| 引用本文： |
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朱旭瑶,杨明柳,潘红平,阎冰,廖馨.红树林土壤环境DNA提取和纯化方法比较[J].广西科学,2020,27(1):65-75. [点击复制]
- ZHU Xuyao,YANG Mingliu,PAN Hongping,YAN Bing,LIAO Xin.Comparison of DNA Extraction and Purification Methods for Environmental Samples from Mangrove Soil[J].Guangxi Sciences,2020,27(1):65-75. [点击复制]
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| 摘要: |
| 探讨适合红树林土壤环境DNA的提取及纯化方法，筛选出可以提取到高质量且物种覆盖度广的环境DNA的方法。本研究通过多种方法提取和纯化红树林土壤环境DNA，并进行物种覆盖度（细菌、真菌、底栖无脊椎动物、植物、线虫）的比较评估。用6种方法提取红树林潮间带林下土壤环境DNA，并使用2种方法对DNA进行纯化；用不同物种类群的通用引物进行PCR扩增，评估各种方法提取和纯化的环境DNA的物种覆盖度。结果显示，不同方法提取DNA的效率有一定的差异，但2种方法纯化前后DNA的质量并没有显著差异；5种试剂盒方法提取的DNA均可以有效扩增出细菌和无脊椎动物的DNA；DNeasy PowerSoil试剂盒提取的DNA扩增植物引物结果最佳；来自不同区域红树林土壤的DNA在扩增真菌对应引物时差异较大；2种试剂盒直接提取的DNA可以有效扩增出线虫的DNA。本研究为今后基于环境DNA技术研究红树林生物多样性工作的顺利开展提供科学依据，奠定了一定的基础。 |
| 关键词: 红树林 环境DNA DNA提取 DNA纯化 生物多样性 |
| DOI：10.13656/j.cnki.gxkx.20200311.008 |
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| 基金项目:国家重点研发计划科技基础资源调查专项（2017FY100704），广西自治区直属公益性科研院所基本科研业务费项目（2018GMRC01）和广西自然科学基金项目（2019GXNSFBA185027）资助。 |
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| Comparison of DNA Extraction and Purification Methods for Environmental Samples from Mangrove Soil |
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ZHU Xuyao1,2, YANG Mingliu2, PAN Hongping1, YAN Bing2, LIAO Xin2
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| (1.College of Animal Science and Technology, Guangxi University, Nanning, Guangxi, 530004, China;2.Guangxi Key Lab of Mangrove Conservation and Utilization, Guangxi Mangrove Research Center, Guangxi Academy of Sciences, Beihai, Guangxi, 536000, China) |
| Abstract: |
| The extraction and purification method suitable for mangrove soil environmental DNA was explored,and a method that can extract environmental DNA with high quality and wide coverage was screened out.In this study,mangrove soil environmental DNA was extracted and purified by various methods,and the species coverage (bacteria,fungi,benthic invertebrates,plants,nematodes) was compared and evaluated.Six methods were used to extract the total DNA of sub-forest soil in the intertidal zone of mangrove and two methods were used to purify the DNA.PCR amplification was performed with universal primers from different species groups to assess the species coverage of environmental DNA extracted and purified by various methods.The results showed that the efficiency of extracting DNA by different methods was different.However,there was no significant difference in DNA quality between the two methods before and after purification.The DNA extracted by the five kit methods could effectively amplify the DNA of bacteria and invertebrates.The DNA amplified plant primers extracted from the DNeasy PowerSoil kit yielded the best results.The DNA from mangrove soils in different regions had a large difference when amplifying the corresponding primers of fungi.The DNA directly extracted by the two kits could effectively amplify the DNA of nematodes.This study provides a scientific basis and lays a foundation for the future study of mangrove biodiversity based on environmental DNA technology. |
| Key words: mangrove environmental DNA DNA extraction DNA purification biodiversity |
