Conserved Arabidopsis response to oviposition and crude egg extract treatment
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ABSTRACT:
PROVIDER: PRJNA602727 | ENA |
REPOSITORIES: ENA
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Project description:Although a seemingly harmless developmental stage of herbivores, insect eggs trigger efficient plant defenses that include necrosis, callus formation, accumulation of ovicidal compounds and release of volatiles to attract egg predators. The large white butterfly Pieris brassicae deposits batches of 20-30 eggs onto Arabidopsis leaves, causing a large transcriptional reprograming that is drastically distinct from the expression profile triggered by larval feeding. Also, P. brassicae eggs induce localized cell death, accumulation of reactive oxygen species (ROS) and salicylic acid (SA), and expression of PTI-related genes, suggesting that egg-associated molecular patterns (EAMPs) activate a response that is similar to the response induced by microbial pathogens. We previously reported that a crude P. brassicae egg extract (EE, soluble fraction from crushed eggs) induced similar responses as oviposition, including ROS and SA accumulation, cell death and defense gene induction. In order to compare oviposition and EE treatment at the transcriptome level, we analyzed changes in transcipt abundance with P. brassicae EE or after natural oviposition. After 5 days, hundreds of genes were significantly upregulated by each treatment and their induction was highly similar between treatments. This conserved transcriptomic signature thus strongly supports our previous observations that oviposition and EE treatment trigger comparable responses in Arabidopsis.
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Project description:Transcriptional plant responses are specially under the scope on herbivore oviposition studies. However, most of the information comes from Lepidopteran models, where egg laying and feeding are separate events, and little is known regarding the plant reaction to those pests where females feed and oviposit simultaneously. The present study unveiled the oviposition-induced transcriptomic response of plants to Tetranychus urticae eggs. Transcriptional evidence indicated that the early events on the plant response to the egg extract involved ATP, Ca2+ and ROS disbalances. The main phytohormones triggered were JA and ET, with which also participated secondary metabolites, volatiles and glucosinolates as defence mechanisms. Many of the regulated genes are also involved during pathogen-defence mechanisms. Negative regulation of defence mechanisms happened at 72 h. Female fertility was significantly reduced when fed on plants pre-exposed to the eggs, while feeding increased on those plants pre-exposed for 72 h.
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Project description:The present study is the first study to identify the involvement of circRNAs in the ovary activation and oviposition regulation processes in honey-bee queens.CircRNAs expresion profiles were examined in ovaries of virgin queens, egg-laying queens, egg-laying inhibited queens and egg-laying recovery queens.
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Project description:crude total soluble protein extract from Arabidopsis thaliana
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Project description:The aim of the project was to analyse the role of DONSON during DNA replication reaction in Xenopus laevis egg extract. To test what is the impact of DONSON on replication we decided to remove DONSON from egg extract by immunodepletion and analyse its impact on DNA replication reaction. In the parallel experiments we have shown that without DONSON egg extract cannot synthesise nascent DNA. Here we wanted to examine the change of replicating chromatin proteome in absence of DONSON. Interphase Xenopus laevis egg extract was immunodepleted with control nonspecific IgG antibodies beads or DONSON-antibodies. The efficiency of immunodepletion was shown to be over 95%. DNA replication was set up in both IgG- and DONSON-depleted egg extracts and chromatin isolated in the middle of S-phase, when replication forks where abundant in chromatin fraction in IgG-depleted extract.
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Project description:Somatic cell reprogramming in vitro prior to nuclear transfer is one strategy expected to improve clone survival during development. In this study, we investigated the reprogramming extent of fish fin somatic cells after in vitro exposure to Xenopus egg extract and subsequent culture. Using a cDNA microarray approach, we observed drastic changes in the gene expression profile of the treated cells. Several actors of the TGFbeta and Wnt/beta-catenin signaling pathways, as well as some mesenchymal markers, were inhibited in treated cells, while several epithelial markers were upregulated. This was associated with morphological changes of the cells in culture, suggesting that egg extract drove somatic cells towards a mesenchymal-epithelial transition (MET), the hallmark of somatic reprogramming in iPSCs. However, treated cells were also characterized by a strong decrease in de novo lipid biosynthesis metabolism, the lack of re-expression of pou2 and nanog pluripotency markers, and absence of DNA methylation remodeling of their promoter region. In all, this study showed that Xenopus egg extract treatment initiated an in vitro reprogramming of fin somatic cells in culture. Although not thorough, the induced changes have primed the somatic chromatin for a better embryonic reprogramming upon nuclear transfer.
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Project description:Bone marrow stromal cells (BMSCs) are multipotent stem cells that preferentially differentiate into mesenchymal cells. If they can be dedifferentiated into embryonic stem cell-like cells, they will be a highly attractive source for cell therapy. Cell and egg extracts have been used in a few studies to evaluate nuclear reprogramming, but these have not examined cell pluripotency in any detail. In this study, we used a cell reversible permeabilization method to treat BMSC with Xenopus laevis mitotic egg extract. We observed an upregulation of the pluripotent protein Oct3/4 in BMSCs treated by this extract. We also further evaluated transcriptional changes with a focused stem cell oligonucleotide array. A number of genes involved in the Notch or Wnt signaling pathways were upregulated in BMSC exposed to Xenopus egg extract. In conclusion, our microarray data from BMSCs exposure to egg extracts may provide interesting clues regarding factors involved in nuclear reprogramming. Our approach is an alternative method towards dedifferentiation of cells without genetic modification, which is preferable in the clinical situation. Keywords: Cell type comparison Expression profiling was performed on murine bonemarrow stromal cells cultured in-vitro versus stromal cells exposed to Xenopus extract and compared with murine embryonic stem cells
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Project description:For many behaviours studied at the phenotypic level, we have little or no idea of where to start searching for “candidate” genes: the transcriptome provides such a starting point. Here we consider transcriptomic changes associated with oviposition in the parasitoid wasp Nasonia vitripennis. Oviposition is a key behaviour, as females are faced with a variety of decisions that will impact offspring fitness. These include choosing between hosts of differing quality, as well as deciding on clutch size and offspring sex ratio. We compared the whole-body transcriptomes of resting or ovipositing female Nasonia using a “DEEP-Sage” gene expression approach on the Illumina sequencing platform. Single 2-day old mated Nasonia vitripennis females (ASymC strain) were isolated in a glass vial and provided with a single host to produce F1 daughters. Eight 2-day old mated F1 females were subsequently provided with three hosts to produce the F2 test females. We randomly selected one host from each F1 female, and isolated 16 2-day old mated F2 test females in glass tubes, of which eight were randomly allocated to the oviposition treatment and eight to the resting treatment. We provided the test females with a single host for 24 hours as pre-treatment to facilitate egg development. We then discarded the pre-treatment hosts and gave each female a piece of chromatography paper soaked in honey solution for a further 24 hours. For the experiment, we transferred the females to 1.5 mL Eppendorf tubes that contained a single host for the oviposition experiment, or were empty for the resting treatment. After 60 mins, females were flash-frozen in liquid nitrogen and stored on dry-ice until the addition of RNAlater-ICE (Ambion, Austin, TX, USA) after which they were transferred to -20C. All females in the oviposition treatment were observed to have commenced ovipositing. We pooled the F2 test females from each F1 mother according to treatment, generating a total of eight pooled samples per treatment (consisting of 8 females per pool) for RNA isolation and sequencing.
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