Muniroh M S, Sariah M, Zainal Abidin M A, Lima N, Paterson R R M
Department of Plant Protection, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
Department of Plant Protection, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
J Microbiol Methods. 2014 May;100:143-7. doi: 10.1016/j.mimet.2014.03.005. Epub 2014 Mar 26.
Detection of basal stem rot (BSR) by Ganoderma of oil palms was based on foliar symptoms and production of basidiomata. Enzyme-Linked Immunosorbent Assays-Polyclonal Antibody (ELISA-PAB) and PCR have been proposed as early detection methods for the disease. These techniques are complex, time consuming and have accuracy limitations. An ergosterol method was developed which correlated well with the degree of infection in oil palms, including samples growing in plantations. However, the method was capable of being optimised. This current study was designed to develop a simpler, more rapid and efficient ergosterol method with utility in the field that involved the use of microwave extraction. The optimised procedure involved extracting a small amount of Ganoderma, or Ganoderma-infected oil palm suspended in low volumes of solvent followed by irradiation in a conventional microwave oven at 70°C and medium high power for 30s, resulting in simultaneous extraction and saponification. Ergosterol was detected by thin layer chromatography (TLC) and quantified using high performance liquid chromatography with diode array detection. The TLC method was novel and provided a simple, inexpensive method with utility in the field. The new method was particularly effective at extracting high yields of ergosterol from infected oil palm and enables rapid analysis of field samples on site, allowing infected oil palms to be treated or culled very rapidly. Some limitations of the method are discussed herein. The procedures lend themselves to controlling the disease more effectively and allowing more effective use of land currently employed to grow oil palms, thereby reducing pressure to develop new plantations.
通过油棕灵芝检测油棕基部茎腐病（BSR）是基于叶片症状和担子果的产生。酶联免疫吸附测定-多克隆抗体（ELISA-PAB）和聚合酶链反应（PCR）已被提议作为该病的早期检测方法。这些技术复杂、耗时且存在准确性限制。开发了一种麦角固醇方法，该方法与油棕的感染程度密切相关，包括种植园中的样本。然而，该方法能够进一步优化。本研究旨在开发一种更简单、快速且高效的麦角固醇方法，该方法在野外实用，涉及使用微波萃取。优化后的步骤包括提取少量灵芝或感染灵芝的油棕，将其悬浮在少量溶剂中，然后在传统微波炉中于70°C和中高功率下辐照30秒，从而实现同时萃取和皂化。通过薄层色谱法（TLC）检测麦角固醇，并使用带二极管阵列检测的高效液相色谱法定量。TLC方法新颖，提供了一种在野外实用的简单、廉价的方法。新方法在从感染油棕中提取高产率麦角固醇方面特别有效，能够对野外样本进行现场快速分析，使感染油棕能够非常迅速地得到处理或淘汰。本文讨论了该方法的一些局限性。这些步骤有助于更有效地控制疾病，并更有效地利用目前用于种植油棕的土地，从而减轻开发新种植园的压力。
