Name: electroantennographic bioassay as a screening tool for host plant volatiles
Uploaded: 2012-05-06T14:00:00
Description: Watch this Scientific Journal Video about Electroantennographic Bioassay as a Screening Tool for Host Plant Volatiles at JoVE.com

Denne forskning blev udført under USDA-ARS CRIS Project 5325-42000-037-00D og med resultater fra CRADA 58-3K95-7-1198 og TFCA 58-5325-8-419. Forfatterne takker Suterra til gave (Z, Z) -11,13-hexadecadienal, B. Higbee for konstruktive drøftelser, og J. Baker for teknisk bistand.

1. Udarbejdelse af flygtige Opdagede fra værtsplanten for Eag Screening <ol><li> Efter en passende identifikation og autentifikation af alle flygtige stoffer via GC-MS, udføre EAG pust analyse af hver tilgængelig flygtigt. Indledende screening kan være et lavt replikat antal antennal responser (N = 3-5) af begge køn for at opnå en indikation af relativ chemoreceptivity i en kort tid (tabel 1). </li><li> Fremstille en opløsning af hver flygtig ved en 5 mg / ml koncentration i pentan. Tæt forseg...

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Anvendelse af electroantennogram optagelser som et bioassay for at bestemme chemoreception responser et mål insekt er ret almindelig, og mange undersøgelser under anvendelse af EAG som en detektor for effluent fra et gaskromatogram (GC-EAD) kan findes i litteraturen. 9,10 Fremgangsmåden vist vil give en hurtig screening af ækvivalente mængder af flygtige komponenter med høje gentagelser for sikker tildeling af den relative lydhørhed. Det AutoSpike programmet i Syntech software er et godt program for sc...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer </td></tr><tr><td> Acetophenon </td><td> Alfa-Aesar </td><td> A12727 </td><td> Kvinde positiv kontrol </td></tr><tr><td> (Z, Z) -11,13-Hexadecadienal </td><td> Suterra </td><td></td><td> Mand positiv kontrol </td></tr><tr><td> α-Humulene </td><td> Aldrich </td><td> 53675 </td><td> Sesquiterpene </td></tr><tr><td> 2-undecanon </td><td> Aldrich </td><td> U1303 </td><td> Fedtsyrederivat </td></tr><tr><td> 2-phenylethanol </td><td> Aldrich </td><td> 77861 </td><td> Benzenoid </td></tr><tr><td> Pentan </td><td> EMD </td><td> PX0167-1 </td><td> Opløsningsmiddel </td></tr><tr><td> 4-kanals erhvervelse controller </td><td> Syntech </td><td> JegDAC-4 </td><td></td></tr><tr><td> EAG probe-forforstærker </td><td> Syntech </td><td> Type PRG-2 </td><td></td></tr><tr><td> Antenne holder </td><td> Syntech </td><td> For PRG-2 </td><td> Fork elektrode </td></tr><tr><td> Stimulus controller </td><td> Syntech </td><td> CS-55 </td><td> Air flow og puffs </td></tr><tr><td> Spectra elektrodegel </td><td> Parker </td><td> 12-02 </td><td></td></tr><tr><td> Bioassays diske </td><td> Whatman </td><td> 2017-006 </td><td> 6 mm </td></tr><tr><td> Pasteur pipetter </td><td> VWR </td><td> 14673-010 </td><td> 5 ¾ &quot;(14,6 cm) </td></tr><tr><td> Parafilm M </td><td> Bemis </td><td> PM-992 </td><td></td></tr></tbody></table>

electroantennographic bioassay as a screening tool for host plant volatiles

Plant flygtige stoffer spiller en vigtig rolle i plante-insekt interaktioner. Planteædende insekter bruger plante flygtige, kendt som kairomones, for at lokalisere deres værtsplante. 1,2 Når en værtsplante er en vigtig agronomisk vare fodring skade ved skadedyr kan påføre alvorlige økonomiske tab for avlerne. Følgelig kan kairomones anvendes som attraktanter at lokke eller forvirre disse insekter og således giver et miljøvenligt alternativ til pesticider til insektkontrol. 3 Uheldigvis kan planterne udsende en lang række flygtige med varierende sammensætninger af og forhold mellem emissionen afhængige af fænologi af varen eller tidspunktet på dagen. Dette gør identifikation af biologisk aktive bestanddele eller blandinger af flygtige bestanddele en besværlig proces. For at hjælpe med at identificere de bioaktive komponenter i værtsplante flygtige emissioner vi anvender laboratoriet-baserede screening bioassay electroantennography (EAG). EAG er et effektivt værktøj til at evaluere og RECORd elektrofysiologisk de olfaktoriske svarene fra et insekt via deres antennal receptorer. Den EAG screening kan medvirke til at reducere antallet af flygtige stoffer testet for at identificere lovende bioaktive komponenter. Men Eag bioassays kun give oplysninger om aktivering af receptorer. Det giver ikke oplysninger om den type insekt opførsel den forbindelse fremkalder, som kan være som et tiltrækkende, frastødende eller anden form for adfærdsmæssig reaktion. Flygtige bevirker en signifikant respons ved EAG i forhold til en passende positiv kontrol, er typisk taget på yderligere testning af adfærdsmæssige responser af skadelige insekt. Det eksperimentelle design forelagt, detaljer anvendte metode til at screene mandel-baserede værtsplante flygtige 4,5 ved måling af de elektrofysiologiske antennal svarene fra en voksen insektskadedyr navlen orangeworm (Amyelois transitella) til de enkelte komponenter og enkle blandinger af komponenter via Eag bioassay. Fremgangsmåden anvender to exbliver offentliggjort antenner placeret på tværs af en &quot;gaffel&quot; elektrode holder. Protokollen vist her udgør en hurtig, high-throughput standardiseret fremgangsmåde til screening af flygtige stoffer. Hver volatile i et sæt, konstant mængde som at standardisere stimulus plan og dermed give antennal reaktioner at være udtryk for den relative chemoreceptivity. Den negative kontrol til at eliminere den elektrofysiologiske svar på både resterende opløsningsmiddel og mekanisk kraft sug. Den positive kontrol (i dette tilfælde acetophenon) er en enkelt forbindelse, der har fremkaldt en konsekvent reaktion fra mandlige og kvindelige navle orangeworm (NU) møl. En yderligere semiochemical standard, der tilvejebringer ensartet reaktion og anvendes til bioassay studier med den mandlige NU møl er (Z, Z) -11,13-hexdecadienal, et aldehyd bestanddel fra det kvindelige producerede kønsferomon 6-8

Watch this Scientific Journal Video about Electroantennographic Bioassay as a Screening Tool for Host Plant Volatiles at JoVE.com

Electroantennographic Bioassay as a Screening Tool for Host Plant Volatiles

En fremgangsmåde til hurtigt at screene værtsplanten flygtige ved måling af elektrofysiologiske respons voksen navlen orangeworm (<em&gt; Amyelois transitella</em&gt;) Antenner til enkelte komponenter og blandinger via electroantennographic analyse er påvist.

Electroantennographic Biotest som et screeningsværktøj for værtsplante Flygtige

Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host ...

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