A molecular linkage map with associated QTLs from a hulless x covered spring oat population.

In spring-type oat ( Avena sativa L.), quantitative trait loci (QTLs) detected in adapted populations may have the greatest potential for improving germplasm via marker-assisted selection. An F(6) recombinant inbred (RI) population was developed from a cross between two Canadian spring oat varieties: 'Terra', a hulless line, and 'Marion', an elite covered-seeded line. A molecular linkage map was generated using 430 AFLP, RFLP, RAPD, SCAR, and phenotypic markers scored on 101 RI lines. This map was refined by selecting a robust set of 124 framework markers that mapped to 35 linkage groups and contained 35 unlinked loci. One hundred one lines grown in up to 13 field environments in Canada and the United States between 1992 and 1997 were evaluated for 16 agronomic, kernel, and chemical composition traits. QTLs were localized using three detection methods with an experiment-wide error rate of approximately 0.05 for each trait. In total, 34 main-effect QTLs affecting the following traits were identified: heading date, plant height, lodging, visual score, grain yield, kernel weight, milling yield, test weight, thin and plump kernels, groat beta-glucan concentration, oil concentration, and protein. Several of these correspond to QTLs in homologous or homoeologous regions reported in other oat QTL studies. Twenty-four QTL-by-environment interactions and three epistatic interactions were also detected. The locus controlling the covered/hulless character ( N1) affected most of the traits measured in this study. Additive QTL models with N1 as a covariate were superior to models based on separate covered and hulless sub-populations. This approach is recommended for other populations segregating for major genes. Marker-trait associations identified in this study have considerable potential for use in marker-assisted selection strategies to improve traits within spring oat breeding programs.

Comparative mapping in grasses. Oat relationships.

The development of RFLP linkage maps in hexaploid and diploid oat allows us to study genetic relationships of these species at the DNA level. In this report, we present the extension of a previously developed diploid oat map (Avena atlantica x A. hirtula) and its molecular-genetic relationships with wheat, rice and maize. Examination of 92-99% of the length of the oat genome map with probes common to Triticeae species, rice or maize showed that 84, 79 and 71%, respectively, was conserved between these species and oat. Generally, the orders of loci among chromosomes homoelogous to oat chromosomes A and D were the most conserved and those of chromosomes homoeologous to oat chromosome G were the least conserved. Conservation was observed for blocks ranging from whole chromosomes 101 cM long to small segments 2.5 cM long containing two loci. Comparison of the homoeologous segments of Triticeae, rice and maize relative to oat indicated that certain regions have been maintained in all four species. The relative positions of major genes governing traits such as seed storage proteins and resistance to leaf rusts have been conserved between cultivated oat and Triticeae species. Also, the locations of three vernalization/or photoperiod response genes identified in hexaploid oat correspond to the locations of similar genes in homoeologous chromosomes of wheat, rice or maize. The locations of the centromeres for six of the seven oat chromosomes were estimated based on the homoeologous segments between oat and Triticeae chromosomes.

A molecular linkage map of cultivated oat.

A molecular linkage map of cultivated oat composed of 561 loci has been developed using 71 recombinant inbred lines from a cross between Avena byzantina cv. Kanota and A. sativa cv. Ogle. The loci are mainly restriction fragment length polymorphisms detected by oat cDNA clones from leaf, endosperm, and root tissue, as well as by barley leaf cDNA clones. The loci form 38 linkage groups ranging in size from 0.0 to 122.1 cM (mean, 39 cM) and consist of 2-51 loci each (mean, 14). Twenty-nine loci remain unlinked. The current map size is 1482 cM and the total size, on the basis of the number of unlinked loci, is estimated to be 2932.0 cM. This indicates that this map covers at least 50% of the cultivated oat genome. Comparisons with an A-genome diploid oat map and between linkage groups exhibiting homoeology to each other indicate that several major chromosomal rearrangements exist in cultivated oat. This map provides a tool for marker-assisted selection, quantitative trait loci analyses, and studies of genome organization in oat.

The identification of random amplified polymorphic DNA markers for daylength insensitivity in oat.

Daylength insensitive accessions of Avena sativa L. are being used to develop cultivars that will flower normally when grown under short or long photoperiods. Field data indicate that the insensitivity trait is under the control of a single dominant gene, designated Di1. The random amplified polymorphic DNA (RAPD) technique and bulk segregant analysis of daylength sensitive and insensitive plants were used to find markers for this gene. Five of 200 random decamer primers tested produced polymorphic bands, which were shown to be linked to the trait using 30 homozygous insensitive and 30 homozygous sensitive F3 individuals. Three of the markers produced a band in the presence of the dominant allele, and two in its absence. Segregation analysis showed that markers 221 and 136 could be mapped to within 9.8 +/- 4.6 and 13.9 +/- 5.4 cM of the trait, respectively; that is, close enough to be useful in a breeding program. A study of different cultivars suggested that the band produced by primer 136 is actually the more closely linked marker and the only one present in the original Di1 gene donor CAV2700. The possibility of using both markers in populations derived from different cultivars is discussed.

Durum wheat haploid production using maize wide-crossing.

While anther culture or pollinations with Hordeum bulbosum have provided suitable methods for haploid production in bread wheat, they have been largely unsuccessful in durum wheat. Pollinations with maize were used in an attempt to produce haploid seedlings and, from these, fertile doubled haploids of durum wheats. Moreover, the effect of various concentrations and combinations of a synthetic auxin, 2, 4-dichlorophenoxyacetic acid (2,4-D), kinetin, and an ethylene inhibitor, silver nitrate (AgNO3), on embryo recovery were also investigated. Haploid seedlings were recovered from Triticum turgidum ssp. turgidum cv 'Rampton Rivet' pollinated with maize following in-vivo treatment of ovaries with 2,4-D for 2 weeks and subsequent embryo culture. The recovery of haploid seedlings from T. turgidum ssp. durum cv. 'Wakona' pollinated with maize necessitated the addition of AgNO3, to the 2,4-D treatment. Overall, haploid seedlings were produced in 1.7% and 3.3% of pollinated florets for "Rampton Rivet" and "Wakona" respectively. The success of the present work represents a significant breakthrough for haploid production in durum wheats. Wide hybridization with maize followed by in-vivo treatment of ovaries with 2,4-D alone, or in combination with AgNO3, may provide a widely-applicable method of haploid production in tetraploid wheats.

Comparative responses of tetraploid wheats pollinated with Zea mays L. and Hordeum bulbosum L.

Ten different tetraploid wheat (Triticum turgidum) genotypes were pollinated with maize (Zea mays). Fertilization was achieved in all ten genotypes and no significant difference in fertilization frequency between the tetraploid wheat genotypes was detected. A mean of 41.1% of pollinated ovaries contained an embryo. All these crosses were characterized by the elimination of the maize chromosomes, and the resulting embryos were haploids. Six of the tetraploid wheat genotypes were also pollinated with Hordeum bulbosum. Fertilization frequencies with H. bulbosum were much lower (mean=13.4%), and significant differences between the tetraploid wheat genotypes were detected. Observation of pollen tube growth revealed that part of the incompatibility reaction between tetraploid wheats and H. bulbosum was due to an effect similar to that of the Kr genes, namely pollen tube growth inhibition. These results indicate that pollinations with maize may have potential as a broad spectrum haploid production system for tetraploid wheats.