Kumar Sandeep, Dhillon Mukesh K
Indian J Exp Biol. 2015 Mar;53(3):170-6.
In order to better understand the biochemical interactions and to identify new biomarkers for plant resistance against insects, we proposed a suitable lipophilic profiling method for insects and their host plants. The critical components of GC-MS based analysis are: sample amount, extraction, derivatization, temperature gradient, run time, and identification of peaks. For lipophilic metabolite profiling of maize and sorghum, and their insect pest, spotted stem borer larvae, we recommend 100 mg sample weight for seeds and insect samples (whole insect body), and 200 mg for seedlings. Maize and sorghum seeds required less time for fat extraction in comparison to their seedlings and the pest fed on these seedlings. GC-MS was standardized for better separation and intensity of peaks using different temperature gradients in the range of 180-300 C. A total of 48 lipophilic compounds encompassing various classes based on their functional groups such as fatty acids, fatty alcohols, hydrocarbons, sterols and terpenoids, vitamin derivative, etc. were separated in the seedlings (30), seeds (14), and the pest (26) in the retention time range of 3.22 to 29.41 min. This method could be useful to study nutritional aspects of different field crops in relation to various stresses apart from the analysis of lipophilic compounds for better understanding of insect-plant interactions.
为了更好地理解生化相互作用并鉴定植物抗虫的新生物标志物,我们提出了一种适用于昆虫及其寄主植物的亲脂性分析方法。基于气相色谱 - 质谱联用(GC-MS)分析的关键组成部分包括:样品量、提取、衍生化、温度梯度、运行时间以及峰的鉴定。对于玉米和高粱及其害虫——斑螟幼虫的亲脂性代谢物分析,我们建议种子和昆虫样品(整个虫体)的样品重量为100毫克,幼苗为200毫克。与它们的幼苗以及取食这些幼苗的害虫相比,玉米和高粱种子提取脂肪所需时间更少。通过在180 - 300℃范围内使用不同温度梯度,对GC-MS进行了标准化,以实现更好的峰分离和峰强度。在3.22至29.41分钟的保留时间范围内,共分离出48种亲脂性化合物,这些化合物根据其官能团分为不同类别,如脂肪酸、脂肪醇、烃类、甾醇和萜类、维生素衍生物等,其中在幼苗中分离出30种,种子中分离出14种,害虫中分离出26种。除了分析亲脂性化合物以更好地理解昆虫与植物的相互作用外,该方法对于研究不同大田作物在各种胁迫下的营养方面也可能有用。