Loss of decreased-rubisco phenotype between generations of wheat transformed with antisense and sense rbcS

The elite UK winter wheat cv. Riband was transformed with constructs containing rbcS in sense and antisense orientations driven by the maize ubiquitin promoter with a transformation efficiency of 1.2%. Of 77 primary transformants 31% of the sense-rbcS transformed lines and 78% of the antisense-rbcS transformed lines had decreased rubisco content compared to wild-type and marker-only controls, with decreases of up to 60%. However, in the T1 progeny which inherited the transgene, only 5% showed significantly decreased rubisco content and these effects were on the margins of significance. Five potential T2 homozygous lines from T1 parents which had transgene segregation consistent with a single locus were identified. There was no significant decrease in rubisco content relative to wild-type in any of these lines (LSD of 8% for P= 0.05). Expression of antisense rbcS transgenes in two of these T2 lines was low but was increased following exposure of the plants to 37°C for 48 h. However this did not induce a significant decrease in rubisco protein content relative to controls. Southern analysis of two antisense lines showed that they had low copy number and 1–2 insertion events. In one of the two lines there was increased methylation of the ubiquitin intron in T2 samples compared to the TO primary transformant. Further work is required to establish whether methylation occurred in all the lines which lost the phenotype, and therefore the likelihood of this being the cause. The disappearance of the decreased rubisco-content phenotype between generations may therefore be attributable to (1) greater activity of the ubiquitin promoter due to greater stress in the T0 generation plants and/or (2) increased methylation of the transgene promoter region between generations.     

The Patchwork Mouse Phenotype: Implication for Melanocyte Replacement in the Hair Follicle

Mice homozygous for the recessive patchwork (pwk) mutation are characterized by a variegated pigment pattern with a mixture of unpigmented and normally pigmented hairs. The pigmented hair bulbs contain functional melanocytes. By contrast, the unpigmented hair bulbs contain no melanocytes. This lack results from the death of melanoblasts in the hair follicle at the end of embryogenesis. Here, we report that melanoblasts and melanocytes are found in the epidermis of pwk/pwk mice. Furthermore, these epidermal pigment cells are able to colonize new hair follicles after skin wounding. Despite the presence of epidermal pigment cells with a colonization potential, a follicle that had produced an unpigmented hair produces a new unpigmented hair during the successive hair growth cycles. This hair color continuity is also true for the pigmented hair follicles. Thus, in normal conditions, the hair acts as an independent functional unit as regards its pigment cells population.     

The Influence of Moisture Stress on Liriodendron tulipifera L. Seedlings

A comparison of yellow poplar seedlings subjected to availablemoisture ranging from 100 per cent to 20 per cent of field capacityin two separate experiments (a soil study and a solution study)indicated that total dry matter production, stem height, totalleaf area and area of individual leaves, decreased with increasedmoisture stress. Differences in unit leaf rate (ULR) were primarilyresponsible for final plant size. Leaf area: weight ratio wasaffected by the method of induced stress. As stress increasedthe ratio increased in the soil study but decreased in the solutionstudy. The number of leaves produced was unaffected by stress,except at the most severe stresses. The number of senescentleaves increased as moisture stress increased. The allocationof new growth was unaffected by either moisture stress or timeand decreased in order of root, foliage, stem for the soil grownplants and foliage, root, stem for the solution grown plants.ULR and relative growth rate (RGR) were reduced by both moisturestress and time. A growth simulation of the soil-grown plantsbased on results from the solution moisture stress experimentpredicted the final plant dry weight within 12 per cent andleaf area within 7 per cent of the actual values.     

Spatial and temporal analysis of non-steady elongation of rice leaves

A precise knowledge of the temporal and spatial distributions of cell division and tissue expansion is essential for appropriate leaf sampling in omics studies and for analyses of plant–environment relations. Elongating leaves of rice were studied during their whole development for elongation rate, distribution of cell length, cell production rate and spatial distribution of growth in the leaf. In seven genotypes, the pattern of leaf elongation rate followed three phases: (1) an exponential increase before leaf appearance; (2) a short phase (2–4 d at 20 °C) with a stable leaf elongation rate around leaf appearance; and (3) a phase of 8–10 d with a progressive decrease in elongation rate. The profile of cell length along the leaf changed with time during the first and last phases, but was time invariant around appearance. We propose a method adapted to non-steady elongation based on anatomical measurements, which was successfully tested by comparing it with the pricking method. It allowed analysis of the change with time in the spatial distribution of growth from initiation to end of leaf growth. The length of leaf zones with cell division and tissue elongation varied with time, with maximums of 21 and 60 mm respectively around leaf appearance.     

Genetic Manipulation of Rubisco: Chromatium vinosum rbcL is expressed in Nicotiana tabacum but does not form a functional protein

N. tabacum lines that lacked functional Rubisco were transformed with plasmids encoding a chloroplast transit peptide in frame with C. vinosum rbcL and stable transformants generated. However, the transgene was transcribed at a low level and no Rubisco activity or C. vinosum large subunits were detectable in any line.     

Phytoplankton communities and antecedent conditions: high resolution sampling in Esthwaite Water

1. The succession of a phytoplankton community was investigated through an intensive period of sampling and related to physical, chemical and biological conditions sampled at an equal, or higher, temporal resolution. 2. Phytoplankton samples were taken on a weekly basis from June to September 2004 and analysed for diversity, species composition, and contribution of different functional groups to total biomass. Physical and chemical data were collected on the sampling days, and physical environmental factors were also logged continuously throughout the period by automatic measuring stations. This continuous logging allowed community structure to be compared with physical data averaged over periods from a day to a week before each sampling date. 3. The Schmidt stability of the lake, a measure of the strength of stratification calculated from thermal data, showed a negative correlation with phytoplankton species diversity. This is consistent with the hypothesis that mixing was preventing exclusion by species that would otherwise dominate in stratified conditions. 4. At a functional level, stress tolerant (S‐type) species dominated during the stratified summer conditions, with small, colonising species (C types) and ruderal, disturbance tolerant species (R types) contributing little to the overall biomass. Of the stress tolerant species, the faster growing (S_C) phytoplankters were significantly favoured by more stable, stratified conditions and higher solar radiation. Increased abundance of this group resulted in decreased species diversity. Correlations were generally strongest when using the 6‐ to 7‐day averaged physical data, stressing the importance of continuous measurements of these drivers in phytoplankton studies.     

Hélène Charniaux-Cotton (1918–1986)

Summary

1-Methyladenine (1-MA) secreted from the follicle cells is the biological signal for meiosis reinitiation of starfish oocytes. The signal of-1-MA is transduced into cytoplasmic formation of maturation-promoting factor (MPF) that eventually induces a germinal vesicle breakdown (GVBD). Microinjection of pertussis toxin (PTX) inhibited 1-MA-induced GVBD in Asterina pectinifera and Asterina (Patina) miniata. PTX-inhibition of GVBD was rescued by the injection of MPF into PTX-preinjected oocytes. Most of the PTX- and MPF-double injected eggs were fertilized and underwent cleavage, suggesting the presence of a GTP-binding protein (G protein) specific for 1-MA signal transduction. Indeed, plasma membrane preparations of A. pectinifera oocytes contained a G protein consisting of 39-kDa α, 37-kDa β, and 8-kDa γ subunits. The α subunit contained a site for ADP-ribosylation catalyzed by PTX. It was also recognized by antibodies specific for a common GTP-binding site of mammalian α subunits or a carboxy-terminal ADP-ribosylation site of mammalian inhibitory G protein (G_i) α subunits. Its gene was 74% and 83.7% identical to the rat G_i-2α gene in nucleotide and deduced amino acid sequences, respectively. The 39-kDa α subunit shared the common GTP-binding site of mammalian G protein α subunits and the PTX-catalyzed ADP-ribosylation site of mammalian G_i α subunits as expected from the immunoreactivity. The oocyte membranes had apparently two forms of 1-MA receptors with high and low affinities. The high-affinity form was converted into the low-affinity one in the presence of a non-hydrolyzable analogue of GTP. The 39-kDa α subunit of starfish G protein was also ADP-ribosylated by cholera toxin only when 1-MA was added to the membranes. These results indicate that in starfish oocyte membranes, 1-MA receptors are functionally coupled with the 39-kDa PTX-substrate G protein that transduces the signal into the formation of a cytoplasmic factor (MPF) and eventually into the reinitiation of meiosis.     

Effects of Partial Defoliation on the Growth of Liriodendron tulipifera L. Seedlings

Partial defoliation of Liriodendron tulipifera L. by removalof leaflets as they emerged from the protective buds affecteda variety of growth parameters. Reductions occurred in totaldry weight production, the percentage of new growth devotedto foliage, leaf area ratio, and the increment of leaf areaper unit of leaf weight increment. Increases occurred in unitleaf rate and the rate of production of new leaves. Weight ofleaves near the base of the seedlings was unaffected by treatmentbut leaves developing at the end of the growing season werelarger on defoliated plants. The net effect of removing one-thirdand two-thirds of the developing leaflets was to reduce totaldry weight production by 14 and 42 per cent respectively.     

RAPID DETECTION OF GERMINATING BACILLUS CEREUS CELLS USING FLUORESCENT IN SITU HYBRIDIZATION

Methods for the specific detection of Bacillus spores are needed in many situations such as the recognition of food poisoning. This study presents an experimental design in order to find the best combination of germination conditions leading to a rapid and detectable fluorescent in situ hybridization (FISH) signal from Bacillus cereus spores present in pure cultures and milk samples.
B. cereus ATCC 14579 and HER 1414 were incubated in 20 different growth media by using a combination of various germinants such as sugars, amino acids and dipicolinic acid. Also, three different germination factors were tested: incubation temperature, inoculum concentration and a heat shock treatment. Permeabilization procedure and hybridization time were optimized on the best germination condition found. B. cereus -specific FISH probes were validated under the optimized condition and in detection of spiked B. cereus spores in 1% ultra heat-treated milk samples. FISH-labeled cells were detected by using flow cytometry, and the results were confirmed by fluorescence microscopy. The optimal condition allows the detection of B. cereus spores in less than 2 h. Overall, a ninefold reduction in total time for detection was achieved when comparing with previous works. Therefore, the permeabilization and hybridization optimizations mentioned in this study are major improvements for the detection time of B. cereus spores.

PRACTICAL APPLICATIONS

By using the optimized conditions of germination/outgrowth, permeabilization and hybridization, the detection of 10³ cfu/mL of Bacillus cereus spores using fluorescent in situ hybridization is possible within 2 h in milk sample.