中麻黄叶绿体DNA序列不同引物扩增条件的筛选
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摘要
目的:对16个不同居群的中麻黄从23对叶绿体编码区及非编码区DNA片段中筛选具有变异的叶绿体DNA片段。方法:通过试剂盒法提取DNA并梯度优化筛选扩增条件,PCR扩增产物经生物工程有限公司测序获得产物序列。结果:引物[trnS(GCU)-trnG(UCC)、atpB-rbcL、psbA-trnH、F71-R1516]能扩增出清晰、单一的条带。结论:16个不同居群中30个麻黄个体的叶绿体DNA扩增成功,初步筛选出具有变异的叶绿体DNA片段F71-R1516。
The Chloroplast DNA fragments with mutations from 23 pairs of chloroplast coding regions and non-coding regions were screened in 16 different populations of Ephedra intermedia. Methods:DNA was extracted and the amplification conditions were selected by gradient optimization. The PCR amplification products were sequenced by Bioengineering Co.,Ltd. to obtain the sequence.Results:The clear,single bands were obtained by primers(trnS(GCU)-trnG( UCC),atpB-rbcL,psbA-trnH,F71-R1516). Conclusion:Thirty individuals from 16 ephedra populations of chloroplast DNA were successfully amplified,and the chloroplast DNA fragments F71-R1516 with mutation were screened.
The Chloroplast DNA fragments with mutations from 23 pairs of chloroplast coding regions and non-coding regions were screened in 16 different populations of Ephedra intermedia. Methods:DNA was extracted and the amplification conditions were selected by gradient optimization. The PCR amplification products were sequenced by Bioengineering Co.,Ltd. to obtain the sequence.Results:The clear,single bands were obtained by primers(trnS(GCU)-trnG( UCC),atpB-rbcL,psbA-trnH,F71-R1516). Conclusion:Thirty individuals from 16 ephedra populations of chloroplast DNA were successfully amplified,and the chloroplast DNA fragments F71-R1516 with mutation were screened.
引文
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[6]朱田田,晋玲,杜弢,等.基于ISSR的甘肃中麻黄遗传多样性研究[J].中草药,2014,45(12):1764-1768.
[7]朱田田,晋玲,杜弢,等.中麻黄ISSR-PCR反应体系的建立和优化[J].中国药房,2013,24(43):4033-4037.
[8]马晓辉,卢有媛,张弦飞,等.中麻黄UPLC指纹图谱及其不同产区化学成分差异研究[J].中药材,2016,39(10):2217-2220.
[9]Hong H,Chen H B,Yang D H,et al.Comparison of contents of five ephedrine alkaloids in three official origins of Ephedra Herb in China by high-performance liquid chromatography[J].J Nat Med,2011,65(3):623-628.
[10]Abdel-Kader MS,Kassem F F,Abdallah RM.Two alkaloids from Ephedra aphylla growing in Egypt[J].Atural Product Sciences,2003,(9):52.
[2]马晓辉,卢有媛,黄得栋,等.中麻黄生态适宜性区划研究[J].中国中药杂志,2017,42(11):2068-2071.
[3]王成信,王耀琳.沙区麻黄人工栽培技术的试验研究[J].甘肃林业科技,1991(1):31-38.
[4]杨自辉,王继和,胡明贵,等.盐碱地麻黄种植试验研究初报[J].中草药,2000,31(5):61-62.
[5]高晓原.麻黄人工繁殖技术研究简报[J].宁夏农林科技,1995(5):41-42.
[6]朱田田,晋玲,杜弢,等.基于ISSR的甘肃中麻黄遗传多样性研究[J].中草药,2014,45(12):1764-1768.
[7]朱田田,晋玲,杜弢,等.中麻黄ISSR-PCR反应体系的建立和优化[J].中国药房,2013,24(43):4033-4037.
[8]马晓辉,卢有媛,张弦飞,等.中麻黄UPLC指纹图谱及其不同产区化学成分差异研究[J].中药材,2016,39(10):2217-2220.
[9]Hong H,Chen H B,Yang D H,et al.Comparison of contents of five ephedrine alkaloids in three official origins of Ephedra Herb in China by high-performance liquid chromatography[J].J Nat Med,2011,65(3):623-628.
[10]Abdel-Kader MS,Kassem F F,Abdallah RM.Two alkaloids from Ephedra aphylla growing in Egypt[J].Atural Product Sciences,2003,(9):52.