Name: herbivore-induced blueberry volatiles and intra-plant signaling
Uploaded: 2011-12-18T14:00:00
Description: Watch this Scientific Journal Video about Herbivore-induced Blueberry Volatiles and Intra-plant Signaling at JoVE.com

The author thanks Robert Holdcraft for technical assistance. This study was funded in part by a USDA CSREES Special Grant (2009-34155-19957) and hatch funds (NJ08192).

1. Local induction of volatiles: herbivore damage
<ol>
 <li>Two branches of blueberry plants are bagged with a spun polyester sleeve.</li>
 <li>Six gypsy moth caterpillars (2nd-3rd instars) are placed inside the bags and allowed to feed on plants for 2 days prior to volatile collection. Control plants do not receive any caterpillars. </li>
 <li>Volatile emissions are collected as described below (protocol &#35;7) on day 3 (Figure 1). Polyester sleeves remain on plants to prevent...

<ol>
	<li>Dicke, M., Van Loon, J. J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multitrophic+effects+of+herbivore-induced+plant+volatiles+in+an+evolutionary+context.">Multitrophic effects of herbivore-induced plant volatiles in an evolutionary context.</a> Entomol. Exp. Appl. 97, 237-237 (2000).</li><li>Mumm, R., Dicke, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variation+in+natural+plant+products+and+the+attraction+of+bodyguards+involved+in+indirect+plant+defense.">Variation in natural plant products and the attraction of bodyguards involved in indirect plant defense.</a> Can. J. Zool. 88, 628-628 (2010).</li><li>Hopke, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herbivore-induced+volatiles:+the+emission+of+acyclic+homoterpenes+from+leaves+of+Phaseolus+lunatus+and+Zea+mays+can+be+triggered+by+a+&#946;-glucosidase+and+jasmonic+acid.">Herbivore-induced volatiles: the emission of acyclic homoterpenes from leaves of Phaseolus lunatus and Zea mays can be triggered by a &#946;-glucosidase and jasmonic acid.</a> FEBS Lett. 352, 146-146 (1994).</li><li>Rodriguez-Saona, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+electrophysiological+responses+of+the+emerald+ash+borer,+Agrilus+planipennis,+to+induced+volatiles+of+Manchurian+ash,+Fraxinus+mandshurica.">Behavioral and electrophysiological responses of the emerald ash borer, Agrilus planipennis, to induced volatiles of Manchurian ash, Fraxinus mandshurica.</a> Chemoecology. 16, 75-75 (2006).</li><li>Rodriguez-Saona, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herbivore+induced+volatiles+in+the+perennial+shrub,+Vaccinium+corymbosum,+and+their+role+in+inter-branch+signaling.">Herbivore induced volatiles in the perennial shrub, Vaccinium corymbosum, and their role in inter-branch signaling.</a> J. Chem. Ecol. 35, 163-163 (2009).</li><li>Karban, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Damage-induced+resistance+in+sagebrush:+volatiles+are+key+to+intra-+and+interplant+communication.">Damage-induced resistance in sagebrush: volatiles are key to intra- and interplant communication.</a> Ecology. 87, 922-922 (2006).</li><li>Frost, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Within-plant+signalling+by+volatiles+overcomes+vascular+constraints+on+systemic+signalling+and+primes+responses+against+herbivores.">Within-plant signalling by volatiles overcomes vascular constraints on systemic signalling and primes responses against herbivores.</a> Ecol. Lett. 10, 490-490 (2007).</li><li>Heil, M., Bueno Silva, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Within-plant+signaling+by+volatiles+leads+to+induction+and+priming+of+an+indirect+plant+defense+in+nature.">Within-plant signaling by volatiles leads to induction and priming of an indirect plant defense in nature.</a> Proc. Natl. Acad. Sci. U.S.A. 104, 5467-5467 (2007).</li><li>Turlings, T. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploitation+of+herbivore-induced+plant+odors+by+host-seeking+parasitic+wasps.">Exploitation of herbivore-induced plant odors by host-seeking parasitic wasps.</a> Science. 250, 1251-1251 (1990).</li><li>Makovic, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volatiles+involved+in+the+nonhost+rejection+of+Fraxinus+pennsylvanica+by+Lymantria+dispar+larvae.">Volatiles involved in the nonhost rejection of Fraxinus pennsylvanica by Lymantria dispar larvae.</a> J. Agric. Food Chem. 44, 929-929 (1996).</li></ol>

The push-pull volatile collection apparatus described here represents a standard method for headspace collections of plant volatiles. This apparatus was used to determine the volatile response of blueberry leaves to herbivory by gypsy moth caterpillars and also allowed me to provide new evidence for the role of HIPVs in intra-plant signaling.
The results presented here show that caterpillar feeding, exogenously-applied MeJA , and mechanical wounding increased volatile emissions at the site of...

<table border="1">
<tr>
<td>Name of the reagent</td>
<td>Company</td>
<td>Catalogue number</td>
<td>Comments </td>
</tr>
<tr>
<td>Volatile collection chambers</td>
<td>Analytical Research Systems, Inc.</td>
<td>VCC-G6X12DT-1P</td>
<td>Gainesville, FL</td>
</tr>
<tr>
<td>Air compressor, 20 gal, oil free, 2 hp</td>
<td>Westward</td>
<td>3JR71 </td>
<td>Sold by Grainger, Inc.</td>
</tr>
<tr>
<td>Air delivery system</td>
<td>Analytical Research Systems, Inc.</td>
<td>VCS-ADS-4AFM4C</td>
<td>Gainesville, FL</td>
</tr>
<tr>
<td>Air collection system</td>
<td>Analytical Research Systems, Inc.</td>
<td>VCS-MVCS-4CX1P</td>
<td>Gainesville, FL</td>
</tr>
<tr>
<td>Vacuum pump 100-150V, &frac14; hp</td>
<td>Gast Manufacturing, Inc.</td>
<td><a href="http://www.grainger.com/Grainger/GAST-Vacuum-Pump-4F740">4F740</a></td>
<td>Sold by Grainger, Inc.</td>
</tr>
<tr>
<td>Methyl jasmonate</td>
<td>Sigma-Aldrich</td>
<td>J2500</td>
<td>St. Louis, MO</td>
</tr>
<tr>
<td>Tween-20 </td>
<td>Sigma-Aldrich</td>
<td>93773</td>
<td>St. Louis, MO</td>
</tr>
<tr>
<td>Rhodamine-B</td>
<td>Sigma-Aldrich</td>
<td>&nbsp;</td>
<td>St. Louis, MO</td>
</tr>
<tr>
<td>Plastic spray bottles, 2 oz</td>
<td>Setco Inc.</td>
<td>&nbsp;</td>
<td>Cranbury, NJ</td>
</tr>
<tr>
<td>Spun polyester sleeves</td>
<td>Rockingham Opportunities Corp.</td>
<td>&nbsp;</td>
<td>Reidsville, NC</td>
</tr>
<tr>
<td>Super-Q volatile collection traps</td>
<td>Analytical Research Systems, Inc.</td>
<td>VCT-1/4X3-SPQ</td>
<td>Gainesville, FL</td>
</tr>
<tr>
<td>Scion Image Software</td>
<td>Scion Corporation</td>
<td>&nbsp;</td>
<td>Frederick, MD</td>
</tr>
<tr>
<td>Dichloromethane</td>
<td>Sigma-Aldrich</td>
<td>270997</td>
<td>&nbsp;St. Louis, MO</td>
</tr>
<tr>
<td>Gas chromatograph HP 6890</td>
<td>Hewlett Packard</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Gas chromatograph Varian 3400 </td>
<td>Varian</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>n-octane</td>
<td>Sigma-Aldrich</td>
<td>296988</td>
<td>St. Louis, MO</td>
</tr>
<tr>
<td>Mass spectrometer MAT 8230 </td>
<td>Finnigan </td>
<td>&nbsp;</td>
<td>San Jose, CA</td>
</tr>
<tr>
<td>HP-1 GC column</td>
<td>Agilent Technologies</td>
<td>&nbsp;</td>
<td>Palo Alto, 
 CA</td>
</tr>
<tr>
<td>MDN-5S GC column</td>
<td>Supelco, Inc.</td>
<td>&nbsp;</td>
<td>Bellefonte, PA</td>
</tr>
</table>

herbivore-induced blueberry volatiles and intra-plant signaling

Herbivore-induced plant volatiles (HIPVs) are commonly emitted from plants after herbivore attack1,2. These HIPVs are mainly regulated by the defensive plant hormone jasmonic acid (JA) and its volatile derivative methyl jasmonate (MeJA)3,4,5. Over the past 3 decades researchers have documented that HIPVs can repel or attract herbivores, attract the natural enemies of herbivores, and in some cases they can induce or prime plant defenses prior to herbivore attack. In a recent paper6, I reported that feeding by gypsy moth caterpillars, exogenous MeJA application, and mechanical damage induce the emissions of volatiles from blueberry plants, albeit differently. In addition, blueberry branches respond to HIPVs emitted from neighboring branches of the same plant by increasing the levels of JA and resistance to herbivores (i.e., direct plant defenses), and by priming volatile emissions (i.e., indirect plant defenses). Similar findings have been reported recently for sagebrush7, poplar8, and lima beans9..
Here, I describe a push-pull method for collecting blueberry volatiles induced by herbivore (gypsy moth) feeding, exogenous MeJA application, and mechanical damage. The volatile collection unit consists of a 4 L volatile collection chamber, a 2-piece guillotine, an air delivery system that purifies incoming air, and a vacuum system connected to a trap filled with Super-Q adsorbent to collect volatiles5,6,10. Volatiles collected in Super-Q traps are eluted with dichloromethane and then separated and quantified using Gas Chromatography (GC). This volatile collection method was used n my study6 to investigate the volatile response of undamaged branches to exposure to volatiles from herbivore-damaged branches within blueberry plants. These methods are described here. Briefly, undamaged blueberry branches are exposed to HIPVs from neighboring branches within the same plant. Using the same techniques described above, volatiles emitted from branches after exposure to HIPVs are collected and analyzed.

Watch this Scientific Journal Video about Herbivore-induced Blueberry Volatiles and Intra-plant Signaling at JoVE.com

Herbivore-induced Blueberry Volatiles and Intra-plant Signaling

A push-pull method for collecting plant volatiles is described. The method allows for a comparison of volatiles induced by herbivore feeding, exogenous methyl jasmonate, and mechanical damage. This technique is also used to investigate the volatile response of undamaged branches to exposure to volatiles from herbivore-damaged branches within blueberry plants.

Herbivore-induced plant volatiles (HIPVs) are commonly emitted from plants after herbivore attack1,2.  These HIPVs are mainly regulated by the defensive ...

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