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Rapid screening of pest resistance genes in maize using a sugarcane mosaic virus vector
bioRxiv - Plant Biology Pub Date : 2021-01-13 , DOI: 10.1101/2021.01.13.425472
Seung Ho Chung , Mahdiyeh Bigham , Ryan R. Lappe , Barry Chan , Ugrappa Nagalakshmi , Steven A. Whitham , Savithramma P. Dinesh-Kumar , Georg Jander

Spodoptera frugiperda (fall armyworm) is a notorious pest that threatens maize production world-wide. Current control measures involve the use of chemical insecticides and transgenic maize expressing Bacillus thuringiensis (Bt) toxins. Although several additional transgenes have confirmed insecticidal activity in other plants, 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 transgenes for the control of S. frugiperda in maize. Four categories of proteins were tested using the SCMV vector: (i) maize defense signaling 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) spider and scorpion toxins: peptides from Urodacus yaschenkoi (UyCT3 and UyCT5) and Hadronyche versuta (Hvt). In most cases, S. frugiperda larval growth on maize was reduced by transient SCMV-mediated overexpression of genes encoding these proteins. Additionally, experiments with some 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 can be exploited as a rapid screening method for testing the efficacy and insecticidal activity of candidate genes in maize.

中文翻译:

使用甘蔗花叶病毒载体快速筛选玉米中的害虫抗性基因

夜蛾(Spodoptera frugiperda)是一种臭名昭著的害虫,它威胁着全世界的玉米生产。当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。尽管几种其他的转基因已证实在其他植物中具有杀虫活性,但对玉米的研究有限,至少部分是由于玉米转化的技术难度。在这里,我们描述了一种甘蔗花叶病毒(SCMV)载体的实现,用于快速测试转基因对玉米中节俭链球菌的控制效果。使用SCMV载体测试了四类蛋白质:(i)玉米防御信号蛋白:肽诱导子(Pep1和Pep3)和茉莉酸结合酶(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证实的抗昆虫活性的蛋白,即抗菌肽(AMP)和凝集素(JAC1);(iii)来自其他植物物种的凝集素:葱属洋葱凝集素(ACA)和雪花莲凝集素(GNA);(iv)蜘蛛和蝎子毒素:来自尤达克(Urodcus yaschenkoi)(UyCT3和UyCT5)和Hadronyche versuta(Hvt)的肽。在大多数情况下,通过瞬时SCMV介导的编码这些蛋白质的基因的过表达,减少了Srug frugiperda幼虫在玉米上的生长。另外,用表达SCMV的某些基因进行的实验表明,它对两种蚜虫(玉米叶蚜)和桃蚜(桃蚜)有有效的作用。一起,
更新日期:2021-01-14
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