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Development of rapid DNA extraction and PCR amplification methods for fungi and parasitic plants

文献类型： 外文期刊

作者： Khoo, Ying Wei 1 ; Li, Shifang 1 ; Chong, Khim Phin 3 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect Beijing, Beijing 100193, Peoples R China

2.Chinese Acad Trop Agr Sci, Environm & Plant Protect Inst, Haikou 571101, Peoples R China

3.Univ Malaysia Sabah, Fac Sci & Nat Resources Jalan UMS, Kota Kinabalu 88400, Sabah, Malaysia

关键词： lysis buffer; pathogenic fungi; mechanical and heat disruption; parasitic plants; PCR amplification

期刊名称：ZEMDIRBYSTE-AGRICULTURE （ 影响因子：1.281； 五年影响因子：1.573 ）

ISSN： 1392-3196

年卷期： 2022 年 109 卷 3 期

页码：

收录情况： SCI

摘要： The main culprits of catastrophic agricultural losses or crop mortality are phytopathogenic fungi and parasitic plants. To manage the plant pathogens, a simple and rapid disease diagnosis is needed. The aim of the experiment was to develop simple and rapid genomic DNA extraction and PCR amplification methods for fungi and parasitic plants. For the DNA extraction, mycelia from 16 fungi species and stems from two parasitic plants species were incubated in a lysis buffer and were homogenised using a sterilised wooden stick. The homogenates were incubated at 95 degrees C temperature for 10 min. Crude extracts served as a template for the PCR amplification containing UKOD polymerase. The application of lysis buffer, mechanical and heat disruption resulted in a fast DNA extraction from fungi and parasitic plants. DNA amplification time is reduced when using Master Mix containing UKOD polymerase.The presented results confirm that these simple and rapid DNA extraction and PCR amplification methods are applicable to diverse fungi species and parasitic plants.

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