trichogramma dendrolimi artificial host interaction and venom extraction

Artificiell värdparasitisering av Trichogramma dendrolimi och giftextraktion

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.

Författare i fokus: Effektiv metod för extraktion av gift från Trichogramma parasitoida getingar 

Extrahera gift från parasitoidgetingen Trichogramma dendrolimi med hjälp av en konstgjord värd

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 Artificiell värdinteraktion och giftextraktion

Börja med att ta en plastfilm av polyeten som är 16 centimeter lång, 12 centimeter bred och 20 mikrometer tjock. Använd ett glasslipverktyg för att pressa ut 30 halvcirkelformade utsprång i en standard 96-håls PCR-layout. Utsätt båda sidor av den pressade polyetenplastfilmen för UV-ljus i en timme för att desinficera den.Tillsätt en liten mängd 10 % polyvinylalkohol till den halvcirkelformade ytan. Lägg sedan cirka 3 000 sövda honsteklar av dendrolimi i en uppsamlingsbössa. Placera den konvexa sidan av filmäggkortet mot uppsamlingslådan och fäst kanterna med ett gummiband.Tillsätt nu fyra mikroliter aminosyralösning till varje utsprång. Täck den sedan med en annan bit polyetenplastfilm. Använd ett gummiband för att täcka uppsamlingslådan ordentligt med två plastark.Låt getingarna parasitera fritt i fyra till åtta timmar. Ge 10 % sackarosvatten genom våt bomull under parasitering. Erhåll sedan den parasitiserade aminosyralösningen från det inre utsprånget av det konstgjorda äggkortet och överför det till locket på 1,45 milliliters rör.Täck rörlocket med ett 10 mikrometer nylonnät med en diameter på 25 millimeter. Fäst sedan nylonnätet lätt i röret. Centrifugera det övre högra röret vid 1,360 g i 10 sekunder.Samla upp den filtrerade lösningen som innehåller giftet. Förvara giftet vid minus 80 grader Celsius för vidare analyser. BCA-analys av giftproverna visade att koncentrationen av giftprotein varierade från 0,35 till 0,46 mikrogram per mikroliter.Den negativa kontrollen av aminosyralösning hade endast en proteinkoncentration på 0,03 till 0,05 mikrogram per mikroliter. SDS-PAGE-analys av giftet visade ett proteinintervall som sträckte sig från under 10 kilodalton till över 130 kilodalton i storlek.

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, ...