Chromosome breakage-fusion-bridge-cycle and phenotypic instability in isochromosome lines of tomato

Summary Cultivated tomato lines with 7–9 copies of extra chromosomes, which were fully heterochromatic, were found to have normal growth and reproduction. Cytological evidence suggested that all extra chromosomes in the 24 plants investigated were derived from a single isochromosome of the short arm of chromosome 2. In spite of their common origin, the individual extra chromosomes within and between plants varied considerably with respect to size and morphology. The morphological variation of these extra chromosomes was found to result from chromosome-or chromatid breakage-fusion-bridge-cycle (BFBC). In order to verify whether BFBC could induce genomic shock and instability, the phenotypes of more than 10,000 seedlings from some of the progeny were examined. There was a 3-to 4-fold increase in the occurrence of the number of chlorophyll variegated sectors in the progeny of plants with BFBC when compared to the control variety.     

ORIGIN OF FRAGARIA POLYPLOIDS. II. UNREDUCED AND DOUBLED-UNREDUCED GAMETES

Offspring from natural hybrids between octoploid Fragaria chiloensis (2n = 56) and diploid F. vesca (2n = 14) backcrossed under natural conditions to F. chiloensis were studied. The natural F₁ hybrids themselves were of three kinds: (1) The expected pentaploids which resulted from the union of normally reduced gametes of diploid F. vesca and octoploid F. chiloensis; (2) A hexaploid F₁ hybrid which resulted from the union of an unreduced gamete from diploid F. vesca with a normally reduced gamete from octoploid F. chiloensis; and (3) A 9-ploid F₁ hybrid which probably arose from the union of an unreduced gamete of the octoploid F. chiloensis with a normally reduced gamete of diploid F. vesca. The progenies that resulted from the natural backcrossing of each of the three sorts of F₁ hybrids to F. chiloensis were as follows: The pentaploid F₁ hybrids (2n = 35) yielded mostly 9-ploid offspring from unreduced 5X gametes; a relatively high percentage of 14-ploid plants arising from doubled-unreduced 10 X gametes and a few 2N = ±46 aneuploids from reduced gametes. The hexaploid F₁ hybrid (2n = 42) on backcrossing yielded over 50% 10-ploid offspring with the rest 2n = ±50 aneuploids from reduced gametes. The 9-ploid F₁ hybrid (2n = 63) on backcrossing yielded mostly aneuploids normally distributed about a modal 2n = 59 chromosome class resulting from a 31 chromosome gamete, with a few 2n = 56 and 2n = 63 euploids. The 9-ploids may facilitate diploid Å octoploid introgression. Screening of the open-pollinated offspring from F. chiloensis revealed almost 2% 12-ploid (2n = 84) offspring from the union of the reduced and unreduced F. chiloensis gametes. The probable genomic constitution of the observed novel ploidy levels and those that theoretically may be generated from the known hybrids are presented. The origin of the existing polyploids from diploids through simple unreduction is postulated.     

Isolation of a series of chromosomal variants (2n=10 to 21) from an open-pollinated population of interspecific hybrids between Coix gigantea and Coix aquatica (Poaceae)

A series of chromosomal variants has been isolated from an open-pollinated progeny of interspecific hybrids between aneuploids of Coix gigantea (2n=18–24) and Coix aquatica (2n=10). The interspecific hybrids (2n=14, 15 and 16) produced several types of gametes not only with different chromosome numbers but also comprised of varied permutations and combinations of gigantea and aquatica chromosomes. This was evident when the open-pollinated progeny obtained from these hybrids was screened chromosomally. Two such open-pollinated experimental progenies were studied in two successive years (1983 and 1984) and plants with from 2n=10 to 2n=21 chromosomes were isolated. Chromosomal configurations at diakinesis in all the variants revealed frequent pairing between the gigantea and aquatica chromosomes. This indicated that the two species are phylogenetically closely related. Restoration of pure parental species from the F₁ hybids and chromosomal variants through genomic segregation and spontaneous back-cross are unique and noteworthy features.     

The segregation pattern of a translocation quadrivalent

The segregation pattern of a translocation quadrivalent was studied in three hybrid families of the tetraploid (2n=28) oat, of the Avena strigosa polyploid complex. The rate of IV formation in F₁ was high and the fertility was normal. The adjacent-alternate orientation of this quadrivalent was very variable. Conspicuous variation in this frequency was found between anthers of the same floret and between florets. The alternate type was usually more common toward the end of MI. The adjacent-alternate ratio was found to be an unreliable measure for calculating the type of gametes produced by the F₁ and the F₂ plants derived from them. An attempt was made to determine the type of viable gametes produced on the F₁ and their frequency by examining the cytology of F₂ plants. The various combinations of the chromosomes of the translocation complex expected in the F₂ plants were derived and their expected frequencies calculated by taking into account chiasma formation at the chromosome ends and various restrictions of gamete viability. In none of the three hybrid families F₂ individuals were found to produce trivalents as the most complex chromosome configuration, indicating that one type of gamete derived from adjacent separation was not formed. The 11 ratio between F₂ plants having only bivalents (2II) and those with F ₁-like quadrivalent configuration R(c), expected when gametes resulting from alternate separation are fully functional, with the exclusion of other types, was not found. That ratio 2II/R(c), was 2-1/3 in the various families. The cytology of selected F₃ individuals basically followed the predictions based on chromosome association in the F₂.     

Chromosomal variation in immature embryo derived calluses of barley (Hordeum vulgare L.)

Summary Chromosome counts of ten morphogenic and seven non-morphogenic immature embryo derived calluses of barley,Hordeum vulgare L. cv. Himalaya, were determined. Morphogenic calluses carried the normal chromosome complement (2n=2x=14) in a majority of the cells. A low frequency of haploid (2n=x=7), triploid (2n=3x=21), tetraploid (2n=4x=28) and octoploid (2n=8x=56) cells were also observed. In contrast, non-regenerability of a callus was attributed to the cells having numerical and structural chromosomal changes. In these calluses, aneuploid cells around diploid, triploid, and tetraploid chromosome numbers predominated. It has been demonstrated that chromosomal changes were induced during the culture and that they did not pre-exist in the cultured barley embryos. Based on this study, it is suggested that chromosome analysis of a non-regenerable callus should be conducted before altering the media composition.     

Interspecific hybrids between Brassica napus L. and B. oleracea L. developed by embryo culture

Summary Interspecific hybrids between Brassica napus and B. oleracea are difficult to produce, and previous attempts to transfer economic characters from one species to the other have largely been unsuccessful. In these studies, oilseed rape cv. Tower (2n38) (B. napus) was crossed with broccoli and kale (2n18) (B. oleracea), and hybrid plants were developed from embryos in culture by either organogenesis or somatic embryogenesis. In rape × broccoli, F₁ plants were regenerated from hybrid embryos and the plants produced viable selfed seeds. F₅ plants (2n38) homozygous for white flower colour were selected for high oil content (47%) and Line 15; a selection from these plants produced fertile hybrids with rape, broccoli and kale without embryo culture. In reciprocal crosses between oilseed rape cv. Tower and an aphid resistant diploid kale, 28 and 56 chromosome F₁ hybrid plants were regenerated from somatic embryos. The 56 chromosome plants were self-fertile and it was concluded from F₂ segregation ratios that a single dominant gene controls resistance to cabbage aphid in kale. The 28 chromosome F₁'s were self-sterile, but these and the 56 chromosome F₁'s could be backcrossed to rape and kale. A cross between the F₁ (2n56) and a forage rape resulted in the selection of a cabbage aphid (Brevicoryne brassicae L.) resistant line (Line 3). Both Line 15 and Line 3 can serve as bridges for gene interchange between B. campestris, B. napus and B. oleracea, which has not been possible hitherto. Hybridisations between rape and tetraploid kale produced F₁ plants with 37 chromosomes. One F₂ plant possessed coronal scales and the inheritance was shown to be controlled by a single recessive gene unlinked to petal colour.This paper is dedicated to Mr. T. P. Palmer, a colleague and close friend who retired from the DSIR as Assistant Director of the Crop Research Division in September 1984     

Meiotic pairing as an indicator of genome composition in polyploid prairie cordgrass (<Emphasis Type="Italic">Spartina pectinata</Emphasis> Link)

The existence of neopolyploidy in prairie cordgrass (Spartina pectinata Link) has been documented. The neohexaploid was discovered coexisting with tetraploids in central Illinois, and has been reported to exhibit competitiveness in the natural environment. It is hypothesized that the natural tetraploid cytotype produced the hexaploid cytotype via production of unreduced gametes. Meiosis I chromosome pairing was observed in tetraploid (2n?=?4x?=?40), hexaploid (2n?=?6x?=?60), and octoploid (2n?=?8x?=?80) accessions and the percentage of meiotic abnormality was determined. Significant differences in meiotic abnormality exist between tetraploid, hexaploid, and octoploid cytotypes. An elevated incidence of abnormal, predominantly trivalent pairing in the neohexaploid suggests that it may possess homologous chromosomes in sets of three, in contrast to the tetraploid and octoploid cytotypes, which likely possess homologous chromosomes in sets of two. Abnormal chromosome pairing in the hexaploid may result in unequal allocation of chromosomes to daughter cells during later stages of meiosis. Chromosome pairing patterns in tetraploid, hexaploid, and octoploid cytotypes indicate genome compositions of AABB, AAABBB, and AABBA′A′B′B′, respectively.     

Development of the synaptonemal complex of two types of pachytene in oocytes and spermatocytes of Carausius morosus Br. (Phasmatodea)

During oogenesis of the parthenogenetic stick insect Carausius morosus (2n =61+XXX) pachytene is followed by a duplication of the desynapsed chromosomes, which results in a second type of pachytene (tetrapachytene) consisting of paired sister chromosomes (autobivalents). Electron microscopic studies on sections revealed that synaptonemal complexes (SCs) are formed during tetrapachytene only. This means that the parthenogenetically produced progeny have the genetic constitution of the mother. During spermatogenesis of rare fatherless males (2n=61 + XX) and intersexes (2n=61 +XXX) either an incomplete chromosome doubling (demonstrated by up to 10% additional DNA synthesis) or a complete chromosome doubling takes place during zygotene. EM studies on sections and spreads of germ cells of the first type of meiosis showed that unpaired lateral components (LCs), pieces of SCs and complete SCs are formed during pachytene only, the sex chromosomes being represented by unpaired thickened LCs. The incomplete SC formation reflects the complex heterozygosity of the chromosome complement. In the duplicated type SCs are found in tetrapachytene nuclei only; they are wider than the SCs in oocytes. The sex chromosome bivalents are represented by unpaired thickened LCs or partially paired LCs, in which localized chiasma formation was found. The idea is discussed that formation of SCs does not take place as long as a germ cell has been programmed either to replicate or to be able to replicate its chromosomes and that consequently SCs can be formed only once per meiosis.     

Cytogenetics of the F1 hybrid between cassava and ceara rubber,and its backcross

Cytogenetical studies of the F₁ hybrid between the commercially cultivated tuber crop, cassava (Manihot esculenta Crantz.) and the closely related wild speciesManihot glaziovii Muell. (Ceara rubber) used as donor specles for Cassava mosaic discase and drought-resistant genes and back crosses (to cassava parent) were made. The contrasting parental characters showed partial to total dominance in the F₁ hybrid, while the back cross plants were similar to cassava in most of their characters. Eleven of the twelve backeross plants exhibited resistance to Cassava mosaic under field conditions. Karyological similarities and differences as resolved on the basis of a comparative study of the karyotypes of the cassava parent and coara rubber were corroborated by the study of chromosomal pairing in the F₁ at pachytene. Major chromosomal differentiation in the two species involved three chromosomes of their haploid complement which were represented by three heteromorphic bivalent associations in F₁ each consisting of a probably basic chromosomal type and a derived type. Pachytene analyses of three back cross plants provided direct proof for random transmission of marker chromosomes of both the parents through male gametes of the F₁ hybrid. An increase in the chiasma frequency in the back cross plants over the F₁ hybrid at metaphase I stage was also observed. Pollen fertility of the backeross plants showed considerable variation.     

Heterogeneity of the α-globin gene defects in German α-thalassemia affected families

Summary Analysis of -thalassemia syndromes in several German families revealed DNA deletion as well as nondeletion forms as the molecular basis for the defects. Thus, the -thalassemia haplotype was identified as the (–)^3.7 rightward deletion form, and the region of the putative recombination process generating such a deletion was further characterized. In addition three different ° haplotypes, (--)^MED, (--)^>26, and ()^T, could be detected using -and -globin gene-specific probes.     

A new mechanism for altering chromosome number during karyotype evolution

Summary A new mechanism for changing chromosome numbers (preserving the fundamental number of long chromosome arms) during karyotype evolution is suggested. It includes: 1) Occurrence of individuals heterozygous for two interchanges between different arms of three chromosomes (a metacentric and two acrocentric ones). 2) Formation in heterokaryotypes of multivalents during meiosis between the chromosomes involved in the interchanges and their unchanged homologues. 3) Mis-segregation of chromosomes from these multivalents resulting in hypoploid (n-1) and hyperploid (n+1) simultaneously instead of euhaploid gametes. 4) Fusion of n-1 or n+1 gametes which gives rise to (zygotes and) individuals representing homokaryotypes with changed number of chromosomes (2n+2 or 2n-2), but preserves (as compared to the parental karyotypes) the number of long chromosome arms. Under definite conditions, chromosome numbers of the progeny may be changed by this process in both directions (upwards and downwards). The mechanism is free of the difficulties associated with the explanation for such changes by direct Robertsonian interchanges (see Discussion), which are usually considered to be responsible for such alterations in chromosome number. The above-mentioned process has been experimentally documented in Vicia faba and it probably also occurred naturally within the Vicia sativa group.     

Genetics of self incompatibility in diploid Ageratum houstonianum mill

Summary Diallels and backcrosses among self-incompatible (SI) clones and progeny of Ageratum houstonianum Mill. could be organized into intra-incompatible classes. Four of 5 progenies segregated in expected ratios of S genotypes. Ageratum expressed a one-locus incompatibility system of the sporophytic type with a linear dominance series of multiple alleles and complete allelic dominance in both pollen and stigma. In the second part of the study, a high percentage of self-seed set was observed during the first flowering of a progeny from a pseudo-self compatible (PSC) seed source. Six progenies were derived from the PSC seed source. Five of the 6 segregated PSC SI plants, 4 of which fit a 3 1 ratio of PSC SI plants. All plants of the sixth progeny were SI. Two F₁ progenies with the same PSC pollen parent produced significantly different segregations of PSC SI plants. It appeared that PSC acted as a major gene when the most recessive S allele was also present, but PSC was not expressed when the most dominant S allele was present. Clones propagated from PSC plants were SI and cross incompatible with a related S-allele tester. Thus, PSC was transient in that it was apparent in seed-propagated plants but not in plants clonally propagated from the PSC individuals.Scientific Journal Series Paper Number 12,299 of the Minnesota Agricultural Experiment Station     

Monotelodisomics in pearl millet

Summary In the progeny of crosses between plants with the chromosome number 2n=13+2 telocentrics as the male parents and the normal diploids of Pennisetum typhoides S. & H., two plants with 2n=13+1 telocentric chromosome were located. These two plants were heterozygous for an interchange, since at diakinesis and metaphase I associations of four chromosomes were observed. These plants had a chromosome constitution of 2n=13+t (or 6+tI); one chromosome of a homologous pair was represented by a telocentric chromosome so was monosomic for one arm, that is, these plants were monotelodisomics (Kimber and Sears, 1968).     

A higher-plant type ζ-carotene desaturase in the cyanobacterium Synechocystis PCC6803

The genomic DNA sequence of Synechocystis was analysed for putative -carotene desaturase genes. Two promising candidates slr0940 and slr0033 were found with similarities to the structurally different -carotene desaturase genes from higher plants and Anabaena, respectively. Only the expression product of the analogue to the plant gene, slr0940, was able to mediate the 2-step desaturation of -carotene via neurosporene to lycopene after complementation of this pathway in Escherichia coli. When enzyme reactions were carried out with this protein, activity was obtained with either -carotene or neuroporene as substrates. The in vitro reaction was inhibited by the pyrimidine derivative J852 which is effective as experimental herbicide in plants. The occurrence of two different types of -carotene desaturases among cyanobacteria and the phylogenetic consequences on chloroplast evolution are discussed.     

Differential expression of GABA<Subscript>A</Subscript> receptor π subunit in cultured rat alveolar epithelial cells

Although type A -aminobutyric acid (GABA) receptors (ligand-gated Cl^– channels) have been extensively studied in the central nervous system, no information is available on this receptor in lung cells. We have examined the expression of GABA_A receptor -subunit (GABRP) during the trans-differentiation between rat alveolar epithelial type II cells and type I cells. Rat alveolar type II cells, when cultured on plastic plates, gradually trans-differentiated into type-I-like cells and lost their GABRP mRNA expression. However, the GABRP mRNA was partially retained in the type II cells cultured on Matrigel. Keratinocyte growth factor (a mitogen of type II cells) increased GABRP expression. A detached collagen gel maintained the GABRP mRNA to a level close to that of the freshly isolated type II cells. An air–liquid interface culture system, mimicking in vivo conditions in the lung, significantly up-regulated the expression of GABRP mRNA and protein. mRNAs of the GABA_A receptor 1-, 3-, 2-, 2-, and 3-subunits were also detected in rat type II cells. These results suggest that GABRP expression is differentially regulated by culture substrata, growth factor, detached gel, and an air-apical surface.This work was supported by NIH R01 HL-52146, R01 NIH-071628, and OCAST HR01-093, and AHA heartland affiliate 0255992Z (to L.L.). N.J. was supported by an AHA heartland affiliate pre-doctoral fellowship (0315256Z).     

Carotenoid composition in milo (Sorghum vulgare) shoots as affected by phytochrome and chlorophyll

The composition of coloured carotenoids in the milo shoot was investigated quantitatively (high performance liquid chromatography) during light-mediated plastidogenesis, including the time span of photodelay as caused by medium and high light fluxes. It was found that as long as only the far-red-absorbing form of phytochrome operates, the carotenoid pattern remains virtually the same as in complete darkness (violaxanthin and lutein as major constituents, traces of -carotene). On the other hand, the pattern changes dramatically in white or red light with increasing amounts of chlorophyll (lutein and -carotene dominate, -carotene showing the strongest relative increase). Photodelay during the early phase of plastidogenesis affects the carotenoid composition strongly. Increase of neoxanthin, violaxanthin and -carotene contents are diminished while lutein accumulation proves resistant towards chlorophyll-mediated photoinhibition. The photodelay can be diminished by an appropriate light pretreatment. The data indicate that light-mediated control over carotenoid accumulation is exerted at three levels: i) a coarse control through phytochrome, ii) fine tuning in connection with chlorophyll accumulation, iii) stabilization of holocomplexes against photodecomposition.Abbreviations GG14 high fluence rate green-yellow light - HPLC high-performance liquid chromatography - Chl chlorophyll - WL_w weak white light (1200 lx) - WL_m medium flux white light (12000 lx)     

Polymorphism of human liver alcohol dehydrogenase: Identification of ADH2 2-1 and ADH2 2-2 phenotypes in the Japanese by isoelectric focusing

Liver homogenate-supernatants from most Japanese exhibit an atypical pH optimum for ethanol oxidation at pH 8.8 instead of 10.5, the typical pH-activity optimum. It has been proposed that atypical livers contain alcohol dehydrogenase isozymes with ₂ subunits while typical livers contain isozymes with ₁ subunits, both produced by the ADH ₂ gene. Because it is difficult to differentiate the atypical ADH₂ 2-2 phenotype from the ADH₂ 2-1 phenotype by starch gel electrophoresis, an agarose isoelectric focusing procedure was developed that clearly separated the atypical Japanese livers into two groups, A1 and A2. The isozymes in A1 and A2 livers were purified. Type A1 livers contained a single isozyme with an atypical pH-rate profile; it was designated ₂₂. Three isozymes were isolated from A2 livers, two of which corresponded to ₁₁ and ₂₂. A third, absent from the typical and the atypical A1 livers, had an intermediate mobility; it was designated ₂₁. Type A1 livers are, therefore, the homozygous ADH₂ 2-2 phenotype, and type A2 livers, the heterozygous ADH₂ 2-1 phenotype. The ADH₂ 2-2 phenotype was found in 53% of 194 Japanese livers, and the ADH₂ 2-1 phenotype, in 31%. Accordingly, the frequency of ADH ₂ ² was 0.68.This study was supported by U.S. Public Health Service Grant AA 02342.     

C-banding and DNA content in three species of Belostoma (Heteroptera) with large differences in chromosome size and number

C-banding was carried out on Belostoma elegans (2n=26+X₁X₂Y) (), B. micantulum (2n=14+XY) () and B. oxyurum (2n=6+XY) () (Belostomatidae, Heteroptera). The C-bands always have a telomeric localization and no interstitial bands were detected. An inverse relationship between chromosome size and chromosome number exists, and besides, an inverse relationship between chromosome size and the size of the C-bands was observed. The DNA content was determined in all three species. B. elegans has a C content of 1.55±0.06 pg, B. micantulum has 0.88±0.04 pg and B. oxyurum had 0.53±0.04 pg.Considering the male meiotic characteristics, the chromosome complement and the results of C-banding and DNA content, the karyotype of B. oxyurum probably originated through autosomal fusions. The karyotype of B. micantulum and B. elegans could have originated through autosomal fusions or fragmentations respectively; with the information available up to now it is not possible to discard any of the two pathways.     

Flagellar surface morphology ofChlamydomonas eugametos

Summary Flagellar surfaces ofChlamydomonas eugametos fixed in OsO₄ were compared to those fixed in glutaraldehyde by scanning and transmission electron microscopical techniques. They appeared to be quite different. Flagella fixed in OsO₄ had a pleated membrane that vesiculated when Tris was present during fixation. Flagella fixed in glutaraldehyde of various concentrations and buffer strengths, appeared to have disc-bearing appendages which were continuous with the membrane. On growing flagella the appendages occurred after 2 hours and they increased in size and complexity during the next 3 to 4 hours. No structural differences were observed between flagella of vegetative cells and those of gametes. Mating competence of gametes appeared to parallel the development of mature appendage-bearing flagella. The possibility that membrane vesicles, found in the cell culture medium, are derived from the flagellar appendages is discussed.     

Influence of Genotype on Endoreduplication of Giant Chromosomes and Expressivity of Character eyelessin Drosophila melanogaster

In Drosophila melanogaster lines LA (low activity), HA (high activity), andOregon-R (wild type), the effect of genetic background on endoreplication in giant chromosomes of salivary glands, fecundity, and expression of mutation eywas studied. The degree of chromosome polyteny and the number of adult flies in saturated lines ey _L, ey _H, and ey _Or were significantly higher than in the original lines. The degree of chromosome polyteny was correlated with fecundity. The expressivity of the ey character was shown to be far lower in lines ey _L, ey _H, and ey _Or than in the original eyelessline. In the saturated lines, the eye facets were reduced to a similar degree. All the lines studied displayed clear-cut sexual distinctions in this parameter. In the ey _L line, the coefficients of variation for the degree of chromosome polyteny, fecundity, and expressivity of the ey mutation were much lower than in lines eyeless, ey _LAandey _Or.