Synaptonemal complex formation in hybrids of Lolium temulentum × Lolium perenne (L.)

Chromosome pairing and synaptonemal complex formation at zygotene and pachytene are described from serial section reconstructions of pollen mother cell nuclei in a triploid hybrid containing two haploid sets of Lolium perenne chromosomes, one of L. temulentum and two acces-sory B chromosomes. At pachytene the homologous L. perenne chromosomes form complete and continuous synaptonemal complexes while the L. temulentum chromosomes show extensive nonhomologous pairing both within and between themselves. At zygotene however, homoeologous pairing in the form of a trivalent and very little non-homologous pairing is observed. Evidently, there exists a mechanism that eliminates homoeologous association during zygotene to ensure strict bivalent formation between homologous chromosomes at pachytene. In Lolium this mechanism is under the influence of the B chromosomes and bears close similarity with that in allohexaploid wheat controlled by the Ph locus.     

The genotypic control of homoeologous chromosome association in Lolium temulentum × Lolium perenne interspecific hybrids

Two contrasting genotypes of Lolium perenne and two inbred lines of L. temulentum were examined with regard to their effect on homoeologous chromosome pairing in interspecific hybrids derived from them. Substantial differences in chiasma frequency were observed between the hybrid progeny of the different parental types. The background genes involved were found to operate in the presence and in the absence of B chromosomes. The combination of A chromosome genes present in some of the 0B hybrids was found to result in a considerable suppression of chiasma formation at the diploid level, and the restriction of pairing to strict homologues at the tetraploid level. It appears, therefore, that genes are present within the diploid species of the genus Lolium which are capable of performing a function similar to that of the Ph locus in wheat.     

黑麦草(Lolium perenne L.)内生真菌的检测、分离及鉴定

从多年生黑麦草(LoliumperenneL.)5个品种———SR4000、Pinnacle、Topgun、CalypsoⅡ、Justus中分离出61个菌株。次培养后,所得形态稳定的菌株可分为4个形态群,依据其形态特征及APPCR的结果,确定其中的57个分离菌株为Neotyphodiumlolii。     

Rapd fingerprinting of diploid Lolium perenne × hexaploid Festuca arundinacea hybrid genomes

We tested the application of RAPD technology for identification of hybrid genomes originated from a maternal clone of Lolium perenne L. (2n = 2x = 14) bearing cytoplasmic male sterility, which was pollinated separately by five clones of Festuca arundinacea Schreb. cv. Barocco (2n = 6x = 42). Six classes of RAPD markers were recognized, specific to: 1) Festuca genome and inherited into F1 hybrid genomes, 2) Lolium genome inherited into F1 hybrid genomes, 3) Lolium-specific bands not found in F1 progeny, 4) Festuca-specific bands not found in F1 progeny, 5) new bands found only in F1 hybrid profiles, 6) bands common to all parental and F1 hybrid genotypes. RAPD data were shown to have full potential a) to serve as an unequivocal proof of genome recombination in perennial ryegrass × tall fescue hybrids, b) to identify hybrid genomes, c) to reveal phenetic relationships of the accessions from crossing families, d) to enhance, by fingerprinting, the selection of superior hybrid material for further breeding. RAPD data were found to be consistent with the festucoid phenotype of F1 hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Synaptonemal complex formation: where does it start?

The synaptonemal complex is a prominent, evolutionarily conserved feature of meiotic prophase. The assembly of this structure is closely linked to meiotic recombination. A recent study in budding yeast reveals an unexpected role in centromere pairing for a protein component of the synaptonemal complex, Zip1. These findings have implications for synaptonemal complex formation.     

Genome balance in six successive generations of the allotetraploid Festuca pratensis × Lolium perenne

In the allotetraploid, Festuca pratensis Huds. (2n = 4x = 28) × Lolium perenne L. (2n = 4x = 28) the balance of chromatin, as determined by GISH, changes over successive generations of open pollination in favour of L. perenne. There is extensive recombination between chromosomes of the two parental genomes, as well as substitution of whole Festuca chromosomes by whole Lolium chromosomes. The total number of Lolium chromosomes increased from a mean 14.36 in the F₂ to 16.26 in the F₆, and the total number of Festuca chromosomes decreased correspondingly from a mean of 13.57 to a value of 11.56. The number of recombinant chromosomes and recombination breakpoints per genotype also increased from generation to generation, although the respective values of both characters were higher for Festuca (0.86–8.41 and 1.14–15.22) than for Lolium (0.68–4.59 and 0.68–6.0). The proportion of total genome length contributed by the L. perenne chromatin increased from about 50% in F₂ to 59.5% in F₆. The results are based on the sample of 134 plants studied (26–28 plants per generation), and are discussed in terms of the dominance of Lolium chromosomes over those of Festuca, and possible mechanisms underlying this phenomenon of chromatin substitution.     

Sympatric Epichloë species and chemotypic profiles in natural populations of Lolium perenne

The distribution of different Epichloë species within eight natural populations of Lolium perenne was studied. In total, 40.2% of the asymptomatic plants were infected by Epichloë festucae var. lolii or by Epichloë typhina. Both species occurred in sympatry in seven grass populations, and some plants had dual infections by both taxa. No hybrid taxa such as Epichloë hybrida were detected. Epichloë festucae strains were classified into two morphotypes, M1 and M3, according to culture characters, both morphotypes occurred in sympatry in seven populations. Plants bearing stromata produced by Epichloë typhina were observed, but asymptomatic plants infected by this species also occurred in seven populations. The alkaloid profile of Lolium perenne plants was related to the morphotype of their infecting strains: most plants infected by M3-strains were characterized by lolitrem, and those with M1-strains contained either ergovaline or lolitrem. Plants infected by Epichloë typhina were characterized by high peramine content.     

Inheritance of Organelle DNA in Rye (Secale cereale L.) with Triticale (×Triticale Thch.) Hybrids

The mode of inheritance of chloroplast and mitochondrial DNA (mtDNA) in rye × triticale intergeneric hybrids has been studied with the use of specific PCR markers for loci 18S/5S and 3rbcL in organelle DNA. In rye × triticale BC₁, mtDNA copies of two types, paternal and maternal, have been found; in BC₂ plants, only paternal mtDNA and chloroplast DNA (cpDNA) have been detected. Mechanisms determining the inheritance and/or differential amplification of organelles of a specific type are discussed.     

Coexpression of myogenic functions in L6 rat × T984 mouse myoblast hybrids

L6 rat myoblasts differ from T984 mouse myoblasts in the expression of several myogenic functions. Simple hybrids between these myogenic lines contained mostly chromosomes and expressed the mouse phenotype. Hybrids containing an approximately balanced set of chromosomes from each parent were constructed by fusing tetraploid L6 cells to T984 myoblasts. These hybrids were allowed to differentiate and their expression of myogenic proteins was compared to the parental phenotypes. The synthesis of creatine phosphokinase isoenzymes is codominantly expressed in the hybrid cells. Myosin light chain synthesis and the levels of acetylcholine receptor are either regulated by the mouse genome or are codominantly expressed.     

Production,morphology, and cytogenetics of Triticum aestivum (L.) Thell × Elymus scabrus (R. Br.) Love intergeneric hybrids obtained by in ovulo embryo culture

Summary Intergeneric hybrids were produced between common wheat, Triticum aestivum (2n=6x=42, AABBDD), and an apomictic Triticeae species, Elymus scabrus (syn. Agropyron scabrum) (2n=6x=42, HHSSSS), the first successful report of this cross. Nine tiny, underdeveloped, and structureless embryos were obtained in vitro only by in ovulo embryo culture at 4 days after pollination, which gave rise to five mature hybrid plants. All the hybrid plants were vigorous and possessed a phenotype intermediate to the two parents. There were 2n=6x=42 (ABDHSS) somatic chromosomes in the hybrids. There was little or no homology between the parental genomes, as shown by an overall meiotic chromosome association of 32.83 I + 4.08 rod II + 0.21 ring II + 0.18 III + 0.02 IV. The hybrids were completely sterile and so far backcrosses to wheat parent have not been successful. Alternate approaches to induce gene transfer(s) from E. scabrus to wheat are being attempted.Agriculture Canada Contribution No. 398.     

“Blind” mapping of genic DNA sequence polymorphisms in Lolium perenne L. by high resolution melting curve analysis

High resolution melting curve analysis (HRM) measures dissociation of double stranded DNA of a PCR product amplified in the presence of a saturating fluorescence dye. Recently, HRM proved successful to genotype DNA sequence polymorphisms such as SSRs and SNPs based on the shape of the melting curves. In this study, HRM was used for simultaneous screening and genotyping of genic DNA sequence polymorphisms identified in the Lolium perenne F2 mapping population VrnA. Melting profiles of PCR products amplified from previously published gene loci and from a novel gene putatively involved in vernalization response successfully discriminated genotypes in absence of allelic sequence information, and allowed to determine allele segregation in VrnA. Here we introduce the concept of “blind” mapping based on HRM as a powerful, fast and cheap method to map any DNA sequence polymorphisms without prior knowledge of allelic sequences in the key grassland species perennial ryegrass (Lolium perenne L.).     

The relative significance for stem elongation and flowering in Lolium temulentum of 3β-hydroxylation of gibberellins

In previous experiments with many gibberellins (GAs) and GA derivatives applied to Lolium temulentum L., quite different structural requirements were evident for stem elongation on the one hand and for the promotion of flowering on the other. Whereas hydroxylation at carbons 12, 13 and 15 enhanced flowering relative to stem growth, the reverse was the case at carbon 3 (L.T. Evans et al. 1990, Planta 182, 97–106). The significance of hydroxylation at carbon 3 is examined in this paper. The application of inhibitors of 3β-hydroxylation, including C/D-ring-rearranged GAs, reduced stem growth but, in the case of the two acylcyclohexanediones, increased the flowering response when applied on the inductive long day. Later applications of the acylcyclohexanediones, made after floral initiation had occurred, were inhibitory to flowering, suggesting that subsequent inflorescence development requires 3β-hydroxylated GAs. Applications of the 3α-hydroxy epimers of GA₁, GA₃ and GA₄ gave slightly less promotion of flowering in comparison with the 3β-hydroxy GAs, but far less promotion of stem elongation, except in the case of 3-epi-GA₄, which was comparable to GA₄. The 3α-hydroxy epimer of 2,2-dimethyl GA₄ gave less promotion of flowering than its 3β-hydroxy epimer but almost no promotion of stem elongation. The 3α-hydroxy epimers of GA₃ and 2,2-dimethyl GA₄ did not act as competitive inhibitors of the stem elongation elicited by GA₃ and 2,2-dimethyl GA₄, respectively. These results extend the differences in GA structure which favour flowering as opposed to stem elongation, and indicate that 3-hydroxylation and its epimeric configuration are of much greater importance to stem elongation than to flower initiation in Lolium.     

Production of trigeneric (barley × wheat) × rye hybrids

Summary Rye (Secale cereale cv. Prolific 2n=14 and 2n =14 + 2B was crossed onto hybrids between barley (Hordeum vulgare 2n = 14) and wheat (Triticum aestivum 2n= 42). Pollinated florets were injected with GA₃ to promote fertilization and hybrid embryo development. At 16 days after pollination the watery caryopses were removed, embryos dissected and cultured on a modified B₅ medium. Approximately 20% of the cultured embryos produced both roots and coleoptile and developed into viable seedlings. Viable seeds were also obtained at a low frequency from the same cross combinations. The hybrids were wheat-like except for the hairy neck characteristic of rye. There were 35 chromosomes in somatic tissue; 21 wheat, 7 barley and 7 rye. The rye chromosomes were distinguishable by their larger size and terminal C-bands. A lower seed set was obtained using pollen from rye plants with 2n=14 + 2B chromosomes than from plants without B chromosomes.Contribution No. 577, Ottawa Research Station     

Cryopreservation of in vitro-grown shoot tips of 'Troyer' citrange [Poncirus trifoliata (L.) Raf. × Citrus sinensis (L.) Osbeck.] by encapsulation-dehydration

. In vitro-grown shoot tips excised from preconditioned stock shoots of 'Troyer' citrange were successfully cryopreserved by encapsulation-dehydration. Optimal survival of cryopreserved shoot tips was achieved when encapsulated shoot tips were dehydrated to 17.1% water content. The sucrose concentration in the preconditioning medium significantly influenced the growth and dry matter percentage of the stock shoots as well as subsequent survival of the cryopreserved shoot tips. Maximal growth of stock shoots was obtained in sucrose concentrations in the range of 0.15 M to 0.29 M, while the dry matter percentage increased as sucrose concentration increased up to 0.44 M. The survival of cryopreserved shoot tips increased from 40% to approximately 80% as the sucrose concentration for stock shoots increased from 0.09 M to 0.22 M or 0.29 M. The benzyladenine concentration in the post-culture medium significantly affected the survival and regrowth of the cryopreserved shoot tips. Survival of the shoot tips was lowest when they were post-cultured on benzyladenine-free medium. However, high benzyladenine concentrations (3-4 µM) induced callus formation. Optimal recovery was obtained in post-culture medium containing 2 µM benzyladenine and 0.05 µM !-naphthalene acetic acid. The extraction of shoot tips from alginate beads greatly improved the regrowth of cryopreserved shoot tips.     

Structural investigations on the lignin–carbohydrate complexes of Lolium perenne

1. Lignin-carbohydrate complexes isolated from leaf blade, leaf sheath and stem tissue of ryegrass by extraction with dimethyl sulphoxide were examined by fractionation procedures. Although the complexes are heterogeneous, heterogeneity is shown only in the ratio of the individual monosaccharide residues and not in the ratio of lignin to carbohydrate. 2. The molecular weight of the complexes is high (>/=150000), but chemical modification by alkaline hydrolysis, borohydride reduction or lead tetra-acetate oxidation does not drastically decrease it. Low-molecular-weight fragments released by alkaline treatment were shown to contain acetic acid, ferulic acid and p-coumaric acid. 3. On the basis of the chemical stability of the complexes, it is postulated that at least three types of bonding may be present between lignin and carbohydrate, namely one cleaved on borohydride reduction, another cleaved by alkali and a linkage resistant to alkali. 4. The carbohydrate portion of the complexes is composed of beta-(1-->4)-linked d-glucose residues (cellulose) and beta-(1-->4)-linked chains of xylose residues. Side chains involving arabinose and galactose residues are linked to C-3 of some of the xylose residues. 5. How the components of the complexes are held together is not certain, but it is suggested that the phenolic acids may act as cross-linking agents.     

Can cell wall peroxidase activity explain the leaf growth response of Lolium temulentum L. during drought?

In two experiments, the effect has been investigated of a mildand a more prolonged drought on the spatial distribution ofgrowth, epidermal cell lengths and cell wall peroxidase activitiesin the leaf elongation zone of the grass species Lolium temulentumL. In both experiments drought reduced the size of the elongationzone and local rates of elongation within it. Abrupt increasesin cell wall-associated peroxidase activity occurred at or closeto the position where elongation ceased in the leaf elongationzones of well-watered and mildly drought-stressed plants. Moreprolonged drought caused a 200–300% increase in the cellwallassociated peroxidase activity in the elongation zone only.The significant increase in the elongation zone cell wall peroxidaseactivity and its spatial variation provides evidence of a potentiallycausal role for cell wall-associated peroxidase in restrictingcell expansion during drought. Key words: Cell wall peroxidase, leaf expansion, drought     

Recovery from drought stress in Lolium perenne (Poaceae): are fungal endophytes detrimental?

Perennial ryegrass (Lolium perenne) is a cool-season, perennial species widely used for forage and turf. It is often infected by a clandestine, endophytic fungus (Neotyphodium lolii) that has the potential to affect host growth responses to abiotically stressful conditions. In some species, the grass-endophyte symbiosis is mutualistic, but the relationship is reported to be contingent on environmental conditions and host genotype in L. perenne. The objective of this research was to determine the potential effects of endophyte infection on recovery from severe drought stress in variable genotypes of a perennial ryegrass cultivar. Sixteen infected (+E) and 16 uninfected (-E) ramets were planted in the greenhouse for each of 10 ryegrass genotypes. Eight +E and eight -E plants per genotype were exposed to three sequential droughts where water was withheld for 11-14 d, resulting in <5% soil moisture; the others (control) were watered as needed. Response variables were tiller numbers 1 wk and 4 wk after drought, and leaf area and dry mass of shoots and roots 7 wk after drought. In both control and drought, -E plants had more tillers, and greater leaf area and total mass, than +E plants, suggesting a detrimental effect of endophytic fungi. Fungal hyphae survived the drought and were abundant in post-drought, +E plants. The effects of endophytes were specific for particular host genotypes, as exemplified by significant genotype × endophyte interactions. Root : shoot ratio and percent of mass allocated to tiller bases (a rough measure of resource storage) showed genotype × endophyte × drought interactions. There was plasticity for root : shoot ratio and genetic variation in the ability to restore root growth during recovery from drought. For 7 of 10 genotypes, -E plants showed an equal or greater allocation to tiller bases than +E plants following drought recovery, illustrating a cost to endophyte infection for some genotypes. The symbiotic relationship between L. perenne and its endophyte primarily benefits the fungus, not the host, under many environmental conditions.     

Changes in behaviour associated with pair formation in the polychaete Harmothoë imbricata (L.)

Spacing between individuals in populations of Harmothoë imbricata has been investigated both on the shore and in the laboratory. Males tend to occur closer together than females, and the mean male‐male individual distance measured was less than the mean distance between females; male—female distances for immature worms were intermediate. When worms mature they pair: the male mounts the female and lies across her dorsal surface. There is evidence that after spawning the members of a pair separate. Contact responses between worms have been investigated in the laboratory. Most encounters between immature worms lead to separation, as a result of one or both worms moving rapidly away from the other or fighting; females show more marked avoidance behaviour than males. The majority of male‐male and female‐female encounters between mature worms also lead to separation but in male‐female encounters the male usually mounts the female. A male which has mounted a female becomes highly aggressive and will attack intruding males but not females.