Hyperactive <em>piggyBac</em> Transposase-mediated Germline Transformation in the Fall Armyworm, <em>Spodoptera frugiperda</em>

Xien Chen; Subba Reddy Palli

doi:10.3791/62714

Hiperaktif piggyBac Transposase aracılı Germline Dönüşümü Sonbahar Ordu Kurdu, Spodo...

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Hyperactive piggyBac Transposase-mediated Germline Transformation in the Fall Armyworm, Spodoptera frugiperda

Hiperaktif piggyBac Transposase aracılı Germline Dönüşümü Sonbahar Ordu Kurdu, Spodoptera frugiperda

Full Text

5,153 Views

05:20 min

September 23, 2021

DOI: 10.3791/62714-v

Xien Chen¹, Subba Reddy Palli¹

¹Department of Entomology, College of Agriculture, Food and Environment,University of Kentucky

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study demonstrates a protocol for successful germline transformation in the fall armyworm, Spodoptera frugiperda, utilizing a piggyBac-based transformation system. The method promises improved transformation efficiency in lepidopteran pests and enhances functional genomics research in these organisms.

Key Study Components

Research Area

Genetic engineering in pest insects
Transformational biology techniques
Functional genomics applications

Background

Fall armyworm is a significant agricultural pest.
The piggyBac transposon system is known for its efficiency in gene delivery.
Improving germline transformation is crucial for pest control strategies.

Methods Used

Microinjection of eggs with piggyBac plasmid and mRNA
Spodoptera frugiperda as the biological model
Fluorescence microscopy for screening transgenic larvae

Main Results

Successful expression of EGFP in transgenic larvae showed functionality of the piggyBac system.
EGFP expression was verified through genomic DNA amplification, distinguishing transgenic from wild-type larvae.
The transformation technique proved efficient for producing genetically modified pest insects.

Conclusions

The study establishes a robust method for germline transformation in Spodoptera frugiperda.
Findings contribute to advancing genetic control measures for agricultural pests.

Frequently Asked Questions

What is the significance of using the piggyBac system?

The piggyBac system is known for its high efficiency and versatility in gene delivery, making it ideal for genetic modifications in insects.

How were the transgenic larvae identified?

Transgenic larvae were identified through fluorescence microscopy, detecting the EGFP protein expressed from the injected construct.

What implications does this research have for pest control?

This research provides new tools for the genetic control of pest insects, potentially reducing their impact on agriculture.

Can this method be applied to other insect species?

Yes, the methods developed can be adapted for use in other lepidopteran insects, aiding in broader pest management efforts.

What are the optimal conditions for the microinjection?

Injecting the eggs under controlled temperature (27 ± 1 °C) and humidity (60 ± 10%) is crucial for successful development.

What role does the hyperactive piggyBac transposase play?

The hyperactive transposase enhances the integration of the piggyBac elements into the host genome, improving transformation efficiency.

Sonbahar ordu kurdunda başarılı germline dönüşümü, Spodoptera frugiperda, hiperaktif piggyBac transposase mRNA kullanılarak elde edildi.

Bu protokol, piggyBac tabanlı dönüşüm sistemi kullanılarak transgenik Sonbahar Ordu kurdu üretimini göstermektedir. Bu yöntemlerin sadece Sonbahar Ordu Kurdu'nda değil, aynı zamanda lepidopteran böceklerde de dönüşümün iyileştirilmesine fayda sağlayacağını umuyoruz. Burada açıklanan yöntem, lepidopteran haşere böceklerinin germline dönüşümini verimli hale getirir.

Prosedürü gösteren dr. Xien Chen olacak, laboratuvarımda doktora sonrası bir arkadaş. Bir kutuya 12 erkek pupa ve 12 dişi pupa yerleştirerek başlayın. Yetişkinleri beslemek için kutuya% 10 sakkaroz çözeltisi yerleştirin.

View the full transcript and gain access to thousands of scientific videos