apple germplasm, SNPs were selected after GBS, and major traits were investigated. Proprietary individuals were confirmed and grouped by association through genetic diversity and population structure analyses of the selected SNPs. Genes highly associated with the target traits were identified, respectively. As the first GWAS report on the apple germplasm, these results will be useful as base data for GWASs on other apple populations and traits.     

A high‐resolution einkorn (Triticum monococcum L.) linkage map involving wild,domesticated and feral einkorn genotypes

A high‐resolution consensus linkage map of Triticum monococcum was assembled from two separate maps involving domesticated, feral and wild einkorn wheat accessions. The genotyping‐by‐sequencing (GBS) approach based on DArTseq markers yielded overstretched maps. Deleting all markers with missing data and then converting dubious singletons to missing data produced two maps of about 1,380 cM, close to the published genome size. The consensus map spanned 1,562 cM, had one bin mapped every 0.92 cM and showed only one gap > 10 cM. Chromosome length varied between 151 cM (chromosome 4) and 270 cM (chromosome 7). The consensus map was compared to other A‐genome maps, and the sequences of genetically mapped DArTseq were used to anchor contigs of the T. monococcum, T. urartu and T. aestivum draft genomes based on sequence homology to assess colinearity and to assign mapped markers to the seven chromosomes of the bread wheat A‐genome. Finally, an in silico functional characterization of the sequences was performed. This high‐resolution map will facilitate QTL and association analysis and assist the genome assembly of the einkorn genome.     

Identification of pre‐ and posthaustorial resistance to rust (Uromyces viciae‐fabae) in lentil (Lens culinaris) germplasm

Lentil (Lens culinaris Medik.) is an important cool‐season legume that can be severely damaged by rust (Uromyces viciae‐fabae (Pers.) Schröt.) in many parts of the world. In this work, a collection of L. culinaris germplasm accessions was screened under growth chamber conditions for response to U. viciae‐fabae. Several sources of incomplete resistance have been identified, both of hypersensitive and of non‐hypersensitive nature. Incomplete hypersensitive resistance was due to a late‐acting host cell necrosis that resulted in an intermediate infection type (IT). Incomplete non‐hypersensitive resistance was due to a hampered cell penetration not associated with host cell necrosis, reducing the number of haustoria per colony and therefore hindering the growth of the colony and reducing and retarding disease development in spite of a high IT.     

Genes for resistance to northern corn leaf blight in diverse maize populations

Turcicum or northern corn leaf blight (NCLB) incited by the ascomycete Setosphaeria turcica, anamorph Exserohilum turcicum, is a ubiquitous foliar disease of maize. Diverse sources of qualitative and quantitative resistance are available but qualitative resistances (Ht genes) are often unstable. In the tropics especially, they are either overcome by new virulent races or they suffer from climatically sensitive expression. Quantitative resistance is expressed independently of the physical environment and has never succumbed to S. turcica pathotypes in the field. This review emphasizes the identification and mapping of genes related to quantitative NCLB resistance. We deal with the consistency of the genomic positions of quantitative trait loci (QTL) controlling resistance across different maize populations, and with the clustering of genes for resistance to S. turcica and other fungal pathogens or insect pests in the maize genome. Implications from these findings for further genomic research and resistance breeding are drawn. Incubation period (IP) and area under the disease progress curve (AUDPC), based on multiple disease ratings, are important component traits of quantitative NCLB resistance. They are generally tightly correlated (r_p? 0.8) and highly heritable (h²? 0.75). QTL for resistance to NCLB (IP and AUDPC) were identified and characterized in three mapping populations (A, B, C). Population A, a set of 121‐150 F₃ families of the cross B52×mo17, represented US Corn Belt germplasm with a moderate level of resistance. It was field‐tested in Iowa, USA, and Kenya, and genotyped at 112 restriction fragment length polymorphism (RFLP) loci. Population B consisted of 194‐256 F₃ families of the cross Lo951×CML202, the first parent being a Corn‐Belt‐derived European inbred line and the second parent being a highly resistant tropical African inbred line. The population was also tested in Kenya and genotyped with 110 RFLP markers. Population C was derived from a cross between two early‐maturing European inbred lines, D32 and D145, both having a moderate level of resistance. A total of 220 F₃ families were tested in Switzerland and characterized with 87 RFLP and seven SSR markers. In each of the three studies, 12‐13 QTL were detected by composite interval mapping at a signifcance threshold of LOD=2.5. The phenotypic and the genotypic variance were explained to an extent of 50‐70% and 60‐80%, respectively. Gene action was additive to partly dominant, as in previous generation means and combining ability analyses with other genetic material. In each population, gene effects of the QTL were of similar magnitude and no putative major genes were discovered. QTL for AUDPC were located on chromosomes 1 to 9. All three populations carried QTL in identical genomic regions on chromosomes 3 (bin 3.06/07), 5 (bin 3.06/07) and 8 (bin 8.05/06). The major genes Ht2 and Htn1 were also mapped to bins 8.05 and 8.06, suggesting the presence of a cluster of closely linked major and minor genes. The chromosomal bins 3.05, 5.04 and 8.05, or adjacent intervals, were further associated with QTL and major genes for resistance to eight other fungal diseases and insect pests of maize. Bins 1.05/07 and 9.05 were found to carry population‐specifc genes for resistance to S. turcica and other organisms. Several disease lesion mimic mutations, resistance gene analogues and genes encoding pathogenesis‐related proteins were mapped to regions harbouring NCLB resistance QTL.     

Comparison of Bt maize hybrids with their non-transgenic counterparts and commercial varieties for resistance to European corn borer and for agronomic traits

The European corn borer (ECB), Ostrinia nubilalis (Hübner), is a major pest of maize (Zea mays L.) in Central Europe. In order to compare transgenic Bt maize hybrids with their non‐transgenic counterparts and commercial hybrids, field trials and a laboratory bioassay were conducted. The field experiments were performed at four locations with natural and manual infestation of ECB larvae in 1998 and 1999. Transgenic Bt hybrids showed significantly lower means than their corresponding non‐transgenic counterparts and commercial hybrids for all resistance traits (damage rating of stalks, number of larvae per plant, and percentage of damaged plants or ears under infestation). Bt hybrids containing the CryIA(b) gene under the control of green tissue and pollen‐specific promoters (event 176) showed a significantly higher percentage of damaged ears than Bt hybrids carrying the CryIA(b) gene under the control of a constitutive promoter (Mon810). Bt and non‐Bt hybrids showed no significant differences for all agronomic traits, except for plant height under insecticide protection and grain yield reduction under infestation, whereas Bt hybrids had significantly lower means than their non‐transgenic counterparts and other commercial hybrids. All resistance traits were significantly correlated with grain yield reduction. The laboratory bioassay confirmed the level of antibiosis of Bt hybrids against neonate ECB larvae. Bt hybrids showed the highest level of ECB resistance and therefore are an attractive method of preventing ECB damage within an integrated pest‐management system.     

QTL mapping of a partial resistance to the corn delphacid‐transmitted viruses in Lepidopteran‐resistant maize line Mp705

A partial resistance to maize mosaic virus (MMV) and maize stripe virus (MStV) was mapped in a RILs population derived from a cross between lines MP705 (resistant) and B73 (susceptible). A genetic map constructed from 131 SSR markers spanned 1399 cM with an average distance of 9.6 cM. A total of 10 QTL were detected for resistance to MMV and MStV, using composite interval mapping. A major QTL explaining 34–41% of the phenotypic variance for early resistance to MMV was detected on chromosome 1. Another major QTL explaining up to 30% of the phenotypic variation for all traits of resistance to MStV was detected in the centromeric region of chromosome 3 (3.05 bin). After adding supplementary SSR markers, this region was found to correspond well to the one where a QTL of resistance to MStV already was located in a previous mapping study using an F₂ population derived from a cross between Rev81 and B73. These results suggested that these QTL of resistance to MStV detected on chromosome 3 could be allelic in maize genome.     

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.     

Marker‐assisted introgression of lpa2 locus responsible for low‐phytic acid trait into an elite tropical maize inbred (Zea mays L.)

Maize is an important food and feed crop worldwide. Phytic acid (PA), in maize kernel, is an antinutritional factor. PA chelates mineral cations and causes mineral deficiency in humans and phosphorous deficiency in animals. The undigested PA excreted by monogastric animals causes phosphorous eutrophication. Therefore, development of low‐phytate maize is indispensable. The low‐phytate locus (lpa2 allele) has been transferred from low‐phytate mutant line ‘EC 659418’ into an elite inbred UMI 395 through marker‐assisted backcross breeding (MABB). The MABB involved three backcrosses followed by two selfing steps, including ‘foreground selection’, that is, selecting lines with lpa2 allele with the help of a codominant SSR marker ‘umc2230’ and ‘background selection’, that is, selecting plants having genetic background similar to that of the recurrent parent using 50 codominant SSR markers. Two low‐phytate lpa2 lines with genome similar (>90% similarity) to that of recurrent parent have been identified. These lines can be used as parent in future hybridization programmes for obtaining low‐phytate high‐yielding maize hybrids.     

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.     

QTL mapping for six ear leaf architecture traits under water‐stressed and well‐watered conditions in maize (Zea mays L.)

Morphological traits for ear leaf are determinant traits influencing plant architecture and drought tolerance in maize. However, the genetic controls of ear leaf architecture traits remain poorly understood under drought stress. Here, we identified 100 quantitative trait loci (QTLs) for leaf angle, leaf orientation value, leaf length, leaf width, leaf size and leaf shape value of ear leaf across four populations under drought‐stressed and unstressed conditions, which explained 0.71%–20.62% of phenotypic variation in single watering condition. Forty‐five of the 100 QTLs were identified under water‐stressed conditions, and 29 stable QTLs (sQTLs) were identified under water‐stressed conditions, which could be useful for the genetic improvement of maize drought tolerance via QTL pyramiding. We further integrated 27 independent QTL studies in a meta‐analysis to identify 21 meta‐QTLs (mQTLs). Then, 24 candidate genes controlling leaf architecture traits coincided with 20 corresponding mQTLs. Thus, new/valuable information on quantitative traits has shed some light on the molecular mechanisms responsible for leaf architecture traits affected by watering conditions. Furthermore, alleles for leaf architecture traits provide useful targets for marker‐assisted selection to generate high‐yielding maize varieties.     

Level of partial resistance to leaf rust,Puccinia hordei,in West-European barley and how to select for it

Summary The partial resistance to leaf rust (Puccinia hordei) of 40 West-European spring barley cultivars was measured in plots isolated from one another to reduce inter plot interference. The leaf area affected by leaf rust was also measured in small plots of 0.5 m² adjacent to each other, and on individual plants. The latent period was measured in the seedling stage and the adult plant stage, the infection frequency in the seedling stage only. The cultivars varied widely for partial resistance, many cultivars carrying a considerable level. Both the small adjacent plots and the single plants showed a marked inter plot interference strongly reducing the difference between cultivars. H wever, the ranking order of the cultivars was hardly, if at all, affected. Both latent period and the infection frequency showed large differences between cultivars, the latent period in the adult plant stage being highly correlated (r=0.82) with partial resistance, infection frequency in the seedling stage only rather weakly (r=–0.33).Selection for partial resistance appeared very effective in all stages tested; the seedling, the single adult plant, and the small plot stage. Selection in the small plot stage was the most effective followed by selection in the seedling stage. Selection for partial resistance therefore appears very well possible at all stages of the selection program.