湘西宏成制药公司不同基地泡桐的分子鉴定

摘要

The genomic DNA was isolated from leaves of Paulownia plants through the CTAB method without liquid nitrogen.The internal transcribed spacer（ITS） of rRNA and two intergenic spacers of psbA-trnH and trnF-trnF were amplified from the genomic DNA by PCR and then sequenced.Sequence analysis of the three DNA markers were conducted with software Clustal X7.0,and their phylogenetic trees were constructed with software MEGA3.1.The results showed three fragments of ITS,psbA-trnH and trnL-trnF was 600 bp,540 bp and 900 bp in length,respectively.Similarity of ITS sequence between the previously reported Paulownia tomentosa and Paulownia No.1 or No.2 was higher than 95%,but that between the P.tomentosa and No.3,No.4 or No.5 of Paulownia was less than 79%.Similarity of the trnL-trnF between the P.tomentosa and No.1 or No.2 of Paulownia both was 99.5%,and that between the P.tomentosa and No.3,No.4 or No.5 of Paulownia was less than 70%.The results indicated that sequence of the psbA-trnH of Paulownia species in different cultivation places was very conservative and it could not provide enough information for the relationship of the species among Paulownia.Molecular identification demonstrated that Paulownia No.1 and No.2 represented P.tomentosa,and Paulownia No.3,No.4 and No.5 represented P.fortunei.%根据核rRNA基因ITS序列、叶绿体trnL-trnF和psbA-trnH序列对湘西宏成制药公司不同基地泡桐进行分子鉴定.用PCR从泡桐基因组DNA中分别扩增出大小约为500bp的ITS序列、540bp的psbA-trnH序列和900bp的trnL-trnF序列.相似性分析结果表明,毛泡桐ITS序列与1号和2号基地泡桐的相似性高于95%,与3号和4号基地和吉首大学校园泡桐的相似性低于79%;毛泡桐trnL-trnF序列与1号和2号基地泡桐的相似性均为99.5%,与3号和4号基地和吉首大学校园泡桐的相似性都低于70%;宏成制药4个基地和吉首大学校园泡桐psbA-trnH序列的相似性都高于99%.说明1号和2号基地泡桐属于毛泡桐,3号和4号基地和吉首大学校园泡桐属于白花泡桐.