槟榔叶表微生物基因组DNA提取方法的比较  

Comparison of Microbial Genomic DNA Extraction Methods from Areca catechu Leaf Surface

作　　者：马瑞 王贞霖 芮凯[3] 徐芮 白蓓蓓 杨顺义 Ma Rui;Wang Zhenlin;Rui Kai;Xu Rui;Bai Beibei;Yang Shunyi(Scientific Observation and Experiment Station of Crop Pests in Haikou Ministry of Agriculture,Key Laboratory of Plant Diseases and Pest Control of Hainan Province,Sciences Research Center of Quality Safety and Standards for Agricultural Products of Hainan Academy of Agricultural Sciences,Institute of Plant Protection,Hainan Academy of Agricultural Sciences,Haikou,571100;Biocontrol Engineering Laboratory of Crop Diseases and Pests of Gansu Province,College of Plant Protection,Gansu Agricultural University,Lanzhou,730070;Sanya Institute of Hainan Academy of Agricultural Sciences,Sanya,572024)

机构地区：[1]海南省农业科学院植物保护研究所,海南省农业科学院农产品质量安全与标准研究中心,海南省植物病虫害防控重点实验室,农业农村部海口作物有害生物科学观测实验站,海口571100 [2]甘肃农业大学植物保护学院,甘肃省农作物病虫害生物防治工程实验室,兰州730070 [3]海南省农业科学院三亚研究院,三亚572024

出　　处：《分子植物育种》2024年第23期7744-7749,共6页Molecular Plant Breeding

基　　金：海南省槟榔产业技术体系建设专项(HNARS);海南省自然科学基金青年基金项目(321QN365)共同资助。

摘　　要：为确定槟榔叶表微生物基因组DNA的最佳提取方法,本研究采用改良CTAB法、MP Biomedicals土壤试剂盒、QIAGEN土壤试剂盒、天根多糖多酚植物试剂盒和MP Biomedicals植物试剂盒分别提取槟榔叶表微生物基因组DNA,通过超微量分光光度计测定、变性梯度凝胶电泳和聚合酶链式反应(PCR)对提取效果进行比较。结果表明,采用MP Biomedicals植物试剂盒提取的DNA效果最好,DNA浓度为81.57μg/mL,A_(260)/A_(280)值为1.80,A_(260)/A_(230)值为2.06,PCR扩增产物亮度高、无杂带,且具有提取时间短、效率高、污染小等特点。本研究为槟榔叶表微生物基因组DNA的提取提供了参考,为后续进行分子生物学实验提供了科学依据。In order to determine the best extraction method of microbial genomic DNA from Areca catechu leaf surface,the modified CTAB method,SPINeasy DNA Kit for Soil(MP Biomedicals),DNeasy PowerSoil Pro Kit(QIAGEN),Super Plant Genomic DNA Kit(Polysacchardes&Polyphenolics-rich)(TIANGEN)and SPINeasy DNA Kit for Plant(MP Biomedicals)were utilized to extract microbial genomic DNA from Areca catechu leaf surface.,The extraction results were compared by ultrafine spectrophotometer,denaturation gradient gel electrophoresis and polymerase chain reaction(PCR).The result showed that SPINeasy DNA Kit for Plant(MP Biomedicals)has the best effect on DNA extraction,with DNA concentration of 81.57μg/mL,A_(260)/A_(280)value of 1.80 and A_(260)/A_(230)value of 2.06.The PCR amplified product has high intensity and no stray bands.It has the characteristics of short extraction time,high efficiency and minimal contamination.This study provides a reference for the extraction of genomic DNA from Areca catechu leaf surface,and provides a scientific basis for the subsequent molecular biology experiments.

关 键 词：槟榔 叶表微生物 基因组DNA 提取方法 

分 类 号：S792.91[农业科学—林木遗传育种]