基于26S rDNA D1D3区和matK基因序列分析的阳春砂分子鉴别_临床医学论文.docVIP

基于26S rDNA D1区和matK基因序列分析的阳春砂分子鉴别_临床医学论文 基于26S rDNA D1区和matK基因序列分析的阳春砂分子鉴别_临床医学论文 作者：黄琼林, 杨锦芬，段中岗，何瑞，詹若挺，徐鸿华，徐晖，陈蔚文 【摘要】 　　【目的】 建立阳春砂不同栽培品种的DNA分子鉴别方法，为阳春砂的优良品种选育提供依据。【方法】以阳春砂栽培种长果、圆果、“春选”及海南砂为材料，用相应引物对26S rDNA D1区和matK基因进行聚合酶链反应（PCR）扩增和测序，对测序结果进行分析，寻找差异位点，并根据序列构建系统发生树。【结果】测序得到的26S rDNA D13序列为739 bp，阳春砂与海南砂在该区域存在4个碱基位点的差异。长果阳春砂与圆果阳春砂序列相同，但它们与“春选”阳春砂存在1个碱基位点的差异，根据26S rDNA D1序列建立的系统发生树揭示了“春选”阳春砂与另外2个栽培品种间的区别。测序得到的matK序列为824 bp，阳春砂3个栽培品种的matK序列相同，与海南砂存在1个碱基位点差异。【结论】从分子水平上可鉴别阳春砂的3个栽培品种，“春选”品种比长果（或圆果）品种与海南砂有着更近的亲缘关系。 【关键词】 阳春砂 基因序列分析 分子鉴定 　　Abstract∶ObjectiveTo establish a molecular identification method for three cultivars of Amomum villosum Lour. (AVL), thus to provide scientific evidence for the identification, selection and breeding of AVL. MethodsThe fragments of 26S rDNA D1D3 region and matK gene of three cultivars of AVL and Amomum longiligulare TLWu were amplified by polymerase chain reaction (PCR) and with corresponding primers, and then their sequences were analyzed, and phylogenetic tree was constructed based on the sequences. ResultsWe obtained 739 bp in 26S rDNA D1D3 sequence. Differences in 4 basic sites of 739 bp were shown between AVL and Amomum longiligulare TLWu. The two cultivars of AVL, Changguo and Yuanguo, had the same sequence, but there was a difference in one basic site of Changguo and Yuanguo from Chunxuan. The phylogenetic tree based on 26S rDNA D1D3 sequence revealed the difference between Chunxuan and the other two cultivars of AVL. We also obtained 824 bp in matK gene sequence. The three cultivars of AVL showed the consistent sequence, but there was a difference in one basic site of three cultivars of AVL from Amomum longiligulare TLWu. ConclusionWe can identify the three cultivars of AVL through the sequence differences at the molecular level, and Chunxuan has a closer genetic relationship with Amomum longiligulare TLWu． 　　Key words: AMOMUM VILLOSUM LOUR.;SEQUENCE ANALYSIS;MOLECULAR IDENTIFICATION; 　　砂仁为我国“四大南药”之一，来源于姜科植物阳春砂Amomu