Real-time quantitative PCR analysis of apple roots and leaves after ARD infection
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PROVIDER: PRJNA557359 | ENA |
REPOSITORIES: ENA
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Project description:Root and leave samples of 4 different apple genotypes were investigated in order to analyse the gene expression after infection with Apple Replant Disease (ARD). All genotypes were cultivated in ARD-infected soil and gamma-irradiated (disinfected) soil in the greenhouse for 7 days. The ARD soil originated from two different orchards representing two different soil compositions. After 7 days root tissue was collected from each plant and used for the subsequent gene expression analysis. This work was part of the project BonaRes-ORDIAmur funded by the German Federal Ministry of Research and Education within the frame of the program BonaRes (grant no. 031B0025B). It was also funded by the German Research Foundation (DFG) via the research training group GRK1798 "Signaling at the Plant-Soil Interface" and a grant to BL and LB (BE 1174/19-1).
2019-12-09 | GSE135081 | GEO
Project description:Real-time quantitative PCR analysis of apple seeds
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Project description:Real-time quantitative PCR analysis of 30 apple cyclophilin genes under abiotic stress
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Project description:Apple seeds were subjected to accelerated aging. After 7, 14, and 21 days of aging, embryos were isolated. Part of the embryos were shortly fumigated with nitric oxide (NO). After 48 h of embryos culture (aged embryos or aged embryos treated with NO), embryonic axes were used to extract the total RNA. RT-qPCR were done to analyze the changes in the expression of genes related to seed aging. Short-term (3 h) treatment of embryos isolated from accelerated aged apple seeds (Malus domestica Borkh.) with NO partially reduced the effects of aging. The aim of the study was to investigate the impact of the short-term NO treatment of embryos isolated from apple seeds subjected to accelerated aging on the expression of genes potentially linked to the regulation of seed aging. Apple seeds were artificially aged for 7, 14, or 21 days. Then the embryos were isolated from the seeds, treated with NO, and cultured for 48 h. Progression of seeds aging was associated with the decreased transcript levels of most of the analyzed genes (Lea1, Lea2a, Lea4, Hsp70b, Hsp20a, Hsp20b, ClpB1, ClpB4, Cpn60a, Cpn60b, Raptor, and Saur). The role of NO in the mitigation of seed aging depended on the duration of the aging. After 7 and 14 days of seed aging, a decreased expression of genes potentially associated with the promotion of aging (Tor, Raptor, Saur) was noted. NO-dependent regulation of seed aging was associated with the stimulation of the expression of genes encoding chaperones and proteins involved in the repair of damaged proteins. After NO application, the greatest upregulation of ClpB, Pimt was noted in the embryos isolated from seeds subjected to 7-day long accelerated aging, Hsp70b, Hsp70c, Cpn in the embryos of seeds aged for 14 days, and Lea2a in the embryos of seeds after 21 days of aging.
2022-07-07 | GSE207591 | GEO
Project description:Plant cyclophilins are widely involved in a variety of abiotic stress regulation processes. Thirty cyclophilin members were identified from apple and their patterns of response to abiotic stress were examined. Apple seedlings with consistent growth were selected, cultivated under drought stress conditions simulated by PEG6000, sampled according to different time periods, and quickly frozen in liquid nitrogen.Total RNAs of apple plantlets were isolated using an RNA extraction kit (Tiangen, Beijing, China).
2022-09-24 | GSE214087 | GEO
Project description:The silkworm (Bombyx mori) has long been considered a source of food and medicine due to its high nutritional, medicinal, and economic value in East Asia. However, in some sensitive individuals, silkworm consumption can cause allergenic reactions such as vomiting, asthma, and anaphylaxis. Therefore, the development of a reliable method for silkworm detection is required to avoid such allergenic incidents. In this study, two different methods (liquid chromatography combined with mass spectrometry [LC-MS/MS] and real-time polymerase chain reaction [PCR]) were developed to determine an efficient technique for silkworm detection in foods. The developed methods demonstrated high sensitivity in detecting the silkworm in processed foods. Silkworm-spiked model cookies were used to confirm the sensitivity of both LC-MS/MS (0.0005%) and real-time PCR (0.001%). These methods were found to be useful for detecting the silkworm in foods and avoiding allergenic reactions. To the best of our knowledge, this is the first study to compare LC-MS/MS and real-time PCR for silkworm detection in complex processed foods.
2024-05-24 | PXD042494 | Pride
Project description:Drought stress is a major problem around the world and although progress in understanding how vegetable crops and model plants adapt to drought have been made, there is still little information about how fruit crops deal with moderate drought stress. In this study, we investigated the response of two apple genotypes: a drought-sensitive genotype (M26) and a drought-tolerant genotype (MBB). Our results of the morphology, physiology and biochemistry under moderate drought stress, indicated that relative water content (RWC) and leaf area (LA) were not significant changes in two genotypes. However, it had larger leaf mass per area (LMA), and accumulated higher free proline (CFP), soluble sugars (CSS) and malonaldehyde (MDA) in the leaves. Thus, it appears that the MBB genotype could produce more osmosis-regulating substances. Phosphoproteomic was analyzed from leaves of both genotypes under moderate drought stress using the isobaric tags for relative and absolute quantification (iTRAQ) technology. A total of 595 unique phosphopeptides, 682 phosphorylated sites and 446 phosphoproteins were quantitatively analyzed in the two genotypes. Motif analyses of the phosphorylation sites showed that six motifs including [PxsP], [sP], [sD], [Rxxs], [sxP] and [sxs] were enriched. We identified 12 and 48 PLSC phosphoproteins in M26 and MBB, respectively. Among these, 9 PLSC phosphoproteins were common to both genotypes, perhaps indicating a partial overlaps of the mechanisms to moderate drought stress. Gene ontology analyses revealed that the PLSC phosphoproteins present a unique combination of metabolism, transcription, translation and protein processing, suggesting that the response in apple to moderate drought stress encompasses a new homeostasis of major cellular processes. The basic trend was an increase in protein abundance related to drought and organic substance upon moderate drought stress between two genotypes. These increases were higher in the drought-tolerant genotype (MBB) than in the drought-sensitive genotype (M26). The 23 differentially expressed mRNA encoding phosphoproteins were analysis by quantitative real-time PCR (qRT-PCR). Our study is the first to address the phosphoproteome of a major fruit crop, apple rootstocks, in response to moderate drought stress, and provide insights into the molecular regulation mechanisms of apple rootstock under moderate drought stress.
2016-11-08 | PXD005238 | Pride
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