摘要
为探讨庙台槭不同组织DNA的高效提取方法,分别以叶片和果实为材料,以完全随机区组试验设计,设置不同浓度的β-巯基乙醇和不同浓度的PVP作为预处理,采用试剂盒法提取基因组DNA,提取后对基因组DNA进行浓度、分光光度值的测定以及PCR扩增,综合各项指标选择出效果最好的预处理方法,并进一步以CTAB法进行验证,之后应用于不同个体庙台槭的DNA提取中。结果发现:以庙台槭果实为材料,不需要预处理,提取的DNA即可满足下游实验需要;以叶片为材料,用0.5%β-巯基乙醇和1%PVP预处理能有效提高提取的DNA浓度和质量,经验证发现该预处理方法适用于CTAB法,且能够极显著地提高DNA提取的浓度和质量;进一步应用于所有132个庙台槭个体的总DNA提取,获得的DNA均能满足后续实验要求。本研究为进一步研究庙台槭遗传多样性等工作提供帮助,同时也能够为槭属其他植物的相关研究提供参考。
In order to study the effecient extraction methods of DNA from different tissues of Acer miaotaiense,DNA was extracted from leaves and fruits using kits,and completed randomized block design was used to set different concentrations ofβ-mercaptoethanol and PVP as pretreatments.Determination of concentrations,spectrophotometric values,and PCR amplification were performed on extracted genomic DNA and then these indicators were used to select the best pretreatment method.This pretreatment method was verified by the CTAB method,and applied in the DNA extraction of different individuals.The results showed that DNA extracted from fruits did not require pretreatment,and the extracted DNA could meet the needs of downstream experiments.Using leaves as material,0.5%β-mercaptoethanol and 1%PVP were considered as effective to improve the concentration and quality of extracted DNA.It was found that the pretreatment method was suitable for CTAB and could significantly improve the concentration and quality of DNA extraction.It was further applied to the extraction of total DNA from all 132 individuals,and the obtained DNA all could meet the requirements of follow-up experiments.This study could lay a foundation for further research on the genetic diversity of Acer miaotaiense,and provide a reference for the related study on other Acer plants.
作者
李萌
李翔
庞晓明
李颖岳
Li Meng;Li Xiang;Pang Xiaoming;Li Yingyue(National Engineering Laboratory for Tree Breeding,Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plant,Ministry of Education,College of Biological Sciences and Biotechnology,Beijing Forestry University,Beijing,100083)
出处
《分子植物育种》
CAS
CSCD
北大核心
2019年第4期1228-1237,共10页
Molecular Plant Breeding
基金
中央高校基本科研业务费专项资金(2017ZY24)
河南省许昌市重大科技专项(20170112006)
国家林业局珍稀濒危野生动植物救护与繁育项目(20170501)共同资助
关键词
庙台槭
多酚
组织
DNA提取
Acer miaotaiense
Polyphenol
Plant tissue
DNA extraction
