Selection and regeneration of toxin-insensitive plants from tissue cultures of oats (Avena sativa) susceptible to Helminthosporium victoriae

Summary Insensitivity to the pathotoxin victorin, which is produced by the fungus Helminthosporium victoriae (Meehan and Murphy), was selected in tissue cultures of oat (Avena sativa L.) lines heterozygous for the dominant sensitive allele Vb. The Vb allele imparts both susceptibility to H. victoriae and resistance to several races of oat crown rust (Puccinia coronata var. avenae, Fraser and E. Led.). None of 84 homozygous Vb Vb oat calli survived when grown on victorin-containing medium. Among 175 calli of heterozygous Vb vb cultures grown on toxin-containing medium, 16 representing 13 separate embryo-derived culture lines produced surviving callus sectors or shoots. Based on leaf bioassays of plants regenerated after toxin selection, nine culture lines gave toxin-insensitive plants and two gave plants showing the toxin sensitivity of the parent. Two selected lines failed to regenerate. Plants regenerated from 30 culture lines which had never been exposed to toxin-containing selection medium were all toxin sensitive. The toxin insensitivity of the regenerants from the toxin-selected culture lines was heritable since progeny of these plants were all insensitive. The toxin-insensitive selected lines all were found to have coincidentally lost the Vb crown rust resistance of the original line. In cytological analysis of meiotic cells of regenerants from the selected cultures, no chromosomal deficiency was found which could be associated with, and thus account for, the loss of sensitivity to the toxin. Somatic recombination and mutation to vb vb are other possible origins of toxin insensitivity in the selections. The victorin selection demonstrates that specific resistance can be selected in tissue cultures of oats. It also provides a highly sensitive scheme to test effects of culture conditions and chemical agents on induction of genetic and chromosomal changes in tissue cultures.Joint contribution of USDA-ARS and the Minnesota and Florida Agric. Exp. Stns. Paper No. 14121, Scientific Journal Series, Minnesota Agric. Exp. Stn     

Mapping of beta-tubulin genomic sequences in hexaploid oat (Arena sativa L.)

Summary The allohexaploid nature of Avena sativa L. (2n=6x=42) and the availability of aneuploid lines was exploited in designing a strategy for mapping beta-tubulin sequences in the oat genome. Evidence for a minimum of eight beta-tubulin genes was obtained by Southern-blot analysis. Three betatubulin sequences were localized to chromosomes using DNA from monosomic and nullisomic lines in the variety Sun II. One sequence was localized to the chromosome missing in nullisome I. Two other sequences were mapped to satellite chromosome 2, the chromosome that is missing in nullisome VIII and to which one ribosomal RNA gene cluster had previously been mapped. Restriction fragments carrying these two beta-tubulin genomic sequences and the cluster of ribosomal RNA sequences were missing in DNA from nullisomics VIII, IX and X, suggesting that all three nullisome classes are deficient for an identical chromosomal segment that includes these three loci. This study demonstrates how molecular analyses can be used to characterize aneuploid stocks and to better define their genetic constitution.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture or the University of Minnesota and does not imply its approval to the exclusion of other products or vendors that may be suitable     

Cytological and molecular characterization of oat x maize partial hybrids

In cereals, interspecific and intergeneric hybridizations (wide crosses) which yield karyotypically stable hybrid plants have been used as starting points to widen the genetic base of a crop and to construct stocks for genetic analysis. Also, uniparental genome elimination in karyotypically unstable hybrids has been utilized for cereal haploid production. We have crossed hexaploid oat (2n=6x=42, Avena sativa L.) and maize (2n=2x=20, Zea mays L.) and recovered 90 progenies through embryo rescue. Fifty-two plants (58%) produced from oatxmaize hybridization were oat haploids (2n=3x=21) following maize chromosome elimination. Twenty-eight plants (31%) were found to be stable partial hybrids with 1–4 maize chromosomes in addition to a haploid set of 21 oat chromosomes (2n=21+1 to 2n=21+4). Ten of the ninety plants produced were found to be apparent chromosomal chimeras, where some tissues in a given plant contained maize chromosomes while other tissues did not, or else different tissues contained a different number of maize chromosomes. DNA restriction fragment length polymorphisms (RFLPs) were used to identify the maize chromosome(s) present in the various oat-maize progenies. Maize chromosomes 2, 3, 4, 5, 6, 7, 8, and 9 were detected in partial hybrids and chromosomal chimeras. Maize chromosomes 1 and 10 were not detected in the plants analyzed to-date. Furthermore, partial self-fertility, which is common in oat haploids, was also observed in some oat-maize hybrids. Upon selfing, partial hybrids with one or two maize chromosomes showed nearly complete transmission of the maize chromosome to give self-fertile maize-chromosome-addition oat plants. Fertile lines were recovered that contained an added maize chromosome or chromosome pair representing six of the ten maize chromosomes. Four independently derived disomic maize chromosome addition lines contained chromosome 4, one line carried chromosome 7, two lines had chromosome 9, one had chromosome 2, and one had chromosome 3. One maize chromosome-8 monosomic addition line was also identified. We also identified a double disomic addition line containing both maize chromosomes 4 and 7. This constitutes the first report of the production of karyotypically stable partial hybrids involving highly unrelated species from two subfamilies of the Gramineae (Pooideae — oat, and Panicoideae — maize) and the subsequent recovery of fertile oat-maize chromosome addition lines. These represent novel material for gene/ marker mapping, maize chromosome manipulation, the study of maize gene expression in oat, and the transfer of maize DNA, genes, or active transposons to oat.Joint contribution of the Minnesota Agricultural Experiment Station and USDA-ARS. Scientific journal series paper No. 21 859 of the Minnesota Agricultural Experiment Station. Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the USDA-ARS or the University of Minnesota and does not imply approval over other products that also may be suitable     

Callus formation and plant regeneration from somatic embryos of oat (Avena sativa L.)

Globular-stage somatic embryos were isolated by vortexing friable, embryogenic callus of oat (Avena sativa L.) followed by fractionation based on size. Somatic embryos were most frequently found in the 300–380 m size fraction. Friable, embryogenic callus was reinitiated from 55% of isolated somatic embryos. Fertile plants were regenerated from 22% of isolated somatic embryos. Reinitiation of callus from somatic embryos and growth of friable, embryogenic callus was inhibited by the selective agents G418 and methotrexate. These results suggest that somatic embryos isolated from friable, embryogenic callus of oat may be useful totipotent targets for particle acceleration-mediated transformation.     

Characterization of the hexaploid oat Avena byzantina cv. Kanota monosomic series using C-banding and RFLPs.

The establishment of a C-banded karyotype of hexaploid oat (Avena spp., 2n = 6x = 42) has facilitated the cytological characterization of a monosomic series in 'Kanota', an A. byzantina (C. Koch) cultivar. The 'Kanota' series of monosomics analyzed in this study consists of only 12 of the 21 different chromosome-deficient lines possible plus potential translocated segments of two or three additional chromosomes. These findings were confirmed by RFLP mapping data from studies in which oat probes were assigned to syntenic groups using the 'Kanota' set of monosomic lines. Among the remaining nine monosomic lines analyzed, eight are missing chromosomes represented in the set of 12 unique lines and one line, monosomic K13, is missing a chromosome from the unique set of 12 that possesses a cytologically detectable translocation. This same translocation, involving chromosomes 7C and 14, is found in 5 of the 21 'Kanota' monosomics. The incompleteness of the set of 'Kanota' monosomics might be due to (i) difficulty in identifying individual oat chromosomes without C-banding, (ii) plant genotypic and phenotypic variability in the original source population of the 'Kanota' monosomics, and (or) (iii) a high frequency of monosomic shifts in progency of the original 'Kanota' monosomic lines.     

Maize centromere mapping: a comparison of physical and genetic strategies

Centromere positions on 7 maize chromosomes were compared on the basis of data from 4 to 6 mapping techniques per chromosome. Centromere positions were first located relative to molecular markers by means of radiation hybrid lines and centric fission lines recovered from oat-maize chromosome addition lines. These centromere positions were then compared with new data from centric fission lines recovered from maize plants, half-tetrad mapping, and fluorescence in situ hybridizations and to data from earlier studies. Surprisingly, the choice of mapping technique was not the critical determining factor. Instead, on 4 chromosomes, results from all techniques were consistent with a single centromere position. On chromosomes 1, 3, and 6, centromere positions were not consistent even in studies using the same technique. The conflicting centromere map positions on chromosomes 1, 3, and 6 could be explained by pericentric inversions or alternative centromere positions on these chromosomes.     

Quantitative trait loci influencing β-glucan content in oat (Avena sativa, 2n=6x=42)

The β-glucan content of oat grain is of inter-est due to its positive human health role as a dietary component influencing serum cholesterol levels and its relation to the energy intake of livestock feed. Two recombinant inbred populations sharing a common parent (Kanota × Ogle and Kanota × Marion), and containing 137 individual lines each, were used to identify genomic regions that influence the β-glucan content in cultivated oat. Single-factor ANOVA, a backward elimination process, simple interval mapping (SIM) and simplified composite interval mapping (sCIM) were used to identify quantitative trait loci (QTLs). Regions on linkage groups 11 and 14 of the hexaploid oat RFLP map influenced β-glucan levels in both populations and over environments. Other genomic regions were identified whose effects varied depending on the genetic background, but were significant over measurements for a given population. Kanota and Ogle exhibit similar β-glucan levels and each parent contributed about the same number of positive β-glucan alleles in the Kanota × Ogle cross. Marion is higher in β-glucan content than Kanota and contributed all of the positive alleles in the Kanota × Marion cross. Three of the β-glucan QTL regions identified have been previously implicated as having a significant influence on the groat oil content in oat. These correlated QTL regions were either in coupling phase, with a region from one parent having the same effect on both traits, or were in repulsion phase. Identification of coupling- and repulsion-phase QTL regions for β-glucan and oil content facilitates the use of markers in manipulating these traits in oat breeding. Received: 8 September 1999 / Accepted: 25 March 2000