The Targeting of the atToc159 Preprotein Receptor to the Chloroplast Outer Membrane is Mediated by its GTPase Domain and is Regulated by GTP

The multimeric translocon at the outer envelope membrane of chloroplasts (Toc) initiates the recognition and import of nuclear-encoded preproteins into chloroplasts. Two Toc GTPases, Toc159 and Toc33/34, mediate preprotein recognition and regulate preprotein translocation. Although these two proteins account for the requirement of GTP hydrolysis for import, the functional significance of GTP binding and hydrolysis by either GTPase has not been defined. A recent study indicates that Toc159 is equally distributed between a soluble cytoplasmic form and a membrane-inserted form, raising the possibility that it might cycle between the cytoplasm and chloroplast as a soluble preprotein receptor. In the present study, we examined the mechanism of targeting and insertion of the Arabidopsis thaliana orthologue of Toc159, atToc159, to chloroplasts. Targeting of atToc159 to the outer envelope membrane is strictly dependent only on guanine nucleotides. Although GTP is not required for initial binding, the productive insertion and assembly of atToc159 into the Toc complex requires its intrinsic GTPase activity. Targeting is mediated by direct binding between the GTPase domain of atToc159 and the homologous GTPase domain of atToc33, the Arabidopsis Toc33/34 orthologue. Our findings demonstrate a role for the coordinate action of the Toc GTPases in assembly of the functional Toc complex at the chloroplast outer envelope membrane

Einfluss von lebenden Mulchen auf die Begleitflora und die Weizenerträge unter Bedingungen des Ökolandbau

For the success of no-tillage in organic farming, new tools have to be developed to control weeds. One possible strategy could be sowing the main crop into an earlier established living mulch of easily controllable cover crops. Field trials were carried out in the Swiss midlands to investigate the impact of different legume cover crops on weed populations and grain yield of directly drilled winter wheat (Triticum aestivum L.) in a living mulch system. In general, weed suppressing effect was best with highly productive legumes. A significant reduction of the weed density of dicotyle, monocotyle, and spring germinating species was observed. Though, effective weed suppression resulted also in strong competition with the winter wheat. In order to improve the practicability of such systems, seeding technique of the main crop and living mulch management should be investigated

The targeting of the atToc159 preprotein receptor to the chloroplast outer membrane is mediated by its GTPase domain and is regulated by GTP

The multimeric translocon at the outer envelope membrane of chloroplasts (Toc) initiates the recognition and import of nuclear-encoded preproteins into chloroplasts. Two Toc GTPases, Toc159 and Toc33/34, mediate preprotein recognition and regulate preprotein translocation. Although these two proteins account for the requirement of GTP hydrolysis for import, the functional significance of GTP binding and hydrolysis by either GTPase has not been defined. A recent study indicates that Toc159 is equally distributed between a soluble cytoplasmic form and a membrane-inserted form, raising the possibility that it might cycle between the cytoplasm and chloroplast as a soluble preprotein receptor. In the present study, we examined the mechanism of targeting and insertion of the Arabidopsis thaliana orthologue of Toc159, atToc159, to chloroplasts. Targeting of atToc159 to the outer envelope membrane is strictly dependent only on guanine nucleotides. Although GTP is not required for initial binding, the productive insertion and assembly of atToc159 into the Toc complex requires its intrinsic GTPase activity. Targeting is mediated by direct binding between the GTPase domain of atToc159 and the homologous GTPase domain of atToc33, the Arabidopsis Toc33/34 orthologue. Our findings demonstrate a role for the coordinate action of the Toc GTPases in assembly of the functional Toc complex at the chloroplast outer envelope membrane

Essential Role of the G-Domain in Targeting of the Protein Import Receptor atToc159 to the Chloroplast Outer Membrane

Two homologous GTP-bindin proteins, atToc33 and atToc159, control access of cytosolic precursor proteins to the chloroplast, at Toc33 is a constitutive outer chloroplast membrane protein, whereas the precusor receptor atToc159 may be able to switch between a soluble and an integral membrane form. By transient expression of GFP fusion proteins, mutant analysis, and biochemical experimentation, we demonstrate that the GTP-binding domain regulates the targeting of cytosolic atToc159 to the chloroplast and mediates the switch between cytosolic and integral membrane forms. Mutant atToc159, unable to bind GTP, does not reinstate a green phenotype in an albino mutant (ppi2) lacking endogenous atToc159, remaining trapped in the cytosol. Thus, the function of atToc159 in chloroplast biogenesis is dependent on an intrinsic GTP-regulated swtich that controls localization of the receptor to the chloroplast envelope

RhoJ/TCL Regulates Endothelial Motility and Tube Formation and Modulates Actomyosin Contractility and Focal Adhesion Numbers

Objective—RhoJ/TCL was identified by our group as an endothelial-expressed Rho GTPase. The aim of this study was to determine its tissue distribution, subcellular localization, and function in endothelial migration and tube formation. Methods and Results—Using in situ hybridization, RhoJ was localized to endothelial cells in a set of normal and cancerous tissues and in the vasculature of mouse embryos; endogenous RhoJ was localized to focal adhesions by immunofluorescence. The proangiogenic factor vascular endothelial growth factor activated RhoJ in endothelial cells. Using either small interfering (si)RNA-mediated knockdown of RhoJ expression or overexpression of constitutively active RhoJ (daRhoJ), RhoJ was found to positively regulate endothelial motility and tubule formation. Downregulating RhoJ expression increased focal adhesions and stress fibers in migrating cells, whereas daRhoJ overexpression resulted in the converse. RhoJ downregulation resulted in increased contraction of a collagen gel and increased phospho–myosin light chain, indicative of increased actomyosin contractility. Pharmacological inhibition of Rho-kinase (which phosphorylates myosin light chain) or nonmuscle myosin II reversed the defective tube formation and migration of RhoJ knockdown cells. Conclusion—RhoJ is endothelial-expressed in vivo, activated by vascular endothelial growth factor, localizes to focal adhesions, regulates endothelial cell migration and tube formation, and modulates actomyosin contractility and focal adhesion numbers

Exosomal cancer immunotherapy is independent of MHC molecules on exosomes

Peptide-loaded exosomes are promising cancer treatment vehicles; however, moderate T cell responses in human clinical trials indicate a need to further understand exosome-induced immunity. We previously demonstrated that antigen-loaded exosomes carry whole protein antigens and require B cells for inducing antigen-specific T cells. Therefore, we investigated the relative importance of exosomal major histocompatibility complex (MHC) class I for the induction of antigen-specific T cell responses and tumour protection. We show that ovalbumin-loaded dendritic cell-derived exosomes from MHCI-/- mice induce antigen-specific T cells at the same magnitude as wild type exosomes. Furthermore, exosomes lacking MHC class I, as well as exosomes with both MHC class I and II mismatch, induced tumour infiltrating T cells and increased overall survival to the same extent as syngeneic exosomes in B16 melanoma. In conclusion, T cell responses are independent of exosomal MHC/peptide complexes if whole antigen is present. This establishes the prospective of using impersonalised exosomes, and will greatly increase the feasibility of designing exosome-based vaccines or therapeutic approaches in humans

Editorial: Responses to Climate Change in the Cold Biomes

publishedVersio

Améliorer la qualité technologique, nutritionnelle et sanitaire du bléiologique. Principaux leviers agronomiques et technologiques

Communication lors du colloque "DinABio 2013", 13 et 14 novembre 2013 à Tours (Fance)The organic bread wheat market has been diversified over time through the emergence of different sale channels. Processors require organic bread wheat of higher quality and safety in order to meet the consumers’ demand. The overall objective of the AGTEC-Org project was to identify agronomical and technological ways to improve the performance of organic wheat and flour. The findings would contribute to enhanced baking quality and nutritional value of organic flour, as well as prevention of mycotoxin contamination. The project involved 9 research centers or universities from 5 European countries for a total budget of about 1.5 million €. More than 400 experimental treatments were analyzed from 23 agronomic trials and 4 lab-experiments on food technology. Choice of cultivar is an efficient way to obtain higher grain quality. Intercropping legumes (grain or forage) improves weed control and N availability for wheat crop or succeeding crop. Green manure can be an effective alternative to farmyard manure. Fertilization with organic fertilizers improves yield and quality when water is available. Reduced tillage affects soil fertility and wheat yield but has little effects on grain quality. Milling process strongly influences flour characteristics. Stone milling improves the nutritive value and flour characteristics remain very stable independently of the milling yield. However, stone milling slightly raises DON levels. Characteristics of flour produced by means of roller milling appear very dependent on milling yield, instead. Increasing milling yield with the aim of enriching nutritional quality has a detrimental effect on either safety (DON) or bread-making quality (bread volume). Debranning before milling has a very positive impact on flour safety by reducing its DON content by 50 %.Les acteurs de la filière blé biologique panifiable font face à divers enjeux complémentaires voire parfois contradictoires. Les agriculteurs et transformateurs souhaitent augmenter les niveaux de production tout en satisfaisant prioritairement les attentes des consommateurs pour une alimentation de meilleure qualité nutritionnelle et sanitaire, plus durable et plus respectueuse de l’environnement. Le projet européen AGTEC-Org a eu pour objectif d’identifier les leviers agronomiques et technologiques permettant d’améliorer conjointement la productivité et la qualité des blés et des farines biologiques. Il a réuni agronomes, technologues et économistes issus de 9 centres de recherche et universités européennes pour un budget total de 1,5 M€. Plus de 400 traitements expérimentaux ont été analysés à partir de 23 expérimentations agronomiques et 4 expérimentations technologiques. Le choix de la variété est un moyen efficace pour obtenir une qualité de grain supérieure. L’insertion de légumineuses (grain ou fourrage) dans le blé améliore la compétition avec les adventices et la nutrition azotée du blé et/ou de la culture de printemps suivante. L’utilisation régulière d’engrais vert peut être une alternative pour compenser l’absence de fumiers sur l’exploitation. La fertilisation organique améliore le rendement et la qualité des blés lorsque la nutrition hydrique est assurée. La suppression du labour peut affecter la fertilité du sol et le rendement du blé alors qu’elle n’intervient pas sur la qualité du grain. Le mode de broyage (meule de pierre vs cylindres) influence fortement les caractéristiques technologiques, la sécurité sanitaire et la valeur nutritionnelle de la farine. Le broyage sur meule améliore la composition nutritionnelle mais augmente aussi la présence de certains contaminants. Les caractéristiques de la farine broyée sur meule de pierre sont moins influencées par le rendement en mouture. Au contraire, les caractéristiques de la farine obtenue par cylindre apparaissent très dépendantes du rendement en mouture. Augmenter le rendement en farine dans l’objectif d’enrichir la qualité nutritionnelle réduit aussi bien la sécurité sanitaire (DON) que la qualité boulangère (volume du pain). Le décorticage des grains avant broyage diminue les risques sanitaires en réduisant le taux de mycotoxines (DON) de 50%

Results of variety trials with buckwheat in Switzerland

Выращивание гречихи (Fagopyrum esculentum Moench) имело давнюю традицию в Швейцарии, которая исчезла в XX веке. В течение нескольких последних лет интерес к гречихе вновь возрос, и среди прочего, необходимо было найти сорта, адаптированные к климатическим условиям страны. Поэтому в 2014 году швейцарская сельскохозяйственная научно-исследовательская станция Agroscope начала полевые испытания с различными сортами гречихи. Среди тестируемых сортов было пять с детерминантным типом роста российской селекции из ФГБНУ ВНИИЗБК. Испытания 14 сортов гречихи были проведены в двух точках (Цюрих и Цолликофен) в 2016 году. Урожайность зерна колебалась от 1,57 т/га (Drollet) до 3,38 т/га (Дружина). Влажность зерна при уборке урожая у российских сортов варьировала от 13,6% до 18% в Цюрихе и от 27% до 29,1% в Цолликофене. Возможность с помощью новых сортов гречихи начать успешное производство швейцарских продуктов из гречихи зависит от потребителя, а также от возможностей переработки зерна гречихи (то есть обрушивания) и разработки других продуктов из гречихи кроме муки.Growing buckwheat (Fagopyrum esculentum Moench) had a long tradition in Switzerland which disappeared in the 20th century. Since a few years interest in buckwheat increased again and among others also varieties adapted to the climatic conditions had to be found. Therefore Agroscope, the Swiss Agricultural Research station, initiated field trials with different buckwheat varieties in 2014. Among the tested varieties there were also five with a determinate growth from a Russian breeding program. Trials at two sites (Zurich and Zollikofen) with 14 varieties have been conducted in 2016. Grain yields varied from 15.7 dt/ha (Drollet) up to 33.8 dt/ha (Drushina). The water content of the Russian varieties at harvest varied between 13.6 % and 18 % at Zurich and 27 % and 29.1 % at Zollikofen. In order to confirm results, trials should be repeated again in 2017. Whether with new buckwheat varieties a successful Swiss buckwheat production can be initiated depends on one side on the consumer but also on possibilities of the processing of buckwheat grains (i.e. dehulling) and the development of other buckwheat products than flour

Improving baking quality, nutritional value and safety of organic winter wheat / Améliorer la qualité technologique, nutritionnelle et sanitaire du blé biologique, Principaux leviers agronomiques et technologiques

The organic bread wheat market has been diversified over time through the emergence of different sale channels. Processors require organic bread wheat of higher quality and safety in order to meet the consumers’ demand. The overall objective of the AGTEC-Org project was to identify agronomical and technological ways to improve the performance of organic wheat and flour. The findings would contribute to enhanced baking quality and nutritional value of organic flour, as well as prevention of mycotoxin contamination. The project involved 9 research centers or universities from 5 European countries for a total budget of about 1.5 million €. More than 400 experimental treatments were analyzed from 23 agronomic trials and 4 lab-experiments on food technology. Choice of cultivar is an efficient way to obtain higher grain quality. Intercropping legumes (grain or forage) improves weed control and N availability for wheat crop or succeeding crop. Green manure can be an effective alternative to farmyard manure. Fertilization with organic fertilizers improves yield and quality when water is available. Reduced tillage affects soil fertility and wheat yield but has little effects on grain quality. Milling process strongly influences flour characteristics. Stone milling improves the nutritive value and flour characteristics remain very stable independently of the milling yield. However, stone milling slightly raises DON levels. Characteristics of flour produced by means of roller milling appear very dependent on milling yield, instead. Increasing milling yield with the aim of enriching nutritional quality has a detrimental effect on either safety (DON) or bread-making quality (bread volume). Debranning before milling has a very positive impact on flour safety by reducing its DON content by 50 %