A study of the effects of paclobutrazol on post-harvest behaviour of apple and tomato fruit

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A Simple and Rapid Determination of ATP, ADP and AMP Concentrations in Pericarp Tissue of Litchi Fruit by High Performance Liquid Chromatography

Razvijena je jednostavna i brza metoda određivanja masenog udjela adenozin trifosfata (ATP), adenozin difosfata (ADP) i adenozin monofosfata (AMP) visokodjelotvornom tekućinskom kromatografijom (HPLC) u perikarpu voća liči. Pri određivanju adenozin fosfata upotrijebljen je acetonitril koji skraćuje vrijeme postupka. Također je postignuta dobra ponovljivost (koeficijent varijacije iznosi 1,28-1,80 %) i obnovljivost (94,7-97,1 %). Koeficijenti korelacije s površinama pika u rasponu od 0 do 80 ng iznosili su 0,9946 za ATP; 0,9994 za ADP i 0,9974 za AMP. Ova je metoda primijenjena za određivanje masenog udjela adenozin fosfata u perikarpu voća liči pri berbi, pa je utvrđeno da je maseni udio ATP-a iznosio 27,4 μg/g, ADP-a 35,4 μg/g i AMP-a 7,9 μg/g, na bazi svježe tvari voća.A simple and rapid method using high performance liquid chromatography (HPLC) was developed to determine levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in litchi fruit pericarp tissue. This HPLC method used acetonitrile gradient elution and shortened the time required for determinations of adenosine phosphates. This analysis exhibited good repeatability (coefficients of variation 1.28–1.80 %) and recovery rate (94.7–97.1 %). The correlation coefficients of ATP, ADP and AMP with their peak areas at a range of 0–80 ng were 0.9946, 0.9994 and 0.9974, respectively. This method was applied to determine levels of adenosine phosphates in pericarp tissue of litchi fruit at harvest. There were 27.4 μg/g of ATP, 35.4 μg/g of ADP and 7.9 μg/g of AMP on a fresh mass basis

Sculpting the maturation, softening and ethylene pathway: The influences of microRNAs on tomato fruits

<p>Abstract</p> <p>Background</p> <p>MicroRNAs (miRNAs), a ubiquitous class of short RNAs, play vital roles in physiological and biochemical processes in plants by mediating gene silencing at post-transcriptional (PTGS) level. Tomato is a model system to study molecular basis of fleshy fruit ripening and senescence, ethylene biosynthesis and signal transduction owing to its genetic and molecular tractability. To study the functions of miRNAs in tomato fruit ripening and senescence, and their possible roles in ethylene response, the next generation sequencing method was employed to identify miRNAs in tomato fruit. Bioinformatics and molecular biology approaches were combined to profile the miRNAs expression patterns at three different fruit ripening stages and by exogenous ethylene treatment.</p> <p>Results</p> <p>In addition to 7 novel miRNA families, 103 conserved miRNAs belonging to 24 families and 10 non-conserved miRNAs matching 9 families were identified in our libraries. The targets of many these miRNAs were predicted to be transcriptional factors. Other targets are known to play roles in the regulation of metabolic processes. Interestingly, some targets were predicted to be involved in fruit ripening and softening, such as Pectate Lyase, beta-galactosidase, while a few others were predicted to be involved in ethylene biosynthesis and signaling pathway, such as ACS, EIN2 and CTR1. The expression patterns of a number of such miRNAs at three ripening stages were confirmed by stem-loop RT-PCR, which showed a strong negative correlation with that of their targets. The regulation of exogenous ethylene on miRNAs expression profiles were analyzed simultaneously, and 3 down-regulated, 5 up-regulated miRNAs were found in this study.</p> <p>Conclusions</p> <p>A combination of high throughput sequencing and molecular biology approaches was used to explore the involvement of miRNAs during fruit ripening. Several miRNAs showed differential expression profiles during fruit ripening, and a number of miRNAs were influenced by ethylene treatment. The results suggest the importance of miRNAs in fruit ripening and ethylene response.</p

Application of a new Structural Joint Inversion Approach to Teleseismic and Gravity Data from Mt.Vesuvius, Italy

A 3-D joint inversion of seismic and gravimetric data is performed to re-investigate the subsurface structure of Mt. Vesuvius (Italy) utilizing an improved joint inversion method. The aim is to derive models of the 3D distribution of velocity and density perturbations that are consistent with both data sets and with local velocity models. Mt. Vesuvius is a strato volcano located within a graben (Campania Plain) formed in Plio-Pleistocene. Campania Plain is bordered by mostly Mesozoic carbonaceous rocks. Mt. Vesuvius is the southernmost and the youngest of a group of Pleistocene volcanoes, three of which (Ischia, Campi Flegrei and Mt. Vesuvius) have erupted in historical times. The most recent eruption of Mt. Vesuvius occurred in 1944 and since then the volcanic activity has been characterized by moderate low magnitude seismicity and low temperature fumaroles at the summit crater. We modified the coupling mechanism between velocity and density models in the JI-3D optimized joint inversion method (Jordan and Achauer, 1999). This method was designed to provide stable and high resolution results and involves iterative optimized parameterization, 3D ray tracing, and the incorporation of a priori information. The coupling of the velocity and density models, vital to the joint inversion, is based on a cross-gradient approach (e.g. Gallardo and Meju, 2004), which has been proven to work very well in a variety of cases involving seismic, magnetic, CSEM, MT and gravity data sets. We implemented the cross-gradient coupling for our 3-D irregular adaptive grid parameterization. In contrast to conventional joint inversion methods this approach encourages structural similarities in the models and does not rely on predefined relationships between velocity and density parameters. As a consequence, the resulting velocity-density relations are not contaminated by a priori assumptions and can be utilized to derive rock physical parameters. We apply this method to data from the TomoVes project (Gasparini et al. 1998), combining seismics and Bouguer gravity and local high resolution velocity models as a priori information. The starting models for the joint inversion are derived by separate inversions of the individual data sets. We show 3D distributions of velocity perturbations and density variations from the joint inversion of teleseismic relative traveltimes and Bouguer anomaly data with the aim of extracting further information about the physical status of the volcano- tectonic system

Establishment of a viable cell detection system for microorganisms in wine based on ethidium monoazide and quantitative PCR

Fermentability and contamination level of wine can be assessed through the detection of viable fermentation-related and spoilage-related microorganisms. Ethidium monoazide in combination with quantitative PCR (EMA-qPCR) has been considered as a promising method to enumerate viable cells. Milling for 80 s by O 500-mu m glass beads is demonstrated to be optimal for DNA extraction from yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB) in wine to be used as a template for PCR. EMA-qPCR results from experiments using DNA extracted by this method correlate well with the results of a plating assay (R-2 > 0.99), and a PCR efficiency between 96% and 105% was obtained. Moreover, for all of these microorganisms, EMA treatment of pure cultures at a low concentration (10 mu g/mL) for 20 min photoactivation resulted in effective differentiation between viable and non-viable cells and had no effect on viable cells. Due to sublethal injury to some cells, underestimation of cell counts was found in most of the wine samples tested using the EMA-qPCR method, and a 40-min incubation in recovery medium could completely offset this error. Our results suggest an optimal glass-bead DNA extraction method and EMA treatment suitable for all of the main microorganisms in wine. The EMA-qPCR method was successfully applied to quantify yeasts. Saccharomyces cerevisiae (S. cerevisiae), LAB, non-Oenococcus oeni LAB (non-O. oeni LAB) and AAB in wine samples. (C) 2012 Elsevier Ltd. All rights reserved

Case report: Microwave ablation is a safe and effective method for primary hyperparathyroidism in pregnancy

Primary hyperparathyroidism (PHPT) is a rare disease in pregnancy and endangers the health of both pregnant women and fetuses. However, the treatments are very limited for PHPT and most of them are unsatisfactory because of the peculiar state in pregnancy. The only curable method is parathyroidectomy which can be safely performed in the second trimester of pregnancy. In this case, we reported a pregnant woman with primary parathyroid adenoma presenting hypercalcemia and severe vomit at the end of first trimester. Finally, she got cured by microwave ablation at the end of first trimester and gave birth to a healthy baby boy

A Novel Universal Primer-Multiplex-PCR Method with Sequencing Gel Electrophoresis Analysis

In this study, a novel universal primer-multiplex-PCR (UP-M-PCR) method adding a universal primer (UP) in the multiplex PCR reaction system was described. A universal adapter was designed in the 5′-end of each specific primer pairs which matched with the specific DNA sequences for each template and also used as the universal primer (UP). PCR products were analyzed on sequencing gel electrophoresis (SGE) which had the advantage of exhibiting extraordinary resolution. This method overcame the disadvantages rooted deeply in conventional multiplex PCR such as complex manipulation, lower sensitivity, self-inhibition and amplification disparity resulting from different primers, and it got a high specificity and had a low detection limit of 0.1 ng for single kind of crops when screening the presence of genetically modified (GM) crops in mixture samples. The novel developed multiplex PCR assay with sequencing gel electrophoresis analysis will be useful in many fields, such as verifying the GM status of a sample irrespective of the crop and GM trait and so on

Transcript and protein profiling analysis of OTA-induced cell death reveals the regulation of the toxicity response process in Arabidopsis thaliana

Ochratoxin A (OTA) is a toxic isocoumarin derivative produced by various species of mould which mainly grow on grain, coffee, and nuts. Recent studies have suggested that OTA induces cell death in plants. To investigate possible mechanisms of OTA phytotoxicity, both digital gene expression (DGE) transcriptomic and two-dimensional electrophoresis proteomic analyses were used, through which 3118 genes and 23 proteins were identified as being up- or down-regulated at least 2-fold in Arabidopsis leaf in response to OTA treatment. First, exposure of excised Arabidopsis thaliana leaves to OTA rapidly causes the hypersensitive reponse, significantly accelerates the increase of reactive oxygen species and malondialdehyde, and enhances antioxidant enzyme defence responses and xenobiotic detoxification. Secondly, OTA stimulation causes dynamic changes in transcription factors and activates the membrane transport system dramatically. Thirdly, a concomitant persistence of compromised photosynthesis and photorespiration is indicative of a metabolic shift from a highly active to a weak state. Finally, the data revealed that ethylene, salicylic acid, jasmonic acid, and mitogen-activated protein kinase signalling molecules mediate the process of toxicity caused by OTA. Profiling analyses on Arabidopsis in response to OTA will provide new insights into signalling transduction that modulates the OTA phytotoxicity mechanism, facilitate mapping of regulatory networks, and extend the ability to improve OTA tolerance in Arabidopsis

A Mitochondria-Dependent Pathway Mediates the Apoptosis of GSE-Induced Yeast

Grapefruit seed extract (GSE), which has powerful anti-fungal activity, can induce apoptosis in S. cerevisiae. The yeast cells underwent apoptosis as determined by testing for apoptotic markers of DNA cleavage and typical chromatin condensation by Terminal Deoxynucleotidyl Transferase–mediated dUTP Nick End Labeling (TUNEL) and 4,6′-diaminidino-2-phenylindole (DAPI) staining and electron microscopy. The changes of ΔΨmt (mitochondrial transmembrane potential) and ROS (reactive oxygen species) indicated that the mitochondria took part in the apoptotic process. Changes in this process detected by metabonomics and proteomics revealed that the yeast cells tenaciously resisted adversity. Proteins related to redox, cellular structure, membrane, energy and DNA repair were significantly increased. In this study, the relative changes in the levels of proteins and metabolites showed the tenacious resistance of yeast cells. However, GSE induced apoptosis in the yeast cells by destruction of the mitochondrial 60 S ribosomal protein, L14-A, and prevented the conversion of pantothenic acid to coenzyme A (CoA). The relationship between the proteins and metabolites was analyzed by orthogonal projections to latent structures (OPLS). We found that the changes of the metabolites and the protein changes had relevant consistency