Differential anaphase-I behaviour between wheat and rye univalents in triticale-wheat hybrid plants

The chromosomes of the D genome of wheat and the genome R of rye can be distinguished at meiosis by C-banding in triticale-wheat hybrid plants. All members of both genomes almost exclusively formed univalents at metaphase I. However, at anaphase I the frequencies of equationally dividing chromosomes were higher for rye than for wheat chromosomes. The differential centromere behaviour at anaphase I is ascribed to differences in the time at which wheat and rye univalents are formed during the first meiotic prophase.     

Most of the homoeologous pairing at metaphase I in wheat-rye hybrids is not chiasmatic

The use of telomeric C-bands in wheat-rye hybrids has made it possible to distinguish three types of wheat-wheat (1B^L) and wheat-rye associations (a, end-to-end extremely distal; b, end-to-ed distal; and c, interstitial) between homoeologous chromosomes at different metaphase I stages (early, middle and late) and also to estimate the actual recombination frequencies for such associations at anaphase I. There was a decrease of the a and b association frequencies during the different metaphase I stages, whereas the c type remained without variation in all stages. A good fit between the frequencies of c associations at metaphase I and the number of recombinant chromosomes at anaphase I, assuming a maximum of one chiasma per bond, was found; however, there was no correspondence between metaphase I and anaphase I data when all associations (a + b + c) were considered. In addition, rye-rye homologous pairing was observed at metaphase I, but no evidence for rye-rye recombination was found at anaphase I. The results indicate that most of end-to-end (a and b) homoeologous and nonhomologous associations are actually nonchiasmatic and are a remnant of prophase pairing.     

Phorbol ester inhibits phosphoinositide hydrolysis and calcium mobilization in cultured astrocytoma cells

In cultured human 1321N1 astrocytoma cells, muscarinic receptor stimulation leads to phosphoinositide hydrolysis, formation of inositol phosphates, and mobilization of intracellular Ca2+. Treatment of these cells with 1 microM 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA) completely blocks the carbachol-stimulated formation of [3H]inositol mono-, bis-, and trisphosphate ( [3H]InsP, [3H]InsP2, and [3H]InsP3). The concentrations of PMA that give half-maximal and 100% inhibition of carbachol-induced [3H]InsP formation are 3 nM and 0.5 microM, respectively. Inactive phorbol esters (4 alpha-phorbol 12,13-didecanoate and 4 beta-phorbol), at 1 microM, do not inhibit carbachol-stimulated [3H]InsP formation. The KD of the muscarinic receptor for [3H]N-methyl scopolamine is unchanged by PMA treatment, while the IC50 for carbachol is modestly increased. PMA treatment also abolishes carbachol-induced 45Ca2+ efflux from 1321N1 cells. The concomitant loss of InsP3 formation and Ca2+ mobilization is strong evidence in support of a causal relationship between these two responses. In addition, our finding that PMA blocks hormone-stimulated phosphoinositide turnover suggests that there may be feedback regulation of phosphoinositide metabolism through the Ca2+- and phospholipid-dependent protein kinase.     

Meiotic behaviour of Un,D and R genomes in the amphiploid Aegilops ventricosa -Secale cereale and the parental species

Summary Meiotic pairing frequencies of the Un and D genomes of Ae. ventricosa and the R of S. cereale could be easily established at metaphase I in Aegilops ventricosa — Secale cereale amphiploid plants as well as in its parental species by using the C-banding technique procedure. The results show a high diminution of chromosome pairing for all genomes in the amphiploid with respect to its parental species probably due to C-heterochromatin content and/or genotypic or cryptic interactions between the three genomes.     

Relationship between the levels of wheat-rye metaphase I chromosomal pairing and recombination revealed by GISH

The metaphase I and anaphase I stages of meiosis of wheat×rye hybrids carrying the ph1b mutation were analyzed by genomic in situ hybridization. This technique allows distinction between three different types of wheat-rye associations in metaphase I configurations as well as detection of wheat-rye recombinant chromosomes in anaphase I cells. The frequency of associations between wheat and rye chromosomes greatly exceeded the level of wheat-rye recombination found in the three hybrids examined. Extremely distal associations, which account for about 50% of the total wheat-rye metaphase I chromosomal pairing, can explain such a discrepancy between metaphase I and anaphase I data. It is further discussed whether these associations reflect very distally located chiasmata or nonchiasmatic pairing. The sizes of the segments exchanged in wheat-rye recombinant chromosomes provide cytological evidence that wheat-rye recombination is restricted to the distal chromosomal regions. Received: 24 August 1995; in revised form: 27 February 1996 / Accepted: 28 March 1996     

Mapping of genes controlling seed storage-proteins and cytological markers on chromosome 1R of rye

Summary Rye secalins, telomeric C-bands, and telocentric chromosomes were used as markers in the progeny of a test-cross in order to determine the position of seed storage-protein genes Glu-R1 and Gli-R1 with respect to the centromere and both telomeres of chromosome 1 R in rye. These genes were linked to the centromere (32.35±3.28% and 36.27±3.37% recombination, respectively). Glu-R1 was loosely linked to the telomere of the long arm (43.63±3.47% recombination), while Gli-R1 was closely linked to the telomere of the short arm (9.80±2.08% recombination). This finding supports the possibility that rye - and -secalin genes may be located on the satellites, as has been described in wheat for genes that code similar proteins. The importance of metaphase-I pairing failure and its consequences for the estimation of the recombination fraction are also discussed.     

OPTIMIZATION OF AN IMMUNOFLUORESCENT ASSAY OF THE INTERNAL ENZYME RIBULOSE-1,5-BISPHOSPHATE CARBOXYLASE (RUBISCO) IN SINGLE PHYTOPLANKTON CELLS1

Immunochemical probes are widely used to identih different species and to quantify and understand the role that different antigens play within cells. We optimized a single-cell immunofluorescent assay for the carbon fixation enzyme ribulose-1,5-bisphosphate carboxylase (Rubisco) in order to quantify the enzyme by flow cytometry in phytoplankton cells. The criteria for optimization of the immunofluorescent assay for Rubisco in single cells included maximization of Rubisco immunogenicity, minimization of Rubisco diffusion out of the cells, minimization of cell breakage, and maximization of the cell labeling. Several fixatives (cross-linkers and denaturing) and permeabilizing agents were tested on 26 species of phytoplankton. The only fixative / permeabilizing agent that fulfilled the criteria established for the assay was 96% ethanol. Phytoplankton cells collected from the field needed further treatment with a strong oxidant to permeabilize ethanolfixed cells and thus allow the antibody probe to access the Rubisco antigen. This study should have a general applicability to the study of other soluble photosynthetic antigens in single phytoplankton cells.     

m-Hydroxyphenylacetic and m-Methoxyphenylacetic Acids of Rhizoctonia solani: Their Effect on Specific Root-Nodule Activity and Histopathology in Soybean

m-Methoxyphenylacetic acid (m-OMePAA), a derivative of m-hydroxyphenylacetic acid (m-OHPAA), having the same chemical composition as that phytotoxic compound produced in culture by Rhizoctonia solani, a fungal pathogen of soybean, reduced growth and symbiotic N₂-fixation activity of ‘Tracy’ soybeans in soil: perlite at 1.5 × 10^-4 M, the lowest concentration used. At twice this concentration m-OMePAA was strongly teratogenic and caused root hypertrophy and root fasciation. At 1.2 × 10^-3 M, m-OMePAA nearly suppressed seed germination. m-OMePAA at the minimum concentration used and equivalent concentrations of the culture filtrate fraction (m-OHPAA and m-OMePAA) caused cytopathological and histopathological disturbances in the nodule central tissue, extrusion of the nucleoli, and abnormal nuclci. The data indicate that these phytotoxic compounds of R. solani are involved in nodule impairment and reduced N₂-fixation in soybean.     

Isolation and nucleotide sequence of the Thiobacillus ferrooxidans genes for the small and large subunits of ribulose 1,5-bisphosphate carboxylase/oxygenase

The genes encoding for the large (rbcL) and small (rbcS) subunits of ribulose-1,5-bisphosphate carboxylase (RuBisCO) were cloned from the obligate autotroph Thiobacillus ferrooxidans, a bacterium involved in the bioleaching of minerals. Nucleotide sequence analysis of the cloned DNA showed that the two coding regions are separated by a 30-bp intergenic region, the smallest described for the RuBisCO genes. The rbcL and rbcS genes encode polypeptides of 473 and 118 amino acids, respectively. Comparison of the nucleotide and amino acid sequences with those of the genes for rbcL and rbcS found in other species demonstrated that the T. ferrooxidans genes have the closest degree of identity with those of Chromatium vinosum and of Alvinoconcha hessleri endosymbiont. Both T. ferrooxidans enzyme subunits contain all the conserved amino acids that are known to participate in the catalytic process or in holoenzyme assembly.