trichogramma dendrolimi artificial host interaction and venom extraction

Parasitation artificielle de l’hôte par Trichogramma dendrolimi et extraction de venin

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.

Pleins feux sur l’auteur : Méthode efficace d’extraction du venin des guêpes parasitoïdes trichogrammes 

Extraction du venin de la guêpe parasitoïde Trichogramma dendrolimi à l’aide d’un hôte artificiel

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 Interaction avec l’hôte artificiel et extraction du venin

Pour commencer, prenez un film plastique polyéthylène de 16 centimètres de long, 12 centimètres de large et 20 micromètres d’épaisseur. Utilisez un outil de meulage du verre pour presser 30 protubérances semi-circulaires dans une disposition PCR standard à 96 puits. Exposez les deux côtés du film plastique de polyéthylène pressé à la lumière UV pendant une heure pour le désinfecter.Ajoutez une petite quantité d’alcool polyvinylique à 10 % sur la surface semi-circulaire. Ensuite, placez environ 3 000 guêpes trichogrammes femelles anesthésiées dendrolimi dans une boîte de collecte. Placez le côté convexe de la carte d’œuf en film vers la boîte de collecte et fixez les bords avec un élastique.Ajoutez maintenant quatre microlitres de solution d’acide aminé à chaque protubérance. Ensuite, couvrez-le d’un autre morceau de film plastique en polyéthylène. Utilisez un élastique pour recouvrir hermétiquement la boîte de collecte de deux feuilles de plastique.Laissez les guêpes parasiter librement pendant quatre à huit heures. Fournir 10% d’eau de saccharose à travers le coton humide pendant le parasitage. Ensuite, obtenez la solution d’acides aminés parasité à partir de la saillie interne de la carte d’œuf artificiel et transférez-la dans le bouchon de tubes de 1,45 millilitre.Recouvrez le bouchon du tube d’un filet en nylon de 10 micromètres, d’un diamètre de 25 millimètres. Fixez ensuite légèrement le filet en nylon dans le tube. Centrifuger le tube supérieur droit à 1, 360 g pendant 10 secondes.Recueillir la solution filtrée contenant le venin. Conservez le venin à moins 80 degrés Celsius pour d’autres analyses. L’analyse BCA des échantillons de venin a montré que la concentration de protéine de venin variait de 0,35 à 0,46 microgramme par microlitre.Le témoin négatif de la solution d’acides aminés n’avait qu’une concentration en protéines de 0,03 à 0,05 microgramme par microlitre. L’analyse SDS-PAGE du venin a montré une gamme de protéines allant de moins de 10 kilodaltons à plus de 130 kilodaltons.

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, ...