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Very little is understood of the structure of mycoplasma promoters, and this limits interpretation of genomic sequence data in these species. In this study the transcriptional start points of 22 genes of Mycoplasma pneumoniae were identified and the regions 5' to the start point compared. Although a strong consensus -10 region could be seen, there was only a weak consensus in the -35 region. A high proportion of transcripts had heterogeneous 5'-ends and characterisation of the sequence of the 5'-ends of two transcripts established that the heterogeneity was derived from initiation of transcription at reduced levels between 1 and 4 bases 5' to the major starting point. In addition to this apparently unique feature, a high proportion of transcripts lacked a 5' untranslated leader region that could contain a ribosomal binding site. Such leaderless transcripts are seen rarely in other bacterial species. Although the promoter regions for a number of members of lipoprotein multigene families were examined, no obvious explanation for regulation of expression was apparent. Using the data from this study an improved matrix for prediction of M.pneumoniae promoters was derived. Application of this matrix to the sequences immediately 3' and 5' to each predicted start codon in the genome suggested that most M. pneumoniae transcriptional start points were likely to occur between 5 and 30 bases 5' to the start codon.

The least understood aspects of the nutritional needs of bees are the elemental composition of pollen and the bees’ need for a stoichiometrically balanced diet containing the required proportions of nutrients. Reduced plant diversity has been proposed as an indirect factor responsible for the pollinator crisis. We suggest stoichiometric mismatch resulting from a nutritionally unbalanced diet as a potential direct factor. The concentrations and stoichiometric ratios of C, N, S, P, K, Na, Ca, Mg, Fe, Zn, Mn, and Cu were studied in the bodies of honeybees of various castes and sexes and in the nectar and pollen of various plant species. A literature review of the elemental composition of pollen was performed. We identified possible co-limitations of bee growth and development resulting mainly from the scarcity of Na, S, Cu, P and K, and possibly Zn and N, in pollen. Particular castes and sexes face specific limitations. Concentrations of potentially limiting elements in pollen revealed high taxonomic diversity. High floral diversity may be necessary to maintain populations of pollen eaters. Single-species crop plantations, even if these species are rich in nectar and pollen, might limit bee growth and development, not allowing for gathering nutrients in adequate proportions. However, particular plant species may play greater roles than others in balancing honeybee diets. Therefore, we suggest specific plant species that may (1) ensure optimal growth and production of individuals by producing pollen that is exceptionally well balanced stoichiometrically (e.g., clover) or (2) prevent growth and development of honeybees by producing pollen that is extremely unbalanced for bees (e.g., sunflower). Since pollen is generally poor in Na, this element must be supplemented using “dirty water”. Nectar cannot supplement the diet with limiting elements. Stoichiometric mismatch should be considered in intervention strategies aimed at improving the nutritional base for bees.

Applying microarray technology, we have investigated the transcriptome of the small bacterium Mycoplasma pneumoniae grown at three different temperature conditions: 32, 37 and 32 degrees C followed by a heat shock for 15 min at 43 degrees C, before isolating the RNA. From 688 proposed open-reading frames, 676 were investigated and 564 were found to be expressed (P < 0.001; 606 with P < 0.01) and at least 33 (P < 0.001; 77 at P < 0.01) regulated. By quantitative real-time PCR of selected mRNA species, the expression data could be linked to absolute molecule numbers. We found M.pneumoniae to be regulated at the transcriptional level. Forty-seven genes were found to be significantly up-regulated after heat shock (P < 0.01). Among those were the conserved heat shock genes like dnaK, lonA and clpB, but also several genes coding for ribosomal proteins and 10 genes of unassigned functions. In addition, 30 genes were found to be down-regulated under the applied heat shock conditions. Further more, we have compared different methods of cDNA synthesis (random hexamer versus gene-specific primers, different RNA concentrations) and various normalization strategies of the raw microarray data.

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The majority of terrestrial biomass is wood, but the elemental composition of its potential consumers, xylophages, differs hugely from that of wood. This causes a severe nutritional imbalance. We studied the stoichiometric relationships of 11 elements (C, N, P, K, Ca, Mg, Fe, Zn, Mn, Cu, Na) in three species of pine-xylem-feeding insects, Stictoleptura rubra, Arhopalus rusticus (Coleoptera, Cerambycidae) and Chalcophora mariana (Coleoptera, Buprestidae), to elucidate their mechanisms of tissue growth and to match their life histories to their dietary constraints. These beetles do not differ from other Coleoptera in their absolute elemental compositions, which are approximately 1000 (N), 100 (P, Cu) and 50 (K, Na) times higher than in dead but undecayed pine wood. This discrepancy diminishes along the wood decay gradient, but the elemental concentrations remain higher by an order of magnitude in beetles than in highly decayed wood. Numerical simulation of the life history of S. rubra shows that feeding on nutrient-poor undecayed wood would extend its development time to implausible values, whereas feeding on highly decomposed wood (heavily infected with fungi) would barely balance its nutritional budget during the long development period of this species. The changes in stoichiometry indicate that the relative change in the nutrient levels in decaying wood cannot be attributed solely to carbon loss resulting from decomposer respiration: the action of fungi substantially enriches the decaying wood with nutritional elements imported from the outside of the system, making it a suitable food for wood-eating invertebrates.

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