HUANG Qionglin,MA Xinye,ZHAN Ruoting,et al.Comparative Analysis and Molecular Identification of matK Gene From Spatholobus suberectus and Its Adulterants[J].Northern Horticulture,2015,39(17):94-98.[doi:10.11937/bfyy.201517024]
鸡血藤及其混伪品matK基因分析和分子鉴定
- Title:
- Comparative Analysis and Molecular Identification of matK Gene From Spatholobus suberectus and Its Adulterants
- Keywords:
- Spatholobus suberectus Dunn.; matK gene; sequence analysis; molecular identification; adulterants
- 文献标志码:
- A
- 摘要:
- 以鸡血藤及其4种常见混伪品为试材,采用植物基因组DNA提取试剂盒提取鸡血藤样品总DNA,并采用matK基因通用引物扩增和测序,从GenBank获取鸡血藤4种混伪品的matK基因序列,采用DNAMAN、MEGA等生物软件比对分析序列差异,计算遗传距离并构建聚类树,研究matK基因对鸡血藤及其混伪品的鉴别效果。结果表明:获得的鸡血藤及其混伪品matK序列长度为690 bp,平均GC含量为31.1%,共发现236个差异位点,占总碱基数的34.2%。8个鸡血藤样品matK序列在41 bp处存在A-G碱基差异。鸡血藤与混伪品之间的种间变异大于鸡血藤样品内的变异。基于matK基因序列建立的聚类树能够明显地鉴别鸡血藤及其混伪品。matK基因可作为鉴别鸡血藤及其混伪品的标准DNA序列。
- Abstract:
- In order to investigate the efficiency of matK gene on identifying Spatholobus suberectus and its adulterants,S.suberectus and four common adulterants were used as materials in this study.Genomic DNA was isolated from fresh leaves and dry materials of S.suberectus,and matK gene was amplified and sequenced using a pair of universal primers.matK sequences of the adulterants were downloaded from GenBank database.The sequences were aligned and analyzed to find the difference and then genetic distance were calculated and the clustering tree was constructed using biological software.The results revealed that matK sequences acquired from S.suberectus and its adulterants contained 690 base pairs with an average GC content of 31.1%.The maximal intraspecific distances among S.suberectus were 0.001,whereas the interspecific distance between S.suberectus and its adulterants ranged from 0.114 to 0.288.The clustering tree based on matK sequences can intuitively discriminate S.suberectus and its adulterants.matK gene can be used as a standard DNA sequence to distinguish Spatholobus suberectus and its adulterants.
参考文献/References:
[1]HEBERT P D N,GREGORY T R.The promise of DNA barcoding for taxonomy [J].Syst Biol,2005,54(5):852-859. [2]黄琼林,梁凌玲,何瑞,等.应用matK基因鉴别青天葵及其常见混伪品[J].广西植物,2014,34(3):299-303. [3]HILU K W,BORSCH T,MLLER K,et al.Angiosperm phylogeny based on matK sequence information[J].Amer J Bot,2003,90(12):1758-1776. [4]HOLLINGSWORTH P M,FORREST L L,SPOUGE J L,et al.A DNA barcode for land plants[J].Proc Natl Acad Sci USA,2009,106(31):12794-12797. [5]国家药典委员会.中华人民共和国药典(一部)[S].北京:人民卫生出版社,2010:134. [6]HUANG Q,DUAN Z,YANG J,et al.SNP typing for germplasm identification of Amomum villosum Lour.based on DNA barcoding markers [J].PLoS ONE,2014,9(12):e114940. [7]HUANG Q,LIANG L,HE R,et al.Applying DNA barcodes to identify Nervilia fordii and six congeneric species [J].Plant Omics,2013,6(5):325-332. [8]张晓芹,刘春生,闫兴丽,等.多基原药材大黄叶绿体matK基因序列分析及鉴定研究[J].药学学报,2013,48(11):1722-1728. [9]安冉,杨锦芬,刘军民,等.基于26S rDNA D1-D3区序列分析的鸡血藤及其混淆品的分子鉴别[J].广州中医药大学学报,2010,27(3):403-406. [10]罗焜,马培,姚辉,等.中药DNA条形码鉴定中的DNA提取方法研究[J].世界科学技术-中医药现代化,2012,14(2):1433-1439. [11]陈士林.中药DNA条形码分子鉴定[M].北京:人民卫生出版社,2012. [12]CHEN S L,YAO H,HAN J P,et al.Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J].PLoS ONE,2012,7(5):e8613.
相似文献/References:
[1]荣广天,何钢,朱丽芳,等.鸡血藤的生理生化研究进展[J].北方园艺,2015,39(06):183.[doi:10.11937/bfyy.201506050]
RONG Guang-tian,HE Gang,ZHU Li-fang,et al.Research Advance in Physiological and Biochemical of Spatholobi caulis[J].Northern Horticulture,2015,39(17):183.[doi:10.11937/bfyy.201506050]
[2]荣广天,何钢,倪尚格,等.鸡血藤的扦插育苗试验[J].北方园艺,2016,40(05):171.[doi:10.11937/bfyy.201605045]
RONG Guangtian,HE Gang,NI Shangge,et al.Experiment of Cutting Seeding-raising of Millettia reticuiata Benth[J].Northern Horticulture,2016,40(17):171.[doi:10.11937/bfyy.201605045]
[3]席飞飞,何钢,刘贤桂,等.不同栽培模式对五年生鸡血藤生理生化指标的影响[J].北方园艺,2017,41(15):124.[doi:10.11937/bfyy.20164265]
XI Feifei,HE Gang,LIU Xiangui,et al.Effects of Different Cultivating Models on Physiological and Biochemical Index of Five-year-old Spatholobus suberectus Dunn.[J].Northern Horticulture,2017,41(17):124.[doi:10.11937/bfyy.20164265]
备注/Memo
第一作者简介:黄琼林(1986-),男,博士,讲师,研究方向为分子生物学。E-mail:perfecthql@163.com.责任作者:陈蔚文(1950-),男,教授,博士生导师,研究方向为创新中药研究与开发。E-mail:chenww@gzucm.edu.cn.基金项目:广东省高等院校学科与专业建设专项资金资助项目(2013CXZDA011);2014年国家工信部中药材生产建设资助项目(工信厅消费函[2014]737号);广东医学院博士学位人员科研启动资助项目(B2013017)。