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不同产区厚朴nrDNA ITS序列分析及亲缘关系鉴定
王洁, 杨旭, 杨志玲*
中国林业科学研究院 亚热带林业研究所, 浙江 富阳 311400
摘要:
以陕西、四川、浙江、重庆、福建、广西主产区的11个种源厚朴为材料,采用PCR直接测序法,测定不同产区厚朴的核糖体DNA ITS区序列,并结合GeneBank中相关植物的ITS序列(以同属近缘物种辛夷为外类群),应用Kimura 2遗传距离与NJ系统树分析法对不同产区厚朴的亲缘关系进行分析。结果表明:11个产区厚朴的ITS全序列长度介于593~600 bp,其中,ITS1长度为214~217 bp,G+C含量为54.42%~5535%,ITS2长度为215~219 bp,G+C含量为59.82%~60.95%;ITS序列共有36个变异位点,均出现在ITS1(7个)和ITS2(29个)序列中,虽然位点变异与部分叶形变化出现吻合,但无必然联系;序列间遗传分化距离为0.000~0.02792,11个产区厚朴构成3个分支,显示出产区间厚朴的亲缘关系,其中陕西洋县(SXYX)、陕西西乡(SXXX)、福建武夷山(FJWYS)及四川彭州(SCPZ)四个产区ITS序列完全一致。nrDNA ITS序列分析可作为厚朴不同产区亲缘关系鉴定的手段。
关键词:  nrDNA ITS序列  厚朴  系统聚类  亲缘关系
DOI:
分类号:Q789
Fund project:
NrDNA ITS sequences analysis and genetic rela tionship identification of Magnolia officinalis from different geographical regions
WANG Jie, YANG Xu, YANG ZhiLing*
Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, China
Abstract:
Magnolia officinalis of 11 main locality of growth (sach as Shanxi,Sichuan,Zhejiang,Chongqing,Fujian and Guangxi and so on) were used in this paper to find the divergence of nrDNA ITS (internal transcribed spacer) sequences for M.officinalis distributed in different areas and to provide references for the identification of the genetic relationship between different geographical regions. Their ITS sequences with direct sequencing were determined and their relationship was analyzed using Kimura 2 genetic distance and NJ(neighbor joining)phylogenetic tree. Meanwhile,M. liliiflora was used as the out group. The results showed that the whole length of ITS sequences of M.officinalis was 593-600 bp,in which the length of ITS1 and ITS2 were 214-217 bp and 215-219 bp,respectively. The contents of G+C were 54.42%-55.35% and 59. 82 %-60.95%,respectively. The length of 58S was 164 bp. A total of 36 variable sites(ITS1,7;ITS2,29) were examined in ITS sequences,and 5.8S sequence was highly conserved. The sites of variation consistent with changes of partial leaf shape,but they had not necessary linkage. The genetic differentiation distance between sequences ranged from 0.000 to 0.02792. M.officinalis of 11 main producing regions constituted three branches,which indicated the genetic relationship of M.officinalis between different producing areas,in which,Yangxian(SXYX),Xixiang(SXXX),Wuyishan(FJWYS)and Pengzhou(SCPZ)ITS sequences were the same. Therefore,nrDNA ITS sequence analysis could be applied to the identification of genetic relationship for M.officinalis in different producing areas.
Key words:  nrDNA ITS sequences  Magnolia officinalis  hierarchical clustering  genetic relationship
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