Accumulation of the Type IV prepilin triggers degradation of SecY and YidC and inhibits synthesis of Photosystem II proteins in the cyanobacterium Synechocystis PCC 6803

Type IV pilins are bacterial proteins that are small in size but have a broad range of functions, including motility, transformation competence and secretion. Although pilins vary in sequence, they possess a characteristic signal peptide that has to be removed by the prepilin peptidase PilD during pilin maturation. We generated a pilD (slr1120) null mutant of the cyanobacterium Synechocystis 6803 that accumulates an unprocessed form of the major pilin PilA1 (pPilA1) and its non‐glycosylated derivative (NpPilA1). Notably, the pilD strain had aberrant membrane ultrastructure and did not grow photoautotrophically because the synthesis of Photosystem II subunits was abolished. However, other membrane components such as Photosystem I and ATP synthase were synthesized at levels comparable to the control strain. Proliferation of the pilD strain was rescued by elimination of the pilA1 gene, demonstrating that PilA1 prepilin inhibited the synthesis of Photosystem II. Furthermore, NpPilA1 co‐immunoprecipitated with the SecY translocase and the YidC insertase, and both of these essential translocon components were degraded in the mutant. We propose that unprocessed prepilins inactivate an identical pool of translocons that function in the synthesis of both pilins and the core subunits of Photosystem II.     

The impact of the 2003–2008 weather variability on intra-annual stem diameter changes of beech trees at a submontane site in central Slovakia

The link between the dynamics of beech (Fagus sylvatica L.) diameter changes, recorded by mechanical dendrometers, and the weather activity was studied at a submontane site in central Slovakia between 2003 and 2008. A new approach was presented by constructing seasonal increment chronologies, dividing them into shorter periods and joining them across several seasons to analyse the main influencing factors. For this purpose, response functions and correlation functions and their intra-seasonal moving variants were calculated. The photoperiod appeared to be synchronising the culmination of the seasonal dynamics of diameter changes; however, the main factors influencing the seasonal dynamics were weather variables. At the beginning of the season, the diameter increase was significantly correlated with temperature and its accumulation over a longer period. In summer, the diameter increase was hampered by long-term heat waves and the positive influence of precipitation became more pronounced. Precipitation towards the end of summer and in autumn significantly positively impacted the variability of the dendrometer signals. In 2003, the unusual seasonal dynamics were caused by a hot May and a hot and dry summer.In view of the expected climate change, including warmer springs and a higher frequency of summer droughts and heat waves, we expect a shift in the culmination of beech seasonal diameter increase towards May. Intensive drought and heat periods during summer will probably stress the beech populations growing close to their lower altitudinal limit.     

A short-term response of macroalgae to potential competitor removal in a mid-intertidal habitat in Korea

Interspecific interactions among three dominant macroalgae, Pterocladia capillacea (Rhodophyta), Hizikia fusiformis (Heterokontophyta) and Chondracanthus intermedius (Rhodophyta), were experimentally investigated on the rocky mid-intertidal zone of Sungsan, Jeju Island, Korea from March 1998 to June 1999. Each of the potentially competing species was removed in permanent plots (20 × 20 cm), and percent covers of non-manipulated species were measured by an image analyzing method using a digital camera. Pterocladia capillacea was the most abundant during all seasons, except for winter. Its abundance was lowered by the removal of the turf-forming alga C. intermedius, indicating that turf had a positive effect on P. capillacea. Conversely, there was a negative effect of P. capillacea on the abundance of C. intermedius. Interactions between C. intermedius and P. capillacea can probably be explained as a consequence of the water-trapping ability of the former and the canopy-forming ability of the latter. There was, however, no apparent effect related to H. fusiformis since the abundance of this alga remained low. This study supports that both negative and positive effects between same pair of species could be common depending on the morphological differences of algae and particular habitat conditions.     

Improved baculovirus vectors expressing barnase using promoters from Cotesia plutellae bracovirus

The goal of this study was to create a novel baculovirus expression system that does not require recombinant virus purification steps. Transfection of insect cells with transfer vectors containing barnase under control of the Cotesia plutellae bracovirus (CpBV) promoters ORF3004 or ORF3005 reduced cell growth. Co-transfection with bApGOZA DNA yielded no recombinant viruses and non-recombinant backgrounds. To further investigate the detrimental effects of barnase on insect cells, two recombinant bacmids harboring the barnase gene under control of the CpBV promoters, namely bAcFast-3004ProBarnase and bAcFast-3005ProBarnase, were constructed. While no viral replication was observed when only the recombinant bacmids were transfected, recombinant viruses were generated when the bacmids were co-transfected with the transfer vector, pAcUWPolh, through substitution of the barnase gene with the native polyhedrin gene by homologous recombination. Moreover, no non-recombinant backgrounds were detected from unpurified recombinant stocks using PCR analysis. These results indicate that CpBV promoters can be used to improve baculovirus expression vectors by means of lethal gene expression under the control of these promoters.     

Molecular characterization of ORFs 2 to 7 of Korean porcine reproductive and respiratory syndrome virus (CA) and its protein expression by recombinant baculoviruses

To determine the characteristics of the Korean porcine reproductive and respiratory syndrome virus (PRRSV), CA, which was isolated from the serum of an infected pig in 2006, we investigated the nucleotide sequence and expression of the structural ORFs (ORFs 2 to 7) using the bApGOZA system. We found that the structural ORFs 2 to 7 of CA consisted of 3188 nucleotides that were the same as those formed from VR-2332. Comparison of the CA with the other strains revealed nucleotide sequence identity ranging from 89.8 to 99.5%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The CA strain was closely related to the other North American genotype strains but formed a distinct branch with high bootstrap support. Additionally, expression levels of the PRRSV proteins in insect cells were strong or partially weak. The results of this study have implications for both the taxonomy of PRRSV and vaccine development.     

Effect of NaCl stress on dihaploid tobacco lines tolerant to Potato virus Y

Salinity is an important abiotic factor that limits plant growth and development. The influence of salt stress induced by sodium chloride on plant growth, proline content, level of lipid peroxidation and activities of antioxidative enzymes was studied in F1 hybrid DH10 and four dihaploid lines (207B, 238C, 239K, 244B) of tobacco (Nicotiana tabacum L.). Dihaploids were obtained from anther-derived haploids of hybrid DH10 and were previously proved to be tolerant to Potato virus Y (PVY). In our study, plants were grown in vitro and exposed to NaCl (100 and 200 mM) for 33 days. All dihaploids and hybrid DH10 showed reduced growth after NaCl treatment. They accumulated significant amounts of sodium and proline in response to salt stress as have already been observed in tobacco and other plant species. In tobacco exposed to NaCl the lipid peroxidation level did not increase and activities of superoxide dismutase (SOD), guaiacol peroxidase (POD), ascorbate peroxidase and catalase (CAT) mostly did not change significantly. The exception was line 239K where salt induced higher activities of SOD, CAT and POD. Two (238C and 244B) out of four dihaploids appeared more susceptible to salt stress as they showed weak growth in correlation with high proline and sodium content. Therefore, it seems that salt tolerance is not associated with tolerance to PVY. Variations in malondialdehyde and proline content as well as in enzymes activities observed among tobacco lines imply that dihaploids have different genetic properties which might result in different sensitivity to NaCl.