Applications of Molecular Diagnostics for Quality Control in Rearing of Spodoptera frugiperda (Lepidoptera: Noctuidae) Larvae for Experimental Use

IF 1.1 4区农林科学 Q3 ENTOMOLOGY

Florida Entomologist Pub Date : 2023-06-22 DOI:10.1653/024.106.0212

Francisco Javier Flores Gallardo, José Luis Hernández Flores, M. A. Ramos López, Jackeline Lizzeta Arvizu Gómez, V. P. Moreno, José Alberto Rodríguez Morales, Erika Álvarez Hidalgo, Jorge Nuñez Ramírez, George H. Jones, Juan Campos Guillén

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引用次数: 0

Abstract

Summary Illumina MiSeq sequencing of 16S rRNA genes was applied in a molecular diagnostic analysis of healthy and dead larvae obtained from a Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) colony. The results indicated the presence of Enterococcus (Lactobacillales: Enterococcaceae) with an average abundance of 89.2% and 94.8% in healthy and dead larvae, respectively. Pseudomonas (Pseudomonadales: Pseudomonadaceae) and Acetobacter (Rhodospirillales: Acetobacteraceae) were present in lower abundance. Additional metagenomic results revealed the presence in dead larvae of viral sequences related to Alphabaculovirus in high relative abundance compared with healthy larvae. Additionally, viral sequences of tomato leaf curl Laos virus, Heliothis zea (Boddie) (Lepidoptera: Noctuidae) nudivirus, Clostera anachoreta (Denis & Schiffermüller) (Lepidoptera: Notodontidae) granulovirus, and Tipula oleracea L. (Diptera: Tipulidae) nudivirus were detected. Our results show that molecular detection of pathogenic agents is a useful tool during insect rearing as a quality control to avoid the spread or acquisition of potential pathogens. Sumario Nuestros resultados demuestran que a través de un diagnóstico molecular en Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), usando secuenciación del 16S rDNA por Illumina MiSeq, en larvas saludables y muertas mostraron una abundancia relativa alta de Enterococcus (Lactobacillales: Enterococcaceae) con un promedio de 89.2% and 94.8% respectivamente. Mientras que Pseudomonas (Pseudomonadales: Pseudomonadaceae) y Acetobacter (Rhodospirillales: Acetobacteraceae) presentaron menor abundancia relativa. Adicionalmente, resultados de metagenomica mostraron en larvas muertas secuencias de virus relacionados con Alphabaculovirus con una abundancia relativa muy alta comparada con las larvas saludables. Se detectaron secuencias virales relacionadas al Encrespamiento amarillo de la hoja del tomate, Heliothis zea (Boddie) (Lepidoptera: Noctuidae) nudivirus, Clostera anachoreta (Denis & Schiffermüller) (Lepidoptera: Notodontidae) granulovirus and Tipula oleracea L. (Diptera: Tipulidae) nudivirus. Nuestros resultados demuestran que una detección molecular de agentes patogénicos es primordial durante la crianza de insectos como un control de calidad para evitar su dispersión o adquisición.

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分子诊断技术在草地贪夜蛾（鳞翅目：夜蛾科）幼虫饲养中的应用

摘要Illumina Miseq对16S rRNA基因进行了总结测序，用于对从斜纹夜蛾（鳞翅目：夜蛾科）群体中获得的健康和死亡幼虫进行分子诊断分析。结果表明，健康和死亡幼虫中存在肠球菌（乳杆菌：肠球菌科），平均丰度分别为89.2%和94.8%。假单胞菌（假单胞菌：假单胞菌科）和醋杆菌（红孢子菌：醋杆菌科）的丰度较低。额外的宏基因组结果显示，与健康幼虫相比，死亡幼虫中与α-球病毒相关的病毒序列相对丰富。此外，还检测到番茄叶卷曲老挝病毒、Heliothis zea（boddie）（鳞翅目：夜蛾科）裸病毒、anachoreta clostera（denis&schiffermüller）（鳞翅目：Notodontide）颗粒病毒和Tipula oleracea L.（双翅目：夜蛾科）裸病毒的病毒序列。我们的结果表明，在昆虫繁殖过程中，对病原体的分子检测是一种有用的工具，可以作为质量控制，以避免潜在病原体的传播或获取。总结我们的结果表明，通过对葡萄夜蛾（Smith）（鳞翅目：夜蛾科）的分子诊断，利用Illumina Miseq对16S rDNA的测序，在健康和死亡的幼虫中，肠球菌（乳杆菌：肠球菌科）的相对丰度较高，平均分别为89.2%和94.8%。而假单胞菌（假单胞菌：假单胞菌科）和醋杆菌（红孢子菌：醋杆菌科）的相对丰度较低。此外，宏基因组结果显示，与健康幼虫相比，死亡幼虫中与α病毒相关的病毒序列相对丰度非常高。检测到与番茄叶黄色卷起有关的病毒序列，即裸病毒、裸病毒（鳞翅目：夜蛾科）、anachoreta梭菌（丹尼斯和希弗穆勒）（鳞翅目：Notodontide）颗粒病毒和Oleracea L.（双翅目：裸病毒科）裸病毒。我们的结果表明，在昆虫繁殖过程中，对病原体的分子检测至关重要，作为质量控制，以防止其传播或获得。

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来源期刊

Florida Entomologist 生物-昆虫学

CiteScore

2.10

自引率

7.10%

发文量

审稿时长

3 months

期刊介绍： Florida Entomologist is the official journal of the Florida Entomological Society. Volumes 1-3 were published under the name The Florida Buggist. The Florida Entomological Society still produces the traditionally printed version of Florida Entomologist, but you can also view, search, or print any article published since June 1917 by accessing online files. Web access is made possible by the Society’s electronic publication project begun in 1993