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连作对穿心莲基因组DNA甲基化的影响
引用本文:李俊仁,陈秀珍,曾湘达,卓一南,何瑞,詹若挺.连作对穿心莲基因组DNA甲基化的影响[J].中国实验方剂学杂志,2018,24(15):65-71.
作者姓名:李俊仁  陈秀珍  曾湘达  卓一南  何瑞  詹若挺
作者单位:广州中医药大学中药资源科学与工程研究中心岭南中药资源教育部重点实验室国家中成药工程技术研究中心南药研发实验室
基金项目:广东省协同创新中心创新科研团队建设项目(A1-AFD01514A04);广州中医药大学“薪火计划”项目(XH20140119)
摘    要:目的:探究连作条件下穿心莲基因组DNA的甲基化水平及模式变化情况。方法:结合甲基化敏感扩增多态性(methylation-sensitive amplified polymorphism,MSAP)技术和液相色谱-质谱(LC-MS/MS)法对3个连作年限(0,1,2年)穿心莲基因组DNA的甲基化水平及模式进行分析。结果:LC-MS/MS测定的连作0~2年穿心莲基因组DNA甲基化水平为13.91%,12.72%,10.26%,MSAP测定的结果则为10.16%,9.64%,8.98%,两者均表明穿心莲基因组DNA的甲基化水平与连作年限呈现负相关,即DNA甲基化水平随着连作时间的延长而逐渐下降。MSAP技术对DNA甲基化模式分析也显示,穿心莲基因组DNA的CCGG位点在连作过程中主要发生了去甲基化。结论:连作条件下,穿心莲基因组DNA的甲基化水平和模式发生改变并以此来响应连作胁迫。

关 键 词:穿心莲  DNA甲基化  连作  甲基化敏感扩增多态性  液相色谱-质谱法
收稿时间:2017/12/30 0:00:00

Effect of Continuous Cropping on DNA Methylation in Andrographis paniculata
LI Jun-ren,CHEN Xiu-zhen,ZENG Xiang-d,ZHUO Yi-nan,HE Rui and ZHAN Ruo-ting.Effect of Continuous Cropping on DNA Methylation in Andrographis paniculata[J].China Journal of Experimental Traditional Medical Formulae,2018,24(15):65-71.
Authors:LI Jun-ren  CHEN Xiu-zhen  ZENG Xiang-d  ZHUO Yi-nan  HE Rui and ZHAN Ruo-ting
Institution:Research Center of Chinese Herbal Resource Science and Engineering, Key Laboratory of Chinese Medicinal Resource from Lingnan, Ministry of Education, Joint Laboratory of National Engineering Research Center for the Pharmaceutics of Traditional Chinese Medicines, Guangzhou University of Chinese Medicine, Guangzhou 510006, China,Research Center of Chinese Herbal Resource Science and Engineering, Key Laboratory of Chinese Medicinal Resource from Lingnan, Ministry of Education, Joint Laboratory of National Engineering Research Center for the Pharmaceutics of Traditional Chinese Medicines, Guangzhou University of Chinese Medicine, Guangzhou 510006, China,Research Center of Chinese Herbal Resource Science and Engineering, Key Laboratory of Chinese Medicinal Resource from Lingnan, Ministry of Education, Joint Laboratory of National Engineering Research Center for the Pharmaceutics of Traditional Chinese Medicines, Guangzhou University of Chinese Medicine, Guangzhou 510006, China,Research Center of Chinese Herbal Resource Science and Engineering, Key Laboratory of Chinese Medicinal Resource from Lingnan, Ministry of Education, Joint Laboratory of National Engineering Research Center for the Pharmaceutics of Traditional Chinese Medicines, Guangzhou University of Chinese Medicine, Guangzhou 510006, China,Research Center of Chinese Herbal Resource Science and Engineering, Key Laboratory of Chinese Medicinal Resource from Lingnan, Ministry of Education, Joint Laboratory of National Engineering Research Center for the Pharmaceutics of Traditional Chinese Medicines, Guangzhou University of Chinese Medicine, Guangzhou 510006, China and Research Center of Chinese Herbal Resource Science and Engineering, Key Laboratory of Chinese Medicinal Resource from Lingnan, Ministry of Education, Joint Laboratory of National Engineering Research Center for the Pharmaceutics of Traditional Chinese Medicines, Guangzhou University of Chinese Medicine, Guangzhou 510006, China
Abstract:Objective: To investigate the DNA methylation level and pattern changes of Andrographis paniculata under continuous cropping conditions. Method: DNA methylation level and patterns of A. paniculata samples continuously cropped for 0, 1, 2 years in sheltered pots were determined by using methylation-sensitive amplified polymorphism (MSAP) and liquid chromatography-mass spectrometry (LC-MS/MS). Result: The DNA methylation level detected by LC-MS/MS was 13.91%, 12.72%, 10.26%, and 10.16%, 9.64%, 8.98% determined by MSAP. Both results indicated that the DNA methylation level of A. paniculata had a gradual decreasing tendency with the increase of the continuous cropping years. MSAP analysis also showed that demethylation was present at CCGG sites of genomic DNA during the continuous cropping process. Conclusion: DNA methylation level and pattern of A. paniculata were changed to response to the continuous cropping stress.
Keywords:Andrographis paniculata  DNA methylation  continuous cropping  methylation-sensitive amplified polymorphism  LC-MS/MS
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