मेजबान संयंत्र वाष्पशील के लिए एक स्क्रीनिंग उपकरण के रूप में electroantennographic bioassay
Summary
वयस्क नाभि orangeworm की electrophysiological प्रतिक्रिया (माप से तेजी से स्क्रीन मेजबान संयंत्र वाष्पशील एक विधि<em> Amyelois transitella</em> एकल घटक और electroantennographic विश्लेषण के माध्यम से मिश्रणों के लिए एंटीना) प्रदर्शन किया है.
Abstract
Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host plant.1,2 When a host plant is an important agronomic commodity feeding damage by insect pests can inflict serious economic losses to growers. Accordingly, kairomones can be used as attractants to lure or confuse these insects and, thus, offer an environmentally friendly alternative to pesticides for insect control.3 Unfortunately, plants can emit a vast number volatiles with varying compositions and ratios of emissions dependent upon the phenology of the commodity or the time of day. This makes identification of biologically active components or blends of volatile components an arduous process. To help identify the bioactive components of host plant volatile emissions we employ the laboratory-based screening bioassay electroantennography (EAG). EAG is an effective tool to evaluate and record electrophysiologically the olfactory responses of an insect via their antennal receptors. The EAG screening process can help reduce the number of volatiles tested to identify promising bioactive components. However, EAG bioassays only provide information about activation of receptors. It does not provide information about the type of insect behavior the compound elicits; which could be as an attractant, repellent or other type of behavioral response. Volatiles eliciting a significant response by EAG, relative to an appropriate positive control, are typically taken on to further testing of behavioral responses of the insect pest. The experimental design presented will detail the methodology employed to screen almond-based host plant volatiles4,5 by measurement of the electrophysiological antennal responses of an adult insect pest navel orangeworm (Amyelois transitella) to single components and simple blends of components via EAG bioassay. The method utilizes two excised antennae placed across a “fork” electrode holder. The protocol demonstrated here presents a rapid, high-throughput standardized method for screening volatiles. Each volatile is at a set, constant amount as to standardize the stimulus level and thus allow antennal responses to be indicative of the relative chemoreceptivity. The negative control helps eliminate the electrophysiological response to both residual solvent and mechanical force of the puff. The positive control (in this instance acetophenone) is a single compound that has elicited a consistent response from male and female navel orangeworm (NOW) moth. An additional semiochemical standard that provides consistent response and is used for bioassay studies with the male NOW moth is (Z,Z)-11,13-hexdecadienal, an aldehyde component from the female-produced sex pheromone.6-8
Protocol
Discussion
Bioassay लक्ष्य कीट chemoreception प्रतिक्रियाओं निर्धारित रूप में electroantennogram रिकॉर्डिंग के प्रयोग काफी आम है और कई गैस वर्णलेख (जीसी EAD) से प्रवाह के लिए एक डिटेक्टर के रूप में ईएजी का उपयोग अध्ययन साहित्य में पाया जा सक?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
इस अनुसंधान USDA-ARS क्रिस 5325-42000-037-00D परियोजना के तहत और 58 3K95-7-1198 CRADA और TFCA 58-5325-8-419 से परिणाम के साथ आयोजित किया गया. लेखक का उपहार के लिए कृतज्ञता Suterra स्वीकार करते हैं (जेड, जेड) -11,13 – hexadecadienal, बी Higbee उत्पादक विचार – विमर्श के लिए, और तकनीकी सहायता के लिए जे बेकर.
Materials
| Name of the reagent | Company | Catalogue number | Comments |
| Acetophenone | Alfa-Aesar | A12727 | Female positive control |
| (Z,Z)-11,13-Hexadecadienal | Suterra | Male positive control | |
| α-Humulene | Aldrich | 53675 | Sesquiterpene |
| 2-Undecanone | Aldrich | U1303 | Fatty acid derivative |
| 2-Phenylethanol | Aldrich | 77861 | Benzenoid |
| Pentane | EMD | PX0167-1 | Solvent |
| 4-Channel acquisition controller | Syntech | IDAC-4 | |
| EAG probe, pre-amplifier | Syntech | Type PRG-2 | |
| Antenna holder | Syntech | For PRG-2 | Fork electrode |
| Stimulus controller | Syntech | CS-55 | Air flow and puffs |
| Spectra Electrode Gel | Parker | 12-02 | |
| Bioassay discs | Whatman | 2017-006 | 6 mm |
| Pasteur pipets | VWR | 14673-010 | 5 ¾” (14.6 cm) |
| Parafilm M | Bemis | PM-992 |
Riferimenti
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