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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 rDNA)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,以1种姜花属(Hedychium)和2种姜属(Zingiber)植物为外类群分析比较其序列变异,用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1810bp,trnK为2696～2703bp。姜黄属植物基因种间变异范围为0～0.05%(18S rDNA)和0～0.19%(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2个trnK外显子与matK内含子之间,共有3个DNA插入/缺失,包括1个8-bp的缺失重复序列和1个4-bp、1个14-bp插入重复序列;5个单核苷酸多态性(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。
Curcuma (Curcuma) Rhizomes or tubers of many plants such as Curcumae Rhizoma, Curcumae Longae Rhizoma and Curcumae Radix are commonly used traditional Chinese medicines, while Curcuma Curcuma phaeocaulis Val is a traditional Chinese medicine. , Guangxi Curcuma C.kwangsinensis SGLee et CFLiang and Curcuma C.wenyujin YHChen et C.Ling rhizome, qi qi and blood, eliminate consumption of food and other effects; turmeric is Curcuma C.longa L. rhizome, with line Qi broken blood, through the analgesic and other effects; turmeric and Curcuma root tuber, with blood circulation, qi and pain, gallbladder and other effects. The three types of herbs are often mixed, and the identification of Curcuma is mainly based on the fresh state, relying on waxy specimens more difficult to identify. In this study, molecular systematics and bioinformatics methods were used to directly sequence the 18s rDNA of ribosomal RNA and chloroplast lysine tRNA gene (trnK) in 18 species of Curcuma, and the integrity of these two genes was obtained Sequence analysis showed that there were 18 species of Curcuma plants in the phylogenetic tree based on the PAUP method, using 1 species of Hedychium and 2 species of Zingiber as outgroups. The results showed that 18S rDNA sequence length 1810bp, trnK 2696 ~ 2703bp. The variation range of turmeric gene was 0 ~ 0.05% (18S rDNA) and 0 ~ 0.19% (trnK), which was confirmed by 100% bootstrap on the first-level single-line differentiation. 18 species of Curcuma 18S rDNA sequence has only one mutation site, in the system tree Guangxi Curcuma C.kwangsiensis and Curcuma C.zedoaria Japanese population and other 16 kinds of differentiation. The trnK gene sequence variable region is between the two trnK exons and the matK intron, with a total of three DNA insertions / deletions, including an 8-bp deletion repeats and a 4-bp, a 14- bp inserted repeat sequences. There were obvious DNA barcoding in 5 single nucleotide polymorphisms (SNPs) sites and trnK sequences. The results show that molecular systematics can be used as an important means to study the genetic relationship of Curcuma and provide the molecular basis for the taxonomic combination of some Curcuma species. The variable region of trnK gene can be used as DNA barcode candidate gene to identify Curcuma plants and Herbal medicine.