LIU Wei,ZHAO Wanyue,ZHOU Changmei,et al.Camparison of Different DNA Extraction Methods for Detection of Meconpsis Vig[J].Northern Horticulture,2017,41(15):97-102.[doi:10.11937/bfyy.20164449]
绿绒蒿属植物不同DNA提取方式的比较
- Title:
- Camparison of Different DNA Extraction Methods for Detection of Meconpsis Vig
- 文献标志码:
- A
- 摘要:
- 以绿绒蒿属（Meconpsis Vig.）3种植物叶片为试材，采用研钵手动研磨和电动匀浆马达研磨器2种研磨方式处理叶片，采用改良的CTAB法、国产天根试剂盒、德国Qiagen试剂盒法3种提取方式从绿绒蒿属植物叶片中提取DNA，研究了不同提取方式对绿绒蒿属植物DNA提取效果的影响，以期为绿绒蒿属植物的分子遗传学研究奠定基础。结果表明：几种不同提取方法均能获得较为完整的DNA，研钵手动研磨的植物叶片提取的DNA无论是从质量、纯度还是得率均好于电动匀浆马达研磨器研磨。3种提取方式提取的DNA均可直接用于下游PCR反应，改良的CTAB法提取的DNA纯度和得率均很高；天根试剂盒法和Qiagen试剂盒法提取的DNA得率不高；Qiagen提取的DNA纯度和质量均比较好，但成本较高。
- Abstract:
- Leaves of three kinds of Meconopsis Vig. were used as materials,two grinding ways were designed to treat the leaves,including manual grinding and small electric motor grinding;the DNA was isolated from Meconopsis Vig.by three methods including modified CTAB method,TianGren kit method and Qiagen kit method.The effects of different extraction methods on DNA yield were studied in order to supply reference for Meconopsis Vig. molecular genetics study.The results showed that several methods all could be used to exact DNA of Meconopsis Vig.,but the quality and quantity of the exacted DNA were different.The DNA extracted by manual grinding method either from the quality or purity rate compared to the small electric motor grinding method was better.The DNA extracted by the method of modified CTAB was high in purity and yield;the rate of the DNA exacted by TianGen kit method and Qiagen kit method was not high,what′s more,the DNA exacted by Qiagen kit method was better than another in purity and quality,but the cost was higher.
参考文献/References:
[1]屈燕,区智.绿绒蒿属植物国内外研究进展[J].北方园艺,2011(3):191-194. [2]吴海峰,丁立生,王环,等.绿绒蒿属植物化学成分及药理活性研究进展[J].天然产物研究与开发,2011,23(1):163-168. [3]吴健梅.绿绒蒿—雪域高原上的美人[J].广东第二课堂（中学生版）,2013(9):53-55. [4]吴健梅.雪域高原上的美人绿绒蒿[J].旅游纵览,2015(9):30-34. [5]姚一麟,徐晔春.中国珍奇植物高山花卉（上）[J].园林,2013(9):64-65. [6]杜军华,张文静,李继荣,等.青藏高原特色植物资源绿绒蒿属植物的开发利用[J].青海师范大学学报（自然科学版）,2011,8(20):52-57. [7]SAMANT S S,JITENDRA S B，AMAN S.Assessment of diversity,distribution,conservation status and preparation of management plan for medicinal plants in the catchment area of Parbati hydroelectric project stage-Ⅲ in northwestern Himalaya[J].Journal of Mountain Science,2007,4(1):34-56. [8]卫生部药典委员会.中华人民共和国卫生部药品标准藏药：1册[M].北京:人民卫生出版社,1995:98. [9]周历萍,肖雯霏,裘劼人,等.一种高效的草莓总核酸提取方法[J].浙江农业科学,2013,6(6):732-733. [10]冉策,陈柳,陆家兰,等.三种 DNA 提取方法对检测草莓镶脉病毒稳定性的研究[J].华北农学报,2015(3):200-204. [11]刘杰,高连明.红豆杉属植物三种不同总DNA提取方法的分析比较[J].广西植物,2011,31(2):244-249. [12]马丽丽,艾力江吐尔逊,张莉,等.石蜡包埋组织提取DNA不同条件下的PCR扩增[J].中国组织工程研究,2012,16(11):1973-1976. [13]XIN Z,VELTEN J P,OLIVER M J,et al.High-throughput DNA extraction method suitable for PCR[J].Biotechniques,2003,34(4):820-825. [14]杨立国,杨晶.5种常见植物DNA提取效率的比较[J].生物学通报,2012,47(9):44-45. [15]应占明,应正河,潘伟华.沙罗科植物DNA提取方法研究[J].安徽农业科学,2010,38(8):3922-3924. [16]MATASYOH L G,WACHIRA F N,KINYUA M G,et al.Leaf storage conditions and genomic DNA isolation efficiency in Ocimum grantissimum from Kenya[J].Afr J Biotechnol,2008,7(5):557-564. [17]ROGERS S O,BENDICH A J.Extraction of DNA from milligram amounts of fresh,herbarium and mummified plant tissues[J].Plant Mol Biol，1985,5(2):69-76. [18]袁长春.濒危植物绿绒蒿属（Meconpsis Vig.）和滇桐属（Craigia）的系统发育及生物地理学研究[D].广州:中山大学,2002. [19]THOMPSON J R,WETZEL S,KLERKS M M,et al.Multiplex RT-PCR detection of four aphid-borne strawberry viruses in Fragaria spp.In combination with a plant mRNA specific internal control[J].Journal of Virological Methods,2003,111(2):85-93. [20]刘伟红,许文涛,商颖,等.果汁DNA提取方法比较及柑橘植物分子生物学检测技术研究[J].中国食品学报,2012,4(12):195-200. [21]张继红,陶能国,张小云,等.三种豆科植物总DNA提取方法的比较[J].湖南农业科学,2007(2):31-33. [22]赵孟良,韩睿,马胜超,等.菊芋基因组DNA提取方法的比较[J].中国蔬菜,2012(14)：41-44. [23]任衍春,张震,毛雪琴,等.水稻纹枯菌总DNA提取方法的比较[J].3版.北京:高等教育出版社,2011,23(4):741-747. [24]孟现东,陈益泰.枫香DNA提取方法与PCR扩增程序的优化[J].林业科学研究,2004,25(6):58-62. [25]王文斗,段英俊,那冬晨.植物总DNA提取方法的改进[J].安徽农业科学,2013,24(16):9913-9923. [26]郭宝林,李家实,阎玉凝.中药材DNA分子标记研究的技术问题[I].植物药基因组DNA的提取[J].中草药,2000,12(31):951-953. [27]金晓玲,乌云塔娜,张智俊,等.樟属植物总DNA提取方法研究[J].植物研究,2004,1(24):107-110. [28]伍艳芳,徐海宁,肖复明,等.陈山红心杉基因组DNA提取方法的比较与分析[J].江西农业大学学报,2012,34(3):517-521. [29]马昕璐,黄俊华.沙枣植物基因组DNA提取方法的比较[J].天津农业科学,2015,21(9):1-5,9. [30]张馨元,赵超越,侯和胜,等.四种中药材DNA提取方法的比较[J].中国生化药物杂志,2015,7(35):17-21. [31]王振华,赵晖,宋祺,等.油菜茎溃疡病菌基因组DNA提取方法的比较[J].植物学报,2011,50(3):511-513. [32]林强,邱长玉,朱方容,等.两种桑树基因组DNA提取方法的比较研究呢[J].广西蚕业,2011,48(2):1-5. [33]仇建标,丁文勇,陈少波,等.几种DNA提取方法对红树植物秋茄叶片DNA提取效果的比较[J].浙江海洋学院学报（自然科学版）,2012,31(5):402-408.
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备注/Memo
第一作者简介：刘伟（1993-），女，硕士研究生，研究方向为分子生物学。E-mail:821411486@qq.com.责任作者：屈燕（1979-），女，博士，副教授，研究方向为园林植物资源保护与开发利用。E-mail:flyersw@163.com.基金项目：国家自然科学基金资助项目(31460218)；云南省高校重点学科（风景园林学）建设资助项目；国家级质量工程园林专业综合改革试点资助项目(50126002)。收稿日期：2017-04-06