Boyce Thompson Institute for Plant Research, Ithaca, NY, USA.
Department of Plant Pathology and Microbiology, Iowa State University, Ames, IA, USA.
Plant Biotechnol J. 2021 Sep;19(9):1713-1724. doi: 10.1111/pbi.13585. Epub 2021 Apr 8.
Spodoptera frugiperda (fall armyworm) is a notorious pest that threatens maize production worldwide. Current control measures involve the use of chemical insecticides and transgenic maize expressing Bacillus thuringiensis (Bt) toxins. Although additional transgenes have confirmed insecticidal activity, limited research has been conducted in maize, at least partially due to the technical difficulty of maize transformation. Here, we describe implementation of a sugarcane mosaic virus (SCMV) vector for rapidly testing the efficacy of both endogenous maize genes and heterologous genes from other organisms for the control of S. frugiperda in maize. Four categories of proteins were tested using the SCMV vector: (i) maize defence signalling proteins: peptide elicitors (Pep1 and Pep3) and jasmonate acid conjugating enzymes (JAR1a and JAR1b); (ii) maize defensive proteins: the previously identified ribosome-inactivating protein (RIP2) and maize proteinase inhibitor (MPI), and two proteins with predicted but unconfirmed anti-insect activities, an antimicrobial peptide (AMP) and a lectin (JAC1); (iii) lectins from other plant species: Allium cepa agglutinin (ACA) and Galanthus nivalis agglutinin (GNA); and (iv) scorpion and spider toxins: peptides from Urodacus yaschenkoi (UyCT3 and UyCT5) and Hadronyche versuta (Hvt). In most cases, S. frugiperda larval growth was reduced by transient SCMV-mediated overexpression of genes encoding these proteins. Additionally, experiments with a subset of the SCMV-expressed genes showed effectiveness against two aphid species, Rhopalosiphum maidis (corn leaf aphid) and Myzus persicae (green peach aphid). Together, these results demonstrate that SCMV vectors are a rapid screening method for testing the efficacy and insecticidal activity of candidate genes in maize.
草地贪夜蛾(秋黏虫)是一种臭名昭著的害虫,它威胁着全世界的玉米生产。目前的控制措施包括使用化学杀虫剂和表达苏云金芽孢杆菌(Bt)毒素的转基因玉米。尽管额外的转基因已经证实具有杀虫活性,但在玉米中的研究有限,这至少部分是由于玉米转化的技术难度。在这里,我们描述了利用甘蔗花叶病毒(SCMV)载体快速测试内源玉米基因和来自其他生物体的异源基因对玉米中 S. frugiperda 的控制效果的方法。使用 SCMV 载体测试了四类蛋白质:(i)玉米防御信号蛋白:肽激发子(Pep1 和 Pep3)和茉莉酸酸结合酶(JAR1a 和 JAR1b);(ii)玉米防御蛋白:先前鉴定的核糖体失活蛋白(RIP2)和玉米蛋白酶抑制剂(MPI),以及两种具有预测但未经证实的抗虫活性的蛋白质,一种抗菌肽(AMP)和一种凝集素(JAC1);(iii)来自其他植物物种的凝集素:葱凝集素(ACA)和雪花莲凝集素(GNA);和(iv)蝎子和蜘蛛毒素:来自 Urodacus yaschenkoi 的肽(UyCT3 和 UyCT5)和 Hadronyche versuta(Hvt)。在大多数情况下,通过瞬时 SCMV 介导的这些基因编码蛋白的过表达,草地贪夜蛾幼虫的生长受到了抑制。此外,对 SCMV 表达的部分基因的实验表明,它们对两种蚜虫,玉米叶蝉(玉米叶蝉)和桃蚜(桃蚜)有效。总之,这些结果表明 SCMV 载体是一种快速筛选方法,可用于测试候选基因在玉米中的功效和杀虫活性。