van Raalte Benjamin, Watrous Kristal, Lujan Miguel, Le Ricky, Sun Penglin, Ellis Benjamin, Mauck Kerry E
Department of Entomology, University of California, Riverside, 165 Entomology Bldg., Citrus Drive, Riverside, CA, 92521, USA.
Department of Ecology and Evolutionary Biology, University of California, Irvine, 321 Steinhaus Hall, Irvine, CA, 92697, USA.
Plant Methods. 2024 May 23;20(1):75. doi: 10.1186/s13007-024-01209-z.
The sweet potato whitefly (Bemisia tabaci) is a globally important insect pest that damages crops through direct feeding and by transmitting viruses. Current B. tabaci management revolves around the use of insecticides, which are economically and environmentally costly. Host plant resistance is a sustainable option to reduce the impact of whiteflies, but progress in deploying resistance in crops has been slow. A major obstacle is the high cost and low throughput of screening plants for B. tabaci resistance. Oviposition rate is a popular metric for host plant resistance to B. tabaci because it does not require tracking insect development through the entire life cycle, but accurate quantification is still limited by difficulties in observing B. tabaci eggs, which are microscopic and translucent. The goal of our study was to improve quantification of B. tabaci eggs on several important crop species: cassava, cowpea, melon, sweet potato and tomato.
We tested a selective staining process originally developed for leafhopper eggs: submerging the leaves in McBryde's stain (acetic acid, ethanol, 0.2% aqueous acid Fuchsin, water; 20:19:2:1) for three days, followed by clearing under heat and pressure for 15 min in clearing solution (LGW; lactic acid, glycerol, water; 17:20:23). With a less experienced individual counting the eggs, B. tabaci egg counts increased after staining across all five crops. With a more experienced counter, egg counts increased after staining on melons, tomatoes, and cowpeas. For all five crops, there was significantly greater agreement on egg counts across the two counting individuals after the staining process. The staining method worked particularly well on melon, where egg counts universally increased after staining for both counting individuals.
Selective staining aids visualization of B. tabaci eggs across multiple crop plants, particularly species where leaf morphological features obscure eggs, such as melons and tomatoes. This method is broadly applicable to research questions requiring accurate quantification of B. tabaci eggs, including phenotyping for B. tabaci resistance.
烟粉虱是一种在全球范围内具有重要影响的害虫，通过直接取食和传播病毒来损害作物。目前对烟粉虱的防治主要围绕杀虫剂的使用，这在经济和环境方面成本高昂。寄主植物抗性是减少粉虱影响的一种可持续选择，但在作物中部署抗性的进展一直缓慢。一个主要障碍是筛选对烟粉虱具有抗性的植物成本高且通量低。产卵率是衡量寄主植物对烟粉虱抗性的常用指标，因为它不需要追踪昆虫整个生命周期的发育情况，但由于烟粉虱的卵微小且半透明，观察困难，准确量化仍然受到限制。我们研究的目的是改进对几种重要作物（木薯、豇豆、甜瓜、甘薯和番茄）上烟粉虱卵的量化。
我们测试了一种最初为叶蝉卵开发的选择性染色方法：将叶片浸入麦克布赖德染色剂（乙酸、乙醇、0.2% 酸性品红水溶液、水；20:19:2:1）中三天，然后在LGW澄清液（乳酸、甘油、水；17:20:23）中加热加压澄清15分钟。由经验较少的人员计数时，所有五种作物上烟粉虱卵的计数在染色后均有所增加。由经验更丰富的计数人员计数时，甜瓜、番茄和豇豆上的卵计数在染色后增加。对于所有五种作物，染色过程后两位计数人员在卵计数上的一致性显著提高。染色方法在甜瓜上效果特别好，两位计数人员计数时，染色后卵的计数普遍增加。
选择性染色有助于在多种作物上观察烟粉虱卵，特别是在叶片形态特征会遮挡卵的物种上，如甜瓜和番茄。该方法广泛适用于需要准确量化烟粉虱卵的研究问题，包括烟粉虱抗性的表型分析。
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