Wageningen UR, Plant Breeding, PO, Box 386, 6700, AJ, Wageningen, the Netherlands.
Plant Methods. 2012 Aug 17;8(1):33. doi: 10.1186/1746-4811-8-33.
Phloem-feeding insects are among the most devastating pests worldwide. They not only cause damage by feeding from the phloem, thereby depleting the plant from photo-assimilates, but also by vectoring viruses. Until now, the main way to prevent such problems is the frequent use of insecticides. Applying resistant varieties would be a more environmental friendly and sustainable solution. For this, resistant sources need to be identified first. Up to now there were no methods suitable for high throughput phenotyping of plant germplasm to identify sources of resistance towards phloem-feeding insects.
In this paper we present a high throughput screening system to identify plants with an increased resistance against aphids. Its versatility is demonstrated using an Arabidopsis thaliana activation tag mutant line collection. This system consists of the green peach aphid Myzus persicae (Sulzer) and the circulative virus Turnip yellows virus (TuYV). In an initial screening, with one plant representing one mutant line, 13 virus-free mutant lines were identified by ELISA. Using seeds produced from these lines, the putative candidates were re-evaluated and characterized, resulting in nine lines with increased resistance towards the aphid.
This M. persicae-TuYV screening system is an efficient, reliable and quick procedure to identify among thousands of mutated lines those resistant to aphids. In our study, nine mutant lines with increased resistance against the aphid were selected among 5160 mutant lines in just 5 months by one person. The system can be extended to other phloem-feeding insects and circulative viruses to identify insect resistant sources from several collections, including for example genebanks and artificially prepared mutant collections.
韧皮部取食昆虫是世界上最具破坏性的害虫之一。它们不仅通过从韧皮部取食来造成损害，从而耗尽植物的光合作用产物，还通过传播病毒。到目前为止，防止此类问题的主要方法是频繁使用杀虫剂。应用抗性品种将是一种更环保和可持续的解决方案。为此，首先需要确定抗性来源。到目前为止，还没有适合高通量表型分析植物种质资源以鉴定抗韧皮部取食昆虫的方法。
本文介绍了一种高通量筛选系统，用于鉴定对蚜虫具有更高抗性的植物。使用拟南芥激活标签突变体系集对此系统的多功能性进行了验证。该系统由绿桃蚜 Myzus persicae（Sulzer）和循环病毒芜菁黄花叶病毒（TuYV）组成。在初步筛选中，用一株植物代表一条突变体系，通过 ELISA 鉴定了 13 条无病毒突变体系。使用这些系产生的种子，重新评估和表征了候选物，结果发现有九条系对蚜虫具有更高的抗性。
这种 M. persicae-TuYV 筛选系统是一种高效、可靠和快速的程序，可以在数千条突变体系中鉴定出对蚜虫具有抗性的系。在我们的研究中，在短短 5 个月内，通过一个人从 5160 条突变体系中选择了九条对蚜虫具有更高抗性的突变体系。该系统可以扩展到其他韧皮部取食昆虫和循环病毒，以从包括基因库和人工制备的突变体库在内的多个收集物中鉴定出抗虫来源。
