樟科16个树种木材的RAPD与ISSR分子鉴别
2017-10-10骆嘉言钟文翰刘海冲蒋火俊
骆嘉言,王 英,钟文翰,刘海冲,蒋火俊
(1.南京林业大学 材料科学与工程学院,江苏 南京210037;2.浙江省家具检测技术研究重点实验室,浙江 杭州 311112)
研究简报
樟科16个树种木材的RAPD与ISSR分子鉴别
骆嘉言1,王 英1,钟文翰2,刘海冲1,蒋火俊1
(1.南京林业大学 材料科学与工程学院,江苏 南京210037;2.浙江省家具检测技术研究重点实验室,浙江 杭州 311112)
采用随机扩增多态性(RAPD)和简单序列重复区间扩增多态性(ISSR)技术鉴别樟科Lauraceae16个树种。用改良十六烷基三甲基溴化铵-十二烷基硫酸钠(CTAB-SDS)法提取樟科16个树种192株(12株·种-1)木质部基因组DNA,用RAPD与ISSR技术对它们进行聚合酶链式反应(PCR)扩增。共筛选出8条RAPD引物与6条ISSR引物,8条RAPD引物共扩增出165条大小为100~2 000 bp的条带,其中多态性条带为153条,多态率为92.7%;6条ISSR引物共扩增出96条大小为100~2 000 bp的条带,其中多态性条带为86条,多态率为89.6%。通过1~2个引物扩增的多态性条带就可鉴别与区分参试的16个树种,为树种鉴定提供技术支持。图3表5参18
植物学;樟科;木材;RAPD;ISSR;分子鉴定
Abstract:Random amplified polymorphic DNA (RAPD)and inter simple sequence repeats(ISSR)techniques were applied to identify 16 species of Lauraceae.The optimized cetyl trimethylammonium bromide (CTAB)-sodium dodecyl sulfate (SDS) method was used to extract wood DNA of 192 trees in the 16 species.RAPD and ISSR techniques were used to amplify DNA of the 16 species.Results showed that eight RAPD primers and six ISSR primers were screened.Through the eight RAPD primers,165 bands were obtained,among which 153 were polymorphic for a species level polymorphism loci rate of 92.7%.96 bands were obtained through the six ISSR primers,among which 86 were polymorphic for a species level polymorphism loci rate of 89.6%.DNA fragment size was 100-2 000 bp.These polymorphism bands amplified by 1 or 2 primers could distinguish the wood of these 16 species in the Lauraceae family,providing technical support for wood identification.[Ch,3 fig.5 tab.18 ref.]
Key words:botany;Lauraceae;wood;random amplified polymorphic DNA (RAPD);inter simple sequence repeats (ISSR);molecular identification
樟科Lauraceae植物广泛分布于热带及亚热带地区,全世界约50属2 500~3 000种[1],中国有24属约430种。樟科树木是中国南方重要的经济林木,在林业、轻工、医药、园林绿化中均占有重要地位。楠属Phoebe和樟属Cinnamomum等木材被认为是优质木材,常用作建筑物的梁柱构件、棺椁、家具、箱盒等。樟科木材普遍含有油细胞或黏液细胞,是樟科木材的识别标识;但樟科各个属间及种间,木材结构特征相似,仅通过木材解剖结构特征很难区分。随机扩增多态性(RAPD)与简单序列重复区间扩……
