塔里木大学学报

针对胡杨、灰叶胡杨分布偏远、新鲜叶片褐化快等特点,探索了新鲜叶片、硅胶干燥后-70℃保存3个月的叶片、硅胶干燥后常温保存15个月的叶片、自然晾干3天的叶片、3个月标本的叶片、15个月的标本的叶片等六种采集保存方法对胡杨、灰叶胡杨总DNA提取质量的影响,并利用分子系统学研究的常用的核糖体RNA基因、线粒体基因、叶绿体基因的保守引物对所提的DNA进行PCR扩增,结果表明:硅胶干燥-70℃保存3个月的叶片和自然晾干3天的叶片均能得到与新鲜叶片相当质量的DNA。考虑到野外样品采集条件的限制,认为野外采集大量样品进行胡杨、灰叶胡杨分子系统学研究的最佳方法为采样后硅胶干燥-70℃长期保存。

It is of great importance that the preservation methods of the leaves of Populus euphratica Oliv.and Populus pruinosa Schrenk.for DNA extraction considering the fresh leave browning quickly and out-of-the-way distributing of the plant.The effect of six different preservation methods(fresh,stored about 3 months at the temperature of-70 ℃ after dried with silica gel,stored about 15months at room temperature after dried with silica gel,dryed about 3 days at room temperature,specimen stored at room temperature 3 months and 15 months) on the quality of total DNA was investigated.PCR amplified with primers of ITS1-5.8s-ITS2,trnL-trnF interval sequences and nad7 intron 2 for appraising the integrality of gDNA,mtDNA,cpDNA.The results showed that the DNA extracted from leaves stored about 3 months at the temperature-70 ℃ after dried with silica gel and drying about 3 days at room temperature could get the same high quality as from the fresh leaves.The method of stored at the temperature-70 ℃ after dried with silica gel could be considered for the best preservation method for study on Molecular Systematics of Populus euphratica oliv.and Populus pruinosa Schrenk.