Polyamines are involved in the gynogenesis process in onion

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699International audienceHaploidization in onion (Allium cepa L.) using immature flower buds simulates zygotic embryogenesis with no fecundation. In order to know the involvement of polyamines (PAs) in this process, we determined the concentration of endogenous PAs in flower buds and experimented the addition of various combinations of PA molecules in the medium. At the inoculation stage, high levels of free and conjugated spermidine and low putrescine + hydroxyputrescine/spermidine + spermine ratio characterized the highest responsive varieties. During in vitro culture, high levels of putrescine and its derivatives characterized the lowest responsive varieties, whereas high levels of spermidine and spermine characterized responsive varieties. The putrescine + hydroxyputrescine + homospermidine/spermidine + spermine ratio remained low in responsive varieties. The addition of spermidine or spermine (2 x 10(-3) M) to the culture medium improved significantly the embryo production. Our results suggest that the arginine decarboxylase pathway is involved in PA biosynthesis during the in vitro culture of flower buds. Our study showed that specific ratios of PAs are required for successful gynogenesis in onio

Ypt31/32 GTPases and their F-Box effector Rcy1 regulate ubiquitination of recycling proteins

Ypt/Rab GTPases are conserved molecular switches that regulate the different steps of intracellular trafficking pathways. In yeast, the Ypt31/32 GTPases are required for exit from the trans-Golgi and for recycling from the plasma membrane (PM), through early endosomes, to the Golgi. We have previously shown that the recycling function of Ypt31/32 is mediated by an effector called Rcy1. Specifically, both Ypt31/32 and Rcy1 are required for recycling the vSNARE Snc1. Rcy1 contains an F-box domain shared by proteins that act in substrate recognition of ubiquitin ligases. Here, we show that both Ypt31/32 and Rcy1 are important for Snc1 ubiquitination and that such ubiquitination plays a role in Snc1 recycling. Direct interaction between Rcy1 and Snc1 was demonstrated using two independent approaches. In vitro interaction was observed using co-precipitation of recombinant proteins, whereas interaction in yeast cells was observed using bimolecular fluorescence complementation. Ubiquitination of Snc1 in vivo at the K63 position was previously shown in a proteomic study. We show that the Snc1-K63R mutant protein is less ubquitinated than wild-type Snc1 and is defective in endosome-to-Golgi transport. Additionally, wild-type Snc1 is ubiquitinated to a lesser extent in ypt31/32ts and rcy1Δ mutant cells and Snc1 recycling is also blocked in endosomes in these mutants. Therefore, ubiquitination plays a role in the recycling of Snc1 from the PM to the Golgi, and Ypt31/32 and Rcy1 regulate this ubiquitination. Together, these results suggest a new role for ubiquitination in cargo recycling. Moreover, we propose that Ypt/Rabs integrate intra-cellular trafficking with ubiquitination