<meta charset="utf8"></head><body>估计三唑类农药在植物中的整合代谢物细胞毒性

<ol>
	<li>Fan, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake+of+halogenated+organic+compounds+(HOCs)+into+peanut+and+corn+during+the+whole+life+cycle+grown+in+an+agricultural+field.">Uptake of halogenated organic compounds (HOCs) into peanut and corn during the whole life cycle grown in an agricultural field.</a> Environ Pollut. 263, 114400 (2020).</li><li>Wu, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+metals+in+e-waste+lead+to+serious+health+risk+through+consumption+of+rice+growing+near+an+abandoned+e-waste+recycling+site:+Comparisons+with+PBDEs+and+AHFRs.">Trace metals in e-waste lead to serious health risk through consumption of rice growing near an abandoned e-waste recycling site: Comparisons with PBDEs and AHFRs.</a> Environ Pollut. 247, 46-54 (2019).</li><li>Liu, A. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tetrabromobisphenol-A/S+and+nine+novel+analogs+in+biological+samples+from+the+Chinese+Bohai+Sea:+Implications+for+trophic+transfer.">Tetrabromobisphenol-A/S and nine novel analogs in biological samples from the Chinese Bohai Sea: Implications for trophic transfer.</a> Environ Sci Technol. 50 (8), 4203-4211 (2016).</li><li>Awasthi, A. K., Zeng, X., Li, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+pollution+of+electronic+waste+recycling+in+India:+A+critical+review.">Environmental pollution of electronic waste recycling in India: A critical review.</a> Environ Pollut. 211, 259-270 (2016).</li><li>Zhang, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plant+accumulation+and+transformation+of+brominated+and+organophosphate+flame+retardants:+A+review.">Plant accumulation and transformation of brominated and organophosphate flame retardants: A review.</a> Environ Pollut. 288, 117742 (2021).</li><li>Sandermann, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Higher+plant+metabolism+of+xenobiotics:+the+'green+liver'+concept.">Higher plant metabolism of xenobiotics: the 'green liver' concept.</a> Pharmacogenetics. 4, 225-241 (1994).</li><li>Zhang, J. J., Yang, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolism+and+detoxification+of+pesticides+in+plants.">Metabolism and detoxification of pesticides in plants.</a> Sci Total Environ. 790, 148034 (2021).</li><li>Debenest, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ecotoxicity+of+a+brominated+flame+retardant+(tetrabromobisphenol+A)+and+its+derivatives+to+aquatic+organisms.">Ecotoxicity of a brominated flame retardant (tetrabromobisphenol A) and its derivatives to aquatic organisms.</a> Comp Biochem Phys C. 152 (4), 407-412 (2010).</li><li>McCormick, J. M., Van Es, T., Cooper, K. R., White, L. A., Haggblom, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbially+mediated+O-methylation+of+bisphenol+A+results+in+metabolites+with+increased+toxicity+to+the+developing+zebrafish+(Danio+rerio)+embryo.">Microbially mediated O-methylation of bisphenol A results in metabolites with increased toxicity to the developing zebrafish (Danio rerio) embryo.</a> Environ Sci Technol. 45 (15), 6567-6574 (2011).</li><li>Zhang, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+metabolic+pathways+of+2,4,6-tribromophenol+in+rice+plants.">Multiple metabolic pathways of 2,4,6-tribromophenol in rice plants.</a> Environ Sci Technol. 53 (13), 7473-7482 (2019).</li><li>Hou, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycosylation+of+tetrabromobisphenol+A+in+pumpkin.">Glycosylation of tetrabromobisphenol A in pumpkin.</a> Environ Sci Technol. 53 (15), 8805-8812 (2019).</li><li>Wu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elucidating+the+metabolism+of+2,4-Dibromophenol+in+plants.">Elucidating the metabolism of 2,4-Dibromophenol in plants.</a> J Vis Exp. (192), e65089 (2023).</li><li>Zucco, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Inter-laboratory+study+to+evaluate+the+effects+of+medium+composition+on+the+differentiation+and+barrier+function+of+Caco-2+cell+lines.">An Inter-laboratory study to evaluate the effects of medium composition on the differentiation and barrier function of Caco-2 cell lines.</a> Atla-Altern Lab Anim. 33, 603-618 (2005).</li><li>Eisenbrand, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+of+in+vitro+toxicology.">Methods of in vitro toxicology.</a> Food Chem Toxicol. 40, 193-236 (2002).</li><li>Sambuy, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Caco-2+cell+line+as+a+model+of+the+intestinal+barrier:+influence+of+cell+and+culture-related+factors+on+Caco-2+cell+functional+characteristics.">The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics.</a> Cell Biol Toxicol. 21, 1-26 (2005).</li><li>Li, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake,+translocation,+and+subcellular+distribution+of+three+triazole+pesticides+in+rice.">Uptake, translocation, and subcellular distribution of three triazole pesticides in rice.</a> Environ Sci Pollut Res. 29 (17), 25581-25590 (2022).</li><li>Satapute, P., Kamble, M. V., Adhikari, S. S., Jogaiah, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+triazole+pesticides+on+tillage+soil+microbial+populations+and+metabolic+changes.">Influence of triazole pesticides on tillage soil microbial populations and metabolic changes.</a> Sci Total Environ. 651, 2334-2344 (2019).</li><li>Xu, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+possible+but+unrecognized+risk+of+acceptable+daily+intake+dose+triazole+pesticides+exposure-bile+acid+disturbance+induced+pharmacokinetic+changes+of+oral+medication.">A possible but unrecognized risk of acceptable daily intake dose triazole pesticides exposure-bile acid disturbance induced pharmacokinetic changes of oral medication.</a> Chemosphere. 322, 138209 (2023).</li><li>Louis, K. S., Siegel, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+viability+analysis+using+trypan+blue:+manual+and+automated+methods.">Cell viability analysis using trypan blue: manual and automated methods.</a> Methods Mol Biol. 740, 7-12 (2011).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2,4-dichlorophenoxyacetic acid</td>
			<td>WAKO</td>
			<td>1 mg/L</td>
			<td></td>
		</tr>
		<tr>
			<td>20% H2O2</td>
			<td>Sinopharm Chemical Reagent Co., Ltd.</td>
			<td>10011218-500ML</td>
			<td></td>
		</tr>
		<tr>
			<td>6-benzylaminopurine</td>
			<td>WAKO</td>
			<td>0.5 mg/L</td>
			<td></td>
		</tr>
		<tr>
			<td>75% ethanol</td>
			<td>Sinopharm Chemical Reagent Co., Ltd.</td>
			<td>1269101-500 mL</td>
			<td></td>
		</tr>
		<tr>
			<td>96-well plate</td>
			<td>Thermo Fisher</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Acetonitrile</td>
			<td>Sigma-Aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Artificial climate incubator</td>
			<td>Ningbo DongNan Lab Equipment Co.,Ltd</td>
			<td>RDN-1000A-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Autoclaves</td>
			<td>STIK</td>
			<td>MJ-Series</td>
			<td></td>
		</tr>
		<tr>
			<td>Caco-2 cells</td>
			<td>Nuoyang Biotechnology Co.,Ltd.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CCK8 reagents</td>
			<td>Nanjing Jiancheng Bioengineering Institute, China</td>
			<td>G021-1-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Thermo Fisher</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CO2 incubator</td>
			<td>Labtrip</td>
			<td>HWJ-3-160</td>
			<td></td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide</td>
			<td>Solarbio Life Sciences</td>
			<td>D8371</td>
			<td></td>
		</tr>
		<tr>
			<td>Diniconazole, 98.7%</td>
			<td>J&#38;K Scientific</td>
			<td>83657-24-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s modified Eagle&#39;s medium</td>
			<td>Solarbio Life Sciences</td>
			<td>11965-500 mL</td>
			<td></td>
		</tr>
		<tr>
			<td>electronic balance</td>
			<td>Shanghai Precision Instrument Co., Ltd</td>
			<td>FA1004B</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Cellmax</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence spectrophotometer</td>
			<td>Tecan</td>
			<td>Infinite M200</td>
			<td></td>
		</tr>
		<tr>
			<td>Flusilazole, 98.5%</td>
			<td>J&#38;K Scientific</td>
			<td>85509-19-9&#160;&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Freeze dryer</td>
			<td>SCIENTZ</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>High-throughput tissue grinder</td>
			<td>SCIENTZ</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>Leica Biosystems</td>
			<td>DMi1</td>
			<td></td>
		</tr>
		<tr>
			<td>Milli-Q system</td>
			<td>Millipore</td>
			<td>MS1922801-4L</td>
			<td></td>
		</tr>
		<tr>
			<td>Murashige &#38; Skoog medium</td>
			<td>HOPEBIO</td>
			<td>HB8469-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrogen blowing concentrator</td>
			<td>AOSHENG</td>
			<td>MD200-2</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Solarbio Life Sciences</td>
			<td>P1022-500 mL</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin Liquid</td>
			<td>Solarbio Life Sciences</td>
			<td>P1400-100 mL</td>
			<td></td>
		</tr>
		<tr>
			<td>Propiconazole, 100%</td>
			<td>J&#38;K Scientific</td>
			<td>60207-90-1&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Research plus</td>
			<td>Eppendorf</td>
			<td>10-1000 &#956;L</td>
			<td></td>
		</tr>
		<tr>
			<td>Seeds of Little Finger carrot (Daucus carota var. sativus)</td>
			<td>Shouguang Seed Industry Co., Ltd</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Shaking Incubators</td>
			<td>Shanghai bluepard instruments Co.,Ltd.</td>
			<td>THZ-98AB</td>
			<td></td>
		</tr>
		<tr>
			<td>Tebuconazole, 100%</td>
			<td>J&#38;K Scientific</td>
			<td>107534-96-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin-EDTA solution</td>
			<td>Solarbio Life Sciences</td>
			<td>T1300-100 mL</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound machine</td>
			<td>ZKI</td>
			<td>UC-6</td>
			<td></td>
		</tr>
		<tr>
			<td>UV-sterilized super clean bench</td>
			<td>AIRTECH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vortex instrument</td>
			<td>Wuxi Laipu Instrument Equipment Co., Ltd</td>
			<td>BV-1010</td>
			<td></td>
		</tr>
	</tbody>
</table>

assessing cytotoxicity of metabolites of typical triazole pesticides in plants

Read this Scientific Article Protocol about Assessing Cytotoxicity of Metabolites of Typical Triazole Pesticides in Plants at JoVE.com

<meta charset="utf8"></head><body>作者聚焦：一种对植物中三唑类农药进行综合代谢物细胞毒性评估的新型高效方法

评估植物中典型三唑类农药代谢物的细胞毒性

Various organic pollutants have been released into the environment because of anthropogenic activities. These pollutants can be taken up by crop plants, ...

assessing-cytotoxicity-metabolites-typical-triazole-pesticides

Research

JoVE Journal

Environment

Assessing Cytotoxicity of Metabolites of Typical Triazole Pesticides in Plants

310 次观看.  Zhejiang University of Technology. 我们的方案描述了一种评估植物中三唑类农药代谢物的整体细胞毒性的新方法。植物愈伤组织培养物用于产生植物中蛋白质的代谢物，然后使用人类细胞系进行毒理学测试，以评估代谢物的整体细胞毒性。植物中外源性物质的代谢产物极其复杂，只能获得少数标准品的代谢物。因此，代谢物的毒理学数据有限，阻碍了全面的毒性评估。该方案提...

视频: 作者聚焦：一种对植物中三唑类农药进行综合代谢物细胞毒性评估的新型高效方法

Expression of Recombinant Cellulase Cel5A from Trichoderma reesei in Tobacco Plants

Cellulose degrading enzymes, cellulases, are targets of both research and industrial interests. The preponderance of these enzymes in difficult-to-culture organisms, such as hyphae-building fungi and anaerobic bacteria, has hastened the use of recombinant technologies in this field. Plant expression methods are a desirable system for large-scale production of enzymes and other industrially useful proteins. Herein, methods for the transient expression of a fungal endoglucanase, Trichoderma reesei Cel5A, in Nicotiana tabacum are demonstrated. Successful protein expression is shown, monitored by fluorescence using an mCherry-enzyme fusion protein. Additionally, a set of basic tests are used to examine the activity of transiently expressed T. reesei Cel5A, including SDS-PAGE, Western blotting, zymography, as well as&#160;fluorescence and dye-based substrate degradation assays. The system described here can be used to produce an active cellulase in a short time period, so as to assess the potential for further production in plants through constitutive or inducible expression systems.

Expression of Recombinant Cellulase Cel5A from Trichoderma reesei in Tobacco Plants

Tobacco plants were used to produce a fungal cellulase, TrCel5A, via a transient expression system. The expression could be monitored using a fluorescent fusion protein, and the protein activity was characterized post-expression.

重组纤维素酶Cel5A从表达<em&gt;里氏木霉</em&gt;在烟草

Cellulose degrading enzymes, cellulases, are targets of both research and industrial interests. The preponderance of these enzymes in difficult-to-culture ...

科研

环境科学

Measuring Fluxes of Mineral Nutrients and Toxicants in Plants with Radioactive Tracers

Unidirectional influx and efflux of nutrients and toxicants, and their resultant net fluxes, are central to the nutrition and toxicology of plants. Radioisotope tracing is a major technique used to measure such fluxes, both within plants, and between plants and their environments. Flux data obtained with radiotracer protocols can help elucidate the capacity, mechanism, regulation, and energetics of transport systems for specific mineral nutrients or toxicants, and can provide insight into compartmentation and turnover rates of subcellular mineral and metabolite pools. Here, we describe two major radioisotope protocols used in plant biology: direct influx (DI) and compartmental analysis by tracer efflux (CATE). We focus on flux measurement of potassium (K+) as a nutrient, and ammonia/ammonium (NH3/NH4+) as a toxicant, in intact seedlings of the model species barley (Hordeum vulgare L.). These protocols can be readily adapted to other experimental systems (e.g., different species, excised plant material, and other nutrients/toxicants). Advantages and limitations of these protocols are discussed.

In planta measurement of nutrient and toxicant fluxes is essential to the study of plant nutrition and toxicity. Here, we cover radiotracer protocols for influx and efflux determination in intact plant roots, using potassium (K+) and ammonia/ammonium (NH3/NH4+) fluxes as examples. Advantages and limitations of such techniques are discussed.

植物与放射性示踪剂测量矿质营养和毒物的通量

Unidirectional influx and efflux of nutrients and toxicants, and their resultant net fluxes, are central to the nutrition and toxicology of plants. ...

Characterization and Application of Passive Samplers for Monitoring of Pesticides in Water

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study provides a protocol for the passive sampler preparation, calibration, extraction method and instrumental analysis. Sampling rates (RS) and passive sampler-water partition coefficients (KPW) were calculated for silicone rubber, polar organic chemical integrative sampler POCIS-A, POCIS-B, SDB-RPS and C18 disk. The uptake of the selected compounds depended on their physicochemical properties, i.e., silicone rubber showed a better uptake for more hydrophobic compounds (log octanol-water partition coefficient (KOW) &#62; 5.3), whereas POCIS-A, POCIS-B and SDB-RPS disk were more suitable for hydrophilic compounds (log KOW &#60; 0.70).

A protocol about the characterization and application of five different passive sampling devices is presented.

表征和被动采样的应用在农药水监测

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment during manufacturing and transport. Because of their environmental persistence, they are widely distributed in the oceans and on beaches all over the world. They can act as a vector of potentially toxic organic compounds (e.g., polychlorinated biphenyls) and might consequently negatively affect marine organisms. Their possible impacts along the food chain are not yet well understood. In order to assess the hazards associated with the occurrence of plastic pellets in the marine environment, it is necessary to develop methodologies that allow for rapid determination of associated organic contaminant levels. The present protocol describes the different steps required for sampling resin pellets, analyzing adsorbed organochlorine pesticides (OCPs) and identifying the plastic type. The focus is on the extraction of OCPs from plastic pellets by means of a pressurized fluid extractor (PFE) and on the polymer chemical analysis applying Fourier Transform-InfraRed (FT-IR) spectroscopy. The developed methodology focuses on 11 OCPs and related compounds, including dichlorodiphenyltrichloroethane (DDT) and its two main metabolites, lindane and two production isomers, as well as the two biologically active isomers of technical endosulfan. This protocol constitutes a simple and rapid alternative to existing methodology for evaluating the concentration of organic contaminants adsorbed on plastic pieces.

Microplastics act as vector of potentially toxic organic contaminants with unpredictable effects. This protocol describes an alternative methodology for assessing the levels of organochlorine pesticides adsorbed on plastic pellets and identifying the polymer chemical structure. The focus is on pressurized fluid extraction and attenuated total reflectance Fourier transform infrared spectroscopy.

从塑料颗粒中提取有机氯农药和塑料类型分析

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

Experimental Protocol for Detecting Mitochondrial Function in Hepatocytes Exposed to Organochlorine Pesticides

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by environmental organochlorine pesticides (OCPs) in hepatocytes. HepG2 cells were exposed to &#946;-hexachlorocyclohexane (&#946;-HCH) for 24 h at an equivalent dose of internal exposure in general population. Ultrastructure in hepatocytes was examined by transmission electron microscopy (TEM) to show the damage of mitochondria. Mitochondrial function was further evaluated by mitochondrial fluorescence intensity, adenosine 5'-triphosphate (ATP) levels, oxygen consumption rate (OCR) and mitochondrial membrane potential (MMP) in HepG2 cells incubated with &#946;-HCH. The mitochondria fluorescence intensity after stained by mitochondrial green fluorescent probe was observed with a fluorescence microscopy. The luciferin-luciferase reaction was used to determine ATP levels. The MMP was detected by the cationic dye JC-1 and analyzed under flow cytometry. OCR was measured with an extracellular flux analyzer. In summary, these protocols were used in detecting mitochondrial function in hepatocytes with to investigate mitochondria damages.

Understanding the influence of environmental organochlorine pesticides (OCPs) on mitochondrial function in hepatocytes is important in exploring the mechanism of OCPs causing metabolic disorders. This paper presents detailed methods on detecting hepatic mitochondrial function.

检测暴露于有机氯农药的肝细胞线粒体功能的实验方案

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by ...

Protocol for Producing Three-Dimensional Infrared Video of Freezing in Plants

Freezing in plants can be monitored using infrared (IR) thermography, because when water freezes, it gives off heat. However, problems with color contrast make 2-dimensions (2D) infrared images somewhat difficult to interpret. Viewing an IR image or the video of plants freezing in 3 dimensions (3D) would allow a more accurate identification of sites for ice nucleation as well as the progression of freezing. In this paper, we demonstrate a relatively simple means to produce a 3D infrared video of a strawberry plant freezing. Strawberry is an economically important crop that is subjected to unexpected spring freeze events in many areas of the world. An accurate understanding of the freezing in strawberry will provide both breeders and growers with more economical ways to prevent any damage to plants during freezing conditions.
The technique involves a positioning of two IR cameras at slightly different angles to film the strawberry freezing. The two video streams will be precisely synchronized using a screen capture software that records both cameras simultaneously. The recordings will then be imported into the imaging software and processed using an anaglyph technique. Using red-blue glasses, the 3D video will make it easier to determine the precise site of ice nucleation on leaf surfaces.

Here, we present a protocol to image a strawberry plant freezing in 3 dimensions. Two infrared cameras positioned at slightly different angles are used to produce a red-blue anaglyph video to observe the freezing of the plant in 3 dimensions.

植物冷冻三维红外视频制作协议

Freezing in plants can be monitored using infrared (IR) thermography, because when water freezes, it gives off heat. However, problems with color contrast ...

Visualizing Efficacy of Pesticides Against Disease Vector Mosquitoes in the Field

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform across ecological zones. To best protect public and veterinary health from these insects, the environmental limitations of pesticides need to be investigated to inform effective use of the most appropriate pesticide formulations and techniques. We have developed a research program to evaluate combinations of pesticides, pesticide application equipment, and application techniques in hot-arid desert, hot-humid tropical, warm and cool temperate, and urban locations to derive pesticide use guidelines specific to target insect and environment. To these ends we designed a system of protocols to support efficient, cost-effective, portable, and standardized evaluation of a diverse range of pesticides and equipment across multiple environments. At the core of these protocols is the use of an array of small cages with colony-reared sentinel mosquitoes (adults and immatures) and sand flies (adults), strategically arranged in natural habitats and exposed to pesticide spray. Spatial and temporal patterns of pesticide efficacy are derived from percent mortality in sentinel cages, then mapped and visualized in a geographic information system. Maps of sentinel mortality data may be statistically compared to evaluate relative efficacy of a pesticide across multiple environments, or to study multiple pesticides in a single environment. Protocols may be modified to accommodate a variety of scenarios, including, for example, the vertical orientation of sentinels in canopy habitats or simultaneous testing of ground and aerial application methods.

The efficacy of public health pesticides targeting nuisance and disease-vector insects is not uniform across different ecological zones. Here we present a system of techniques using captive vector insects as sentinels for pesticide efficacy to derive electronic maps supporting the standard evaluation of pesticides across multiple environments.

农药在田间对病媒蚊子的影响

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform ...

A Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci.

A Loop-mediated Isothermal Amplification LAMP Assay for Rapid Identification of Bemisia tabaci

This paper reports the protocol for a rapid identification assay for Bemisia tabaci based on loop-mediated isothermal amplification (LAMP) technology. The protocol requires minimal laboratory training and can, therefore, be implemented on-site at points of entry for plant imports such as seaports and airports.

循环介导等温扩增 (灯管) 法快速识别烟粉虱

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many ...

Cytotoxicity Assays with Zebrafish Cell Lines

Fish cell lines have become increasingly used in ecotoxicity studies, and cytotoxicity assays have been proposed as methods to predict fish acute toxicity. Thus, this protocol presents cytotoxicity assays modified to evaluate cell viability in zebrafish (Danio rerio) embryo (ZEM2S) and liver (ZFL) cell lines in 96-well plates. The cytotoxicity endpoints evaluated are mitochondrial integrity (Alamar Blue [AB] and MTT assays), membrane integrity via esterase activity (CFDA-AM assay), and lysosomal membrane integrity (Neutral Red [NR] assay). After the exposure of the test substances in a 96-well plate, the cytotoxicity assays are performed; here, AB and CFDA-AM are carried out simultaneously, followed by NR on the same plate, while the MTT assay is performed on a separate plate. The readouts for these assays are taken by fluorescence for AB and CFDA-AM, and absorbance for MTT and NR. The cytotoxicity assays performed with these fish cell lines can be used to study the acute toxicity of chemical substances on fish.

This protocol presents commonly used cytotoxicity assays (Alamar Blue [AB], CFDA-AM, Neutral Red, and MTT assays) adapted for the assessment of cytotoxicity in zebrafish embryo (ZEM2S) and liver (ZFL) cell lines in 96-well plates.

斑马鱼细胞系的细胞毒性测定

Fish cell lines have become increasingly used in ecotoxicity studies, and cytotoxicity assays have been proposed as methods to predict fish acute ...

Elucidating the Metabolism of 2,4-Dibromophenol in Plants

Crops can be extensively exposed to organic pollutants, since soil is a major sink for pollutants discarded into the environment. This creates potential human exposure through the consumption of pollutant-accumulated foods. Elucidating the uptake and metabolism of xenobiotics in crops is essential for the assessment of dietary exposure risk in humans. However, for such experiments, the use of intact plants requires long-term experiments and complex sample preparation protocols that can be affected by various factors. Plant callus cultures combined with high-resolution mass spectrometry (HRMS) may provide a solution for the accurate and time-saving identification of metabolites of xenobiotics in plants, as it can avoid interference from the microbial or fungal microenvironment, shorten the treatment duration, and simplify the matrix effect of intact plants. 2,4-dibromophenol, a typical flame retardant and endocrine disrupter, was chosen as the model substance due to its widespread occurrence in soil and its uptake potential by plants. Herein, plant callus was generated from asepsis seeds and exposed to sterile 2,4-dibromophenol-containing culture medium. The results showed that eight metabolites of 2,4-dibromophenol were identified in the plant callus tissues after 120 h of incubation. This indicates that 2,4-dibromophenol was rapidly metabolized in&#160;the plant callus tissues. Thus, the plant callus culture platform is an effective method to evaluate the uptake and metabolism of xenobiotics in plants.

The present protocol describes a simple and efficient method for the identification of 2,4-dibromophenol metabolites in plants.

阐明植物中2,4-二溴苯酚的代谢

Crops can be extensively exposed to organic pollutants, since soil is a major sink for pollutants discarded into the environment. This creates potential ...