Ribosomal DNA magnification in Saccharomyces cerevisiae.

Strains monosomic for chromosome I of Saccharomyces cerevisiae contain 25 to 35% fewer rRNA genes than do normal diploid strains. When these strains are repeatedly subcultured, colonies are isolated that have magnified their number of rRNA genes to the diploid amount while remaining monosomic for chromosome I. We have determined the amount of DNA complementary to rRNA in viable haploid spores derived from a magnified monosomic strain. Some of these haploids contained 24 to 48% more rRNA genes than a normal euploid strain. These extra genes may be responsible for the increased number of rRNA genes in the strain monosomic for chromosome I. Genetic analysis of the haploids containing extra rRNA genes suggested that these genes are linked to chromosomal DNA and are heterozygous. They were not closely linked to any centromere and were not located on chromosome I. Furthermore, all the DNA complementary to rRNA in one of these haploid strains with magnified rRNA genes sedimented at a chromosomal molecular weight, consistent with chromosomal linkage. In addition, several new mutations mapping on chromosome I were used to show that ribosomal DNA magnification was not due to a chromosome I duplication.     

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     

A complete set of maize individual chromosome additions to the oat genome

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.     

The genetics of resistance to powdery mildew in cultivated oats (Avena sativa L.): current status of major genes

The genetics of resistance to powdery mildew caused by Blumeria graminis f. sp. avenae of four cultivated oats was studied using monosomic analysis. Cultivar ‘Bruno’ carries a gene (Pm6) that shows a recessive mode of inheritance and is located on chromosome 10D. Cultivar ‘Jumbo’ possesses a dominant resistance gene (Pm1) on chromosome 1C. In cultivar ‘Rollo’, in addition to the gene Pm3 on chromosome 17A, a second dominant resistance gene (Pm8) was identified and assigned to chromosome 4C. In breeding line APR 122, resistance was conditioned by a dominant resistance gene (Pm7) that was allocated to chromosome 13A. Genetic maps established for resistance genes Pm1, Pm6 and Pm7 employing amplified fragment length polymorphism (AFLP) markers indicated that these genes are independent of each other, supporting the results from monosomic analysis.     

rDNA and acrocentric chromosomes in man

Summary A malformed female infant was found to have a 46,XX complement with a chromosome 8 shorter than normal with a secondary constriction and satellites on the short arm. Chromosome studies on the clinically normal father showed a balanced translocation between chromosome 8 and 13, i.e., 46,XY,t(8;13) (p21 p12). The proposita, carrier of the unbalanced form of the translocation, resulted partially monosomic for short arm of chromosome 8 (8p-) and partially trisomic for short arm of chromosome 13.The levels of DNA complementary to rRNA (normal in the father who had 10 NOR and increased in the proposita who had 11 NOR) confirmed our interpretation of the rearrangement.     

Genes in diploid triticinae compensating for the low temperature regulating gene Ltp in chromosome 5D of Triticum aestivum

Summary Hybrids of Triticum aestivum (monosomic 5D or ditelosomic 5DL) x T. speltoides (= Ae. speltoides) showed that the genotype of T. speltoides carries gene(s) which can partially compensate for the expected decrease in chromosome association at low temperatures (10°C) in the absence of chromosome 5D. In hybrids of T. aestivum (normal, ditelosomic 3DL or ditelosomic 3DL-monosomic 5D) x T. longissimum (= Ae. sharonensis), this compensation was not observed.In normal F₁ hybrids of T. durum x T. longissimum partial chromosome association occurred at 10°C and this stabilizer effect may be explained by the presence of a Ltp-like gene on chromosome 5A. When a line of T. durum carrying a homozygous translocated 5B-5D chromosome was used in the crosses an even higher chromosome association was observed.These results suggest either the existence of a promoter gene for chromosome association in the 5D translocated segment or the loss of a weak suppressor gene in the removed segment of 5B. It was concluded that the translocated 5D segment did not carry the Ltp stabilizer gene.The work was supported by a fellowship of the Gulbenkian Foundation and partly carried out while the author was at the Department of Genetics, Agricultural University, Wageningen, The Netherlands.     

Localization of Structural and Regulatory Genes for Phosphodiesterase in Wheat (TRITICUM AESTIVUM)

Genes (Pde-A3; Pde-B3; Pde-D3) for phosphodiesterase (PDE; E.C. 3.1.4.1.) isoenzymes in hexaploid wheat were located on the three homoeologous chromosomes of group 3 by testing the electrophoretic banding pattern of monosomic, nullisomic and nullisomic/tetrasomic compensation lines of "Chinese Spring" variety. In plants nullisomic for chromosome 5B, the 3D structural gene is not expressed and this lack of expression can be overcome by four doses of either homoeologous chromosome 5A or 5D. Our data conclusively indicate that there are genes on group 5 chromosomes which positively control the expression of the 3D structural gene. In addition, the expression of the "regulatory genes" is dosage dependent. Thus, our study reveals a complex interaction of the three genomes of wheat for regulation of PDE gene expression.     

Regulation of Ribosomal RNA Gene Multiplicity in DROSOPHILA MELANOGASTER

The ribosomal RNA (rRNA) genes of Drosophila melanogaster can undergo a disproportionate replication of their number. This occurs when the cluster of rRNA genes (rDNA) of one chromosome is maintained with a homologous chromosome that is completely or partially deficient in its rDNA. Under appropriate genetic conditions, it appears that disproportionate rDNA replication can be generated at the level of both somatic and germ line cells. In the latter case, mutants partially deficient for rDNA can increase their rRNA gene number to the wild type level and transmit this new genotype to successive generations.     

Selection of monosomic addition plants in offspring families using repetitive DNA probes in Beta L.

The distribution of two repetitive DNA probes Sat-121 and PB6-4, specific for the section Procumbentes of the genus Beta, was tested in 16 B. patellaris monosomic addition families using a dot-blot hybridization procedure. All monosomic additions were accurately distinguished from diploid sib plants with both DNA probes. The probe PB6-4, with the strongest signal after hybridization, was selected for rapid screening of an extensive number of putative monosomic additions in B. patellaris or B. procumbens addition families using a squash-blot hybridization procedure. The probe PB6-4 detected 118 monosomic additions in 640 plants (18.4%) in eight different B. procumbens addition families. The addition family with chromosome 4 of B. procumbens was semi-lethal and could not be tested. The distribution of PB6-4 in B. patellaris addition families was confirmed in 63 addition families using the squash-blot procedure. In 4580 plants of these addition families, 628 individual monosomic additions (13.7%) were found. The relationship of the morphological characteristics of monosomic addition plants to the results of the squash-blot hybridization (plants with signal) using probe PB6-4 is quite rigorous but not complete. The correlation between plants with a signal and chromosome number (2n=19) is complete. These results indicate that sequences present on PB6-4 are probably present on all chromosomes of B. patellaris and B. procumbens. The possibility of utilizing the sequence information of Sat-121 for a PCR-based assay to screen for putative monosomic addition plants was also investigated as an alternative to chromosome counting. The DNA-amplification profiles using the primers REP and REP.INV clearly distinguished monosomic addition plants from their diploid sibs.     

Further study of histone structural gene multiplication in Drosophila melanogaster

Summary Heterozygous flies with a deficiency of histone genes show a gradual increase in the number of these genes reaching 90% of the normal level during eight generations. After removing the deficient chromosome the increased histone gene number is not stably inherited and reverts to normal in the course of 5–7 generations. Males heterozygous for the deficient chromosome show extrachromosomal histone genes in the first generation and have a changed ratio of the two histone gene repeat units. The multiplication of histone genes is compared with compensation and magnification of rRNA.     

采用玉米Ubi-1启动子获得低拷贝转基因玉米植株

通过基因枪粒子轰击和草丁膦（PPT）选择获得可育的玉米转基因植株,并分析了外源基因在转化体中的拷贝数与启动子之间的关系。用玉米Ubi-1启动子驱动外源基因,玉米转化体中外源基因的拷贝数较低;可能的原因为Ubi-1启动子通过与其内部同源序列发生重组而被定点整合进玉米基因组,共转化的两种质粒DNA在整合至玉米染色体DNA之前已重构成为一个整体。结果显示使用某一植物自身基因的启动子可以降低外源基因在该物种转基因个体中的拷贝数,进而避免基因沉默现象的发生。目前已得到第二代转基因玉米种子。     

The origin, identification, and plant morphology of five rice monosomics.

Five different monosomics of rice (Oryza sativa L.) were obtained by treatment of pollen with gamma irradiation, as a by-product of attempts to determine the cytological loci of certain marker genes, i.e., mature pollen carrying normal alleles at all loci was given gamma rays and used for pollinating strains that were homozygous for recessive marker genes. The monosomics showed distinguishable morphological features and had complete seed sterility. Cytological studies revealed that one monosomic was tertiary, the others primary. The tertiary monosomic was related to chromosome 10. Two primary monosomics for chromosomes 10 and 11 were identified. At metaphase I, the tertiary monosomic showed the chromosome configurations 1 III + 10 II, 11 II + 1 I, and 10 II + 3 I, and all primary ones showed the configuration 11 II + 1 I. All five monosomics showed very poor crossing ability and were not transmitted to the few progenies observed. A few trisomic plants were found in the progenies of a cross between monosomic and normal pollen in one monosomic. This is the first time that many monosomics in rice have been characterized. This information will be useful in studies of rice aneuploidy and cytogenetics. Key words : rice, monosomics, morphology, cytology, transmission, trisomics.     

A full set of monosomic addition lines in Beta vulgaris from Beta webbiana: morphology and isozyme markers

Summary Nine different monosomic additions in Beta vulgaris from Beta webbiana were characterized through morphological characters and isozyme markers. The effect of the alien chromosome on the morphology of the recipient species is chromosome specific, and nine morphotypes could be distinguished. The added chromosome caused a growth reduction in the recipient plants. Eleven isozyme systems were used as marker systems. A 6PGDH band was found as a marker for chromosome 7, which contains a resistance gene for the beet cyst nematode in monosomic additions from Beta procumbens and Beta webbiana. A difference in the 6PGDH zymogram pattern between the two species with respect to this chromosome has been noted.     

Localization of rRNA genes in the nuclear space of Calliphora erythrocephala Mg. nurse cells during polytenization

Multicolor 3D fluorescence in situ hybridization was used to study arrangement of rRNA genes in Calliphora erythrocephala nurse cell nuclei with different levels of polyteny. It has been shown that the rRNA genes are exclusively localized to chromosome 6, suggesting that chromosome 6 is the only C. erythrocephala chromosome responsible for nucleolar formation. We have also described changes in localization of ribosomal genes within the chromosome territory during polytenization, namely, that rDNA signals are detected in the peripheral region of chromosome territory starting from the stage of polytene chromosomes. In addition, it has emerged that large nucleolus associated with chromosome 6 starts to develop in the central nuclear region in the C. erythrocephala nurse cell nuclei at the stage of a primary reticular structure. The central position and nucleolar structure are retained at the stages when chromosome 6 occupies the central position, that is, at the stages of polytene and bloblike chromosomes. When the nucleus restores a reticular structure but at a higher polyteny level, the displacement of chromosome 6 to the nuclear periphery is accompanied by disruption of the large nucleolus into micronucleoli. The micronucleoli are distributed in the nuclear space retaining their association with the nucleolar-organizing regions of chromosome 6. Thus, our data suggest that the large-scale alterations in the organization of chromosome 6 and the nucleolus during polytenization are the correlated processes directly dependent on the rRNA gene activity. The earlier described dynamics of nucleolar-organizing chromosome territory and nucleolus in the nuclear space is likely to be associated with the change in the total expression activity of the nucleus, which complies with the hypothesis on the correlation between spatial nuclear organization and expression regulation of genetic material.