曹 晖 佐佐木阳平 伏见裕利 小松かつ子. 基于核18S rDNA和叶绿体trnK序列鉴定姜黄属植物(英文)J. 药学学报, 2010,45(7): 926-933.
引用本文: 曹 晖 佐佐木阳平 伏见裕利 小松かつ子. 基于核18S rDNA和叶绿体trnK序列鉴定姜黄属植物(英文)J. 药学学报, 2010,45(7): 926-933.
CAO Hui, ZUO Zuo-Mu-Yang-Beng, FU Jian-Yu-Li, XIAO Song-かつZi. Authentication of Curcuma species (Zingiberaceae) based on nuclear 18S rDNA and plastid trnK sequencesJ. 药学学报, 2010,45(7): 926-933.
Citation: CAO Hui, ZUO Zuo-Mu-Yang-Beng, FU Jian-Yu-Li, XIAO Song-かつZi. Authentication of Curcuma species (Zingiberaceae) based on nuclear 18S rDNA and plastid trnK sequencesJ. 药学学报, 2010,45(7): 926-933.

基于核18S rDNA和叶绿体trnK序列鉴定姜黄属植物(英文)

Authentication of Curcuma species (Zingiberaceae) based on nuclear 18S rDNA and plastid trnK sequences

  • 摘要:

    姜科 (Zingiberaceae) 姜黄属 (Curcuma) 多种植物的根茎或块根如莪术 (Curcumae Rhizoma)、姜黄 (Curcumae Longae Rhizoma)、郁金 (Curcumae Radix) 等为常用中药, 莪术为蓬莪术Curcuma phaeocaulis Val.、广西莪术C. kwangsinensis S.G. Lee et C.F. Liang和温莪术C. wenyujin Y.H. Chen et C. Ling根茎, 具有行气破血、消积化食等功效; 姜黄为姜黄C. longa L.根茎, 具有行气破血、通经止痛等功效; 郁金为莪术及姜黄块根, 具有活血化瘀、行气止痛、利胆等功效。此3类药材常有混用, 而且姜黄属植物鉴别主要基于新鲜状态, 依赖腊叶标本较难鉴别。本研究通过分子系统学和生物信息学手段对姜黄属18种植物核糖体RNA小亚基基因 (18S rRNA) 和叶绿体赖氨酸tRNA基因 (trnK) 进行PCR直接测序, 获得这2个基因的完整序列, 1种姜花属 (Hedychium) 2种姜属 (Zingiber) 植物为外类群分析比较其序列变异, PAUP法建立了18种姜黄属植物的亲缘关系。所得结果显示, 18S rDNA序列长度1 810 bp, trnK2 6962 703 bp姜黄属植物基因种间变异范围为00.05% (18S rDNA) 00.19% (trnK), 在种一级水平单系分化上得到100% bootstrap确证。18种姜黄属植物18S rDNA序列仅有1个变异位点, 在系统树上广西莪术C. kwangsiensis 和莪术C. zedoaria日本居群与其他16种形成分化。trnK基因序列可变区在2trnK外显子与matK内含子之间, 共有3DNA插入/缺失, 包括18-bp的缺失重复序列和14-bp114-bp插入重复序列; 5单核苷酸多态性 (SNPs) 位点, trnK序列存在明显的种间基因条形码 (DNA barcoding)。研究结果表明分子系统学可以作为姜黄属植物亲缘关系研究的重要手段, 为部分姜黄属植物的种属归并提供了分子依据, trnK基因序列可变区作为DNA条形码候选基因可用于鉴定姜黄属植物及其药材。

     

    Abstract:

    Curcuma drugs have been used discriminatingly for invigorating blood circulation, promoting  digestion, and as a cholagogic in China.  However, there is confusion about the drug’s botanical origins and clinical uses because of morphological similarity of Curcuma plants and drugs.  Comparative sequencing of  the 18S rRNA gene in nuclear ribosomal DNA (rDNA) and trnK gene in chloroplast DNA (cpDNA) was carried out in order to examine interspecies phylogeny and to identify ultimately Curcuma species.  A total of a hundred of accessions of eighteen species were analyzed.  This resulted in an aligned matrix of 1 810 bp for 18S rDNA and 2 800 bp for trnK.  18S rDNA sequence divergence within the ingroup ranged from 00.05%, trnK ranged from 00.19%.  One base transversion-substituted site (from cytosine to thymine) was observed from the    upstream of 18S rDNA at nucleotide position 234 in C. kwangsiensis and Japanese population of C. zedoaria which have separated genetic distance to other Curcuma taxa.  Two noncoding regions embedded in trnK intron showed higher variability, including nucleotide substitutions, repeat insertion and deletions.  Based on consensus of relationship, eighteen major lineages within Curcuma are recognized at the species level.  The results suggest that Curcuma is monophyletic with 100% bootstrap support and sister to the genera Hedychium and Zingiber.  The trnK sequences showed considerable variations between Curcuma species and thus were revealed as a  promising candidate for barcoding of Curcuma species, which provide valuable characters for inferring relationship within species but are insufficient to resolve relationships among closely related taxa.

     

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