Epicuticular waxes from Agropyron dasystachyum,agropyron riparium and Agropyron elongatum

Epicuticular waxes from whole plants of Agropyron dasystachyum var. psammophylum, A. riparium and A. elongatum contain hydrocarbons (5–8 %), long chain esters (12–15%) and free acids (2–5%). The major esters are C₃₄C₅₆ esters derived from C₁₆C₃₀ acids and alcohols (1-hexacosanol is the major alcohol) but C₃₁, C₃₃ and C₃₅ esters (3–11%) are also present. The latter esters are C₁₈ and C₂₀ acid esters of C₁₃ and C₁₅ 2-alkanols. A. dasystachyum wax contains 2% free alcohols, that of A. riparium contains 17% and that of A. elongatum 11% (1-hexacosanol is the major alcohol in each). Diesters (2%), C₈C₁₂ diols esterified by (E)-2-alkenoic acids, are present in A. riparium wax. Hentriacontane-14,16-dione is present: 29% in A. dasystachyum wax and 32% in A. riparium wax, but only 5% in A. elongatum wax. 25-Oxohentriacontane-14,16-dione forms 14% of A. dasystachyum wax and 27% of A. elongatum wax but the oxo β-diketones of A. riparium wax (5%) consist of both 10-oxo- and 25-oxohentriacontane-14,16-diones in the ratio 4:1. Hydroxy β-diketones of the waxes are 25- and 26-hydroxyhentriacontane-14,16-diones; in A. dasystachyum (20%) the ratio is 3:1, in A. elongatum (20%) the ratio is 9:1 but in A. riparium (5%) it is ca 1:2. The configuration of the hydroxyl group in the 26-hydroxy β-diketone is opposite to that in the 25-hydroxy derivative. The unusual composition of the oxygenated β-diketones of A. riparium confirms that this species should be regarded as separate from A. dasystachyum. Wax from A. elongatum also contains 4-hydroxy-25-oxohentriacontane-14,16-dione (4%) and an unusual oxo-β-ketol, 18-hydroxy-7,16-hentriacontanedione (2%), both these components are probably derived biosynthetically from the 25-oxo β-diketone which is the major component of this wax. Syntheses of racemic 18-hydroxy-7,16-hentriacontanedione and of a model β-ketol, 12-hydroxy-10-pentacosanone, are described.     

Production and identification of primary trisomics in diploid Agropyron cristatum (crested wheatgrass).

Six of the seven possible primary trisomics in Agropyron cristatum were produced. Based on morphology, arm length ratios, and C-banding patterns, they were identified as primary trisomics for chromosomes A, B, C, D, E, and G. Agropyron cristatum is one of several species constituting the crested wheatgrass complex. All species in this complex contain one basic genome (P). A study was conducted to produce and identify a primary trisomic series that will be used to map genes to individual chromosomes. A population of 157 plants were generated by crossing autotriploids (PPP) with diploid (PP) A. cristatum: 58 were diploid (2n = 14), 76 were primary trisomies (2n = 15), 17 were double trisomic (2n = 16), 4 were triple trisomics (2n = 14 + 3), 1 was telocentric trisomic (2n = 14 + 1 telo), and 1 was tetratrisomic (2n = 14 + 4). Karyotype analysis of acetoorcein-stained chromosomes was carried out using the CHROMPAC III computer program; for analysis of C-banded karyotypes, the computer imaging analysis program PCAS (Plant Chromosome Analysis System) was used to identify the primary trisomics. Of the 47 primary trisomics analyzed, 21 plants had one extra satellited chromosome E, 18 with the satellited D chromosome, 3 each for chromosomes B and G, and 1 each for chromosomes C and A. Chromosome pairing was studied in trisomies B, D, E, and G. Trisomics for chromosomes B and G were similar in their mieotic behavior. Each had a trivalent frequency of about 60% and pollen stainability of less than 40%. Trisomics for chromosomes D and E had a trivalent frequency of about 30% and pollen stainability of over 70%.     

Regeneration of albino shoots in cultured anthers of intermediate wheatgrass (Agropyron intermedium (Host) Beauv.)

Shoots were regenerated from Oahe intermediate wheatgrass anthers cultured on Tsay's, N6, Yu-pei and 85D12 basal media supplemented with kinetin and 2,4-D or NAA. Androgenesis mainly started with symmetrical divisions of pollen nuclei immediately followed by cytokinesis. Formation of tetranucleate pollen grains resulting from asymmetrical divisions of the pollen nuclei was also noted. Tsay's medium was more effective for callus induction, while N6, Tsay's and Yu-pei differentiation media were equally effective for shoot regeneration in the calluses. All regenerants were albino.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - NAA naphthaleneacetic acid     

The origin of spelt and free-threshing hexaploid wheat

It is widely believed that hexaploid wheat originated via hybridization of hulled tetraploid emmer with Aegilops tauschii (genomes DD) and that the nascent hexaploid was spelt, from which free-threshing wheat evolved by mutations. To reassess the role of spelt in the evolution of Triticum aestivum, 4 disomic substitution lines of Ae. tauschii chromosome 2D in Chinese Spring wheat were developed and one of them was used to map the Tg locus, which controls glume tenacity in Ae. tauschii, relative to simple sequence repeat (SSR) and expressed sequence tag loci on wheat chromosome 2D. The segregation of SSR markers was used to assess the presence of Tg alleles in 11 accessions of spelt, both from Europe and from Asia. Ten of them had an inactive tg allele in the D genome and most had an active Tg allele in the B genome. This is consistent with spelt being derived from free-threshing hexaploid wheat by hybridization of free-threshing wheat with hulled emmer. It is proposed that the tetraploid parent of hexaploid wheat was not hulled emmer but a free-threshing form of tetraploid wheat.     

Evolutionary dynamics of Waxy and the origin of hexaploid Spartina species (Poaceae)

We investigated the evolutionary dynamics of duplicated copies of the granule-bound starch synthase I gene (GBSSI or Waxy) within polyploid Spartina species. Molecular cloning, sequencing, and phylogenetic analyses revealed incongruences between the expected species phylogeny and the inferred gene trees. Some genes within species were more divergent than expected from ploidy level alone, suggesting the existence of paralogous sets of Waxy loci in Spartina. Phylogenetic analyses indicate that this paralogy originated from a duplication that occurred prior to the divergence of Spartina from other Chloridoideae. Gene tree topologies revealed three divergent homoeologous sequences in the hexaploid S. alterniflora that are consistent with the proposal of an allopolyploid origin of the hexaploid clade. Waxy sequences differ in insertion–deletion events in introns, which may be used to diagnose gene copies. Both paralogous and homoeologous coding regions appear to evolving under selective constraints.     

Leaf water potential,component potentials and relative water content in a xeric grass,Agropyron dasystachyum (Hook.) Scribn.

Summary Leaf water potential ( _l ), osmotic potential ( _s ), pressure potential ( _p , turgor pressure), relative water content (R) and their interrelationships were determined for a xeric grass (Agropyron dasystachyum) found in the grasslands of Canada. Thermocouple psychrometers were used to measure _l and _s ; _p was obtained by subtraction. _l dropped from near 0 bars to about-28 bars as R went from 90% to 75%. R greater than 90% was not observed, perhaps because of a systematic error in determination of turgid water content. R remained relatively high in A. dasystachyum, even at low _l . The slope of the _l -R relationship was similar to other species which are generally considered to be drought tolerant. _p as high as 14 bars was observed. Most of the decrease in _l was accounted for by a decline in _p . The ability of A. dasystachyum to adjust to fluctuating water stress over the growing season is probably as much related to changes in tissue structure and turgor relationships as to simple changes in osmotic potential.     

The meiotic behaviour of the XY pair in Lutreolina crassicaudata (Marsupialia: Didelphoidea)

The meiotic behaviour of the XY pair of the didelphid Lutreolina crassicaudata is analyzed by microspreading of spermatocytes for visualization of chromosomal axes and by three-dimensional reconstruction of spermatocyte nuclei from EM thin sections. The delay in pairing of sex chromosomes compared to autosomes and the absence of a synaptonemal complex between the axes of the X and Y chromosomes, already described for South American marsupials by three-dimensional reconstruction and for Australian species with synaptonemal complex microspreadings, is confirmed for this species. Sections demonstrate that at the diffuse stage and diplotene the dense plate occupies the region of the inner face of the nuclear envelope in contact with the XY body. Spreads show an structure similar in staining to the axes that becomes apparent simultaneously with the dense plate, called a balloon. The mechanism of XY pairing during meiotic prophase appears to be common to American and Australian marsupials as the same morphological pattern is found in all the species described. This mechanism is different from the way of pairing and segregation known for eutherian sex chromosomes.     

Tetraploids inLuzula multiflora (Juncaceae) in Ireland: Karyology and meiotic behaviour

Populations ofLuzula multiflora s.l. in Ireland were examined karyologically. Plants from 14 populations were invariably tetraploid with 2n=24. Chromosomes of the tetraploid are of AL type (true tetraploidy). Meiosis of the tetraploids is of the same type as described for otherLuzula taxa in the literature. In meiosis, 12 bivalents are regularly formed. A hypothesis based on the morphological and allozyme data, that the tetraploids are of alloploid origin, is supported by the present results. Meiosis in an artificial hybrid between the presumed parental taxa,L. campestris andL. pallidula, was studied; a certain tendency towards chromosome doubling was observed. The geographical distribution of theL. multiflora cytotypes is also discussed.     

The meiotic behaviour of a haploid pearl millet

Meiosis was studied in one haploid plant of pearl millet, obtained from twin seedlings. Apparent pairing resulted in up to three bivalent associations at pachytene. At diakinesis and metaphase I associations of two, three or four chromosomes were observed. The frequency distribution of bivalents at metaphase followed a truncated Poisson distribution, suggesting that the bivalents were random pairs. They were considered to be pseudo-bivalents. Univalents varied in number from three to seven and they formed s-s and e-g associations. The s-s and e-s associations were random associations since their frequency distributions also followed a truncated Poisson distribution. A bipolar spindle was observed in a large number of PMC's but in a few cases two unipolar spindles were observed. The anaphase I distribution of the chromosomes deviated from abinomial distribution. Laggards were observed at telophase I. The dyads varied in size and in number of chromosomes. After the second division cell wall formation often failed to take place in one or in both the dyads, resulting in the formation of 2 to 4 microspores and microspores with two nuclei. The pollen grains varied in size and number of chromosomes. The plant was completely sterile.     

The meiotic behaviour of inversions in polyploid aloineae

In addition to the four classical inversion phenomena, meiosis in tetraploid paracentric inversion heterozygotes produces multiple dicentric and complex tricentric bridges which were previously little understood. Also formed are open loop chromatids which can give rise to dicentric chromosomes in the progeny. A qualitative and quantitative study of the first and second meiotic division in Gasteria nigricans var. crassifolia (Liliaceae, Aloineae) agrees closely with theoretical considerations. Breakage of dicentric bridges results in the formation of chromosomes carrying large terminal deletions. These are shown to be viable in the diploid gametes produced by tetraploids because of the buffering effect of the second haploid set of chromosomes.     

C-banding pattern and meiotic pairing in five rye chromosomes of hexaploid triticale

Summary The meiotic behaviour of rye chromosomes 1R, 2R, 3R, 6R and 7R/4R of hexaploid triticale Cachirulo is analyzed using the C-banding technique. These chromosomes show different C-banding patterns and present different pairing levels at metaphase I. A decreasing effect of large telomeric heterochromatin bands on pairing is deduced from the following two main facts: i) The chromosome 7R/4R shows the highest pairing associated with the smallest amount of heterochromatin, ii) pairing levels of 2 R short arm and 3 R long arm which carry large telomeric bands are less than their respective long and short arms lacking telomeric heterochromatin. Possible desynaptic effects of heterochromatin are discussed although an asynaptic effect cannot be rejected.     

The structure,meiotic behaviour and effects of B chromosomes in Briza humilis Bieb. (Gramineae)

A single population of Briza humilis contained two types of B chromosome, one a large (B^L) and the other a small (B^S) acrocentric. DNA measurements show that the B^L chromosome contains approximately twice as much DNA per unit length as the members of the regular complement. The meiotic pairing behaviour of the Bs is variable and B^L and B^S are seen to pair in some cells. The presence of B^L depresses the chiasma frequency of the regular complement but the chiasma frequency of A and B chromosomes does not appear to be related. The transmission rate of the B chromosomes is variable and the B^L shows a non-disjunction mechanism during microsporogenesis that is absent during megasporogenesis. For the B^S chromosome the transmission rate is very low and there is no evidence of a non-disjunction mechanism. In general seeds containing B^L chromosomes germinate more slowly than those without B chromosomes.     

The meiotic behaviour of autosomal heterochromatic segments in hedgehogs

Male meiosis in the two species of hedgehogs Erinaceus europaeus and Aethechinus algirus, possessing respectively three and two pairs of autosomes with large blocks of heterochromatin, has been studied. The heterochromatic segments pair homologously till the end of pachytene, but separate during diplotene, owing to lack of chiasmata in these regions. They also organize the nucleolus in both species. The sex chromosomes (sex vesicle) are not associated with the nucleolus. The lack of chiasmata in the heterochromatic segments is interpreted as possible mechanism for the conservation of vital genes, such as ribosomal cistrons.     

Molecular origin of female meiotic aneuploidies

Chromosome aneuploidy is a major cause of pregnancy loss, abnormal pregnancy and live births following both natural conception and in vitro fertilisation (IVF) and increases exponentially with maternal age in the decade preceding the menopause. Molecular genetic analysis has shown that these are predominantly maternal in origin and trisomies most frequently occur through errors in the first meiotic division. Analysis of chromosome copy number in the three products of female meiosis, the first and second polar bodies and the corresponding zygote by microarray comparative genomic hybridisation (array CGH), in women of advanced maternal age undergoing IVF, has recently revealed a pattern of frequent multiple meiotic errors, caused by premature predivision of sister chromatids in meiosis I and a high incidence of errors in meiosis II. This pattern is similar to those observed in various mouse models which implicate the gradual depletion of cohesins, which are essential for cohesion of sister chromatids, as the primary cause of age related aneuploidy in female meiosis. However, defects in other aspects of meiosis including the formation and stabilisation of chiasmata and the spindle assembly checkpoint (SAC) may also contribute. The challenge remains to explain the molecular basis of ‘physiological’ rather than ‘chronological’ female ageing and the contribution of multifactorial causes from the fetal to adult ovary. This article is part of a Special Issue entitled: Molecular Genetics of Human Reproductive Failure.     

The origin of natural tetraploid loach Misgurnus anguillicaudatus (Teleostei: Cobitidae) inferred from meiotic chromosome configurations

In the loach, or Oriental weatherfish Misgurnus anguillicaudatus (Teleostei: Cobitidae), diploid (2n = 50) and tetraploid individuals (4n = 100) are often sympatric in central China. The evolutionary mechanism of this tetraploidization was analyzed with the observation of meiotic behavior of chromosomes in both the germinal vesicles of mature oocytes and the primary spermatocytes in diploid and tetraploid loaches. Whereas diploid specimens usually showed 25 bivalents in meiotic cells, tetraploid loaches exhibited 0-6 quadrivalents and 38-50 bivalents in both sexes, with the modal number of quadrivalents as three in females and four in males. In the diploid specimens, the two largest metacentric chromosomes bearing nucleolar organizing regions (NORs) identified by chromomycin A(3) staining and fluorescence in situ hybridization with a 5.8S + 28S rDNA probe formed one bivalent with terminal association. In the tetraploids, four NOR-bearing chromosomes never formed a quadrivalent, but were organized into two terminally-associated bivalents. These findings suggest an autotetraploid origin of the natural tetraploid loach and subsequent rediploidization of whole genome. The latter process, however, seems still in progress as inferred from the concurrence of up-to several quadrivalents and the majority of bivalents.     

Origin of triploid Arachis pintoi (Leguminosae) by autopolyploidy evidenced by FISH and meiotic behaviour

Background and Aims

Polyploidy is a dominant feature of flowering-plant genomes, including those of many important crop species. Arachis is a largely diploid genus with just four polyploid species. Two of them are economically important: the cultivated peanut and A. glabrata, a tropical forage crop. Even though it is usually accepted that polyploids within papilionoid legumes have arisen via hybridization and further chromosome doubling, it has been recently suggested that peanut arose through bilateral sexual polyploidization. In this paper, the polyploid nature of the recent, spontaneously originated triploid cytotype of the tropical lucerne, A. pintoi, was analysed, and thereby the mechanism by which polyploids may arise in the genus.

Methods

Chromosome morphology of 2x and 3x A. pintoi was determined by the Feulgeńs technique and the rDNA sites were mapped by FISH. To investigate whether polyploidization occurred by means of unreduced gametes, a detailed analysis of the microsporogenesis and pollen grains was made.

Key Results

The 2x and 3x plants presented 9m + 1sm and a satellited chromosome type 2 in each haploid genome. Physical mapping revealed a cluster of 18S–26S rDNA, proximally located on chromosome 6, and two 5S rDNA loci on chromosomes 3 and 5. Diploid plants presented 10II in meiosis while trivalents were observed in all triploids, with a maximum of 10III by cell. Diploid A. pintoi produced normal tetrads, but also triads, dyads and monads. Two types of pollen grains were detected: (1) normal-sized with a prolate shape and (2) large ones with a tetrahedral morphology.

Conclusions

Karyotype and meiotic analysis demonstrate that the 3x clone of A. pintoi arose by autopolyploidy. The occurrence of unreduced gametes strongly supports unilateral sexual polyploidization as the most probable mechanism that could have led to the origin of the triploid cytotype. This mechanism of polyploidization would probably be one of the most important mechanisms involved in the origin of economically important species of Arachis, either by triploid bridge or bilateral sexual polyploidization.     

Hybridization in the genera Vulpia and Festuca (Poaceae): meiotic behaviour of artificial hybrids

The course of meiosis, including an analysis of chromosome configurations, is described for five diploid × diploid Vulpia crosses, five tetraploid × diploid Vulpia crosses, one hexaploid × diploid Festuca × Vulpia cross, one tetraploid × hexaploid Vulpia × Festuca cross, and one hexaploid × hexaploid Vulpia × Festuca cross. In most cases there was 97.5% or more pollen sterility, but two heptaploid plants obtained (presumably by non-reduction) from a hexaploid × diploid cross had about 60% stainable pollen. In the diploid hybrids pairing was quite extensive, and in V. ligustica × V. geniculata it was more or less as in the parent species (mode 7 bivalents, with regular separation). In the triploid hybrids the modal situation was 7 bivalents + 7 univalents, but evidence concerning the genomes which were pairing was equivocal. Evidence from the crosses at higher ploidy levels shows that both homogenetic and heterogenetic pairing does occur, although the relative amounts are uncertain. The results in general support the current classsification of Vulpia , except that they suggest the removal of V. alopecuros from section Loretia.     

The role of sister chromatid cohesiveness and structure in meiotic behaviour

Sister chromatid cores, kinetochores and the connecting strand between sister kinetochores were differentially silver stained to analyse the behaviour of these structures during meiosis in normal and two spontaneous desynaptic individuals of Chorthippus jucundus (Orthoptera). In these desynaptic individuals most of the chromosomes appear as univalents and orient equationally in the first meiotic division. Despite this abnormal segregation pattern, the changes in chromosome structure follow the same timing as in normal individuals and seem to be strictly phase dependent. Chromosomes in the first prometaphase have associated sister kinetochores and sister chromatid cores that lie in the chromosome midline; we propose that this promotes the initial monopolar orientation of chromosomes. However, the requirements of tension for stable attachment to the spindle force the autosomal univalents to acquire amphitelic orientation. Sister kinetochores behave in a chromosome orientation-dependent manner and, in the first metaphase, they appear to be interconnected by a strand that can be detected by silver impregnation, as seen in the second metaphase of wild-type individuals. The disappearance of the sister kinetochore-connecting strand, needed for equational chromatid segregation, however, can only take place in the second meiotic division. This connecting strand is ultimately responsible for the inability of chromosomes to segregate sister chromatids in the first anaphase. Received: 25 March 1997; in revised form: 14 July 1997 / Accepted: 22 August 1997