trichogramma dendrolimi artificial host interaction and venom extraction

Kunstmatige parasitering van de gastheer door Trichogramma dendrolimi en gifextractie

Parasitoid wasps are parasitic hymenopteran insects that are important resources for biological control1. There is a wide variety of parasitoid wasps, with over 600,000 estimated species2. The diversity of parasitoid wasps far exceeds that of other venomous arthropods, such as snakes, cone snails, spiders, scorpions, and bees. Venom is an important parasitic factor in parasitoid wasps. For successful parasitism, venom is injected into the host, modulating the host&#39;s behavior, immunity, development, and metabolism3. Moreover, the venom of parasitoid wasps displays remarkable diversity in its molecular structures, targets, and functions, reflecting complex coevolution with their hosts. Thus, parasitoid venom is a valuable and underappreciated resource of active molecules for insecticidal or medical purposes4. Unlike the venom of snakes, cone snails, spiders, scorpions, and bees, parasitoid wasp venom cannot be collected by direct stimulation or electrical stimulation5. The current method of extraction of parasitoid wasp venom is to dissect the venom reservoir. However, parasitoid wasps are often small, and dissection of parasitoid wasps requires high technical skills. Therefore, if we can find a way to collect the venom of parasitoid wasps efficiently and conveniently, it will be of great help to research the venom of parasitoid wasps.
Trichogramma (Hymenoptera: Trichogrammatidae) is a genus of tiny (~0.5 mm long) parasitoid wasps6. These wasps are among the most widely used biocontrol agents, particularly targeting eggs of various lepidopteran pests in both agriculture and forests. For example, T. dendrolimi, one of the most widely used Trichogramma species in China, has been extensively applied to control a variety of agricultural and forestry pests, such as Dendrolimus superans, Ostrinia furnacalis, and Chilo suppressalis. Previous studies showed that Trichogramma wasps could inject their eggs into artificial hosts7. Artificial hosts can be created using materials such as wax8, agar9, Parafilm10, and plastic film11. The solution in artificial hosts that induces sufficient oviposition for Trichogramma can be simple, such as amino acids or inorganic salts12. Based on the characteristic that T. dendrolimi can parasitize artificial hosts, this study provides a new method for extracting venom from parasitoid wasps using artificial hosts. This approach aims to address the shortcomings of low yield, low purity, and susceptibility to contamination in current extraction techniques. By using this method, a large amount of high-purity venom from T. dendrolimi can be extracted, which meets the needs of scientific research and screening of bioactive molecules for insecticidal or medical purposes.

Auteur in de schijnwerpers: Efficiënte gifextractiemethode van Trichogramma sluipwespen 

Gif extraheren uit de sluipwesp Trichogramma dendrolimi met behulp van een kunstmatige gastheer

The lack of efficient venom extraction methods often limits the research on the venom of parasitoid wasps, especially in tiny parasitoid wasps. This study provides an efficient method to extract the venom of Trichogramma with follow up post study of Trichogramma venom, such as the protein compensation and the venom function analysis. Previously, no methods of exchanging of parasitoid wasp venom requires the distinguishing of venom reserve.However, parasitoid wasps are tiny. Not only are the technical requirements of distinguishing venom was high, but continuation of other tissues during dissection is also common. Our normal method uses artificial host, thus avoiding such problems.In the future, we will focus on the agriculture and the medical applications of Trichogramma venom. We aim to utilize specific insecticidal venom genes for pest control. Meanwhile, we plan to investigate the untapped potential of Trichogramma venom in methodology, particularly its prospective utility in enhancing immunotherapy techniques, and in the discovery of novel antibiotic molecules.

Watch this Scientific Journal Video about Trichogramma dendrolimi  Artificial Host Interaction and Venom Extraction at JoVE.com

Trichogramma dendrolimi  Artificial Host Interaction and Venom Extraction  

Dendrolimi van Trichogramma Kunstmatige gastheerinteractie en gifextractie

Neem om te beginnen een polyethyleen plastic film van 16 centimeter lang, 12 centimeter breed en 20 micrometer dik. Gebruik een glasslijpgereedschap om 30 halfronde uitsteeksels uit te drukken in een standaard PCR-lay-out met 96 putjes. Stel beide zijden van de geperste polyethyleen plastic folie gedurende een uur bloot aan UV-licht om deze te desinfecteren.Voeg een kleine hoeveelheid 10% polyvinylalcohol toe aan het halfronde oppervlak. Plaats vervolgens ongeveer 3.000 verdoofde vrouwelijke trichogramma dendrolimi wespen in een verzameldoos. Plaats de bolle kant van de folie-eierkaart in de richting van de opvangbak en zet de randen vast met een elastiekje.Voeg nu vier microliter aminozuuroplossing toe aan elk uitsteeksel. Bedek het vervolgens met een ander stuk polyethyleen plastic folie. Gebruik een elastiekje om de opvangbak goed af te dekken met twee vellen plastic.Laat de wespen vier tot acht uur vrij rondsluipen. Geef 10% sucrosewater door nat katoen tijdens parasitisatie. Verkrijg vervolgens de geparasiteerde aminozuuroplossing uit het binnenste uitsteeksel van de kunstmatige eierkaart en breng deze over naar de dop van buizen van 1,45 milliliter.Bedek de buisdop met een nylon net van 10 micrometer met een diameter van 25 millimeter. Bevestig vervolgens het nylon net lichtjes in de buis. Centrifugeer de buis rechtsboven gedurende 10 seconden bij 1.360 g.Vang de gefilterde oplossing met het gif op. Bewaar het gif bij min 80 graden Celsius voor verdere analyses. BCA-analyse van de gifmonsters toonde aan dat de concentratie gifeiwit varieerde van 0,35 tot 0,46 microgram per microliter.De negatieve controle van de aminozuuroplossing had slechts een eiwitconcentratie van 0,03 tot 0,05 microgram per microliter. SDS-PAGE-analyse van het gif toonde een eiwitbereik van minder dan 10 kilodalton tot meer dan 130 kilodalton.

Research

JoVE Journal

Biochemistry

Trichogramma dendrolimi Artificial Host Interaction and Venom Extraction

Extracting Venom from the Parasitoid Wasp Trichogramma dendrolimi Using an Artificial Host

Parasitoid wasps are a diverse group of hymenopteran insects that serve as invaluable resources for pest biocontrol. To ensure successful parasitism, ...