ABSTRACT
Hybrid (oat×maize) zygotes developed into euhaploid plants with complete oat chromosome complements without maize chromosomes and into aneuhaploid plants with complete oat chromosome complements and different numbers of retained individual maize chromosomes. The elimination of maize chromosomes in the hybrid embryo is caused by uniparental genome loss during early steps of embryogenesis. Some of these haploid plants set seed in up to 50% of their self-pollinated spikelets. The high fertility was found to be mainly caused by formation of numerically unreduced female and male gametes (nunreduced=3x+0 3=21 24 chromosomes). Gamete formation involves meiotic nuclear restitution. The restitution process is caused by an alternative type of meiosis. It follows the model of levigatum-type semi-heterotypic divisions, but with a formation of the nuclear membrane at the transition from telophase I to interkinesis, which resembles the model of pygaera-type pseudo-homotypic divisions. We propose the name haploid meiotic restitution for this particular process combination. We discuss the use and implications of the specific process of gamete formation in F1 (oat×maize) plants.
Subject(s)
Avena/genetics , Cell Nucleus/genetics , Crosses, Genetic , Germ Cells, Plant/growth & development , Haploidy , Meiosis , Zea mays/genetics , Avena/cytology , Chromosome Pairing/genetics , Chromosomes, Plant/genetics , Fertility/genetics , Genome, Plant/genetics , Germ Cells, Plant/cytology , Hybridization, Genetic , Meiosis/genetics , Parthenogenesis/genetics , Pollination/physiology , Zea mays/cytologyABSTRACT
Management of oat crown rust disease with host resistance is challenging because major gene resistance is generally short lived. Partially resistant oat cultivars could benefit oat growers by providing more durable resistance. The objective of this study was to validate and discover quantitative trait loci (QTL) affecting crown rust resistance in the partially resistant oat line MN841801-1 using conventional and molecular assessments of disease produced in single-race greenhouse inoculations, single-race polycyclic field tests, and under natural infection in disease-conducive environments. Crown rust was assessed on 150 F(6:9) MN841801-1/'Noble-2' recombinant inbred lines. In total, eight QTL associated with MN841801-1 alleles were detected. Of these, seven matched QTL previously identified while a new QTL (Prq8) was detected on linkage group MN13. Four QTL (Prq1a, Prq2, Prq7, and Prq8) were consistently detected and predicative genetic assays for these QTL should be developed for future validation in additional genetic backgrounds.
Subject(s)
Avena/genetics , Basidiomycota/physiology , Plant Diseases/genetics , Plant Proteins/metabolism , Quantitative Trait Loci , Avena/microbiology , Chromosome Mapping , Chromosomes, Plant , Gene Expression Regulation, Plant/physiology , Genetic Predisposition to Disease , Plant Diseases/microbiology , Plant Proteins/geneticsABSTRACT
To facilitate the detection of quantitative trait loci (QTLs) for partial resistance to oat crown rust, Puccinia coronata f. sp. avenae Eriks., a genetic map was generated in a population of 158 F(6)-derived oat recombinant inbred lines from a cross of a partial resistance line MN841801-1 by a susceptible cultivar selection 'Noble-2'. The map, developed using 230 marker loci, mostly restriction fragment length polymorphism and amplified fragment length polymorphism markers, spanned 1,509 cM (Haldane) arranged into 30 linkage groups of 2-18 markers each. Four consistently detected major QTLs for partial rust resistance, Prq1a, Prq1b, Prq2, and Prq7, and three minor QTLs, Prq3, Prq5, and Prq6, were found in tests involving three field and two greenhouse environments. In addition, two major QTLs for flowering time, Ftq1 and Ftq7, and five weaker QTLs, Ftq2, Ftq3, Ftq4, Ftq5, and Ftq6, were revealed. Overlapping of the map segments of Ftq1 and Prq1 and of Ftq7 and Prq7 suggested either linkage between the flowering time QTLs and resistance QTLs or a pleiotropic effect of the Ftq QTLs on rust resistance. Relatively low heritability estimates (0.30) obtained for partial resistance to crown rust in the field indicate a potential value for marker-assisted selection.
Subject(s)
Avena/genetics , Basidiomycota , Chromosome Mapping , Immunity, Innate/genetics , Plant Diseases/microbiology , Quantitative Trait Loci , Crosses, Genetic , Flowers/genetics , Flowers/physiology , Nucleic Acid Amplification Techniques , Plant Diseases/genetics , Polymorphism, Restriction Fragment LengthABSTRACT
The duplicated and rearranged nature of plant genomes frequently complicates identification, chromosomal assignment and eventual manipulation of DNA segments. Separating an individual chromosome from its native complement by adding it to an alien genetic background together with the generation of radiation hybrids from such an addition line can enable or simplify structural and functional analyses of complex duplicated genomes. We have established fertile disomic addition lines for each of the individual maize chromosomes, except chromosome 10, with oat as the host species; DNA is available for chromosome 10 in a haploid oat background. We report on instability and transmission in disomic additions of maize chromosomes 1, 5, and 8; the chromosome 2, 3, 4, 6, 7, and 9 additions appear stable. The photoperiodic response of the two recovered maize chromosome 1 addition lines contrasts to the long-day flowering response of the oat parents and the other addition lines. Only when grown under short days did maize chromosome 1 addition lines set seed, and only one line transmitted the maize chromosome 1 to offspring. Low resolution radiation hybrid maps are presented for maize chromosomes 2 and 9 to illustrate the use of radiation hybrids for rapid physical mapping of large numbers of DNA sequences, such as ESTs. The potential of addition and radiation hybrid lines for mapping duplicated sequences or gene families to chromosome segments is presented and also the use of the lines to test interactions between genes located on different maize chromosomes as observed for ectopic expression of cell fate alterations.
Subject(s)
Zea mays/genetics , Avena/genetics , Chromosomes , Genetic Markers , Genomics , In Situ Hybridization , Phenotype , Polymerase Chain Reaction , Radiation Hybrid MappingABSTRACT
Oat- (Avena sativa) maize (Zea mays) chromosome additions are produced by crossing maize and oat. During early embryo development maize chromosomes are preferentially eliminated, and oat plants are often recovered that retain a single maize chromosome. Each of the 10 maize chromosomes recently has been isolated as a separate oat-maize addition. We describe here the mapping of 400 maize sequences to chromosomes using polymerase chain reaction and DNA from the oat-maize addition material. Fifty of the sequences were from cloned markers that had been previously mapped by linkage analysis, and our results were consistent with those obtained using Southern-blot analysis. Previously unmapped expressed sequence tags and sequence tagged sites (350) were mapped to chromosomes. Maize gene sequences and expression data are rapidly being accumulated. Coupling this information with positional information from high throughput mapping programs provides plant biologists powerful tools for identifying candidate genes of interest.
Subject(s)
Avena/genetics , Chromosomes , Zea mays/genetics , Expressed Sequence Tags , Sequence Tagged SitesABSTRACT
All 10 chromosomes of maize (Zea mays, 2n = 2x = 20) were recovered as single additions to the haploid complement of oat (Avena sativa, 2n = 6x = 42) among F(1) plants generated from crosses involving three different lines of maize to eight different lines of oat. In vitro rescue culture of more than 4,300 immature F(1) embryos resulted in a germination frequency of 11% with recovery of 379 F(1) plantlets (8.7%) of moderately vigorous growth. Some F(1) plants were sectored with distinct chromosome constitutions among tillers of the same plant and also between root and shoot cells. Meiotic restitution facilitated development of un-reduced gametes in the F(1). Self-pollination of these partially fertile F(1) plants resulted in disomic additions (2n = 6x + 2 = 44) for maize chromosomes 1, 2, 3, 4, 6, 7, and 9. Maize chromosome 8 was recovered as a monosomic addition (2n = 6x + 1 = 43). Monosomic additions for maize chromosomes 5 and 10 to a haploid complement of oat (n = 3x + 1 = 22) were recovered several times among the F(1) plants. Although partially fertile, these chromosome 5 and 10 addition plants have not yet transmitted the added maize chromosome to F(2) offspring. We discuss the development and general utility of this set of oat-maize addition lines as a novel tool for maize genomics and genetics.
Subject(s)
Avena/genetics , Chromosomes , Genome, Plant , Zea mays/genetics , Base Sequence , DNA Primers , Hybridization, Genetic , In Situ Hybridization, FluorescenceABSTRACT
In maize (Zea mays L., 2n = 2x = 20), map-based cloning and genome organization studies are often complicated because of the complexity of the genome. Maize chromosome addition lines of hexaploid cultivated oat (Avena sativa L., 2n = 6x = 42), where maize chromosomes can be individually manipulated, represent unique materials for maize genome analysis. Maize chromosome addition lines are particularly suitable for the dissection of a single maize chromosome using radiation because cultivated oat is an allohexaploid in which multiple copies of the oat basic genome provide buffering to chromosomal aberrations and other mutations. Irradiation (gamma rays at 30, 40, and 50 krad) of a monosomic maize chromosome 9 addition line produced maize chromosome 9 radiation hybrids (M9RHs)-oat lines possessing different fragments of maize chromosome 9 including intergenomic translocations and modified maize addition chromosomes with internal and terminal deletions. M9RHs with 1 to 10 radiation-induced breaks per chromosome were identified. We estimated that a panel of 100 informative M9RHs (with an average of 3 breaks per chromosome) would allow mapping at the 0. 5- to 1.0-Mb level of resolution. Because mapping with maize chromosome addition lines and radiation hybrid derivatives involves assays for the presence or absence of a given marker, monomorphic markers can be quickly and efficiently mapped to a chromosome region. Radiation hybrid derivatives also represent sources of region-specific DNA for cloning of genes or DNA markers.
Subject(s)
Zea mays/genetics , Avena/genetics , Chromosomes/genetics , Gamma Rays , Gene Rearrangement , Genetic Markers , Genome, Plant , Germination/radiation effects , Hybridization, Genetic , In Situ Hybridization, Fluorescence , Seeds/radiation effects , Zea mays/radiation effectsABSTRACT
The recovery of maize (Zea mays L.) chromosome addition lines of oat (Avena sativa L.) from oat x maize crosses enables us to analyze the structure and composition of individual maize chromosomes via the isolation and characterization of chromosome-specific cosmid clones. Restriction fragment fingerprinting, sequencing, and in situ hybridization were applied to discover a new family of knob associated tandem repeats, the TR1, which are capable of forming fold-back DNA segments, as well as a new family of centromeric tandem repeats, CentC. Analysis of knob and centromeric DNA segments revealed a complex organization in which blocks of tandemly arranged repeating units are interrupted by insertions of other repeated DNA sequences, mostly represented by individual full size copies of retrotransposable elements. There is an obvious preference for the integration/association of certain retrotransposable elements into knobs or centromere regions as well as for integration of retrotransposable elements into certain sites (hot spots) of the 180-bp repeat. DNA hybridization to a blot panel of eight individual maize chromosome addition lines revealed that CentC, TR1, and 180-bp tandem repeats are found in each of these maize chromosomes, but the copy number of each can vary significantly from about 100 to 25,000. In situ hybridization revealed variation among the maize chromosomes in the size of centromeric tandem repeats as well as in the size and composition of knob regions. It was found that knobs may be composed of either 180-bp or TR1, or both repeats, and in addition to large knobs these repeated elements may form micro clusters which are detectable only with the help of in situ hybridization. The association of the fold-back elements with knobs, knob polymorphism and complex structure suggest that maize knob may be consider as megatransposable elements. The discovery of the interspersion of retrotransposable elements among blocks of tandem repeats in maize and some other organisms suggests that this pattern may be basic to heterochromatin organization for eukaryotes.
Subject(s)
Centromere/ultrastructure , Chromatids/ultrastructure , Chromosomes/ultrastructure , Zea mays/ultrastructure , Avena/genetics , Avena/ultrastructure , Centromere/genetics , Chromatids/genetics , Chromosomes/genetics , DNA, Plant/genetics , DNA, Plant/ultrastructure , Heterochromatin/genetics , Heterochromatin/ultrastructure , Hybridization, Genetic , Retroelements/genetics , Tandem Repeat Sequences/genetics , Zea mays/geneticsABSTRACT
To improve knowledge of the prerequisites for meiotic chromosome segregation in higher eukaryotes, we analyzed the spatial distribution of a pair of homologs before and during early meiotic prophase. Three-dimensional images of fluorescence in situ hybridization (FISH) were used to localize a single pair of homologs in diploid nuclei of a chromosome-addition line of oat, oat-maize9b. The system provided a robust assay for pairing based on cytological colocalization of FISH signals. Using a triple labeling scheme for simultaneous imaging of chromatin, telomeres and the homolog pair, we determined the timing of pairing in relation to the onset of three sequential hallmarks of early meiotic prophase: chromatin condensation (the leptotene stage), meiotic telomere clustering (the bouquet stage) and the initiation of synapsis (the zygotene stage). We found that the two homologs were mostly unpaired up through middle leptotene, at which point their spherical cloud-like domains began to transform into elongated and stretched-out domains. At late leptotene, the homologs had completely reorganized into long extended fibers, and the beginning of the bouquet stage was conspicuously marked by the de novo clustering of telomeres at the nuclear periphery. The homologs paired and synapsed during the bouquet stage, consistent with the timing of pairing observed for several oat 5S rDNA loci. In summary, results from analysis of more than 100 intact nuclei lead us to conclude that pairing and synapsis of homologous chromosomes are largely coincident processes, ruling out a role for premeiotic pairing in this system. These findings suggest that the genome-wide remodeling of chromatin and telomere-mediated nuclear reorganization are prerequisite steps to the DNA sequence-based homology-search process in higher eukaryotes.
Subject(s)
Meiosis , Telomere , Avena , Base Pairing , In Situ Hybridization, Fluorescence , Telomere/ultrastructure , Zea maysABSTRACT
Maize chromosome addition lines of oat offer the opportunity to study maize gene expression in oat and the resulting phenotypes. Morphological examination of a maize chromosome 3 addition line of oat showed that this line exhibited several morphological abnormalities including a blade-to-sheath transformation at the midrib region of the leaf, a hook-shaped panicle, and abnormal outgrowth of aerial axillary buds. Dominant mutations in the maize liguleless3 (lg3) homeobox gene result in a blade (distal)-to-sheath (proximal) transformation at the midrib region of the leaf. Ectopic expression of the dominant mutant Lg3 allele is believed to cause the phenotype. Therefore, we suspected that the maize lg3 gene, which is located on maize chromosome 3, was involved in the phenotypes observed in the maize chromosome 3 addition line of oat. Genetic analyses of an oat BC1F2 family segregating for maize chromosome 3 showed that the presence of a stable maize chromosome 3 was required for the expression of these cell fate abnormalities. RNA expression analysis of leaf sheath tissue from oat plants carrying maize chromosome 3 demonstrated that maize LG3 transcripts accumulated in oat, indicating that this expression is associated with the blade-to-sheath transformation, hook-shaped panicle and outgrowth of aerial axillary bud phenotypes. Our results demonstrate that the maize chromosome addition lines of oat are useful genetic stocks to study expression of maize genes in oat.
Subject(s)
Avena/genetics , Gene Expression , Genes, Homeobox , Homeodomain Proteins/genetics , Plant Proteins , Zea mays/genetics , Chromosome Mapping , Chromosomes , Genome, Plant , Homeodomain Proteins/biosynthesis , In Situ Hybridization , Karyotyping , Nucleic Acid Hybridization , Phenotype , Zea mays/anatomy & histologyABSTRACT
Anther culture and maize hybridization are two frequently used techniques for doubled haploid production in wheat (Triticum aestivum L.). Information on the field performance of lines derived from these techniques is limited. This study was conducted to compare the performance of F(4:6) lines obtained by single-seed descent with lines obtained by anther culture and maize (Zea mays L.) pollination from the same cross of spring wheat, 'Chris'/MN 7529. Thirty-three lines derived from each of those techniques were evaluated in six environments for grain yield, protein content, test weight, heading date, kernel weight and plant height. Mean performance of the single-seed descent lines exceeded performance of the anther culture lines for grain yield, kernel weight and plant height with no apparent differences for grain protein content, test weight and heading date. No differences between trait means for the single-seed descent and maize pollination lines were found except for plant height. The best 5 lines from each method for grain yield, protein content and test weight were similar in performance except that the protein content was higher for the maize pollination lines than for the single-seed descent lines. Acceptable levels of agronomic performance could be found among lines from each method. Wide acceptance of the doubled haploid technique for pure line production in breeding programs may, however, be limited by the often poor efficiency of doubled haploid line production, resulting in smaller population sizes for selection of desirable traits in comparison to the single-seed descent method.
ABSTRACT
A set of oat-maize chromosome addition lines with individual maize (Zea mays L.) chromosomes present in plants with a complete oat (Avena sativa L.) chromosome complement provides a unique opportunity to analyze the organization of centromeric regions of each maize chromosome. A DNA sequence, MCS1a, described previously as a maize centromere-associated sequence, was used as a probe to isolate cosmid clones from a genomic library made of DNA purified from a maize chromosome 9 addition line. Analysis of six cosmid clones containing centromeric DNA segments revealed a complex organization. The MCS1a sequence was found to comprise a portion of the long terminal repeats of a retrotransposon-like repeated element, termed CentA. Two of the six cosmid clones contained regions composed of a newly identified family of tandem repeats, termed CentC. Copies of CentA and tandem arrays of CentC are interspersed with other repetitive elements, including the previously identified maize retroelements Huck and Prem2. Fluorescence in situ hybridization revealed that CentC and CentA elements are limited to the centromeric region of each maize chromosome. The retroelements Huck and Prem2 are dispersed along all maize chromosomes, although Huck elements are present in an increased concentration around centromeric regions. Significant variation in the size of the blocks of CentC and in the copy number of CentA elements, as well as restriction fragment length variations were detected within the centromeric region of each maize chromosome studied. The different proportions and arrangements of these elements and likely others provide each centromeric region with a unique overall structure.
Subject(s)
Centromere/genetics , Chromosome Mapping , Zea mays/genetics , Base Sequence , Centromere/chemistry , Cosmids , DNA, Plant/chemistry , DNA, Plant/genetics , Genomic Library , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Oligonucleotide Probes , Restriction MappingABSTRACT
A class of tandemly repeated DNA sequences (TR-1) of 350-bp unit length was isolated from the knob DNA of chromosome 9 of Zea mays L. Comparative fluorescence in situ hybridization revealed that TR-1 elements are also present in cytologically detectable knobs on other maize chromosomes in different proportions relative to the previously described 180-bp repeats. At least one knob on chromosome 4 is composed predominantly of the TR-1 repeat. In addition, several small clusters of the TR-1 and 180-bp repeats have been found in different chromosomes, some not located in obvious knob heterochromatin. Variation in restriction fragment fingerprints and copy number of the TR-1 elements was found among maize lines and among maize chromosomes. TR-1 tandem arrays up to 70 kilobases in length can be interspersed with stretches of 180-bp tandem repeat arrays. DNA sequence analysis and restriction mapping of one particular stretch of tandemly arranged TR-1 units indicate that these elements may be organized in the form of fold-back DNA segments. The TR-1 repeat shares two short segments of homology with the 180-bp repeat. The longest of these segments (31 bp; 64% identity) corresponds to the conserved region among 180-bp repeats. The polymorphism and complex structure of knob DNA suggest that, similar to the fold-back DNA-containing giant transposons in Drosophila, maize knob DNA may have some properties of transposable elements.
Subject(s)
Chromosome Mapping , DNA Transposable Elements , DNA, Plant/genetics , Repetitive Sequences, Nucleic Acid , Zea mays/genetics , Base Sequence , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Restriction MappingABSTRACT
An irregular pattern of transgene silencing was revealed in expression and inheritance studies conducted over multiple generations following transgene introduction by microprojectile bombardment of allohexaploid cultivated oat (Avena sativa L.). Expression of two transgenes, bar and uidA, delivered on the same plasmid was investigated in 23 transgenic oat lines. Twenty-one transgenic lines, each derived from an independently selected transformed tissue culture, showed expression of both bar and uidA while two lines expressed only bar. The relationship of the transgenic phenotypes to the presence of the transgenes in the study was determined using (1) phenotypic scoring combined with Southern blot analyses of progeny, (2) coexpression of the two transgenic phenotypes since the two transgenes always cosegregated, and (3) reactivation of a transgenic phenotype in self-pollinated progenies of transgenic plants that did not exhibit a transgenic phenotype. Transgene silencing was observed in 19 of the 23 transgenic lines and resulted in distorted segregation of transgenic phenotypes in 10 lines. Silencing and inheritance distortions were irregular and unpredictable. They were often reversible in a subsequent generation of self-pollinated progeny and abnormally segregating progenies were as likely to trace back to parents that exhibited normal segregation in a previous generation as to parents showing segregation distortions. Possible causes of the irregular patterns of transgene silencing are discussed.
Subject(s)
Avena/genetics , Genes, Plant , Acetyltransferases/genetics , Avena/enzymology , Gene Expression , Genes, Reporter , Glucuronidase/genetics , Phenotype , Plants, Genetically Modified , PolyploidyABSTRACT
The recovery of maize (Zea mays L.) chromosome addition lines of oat (Avena sativa L.) from oat x maize crosses enables us to analyze the structure and composition of specific regions, such as knobs, of individual maize chromosomes. A DNA hybridization blot panel of eight individual maize chromosome addition lines revealed that 180-bp repeats found in knobs are present in each of these maize chromosomes, but the copy number varies from approximately 100 to 25, 000. Cosmid clones with knob DNA segments were isolated from a genomic library of an oat-maize chromosome 9 addition line with the help of the 180-bp knob-associated repeated DNA sequence used as a probe. Cloned knob DNA segments revealed a complex organization in which blocks of tandemly arranged 180-bp repeating units are interrupted by insertions of other repeated DNA sequences, mostly represented by individual full size copies of retrotransposable elements. There is an obvious preference for the integration of retrotransposable elements into certain sites (hot spots) of the 180-bp repeat. Sequence microheterogeneity including point mutations and duplications was found in copies of 180-bp repeats. The 180-bp repeats within an array all had the same polarity. Restriction maps constructed for 23 cloned knob DNA fragments revealed the positions of polymorphic sites and sites of integration of insertion elements. Discovery of the interspersion of retrotransposable elements among blocks of tandem repeats in maize and some other organisms suggests that this pattern may be basic to heterochromatin organization for eukaryotes.
Subject(s)
DNA, Plant/chemistry , DNA, Plant/genetics , Heterochromatin/chemistry , Heterochromatin/genetics , Retroelements , Zea mays/genetics , Avena/genetics , Base Sequence , DNA Fingerprinting , DNA Primers/genetics , Hybridization, Genetic , Molecular Sequence Data , Nucleic Acid Conformation , Polymerase Chain Reaction , Repetitive Sequences, Nucleic AcidABSTRACT
Nullisomic lines of hexaploid oat Avena sativa L. (2n = 6x - 2 = 40, AACCDD) cultivar Sun II were used to assign 134 DNA sequences to 10 chromosome-associated syntenic groups. A limited set of ditelosomic lines allowed localization of subsets of these sequences to six chromosome arms. Advantages of using such aneuploids in mapping are in the assignment of gene families, monomorphic RFLP sequences, and oat linkage groups to chromosomes. The published hexaploid oat RFLP linkage map has 38 linkage groups, 17 more than expected on the basis of the haploid chromosome number. Using nullisomics, eight linkage groups were assigned to five physical chromosomes; using ditelosomics, three of these linkage groups were assigned to their respective chromosome arms. The A- and D-genome chromosome sets of oat are indistinguishable from each other based on different staining and genomic in situ hybridization techniques, while C-genome chromosomes are distinct. Because chromosomal rearrangements such as translocations and inversions have played an important role in the evolution of hexaploid oat, the distinction of C-genome chromosomes can be used to determine remnant homoeologous segments that exist in the other two genomes. Among the 10 syntenic groups identified, six chromosomes showed sequence homoeology believed to represent segmental homoeologous regions. Owing to various evolutionary forces, segmental homoeology instead of whole chromosome homoeology appears to best describe the genome organization in hexaploid oat.
Subject(s)
Aneuploidy , Avena/genetics , Chromosome Mapping/methods , Genome, Plant , Polyploidy , Genetic Markers , Polymorphism, Restriction Fragment LengthABSTRACT
Novel plants with individual maize chromosomes added to a complete oat genome have been recovered via embryo rescue from oat (Avena sativa L., 2n = 6x = 42) x maize (Zea mays L., 2n = 20) crosses. An oat-maize disomic addition line possessing 21 pairs of oat chromosomes and one maize chromosome 9 pair was used to construct a cosmid library. A multiprobe (mixture of labeled fragments used as a probe) of highly repetitive maize-specific sequences was used to selectively isolate cosmid clones containing maize genomic DNA. Hybridization of individual maize cosmid clones or their subcloned fragments to maize and oat genomic DNA revealed that most high, middle, or low copy number DNA sequences are maize-specific. Such DNA markers allow the identification of maize genomic DNA in an oat genomic background. Chimeric cosmid clones were not found; apparently, significant exchanges of genetic material had not occurred between the maize-addition chromosome and the oat genome in these novel plants or in the cloning process. About 95% of clones selected at random from a maize genomic cosmid library could be detected by the multiprobe. The ability to selectively detect maize sequences in an oat background enables us to consider oat as a host for the cloning of specific maize chromosomes or maize chromosome segments. Introgressing maize chromosome segments into the oat genome via irradiation should allow the construction of a library of overlapping fragments for each maize chromosome to be used for developing a physical map of the maize genome.
Subject(s)
Chromosome Mapping/methods , Genes, Plant , Genome, Plant , Zea mays/geneticsABSTRACT
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.
ABSTRACT
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.
ABSTRACT
A collection of 19 wheat (Triticum aestivum) probes, detecting sequences in the seven homoeologous groups of chromosomes, were hybridized to DNA from the 'Kanota' series of oat monosomic lines (Avena byzantina) to investigate their use for identifying groups of homoeologous oat chromosomes. Three probes from homoeologous group 1 of wheat, psr161, psr162, and psr121, mapped among the set of oat chromosomes 1C, 14, and 17. One homoeologous group 6 probe, psr167, mapped to oat chromosomes 1C and 17. Two oat probes that had previously been shown to map to oat chromosomes 1C, 14, and 17 were then hybridized to DNA from the 'Chinese Spring' wheat ditelosomics. They localized to homoeologous group 1 wheat chromosomes, one to the short arm and one to the long arm. These results reveal that in hexaploid oat there is a group of three chromosomes that correspond at least in part to homoeologous group 1 of wheat. The remaining wheat probes identifying other wheat homoeologous sets did not detect a complete series of homoeologous chromosomes in oat. This was presumably due to the incomplete status of the 'Kanota' monosomic series, chromosomal rearrangement in Avena, weak hybridization signals owing to low probe-target sequence homology, and (or) detection of only two hybridization bands by the wheat probe.