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| Assay of DNA Methylation in Arabidopsis Under Cd Stress Using Improved MSAP-PCR Technique |
| Received:March 16, 2015 |
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| KeyWord:methylation;MSAP-PCR;Arabidopsis;biomarker;epigenetic damage |
| Author Name | Affiliation | E-mail | | WANG He-tong | Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China | | | HE Lei | Liaoning University, Shenyang 110036, China | | | SONG Jie | Liaoning University, Shenyang 110036, China | | | SUN Li-zong | Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China | | | CUI Wei-na | Shanghai Institute of Technology, Shanghai 201418, China | | | TAI Pei-dong | Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China | | | JIA Chun-yun | Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China | | | LIU Wan | Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China | liuwan63@hotmail.com |
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| Abstract: |
| Global genomic DNA methylation assay is a hot research topic in plant epigenetic damages induced by stress. However, there is no method used for a rapid diagnosis of epigenetic damages. In this study, Arabidopsis was used as the test plant to examine the applicability of the methylation-sensitive arbitrarily primed PCR(MSAP-PCR) in plant methylation assay. The experimental condition optimization, primer screening, and polymorphism and sensitivity analysis were performed. A higher methylation polymorphism of MSAP-PCR products could be obtained with the optimum template DNA of 150~250 ng and 4% PAGE gel with 50% urea. MSAP-PCR had higher sensitivity to Cd stresses, with 30 sites of methylation polymorphism detected under 0.2 mg·L-1 Cd2+. The primer AP-4 was sensitive to methylation in CpG sites whereas the primer AP-3 was sensitive in CHG sites. In summary, MSAP-PCR was a method with simple procedure, low cost, high accuracy, and superior sensitivity. It could be an ideal method for studying the plant global genomic methylation, the plant stress resistance, and the early diagnosis of environmental contamination. |
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