Comparative evaluation on different methods to extract endophytic bacteria DNA from Diaphorina citri Kuwayama
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摘要: 通过分析细菌16SrDNA基因序列的PCR扩增与克隆测序结果,比较传统方法和试剂盒法对柑橘木虱Diaphorina citri Kuwayama内生菌DNA模板的提取效果。试验表明,4种方法中,FastDNA试剂盒方法提取的DNA模板较适合于后续柑橘木虱内生菌16SrDNA的扩增,且扩增产物可以很好地用于克隆测序;传统提取方法经济节约,但提取的DNA量及纯度较低;试剂盒提取方法,操作简单,质量稳定,易标准化,但成本较高。4种方法各具优缺点,用FastDNA试剂盒提取的DNA模板,其PCR产物能满足克隆需要。Abstract: To find a better method for the extraction of endophytic bacteria DNA from Asian Citrus Psyllid,Diaphorina citri Kuwayama,we compared the traditional methods and FastDNA Kits.The result showed that the DNA template extracted with the method of FastDNA Kits(MP Biomedical)was more suitable for PCR amplification,and the amplification product could be well used for cloning and sequencing.In the contrast,although the other three methods were cheaper,the amount and purity of DNA extracted with them were decreased.Our results suggested that the extraction with the FastDNA kits was stable and convenient for operation and standardization but expensiveness.In summary,four methods had respective advantages and disadvantages,and FastDNA kits provided more stable DNA for PCR and sequence cloning.
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Key words:
- Diaphorina citri Kuwayama /
- endophytic bacteria /
- total DNA /
- extraction
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[1] 陈建利,阮传清,刘波,等.柑橘木虱对柑橘不同品种的趋性[J].福建农业学报,2011,26(2):280-283. [2] 袁辉,李安国.桂林柑橘木虱种群分布特点及防治对策探讨[J].广西植保,2007,20(4):31-32. [3] 杜丹超,鹿连明,张利平,等.柑橘木虱的防治技术研究进展[J].中国农学通报,2011,27(25):178-181. [4] 殷幼平,刘婷婷,田圣超,等.16SrDNA序列的柑桔木虱体内共生菌多样性研究[J].昆虫学报,2011,54(6):664-674. [5] DOUGLAS A E.Nutritional interactions insect-microbialsym-bioses:aphids and their symbiotic bacteria Buchmera[J].Annu Rev Entomol,1998,43:17-37. [6] 林林,吴云锋,崔晓峰.玉米蚜体内参与传毒的共生菌groEL基因的克隆和原核表达[J].中国病毒学,2003,18(1):54-57. [7] 许长藩,夏雨华,李开,等.柑桔黄龙病的病原在木虱体内循回期的研究[J].植物病理学报,1990,20(2):25-30. [8] SUBANDIYAH S,NIKOH N,TSUYUMU S,et al.Complexendosymbiotic microbiota of the Citrus Psyllid Diaphorina citri(Homoptera:Psylloidea)[J].Zoological Science,2000,17:983-989. [9] 石晶,谢映平,薛皎亮,等.蚧虫基因组DNA不同提取方法的比较[J].昆虫知识,2005,42(2):207-211. [10] 贺虹,魏琮,袁向群,等.一种提取麦红吸浆虫基因组DNA的方法[J].昆虫知识,2008,45(6):984-986. [11] 李正西,李定旭.桃蚜自然种群初级和次级共生菌的分子鉴定[J].昆虫学报,2005,48(5):810-814. [12] 田英芳,黄刚,郑哲民,等.一种简易的昆虫基因组DNA提取方法[J].陕西师范大学学报,1999,27(4):82-84. [13] 苗雪霞,丁德诚.蚜虫与其胞内共生细菌的相互作用[J].生命科学,2003,15(4):242-247. [14] DILLON R J,CHARNLEY A K.Mutualism between thedesert locust Schistocerca gregaria and its gut microbiota[J].Research in Microbiology,2002,153(8):503-509. -
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