trichogramma dendrolimi artificial host interaction and venom extraction

Kunstig værtsparasitering ved Trichogramma dendrolimi og giftekstraktion

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.

Forfatter Spotlight: Effektiv giftekstraktionsmetode fra Trichogramma parasitoid hvepse 

Ekstraktion af gift fra parasitoid hveps Trichogramma dendrolimi ved hjælp af en kunstig vært

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  

Trichogramma dendrolimi Kunstig værtsinteraktion og giftekstraktion

Til at begynde med skal du tage en polyethylenplastfilm 16 centimeter lang, 12 centimeter bred og 20 mikrometer tyk. Brug et glasslibeværktøj til at presse 30 halvcirkelformede fremspring ud i et standard 96-brønds PCR-layout. Udsæt begge sider af den pressede polyethylenplastfilm for UV-lys i en time for at desinficere den.Tilsæt en lille mængde 10% polyvinylalkohol til den halvcirkelformede overflade. Derefter placeres ca. 3.000 bedøvede kvindelige trichogramma dendrolimi hveps i en opsamlingsboks. Placer den konvekse side af filmægkortet mod opsamlingsboksen og fastgør kanterne med et gummibånd.Tilsæt nu fire mikroliter aminosyreopløsning til hvert fremspring. Dæk det derefter med et andet stykke polyethylenplastfilm. Brug et gummibånd til tæt at dække opsamlingsboksen med to ark plast.Lad hvepsene parasitere frit i fire til otte timer. Giv 10% saccharosevand gennem våd bomuld under parasitering. Derefter opnås den parasiterede aminosyreopløsning fra det indre fremspring af det kunstige ægkort og overføres til hætten på 1, 45 ml rør.Dæk rørhætten med et 10 mikrometer nylonnet med en diameter på 25 millimeter. Fastgør derefter nylonnettet let i røret. Centrifuger det øverste højre rør ved 1.360 g i 10 sekunder.Opsaml den filtrerede opløsning indeholdende giften. Opbevar giften ved minus 80 grader Celsius til yderligere analyser. BCA-analyse af giftprøverne viste, at koncentrationen af giftprotein varierede fra 0,35 til 0,46 mikrogram pr. mikroliter.Den negative kontrol af aminosyreopløsningen havde kun en proteinkoncentration på 0,03 til 0,05 mikrogram pr. Mikroliter. SDS-PAGE-analyse af giften viste et proteinområde, der spænder fra under 10 kilodalton til over 130 kilodalton i størrelse.

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, ...