四川农业大学学报 ›› 2016, Vol. 34 ›› Issue (04): 450-455,477.doi: 10.16036/j.issn.1000-2650.2016.04.010

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3种楠木DNA提取及其ISSR标记鉴别

王英1, 骆嘉言1, 钟文翰2, 石江涛1, 樊帅1   

  1. 1. 南京林业大学材料科学与工程学院, 南京 210037;
    2. 浙江省家具检测技术研究重点实验室, 杭州 311112
  • 收稿日期:2016-05-13 出版日期:2016-12-31 发布日期:2017-02-15
  • 通讯作者: 骆嘉言,副教授,主要从事木材学方面的研究,E-mail:luojiayan@njfu.edu.cn。 E-mail:luojiayan@njfu.edu.cn
  • 作者简介:王英,硕士研究生。
  • 基金资助:
    江苏高校优势学科建设工程资助项目(PAPD)

DNA Extraction and Wood ISSR Identification from Three Phoebe Species

WANG Ying1, LUO Jia-yan1, ZHONG Wen-han2, SHI Jiang-tao1, FAN Shuai1   

  1. 1. College of Materials Science and Engineering, Nanjing Forestry University, Nanjing 210037, China;
    2. Key Laboratory of Furniture Inspection Technology of Zhejiang Province, Hangzhou 311112, China
  • Received:2016-05-13 Online:2016-12-31 Published:2017-02-15

摘要: [目的] 采用ISSR分子标记鉴别楠属3个树种。[方法] 以紫楠、浙江楠和桢楠木材为对象,比对改良CTABSDS法、CTAB法和试剂盒法3种方法提取的木材基因组DNA质量,采用ISSR-PCR技术对3个树种24个样本基因组DNA进行扩增。[结果] 改良CTAB-SDS法和CTAB法较适合这3种木材的DNA提取。序列扩增筛选出7条稳定、条带清晰且多态性好的引物,确定其最佳退火温度。7条引物共扩增出67条大小为100~2000 bp的条带,其中多态性条带43条,多态率64.2%。[结论] 这7条引物扩增的多态性条带都可用于鉴别与区分这3种楠木。

关键词: 紫楠, 浙江楠, 桢楠, ISSR-PCR, 分子鉴定

Abstract: [Objective] The aim of the study was to identifying 3 phoebe species by inter simple sequence repeat(ISSR) technique. [Method] The optimized CTAB-SDS, CTAB and kit for DNA extraction methods were used to extract wood DNA from P. sheareri, P. chekiangensis and P. zhennan. Three species were amplified with ISSR-PCR technique. [Results] The optimized CTAB-SDS and CTAB methods were better for DNA extraction of 3 woods. 7 ISSR primers were screened and their amplification were stable, clear and high polymorphic. A total of 67 bands were obtained through 7 primers, 43, 64.2% of which were polymorphic. DNA fragment size were ranged from 100 bp to 2 000 bp. [Conclusion] These polymorphism bands amplified by the 7 primers can distinguish the woods of 3 species.

Key words: P. sheareri, P. chekiangensis, P. zhennan, ISSR-PCR, molecular identification

中图分类号: 

  • S792.24