Optimization of microbiological parameters for enhanced griseofulvin production using response surface methodology

Central composite design was used to determine the optimal levels of microbiological parameters, viz., slant age, seed age and inoculum level, for enhanced griseofulvin production by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004 in shake flask fermentation. The optimal levels of slant age, seed age and inoculum level for Penicillium griseofulvum MTCC 1898 were found to be 8.8772 days, 4.2093 days, 12% (v/v) (᷁.56 kg dry cell mass/m³) and for Penicillium griseofulvum MTCC 2004, 8.221 days, 3.4875 days and 9% (v/v) (̀.09 kg dry cell mass/m³) respectively. The yield of griseofulvin under optimal conditions was found to be 1.65 times for Penicillium griseofulvum MTCC 1898 and 1.07 times for Penicillium griseofulvum MTCC 2004 higher than that obtained using unoptimized conditions. The fermentation time for maximum production of griseofulvin by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004 decreased by 4 days and 2 days respectively.     

Cryopreservation of alfalfa (Medicago sativa L.) cells by encapsulation-dehydration

Our objective was to establish a cryopreservation protocol for alfalfa (Medicago sativa L.) cells and study the physiological changes occurring in cells during cryopreservation treatment. Cell cultures of Pioneer cvs. 5262 (fall-dormant, winter-hardy) and 5929 (non-dormant, non-hardy) plants initiated regrowth after cryopreservation by encapsulation-dehydration (ED). Pre-treatment of the encapsulated cells for 4 days in B5 medium containing 0.75 M sucrose and dehydration for 4 h in a laminar flow hood were necessary to achieve maximum cell viability after ED and cryopreservation in liquid N₂ (EDN). Viability (measured as triphenyl tetrazolium chloride reduction) of the cv. 5262 cells after cryopreservation was two- to three-fold greater than that of the cv. 5929 cells. Cold acclimation of the cells (10 days at 2°C) improved viability after cryopreservation. The addition of 7.6 µM ABA to the B5 medium enhanced viability in ED but did not improve cell cryopreservability. Cold-acclimated cells had higher protein concentrations, but neither ABA nor cold acclimation influenced protein composition of cold-acclimated cells determined using SDS-PAGE. Encapsulated cells pre-treated for 4 days in B5 medium containing 0.75 M sucrose showed an increased concentration of cell protein and an altered protein composition. Suspension cultures were re-initiated from both ED and EDN treatments by transferring beads sequentially to B5 media containing 0.75, 0.5, 0.25 M sucrose and then to fresh B5 medium. The ED cells resumed rapid growth after two subcultures, whereas EDN cells needed four or five subcultures to resume rapid growth.     

Lipid biomarkers indicate different ecological niches and trophic relationships of the Arctic hyperiid amphipods Themisto abyssorum and T. libellula

The hyperiid amphipods Themisto libellula and T. abyssorum are important components of Arctic pelagic ecosystems. Both species are carnivorous and prey on mesozooplankton. They represent a substantial food source for marine vertebrates and are a key link between zooplankton secondary production and higher trophic levels. We present data on the total lipid content, lipid class and fatty acid composition of T. libellula and T. abyssorum from northern Fram Strait and the central Arctic Ocean. Both species had moderate to high lipid contents of 14-42% of body dry mass. In T. abyssorum, total lipid content was correlated to body mass, while T. libellula showed sex-related differences in lipid content. Despite their smaller body size, females of T. libellula had higher lipid contents than males. Wax esters represented the major lipid class in both species with 41-43% of total lipid, while triacylglycerols contributed 23-32%. The fatty acid composition was dominated by the long-chain polyunsaturated moieties 20:5(n-3) and 22:6(n-3), short-chain saturated compounds (16:0 and 14:0) and monounsaturated fatty acids of varying length, i.e. 16:1(n-7), 20:1(n-9), 18:1(n-9) and 22:1(n-11). Species-specific and geographic variations in the fatty acid and alcohol patterns were apparently linked to differences in diet and life-cycle. High amounts of the fatty acids and alcohols 20:1(n-9) and 22:1(n-11) in T. libellula indicate predation on herbivorous Calanus copepodids. In addition, elevated levels of 20:5(n-3) in T. libellula indicate a close connection with ice-algal production and the importance of cryo-pelagic coupling processes (i.e. exchange processes between the sea ice and the pelagic communities) for the nutrition of this high-Arctic epipelagic species. In contrast, T. abyssorum is characterised by lower amounts of 20:5(n-3) and its biomarker ratios indicate a higher trophic level. This observation is consistent with the subarctic-boreal origin of T. abyssorum and its occurrence in deeper layers of the Arctic Ocean, where it may feed on omnivorous and/or carnivorous prey.     

Growth rate and body weight of foundress-reared offspring in a paper wasp, <Emphasis Type="Italic">Polistes chinensis</Emphasis> (Hymenoptera,Vespidae): no influence of food quantity on the first offspring

Summary. Laboratory experiments with Polistes chinensis (Pérez) showed that growth rates and weights of the first-hatched offspring were not different between food-available and food-deficient conditions. In food-deficient colonies, some first-hatched offspring grew quickly, but their dry weights were not different from that of later offspring. The present results suggest that foundresses of P. chinensis may intensively feed some first-hatched larvae to produce the first workers quickly under food-deficient conditions. Since food availability of field foundresses of this species has been estimated as very low, it seems likely that they may produce the first offspring as early as possible by such feeding. On the other hand, heavier offspring were produced when extra food is available, especially in the latter half of the order of emergence. Foundresses of this species may change the amount of prey fed to each larva in response to food availability.     

Phytoplankton dynamics at the ice-edge zone of the Lazarev Sea (Southern Ocean) during the austral summer 1994/1995 drogue study

Phytoplankton dynamics in the open waters of the ice-edge zone of the Lazarev Sea were investigated over 12 consecutive days during a drogue study conducted in austral summer (December/January) 1994/1995. Throughout the study, the upper water column (<30 m) was stratified with a well-defined pycnocline evident. Although a subsurface intrusion of colder, more saline water into the region was recorded on days 5-9 of the experiment, its effect on the water column structure was negligible. Total surface chlorophyll a biomass doubled between days 1 and 5 (from 0.82 to 1.62 mg m^-3), and then showed a tendency to stabilise, while the depth-integrated chlorophyll a standing stock displayed an increasing trend during the entire experiment. All changes in biomass were associated with an increase in microphytoplankton. Flagellates and picoplankton dominated cell counts, while diatoms composed most of the phytoplankton biovolume. Results of the study indicate that, during the period of investigation, average cell abundance decreased. Coupled with this decrease was an increase in the biovolume and average size of the phytoplankton. Phytoplankton succession was observed in the ice edge during the drogue study. Typical ice-associated species of genera Haslea, Fragilariopsis and Chaetoceros, which dominated at the beginning of study, were replaced by open-water species of genera Corethron, Dactyliosolen and Rhizosolenia. The shift in phytoplankton species composition and size can likely be related to high light intensities and grazing by microzooplankton. The intrusion of colder, more saline water on day 5 appeared to modify the diatom succession, indicating extreme variability in phytoplankton dynamics in the near ice-edge zone of the Lazarev Sea.     

Host-symbiont specificity during onset of symbiosis between the dinoflagellates Symbiodinium spp. and planula larvae of the scleractinian coral Fungia scutaria

Many corals which engage in symbioses with dinoflagellates from the genus Symbiodinium (zooxanthellae) produce offspring which initially lack zooxanthellae. These species must choose their symbionts from numerous genetically distinct strains of zooxanthellae co-occurring in the environment. In most cases, symbiosis onset results in an association between a specific host coral and a specific strain of algal symbiont. This is the first study to examine host-symbiont specificity during symbiosis onset in a larval cnidarian, and the first to examine such events in a scleractinian of any life stage. We infected planula larvae of the solitary Hawaiian scleractinian Fungia scutaria with both homologous zooxanthellae, freshly isolated from F. scutaria adults, and heterologous zooxanthellae, isolated from Montipora verrucosa, Porites compressa, and Pocillopora damicornis, three species of scleractinians which co-occur with F. scutaria. We found that homologous zooxanthellae were better able to establish symbioses with larval hosts than were heterologous isolates, by two separate measures: percent of a larval population infected, and densities of zooxanthellae per larva. We also measured algal densities in larvae over a 4-day period until the onset of settlement and metamorphosis. We found no changes in zooxanthella population densities, regardless of zooxanthella type or the light environment in which they were incubated. Strong infection of host larvae with homologous algae compared to heterologous algae suggests that there is a specificity process which occurs sometime during the early stages of infection between the partners, and which results in the establishment of a specific symbiosis.     

A genetic and physical map of the region containing PLASTOCHRON1, a heterochronic gene,in rice ( Oryza sativa L.)

The rice heterochronic gene plastochron1, pla1, shows shorter plastochron and ectopic expression of the vegetative program during the rice reproductive phase resulting in aberrant panicle formation. A genetic and physical map was constructed to isolate the causal gene for the pla1 syndrome. Small-scale mapping was carried out to determine the approximate map position of the pla1 locus, and then a high-resolution genetic map was made for pla1-1, one of the pla1 alleles, using an F₂ population comprising 578 pla1-1 homozygous plants. In a high-resolution genetic map, the pla1-1 locus was found to map between RFLP markers C961 and R1738A on chromosome 10, within a 3.6-cM genetic distance. A physical map encompassing the pla1-1 locus was constructed by overlapping Bacterial Artificial Chromosome (BAC) clones through chromosome walking. PCR-based RFLP markers from BAC-end clones were developed and mapped relative to the pla1 locus. Physical map construction using BAC clones indicated that a BAC clone, B44A10 (167-kb), contained the pla1 locus within 74-kb corresponding to a 0.52-cM genetic distance. Gene prediction of 74-kb region carrying the pla1 locus suggested several candidate genes for the pla1 gene. Identification of a candidate gene for pla1 will be made by sequence analysis of allele variation and cDNA screening.     

The effects of desiccation on seeds of Acer saccharinum and Aesculus pavia: recalcitrance in temperate tree seeds

This study was undertaken to determine how the results from lipid, moisture, and differential scanning calorimetry analyses conducted on silver maple (Aceraceae: Acer saccharinum L.) and red buckeye (Hippocastanaceae: Aesculus pavia L.) compared with those obtained from previous studies on white and water oaks (Fagaceae: Quercus alba and Q. nigra), and the tropical zone species American muskwood (Meliaceae: Guarea guidonia) and carapa (Meliaceae: Carapa guianensis). Seeds were air-dried at room temperature for 9-11 days. At intervals, germination was tested, moisture determined, and lipids extracted. It was found that, like the other recalcitrant seeds, (1) viability was greatly reduced or lost after 11 days of drying, (2) percentage changes in individual fatty acids were not related to seed viability, and (3) results from the differential scanning calorimetry studies revealed a strong relationship between enthalpy/onset data from the embryo and cotyledon tissues and loss of viability. Also, silver maple seeds experienced a 50% reduction in viability by day 5 of drying and retained an axis moisture content over 25% throughout the experiment. However, unlike the other recalcitrant seeds surveyed, both silver maple and red buckeye had a significant reduction in the total amount (mg/g) of cotyledon lipids as the experiment progressed. However, no decrease in the unsaturated/saturated fatty acid ratio was found, so we conclude that in these species lipid peroxidation is not a marker of declining seed viability. Also, red buckeye seeds did not lose 50% viability until after day 8 of the experiment, and axis moisture content fell well below 20% as the seeds dried.     

Optimization of in vitro micropropagation and regeneration for Populus × interamericana and Populus × euramericana hybrids (P. deltoides, P. trichocarpa, and P. nigra)

A rapid and efficient micropropagation method has been established for six European poplar cultivars of economic interest - four Populus 2 interamericana and two Populus 2 euramericana. Using a three-step procedure, we were able to regenerate plantlets from callus and acclimate them within 4 months. In the first step, callogenesis was induced when explants were cultured for 25 days on culture medium supplemented with 10 µM !-naphthaleneacetic acid and 5 µM N⁶(2-isopentenyl)adenine. Bud regeneration followed by shoot elongation was then obtained from callus tissue by combining the cytokinin-like compound thidiazuron with the surfactant Pluronic F-68 at concentrations adjusted for each cultivar. The usefulness of this procedure in the area of genetic engineering is discussed.     

Production of inulo-oligosaccharides from inulin by recombinant E. coli containing endoinulinase activity

To utilize intracellular endoinulinase for inulo-oligosaccharide (IOS) production from inulin, the endoinulinase gene (inu1) of Pseudomonas sp. was successfully cloned into the plasmid pBR322 by using EcoRI restriction endoinulinase and E. coli HB101 as a host strain. The endoinulinase from E. coli HB101/pKMG50 was constitutively expressed, showing similar reaction modes as compared to those of the original strain. However, some critical differences existed in optimal reaction conditions and oligosaccharide compositions between the two products catalyzed by the native enzyme of original strain and those by intact cells from recombinant cells. The IOS compositions produced by recombinant E. coli were quite different due to the diffusional restriction of the substrate and products within the cell wall. Optimal reaction conditions for batchwise production of IOS were as follow : optimum temperature, 55v°C; pH, 7.5; substrate concentration, 100 g/l inulin; enzyme dosage, 20 units/g substrate. Continuous production of IOS from inulin was also carried out at 50v°C using a bioreactor packed with the recombinant cells immobilized on calcium alginate gel. The optimal feed concentration and the feed flow rate were 100 g/l inulin and 0.6 h^у as a superficial space velocity, respectively. Under the optimum operation conditions, continuous production of IOS was successfully performed with productivity of 166.7 g/l·h for 15 days at 50v°C without significant loss of initial activity.     

Single and dual parasitic mite infestations on the honey bee, Apis mellifera L.

Summary: The onset of foraging, proportion of pollen collectors, and weight of pollen loads were compared in individual honey bees (Apis mellifera) infested by zero, one (Acarapis woodi, the honey bee tracheal mite, or Varroa jacobsoni,varroa), or both species of parasitic mites. Phoretic varroa host choice also was compared between bees with and without tracheal mites, and tracheal mite infestation of hosts was compared between bees parasitized or not by varroa during development. The proportion of pollen collectors was not significantly different between treatments, but bees parasitized by both mites had significantly smaller pollen loads than uninfested bees. Mean onset of foraging was earliest for bees parasitized by varroa during development, 15.9 days. Bees with tracheal mites began foraging latest, at 20.5 days, and foraging ages were intermediate in bees with no mites and both, 17.6 and 18.0 days respectively. Phoretic varroa were found equally on bees with and without tracheal mite infestations, but bees parasitized by varroa during development were almost twice as likely to have tracheal mite infestations as bees with no varroa parasitism, 63.9 % and 35.5 %, respectively. These results indicate that these two parasites can have a biological interaction at the level of individual bees that is detrimental to their host colonies.     

Assessment of cell alignment by fibronectin multi-fibre cables capable of large scale production

The repair of damaged human tissue will be enhanced greatly by a capacity to organise the arrangement of the cells. We have demonstrated an approach to quantifying the capacity of fibronectin multi-fibre cables to align human cells. It is based on staining and subsequent image analysis of cells in the neighbourhood of the cables, and the application of an Orientation Index (S), to give a quantitative measure of alignment of the cells. Alignment of human dermal fibroblasts, parallel to the fibronectin cable axis, was observed by light microscopy after 3 days in culture. At 7 days, alignment was determined by image analysis after hematoxylin and eosin staining. This showed that the lateral extent of cell alignment i.e. the number of cell layers lined up beside a cable, termed the cell docking band width, was independent of cable diameter. Scanning electron microscopy confirmed cell alignment and the deposition of fine collagen fibrils by the cells. Orientation of cells to the cable, as measured by an orientation index (S), was S = 0.97 - 0.05 in cell docking bands denoting almost perfect alignment. Cell seeding levels ranging from 14 K cells/cm² up to 54 K cells/cm² resulted in cell docking band widths of aligned cells from 335 wm to 890 wm respectively, increasing as a function of cell seeding levels. This method represents a quantitative measure of quality for potential contact guidance materials. The guidance system is capable of large-scale manufacture.     

Predation risk and competition effects on the life-history characteristics of larval Oregon spotted frog and larval red-legged frog

We conducted an artificial pond experiment to test hypotheses about the effects of competition and non-lethal predator cues on metamorphic characteristics of sympatric Oregon spotted frogs (Rana pretiosa) and red-legged frogs (Rana aurora) in southwestern British Columbia. Tadpoles were exposed to the presence or absence of one another, two density levels and to the presence or absence of predacious odonate larvae (Aeshna palmata) isolated in enclosures. In the artificial pond study, R. aurora were significantly larger at metamorphosis (12%) and exhibited only slightly longer larval periods when exposed to Aeshna. In the presence of R. pretiosa, they significantly decreased time to metamorphosis, and were significantly larger at metamorphosis (12%) than those reared alone. Rana pretiosa in treatments with R. aurora were somewhat larger at metamorphosis when a non-lethal predator was present, and in treatments where R. pretiosa were alone with a predator tadpole mass at metamorphosis was smaller than those in the absence of Aeshna, but these results were not statistically significant. Both species reduced activity and moved away from the predator in the presence of an enclosed dragonfly larva in the laboratory. Most tadpole mesocosm experiments have found that the trade-off between size and timing of metamorphosis is extremely important to amphibians, but we suggest that the trade-off discussed in traditional amphibian models may not apply to species like R. pretiosa that are exposed to the same gape-limited predators upon reaching metamorphosis.     

Assessing the quality of different ant species as partners of a myrmecophilous butterfly

We assessed the quality of different ant species as partners of the facultatively myrmecophilous lycaenid butterfly Glaucopsyche lygdamus. We compared disappearance and parasitism rates of G. lygdamus larvae in the field, and development of non-feeding prepupae in the laboratory, when individuals were untended or tended by one of four ant species. Formica podzolica was the only ant species to provide a clear benefit to G. lygdamus, in the form of reduced larval parasitism relative to untended larvae. F. 'neogagates' (F. neogagates + F. lasioides) and Tapinoma sessile were essentially neutral partners, providing no significant cost or benefit for any of the parameters measured. Relative to untended individuals, association with F. obscuripes significantly increased larval disappearance and significantly decreased pupal mass. Thus, F. obscuripes may act as a parasite of the general association between G. lygdamus and ants under certain conditions. Ant species also differed in their persistence as tenders of G. lygdamus larvae once an interaction was established. Over the lifetime of a larva, F. podzolica and F. obscuripes usually remained as the attendant ant species on plants over consecutive census dates, while F. 'neogagates' and T. sessile were frequently replaced, most commonly by F. obscuripes. It remains to be determined if disappearance and developmental outcomes reported here reflect true fitness costs (i.e. reduced survivorship and lower reproductive success) for G. lygdamus. The potential and limitations for specialization in association between G. lygdamus and high quality ant partners are discussed.     

Pigment-dispersing hormone-like immunoreactive neurons in the central nervous system of the gastropods, Helix pomatia and Lymnaea stagnalis

By using an antiserum raised against a crustacean #-pigment-dispersing hormone (PDH), the distribution and chemical neuroanatomy of PDH-like immunoreactive neurons was investigated in the central nervous system of the gastropod snails, Helix pomatia and Lymnaea stagnalis. The number of immunoreactive cells in the Helix central nervous system was found to be large (700-900), whereas in Lymnaea, only a limited number (50-60) of neurons showed immunoreactivity. The immunostained neurons in Helix were characterized by rich arborizations in all central ganglia and revealed massive innervation of all peripheral nerves and the neural (connective tissue) sheath around the ganglia and peripheral nerve trunks. A small number of Helix nerve cell bodies in the viscero-parietal ganglion complex were also found to be innervated by PDH-like immunoreactive processes. Hence, a complex central and peripheral regulatory role, including neurohormonal actions, is suggested for a PDH-like substance in Helix, whereas the sites of action may be more limited in Lymnaea.     

In vivo study of microsclere formation in sponges of the genus <Emphasis Type="Italic">Mycale</Emphasis> (Demospongiae,Poecilosclerida)

The process of microsclere secretion was examined in vivo through glass coverslip implants in three species of the genus Mycale from São Sebastião channel, southeastern Brazil: Mycale (Aegogropila) angulosa, Mycale (Arenochalina) laxissima, and Mycale (Carmia) microsigmatosa. All three species adhered well to coverslips and developed normally through at least 2 weeks. Similar experiments with different species (Cinachyrella alloclada, Amphimedon viridis, Haliclona melana, and Aplysina caissara) were also successful with one exception (the cartilaginous Chondrilla nucula), indicating that the method can be applied to most demosponges. Microsclerocyte size varied according to the type of microsclere secreted, but all were elongated to fusiform and had small, anucleolated nuclei. Spicules were transported by microsclerocytes alone, without any other cell type ("helper cells") involved. Secretion of a microsclere was performed by a single sclerocyte. Although some axial filaments were found free in the mesohyl, all microsclere secretion in these animals was fully intracellular. Normal axial filaments were observed in most types of microscleres of the Mycale species (sigmas, toxas, and microxeas). Timed observations of sclerocytes suggest that immature spicules with the aspect of short straight rods with thick ends might be the precursors of the anisochelae. Observed differences in the size versus number of toxa secreted may indicate either the presence of two distinct subpopulations of toxa-producing microsclerocytes or that the initial number of axial filaments at the beginning of silica deposition may determine the final size of the spicules. Although other microscleres such as sigmas and chelae are secreted in a one cell-one spicule basis, several toxas and microxeas can be secreted simultaneously in a single cell.     

Responses of bullfrog tadpoles to hypoxia and predators

Low dissolved oxygen concentrations present numerous challenges for non-air-breathing aquatic organisms. Amphibian larvae and their predators can respond to oxygen levels by altering their behavior and physiology, but the ecological consequences of these responses are generally unknown. We conducted two laboratory experiments to study the effects of dissolved oxygen on respiratory behavior and susceptibility to predation of larval bullfrogs (Rana catesbeiana). In the first, we exposed small, lungless tadpoles to a predatory salamander larva (Ambystoma tigrinum) under high and low oxygen conditions. More tadpoles were consumed in high oxygen tanks than in low ones, presumably because salamanders remained near the surface in the low oxygen tanks while most tadpoles rested on the bottom. Tadpole activity depended on both oxygen and predator presence: swimming decreased after addition of salamanders under high oxygen, but increased under low oxygen. In the second experiment, we examined the effect of predator chemical cues on the air-breathing rate of large tadpoles with well-developed lungs under low oxygen conditions. In the presence of chemical cues produced by dragonfly larvae consuming bullfrog tadpoles, air-breathing and swimming were significantly reduced relative to controls. These experiments demonstrate the potential impact of dissolved oxygen on predator-prey interactions, and suggest that outcomes depend on the respiratory ecology of both predator and prey.     

Morphology, wood structure and cell wall composition of rolC transgenic and non-transformed aspen trees

Morphology, wood structure and cell wall composition of 35S-rolC transgenic hybrid aspen (P. tremula2tremuloides) were compared with non-transformed control trees. The transgenics are characterised by stunted growth, altered physiological parameters and light green leaves of reduced size. Histometric measurements revealed thinner fibre walls as compared to the controls. UV microspectrophotometry of individual wall layers did not reveal distinctive differences in the lignification of xylem cells, but in the extremely thin-walled fibres of the transgenics the secondary walls were less lignified as revealed by KMnO₄ staining in transmission electron microscopy. In the transgenics the formation of xylem cells was delayed and the differentiation zone reduced to only a few rows. Immunocytochemical analyses revealed the deposition of lignins in less differentiated xylem cells as compared to the controls. The first labelling of condensed lignin appeared in cell corners and of non-condensed lignin in secondary walls near cell corners during the deposition of S1 polysaccharides. Because of alterations in the formation and differentiation of xylem cells, 35S-rolC transgenic aspen may be useful for studies on molecular factors controlling the differentiation continuum.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of a doubled haploid line of cabbage

Doubled haploid lines, which have high levels of genetic uniformity, are suitable for physiological or genetic studies. We established a transformation system using a doubled haploid line of cabbage. Hypocotyl explants that had been precultured for 3 days on Murashige and Skoog medium containing 50 µM acetosyringone were inoculated and cocultured with Agrobacterium tumefaciens strain LBA4404 (pIG121Hm) for 3 days. Kanamycin-tolerant shoots were regenerated onto shoot induction medium 3 months after Agrobacterium inoculation. The transformation efficiency was about 3% under optimal conditions. The segregation pattern of the self-pollinated transformant that carried one copy of the introduced transgene revealed that the #-glucuronidase gene was inherited in a 3:1 ratio.     

Mapping of a thermo-sensitive earliness <Emphasis Type="Italic">per se</Emphasis> gene on <Emphasis Type="Italic">Triticum monococcum</Emphasis> chromosome 1A<Superscript>m</Superscript>

An earliness per se gene, designated Eps-A^m1, was mapped in diploid wheat in F₂ and single-seed descent mapping populations from the cross between cultivated (DV92) and wild (G3116) Triticum monococcum accessions. A QTL with a peak on RFLP loci Xcdo393 and Xwg241, the most distal markers on the long arm of chromosome 1A^m, explained 47% of the variation in heading date (LOD score 8.3). Progeny tests for the two F_2:3 families with critical recombination events between Xcdo393 and Xwg241 showed that the gene was distal to Xcdo393 and linked to Xwg241. Progeny tests and replicated experiments with line #3 suggested that Eps-A^m1 was distal to Xwg241. This gene showed a large effect on heading date in the controlled environment experiments, and a smaller, but significant, effect under natural conditions. Eps-A^m1 showed significant epistatic interactions with photoperiod and vernalization treatments, suggesting that the different classes of genes affecting heading date interact as part of a complex network that controls the timing of flowering induction. Besides its interactions with other genes affecting heading date, Eps-A^m1 showed a significant interaction with temperature. The effect of temperature was larger in plants carrying the DV92 allele for late flowering than in those carrying the G3116 allele for early flowering. Average differences in heading date between the experiments performed at 16 °C and 23 °C were approximately 11 days (P < 0.001) for the lines carrying the Eps-A^m1 allele for early flowering but approximately 50 days (P < 0.0001) for the lines carrying the allele for late flowering. The large differences in heading time (average 80 days) observed between plants carrying the G3116 and DV92 alleles when grown at 16 °C, suggest that it would be possible to produce very detailed maps for this gene to facilitate its future positional cloning.