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一种快速高效提取柑橘黄龙病植物样品DNA的方法
引用本文:温玉婷,雷翠云,王任翔,宋雅琴,张松,娄兵海.一种快速高效提取柑橘黄龙病植物样品DNA的方法[J].中国果业信息,2023,52(2).
作者姓名:温玉婷  雷翠云  王任翔  宋雅琴  张松  娄兵海
作者单位:广西师范大学、广西特色作物研究院,广西特色作物研究院,广西师范大学,广西特色作物研究院,广西特色作物研究院,广西特色作物研究院
基金项目:国家重点研发计划支持项目(2021YFD1400800)
摘    要:为快速获得高质量的柑橘黄龙病(HLB)植物样品DNA,本研究在传统CTAB法基础上,结合热萃取技术,建立了一种简便、快速提取HLB植物样品DNA的CTAB简化法。与Solarbio植物基因组DNA提取试剂盒和传统CTAB提取法相比,该方法所需时间短,1~2 h即可完成整个提取过程,获得的HLB植物样品DNA质量高、产量多。同时比较了三种方法制备模板时黄龙病病原菌和其他柑橘病原菌的PCR检出率,CTAB简化法和传统CTAB法检出率一致,试剂盒稍低,表明本研究建立的CTAB简化法可用于柑橘DNA病害PCR检测DNA模板的制备。本研究研发的HLB植物样品DNA提取方法优势明显,适用于大量病样的快速检测。

关 键 词:柑橘  DNA提取  黄龙病
收稿时间:2022/9/1 0:00:00
修稿时间:2022/9/19 0:00:00

Development of A Rapid and Efficient DNA Extraction Method for Citrus Huanglongbing plant samples Detection
Abstract:To rapidly extract high quality bacteria DNA from citrus greening disease (HLB) plant samples, a simple, rapid, and efficient DNA extraction method was established based on the conventional cetyltrimethylammonium bromide (CTAB) method, combined with thermal extraction. The result showed that shorter time, the whole extraction process can be completed in 1 to 2 hours, and good-quality DNA and higher DNA yield were obtained through this extraction method, comparing with the conventional CTAB extraction method and Plant Genomic DNA Extraction Kit (Solarbio). Meanwhile, the total DNA extracted by three methods was used for citrus Huanglongbing (HLB) and other citrus DNA pathogens detection, the simplified CTAB method and the conventional CTAB extraction method results were consistent, but the kit was slightly lower, indicating that simplified CTAB method can be used to prepare DNA template for PCR detection of citrus DNA diseases.Thus, the new DNA extract method has significant advantages on HLB detection.
Keywords:citrus  DNA extraction  Huanglongbing
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