珍稀濒危鸟类微量取样及DNA提取的研究Microsampling and DNA Extraction of Rare and Endangered Birds
田秀华,刘铸,白素英
摘要(Abstract)：
为解决珍稀濒危鸟类分子生物学研究中存在的取样局限性,采用非伤害性取样和非损伤性取样,对大鸨10~70μL的血液和羽毛的不同部位(羽根、羽髓、羽鞘、实心羽茎和羽毛上脐周围绒羽)的DNA提取进行了研究。结果表明,鸟类血液样本与哺乳动物样本极为不同,两者相同体积的血液中,鸟类样本DNA含量是哺乳动物样本的几百倍,研究发现,大鸨可以从微量的血液中(10μL)提取出较多的DNA(45.75μg)。对于羽毛的不同部位,羽髓、羽鞘和羽根可以提出一定纯度的核DNA和mtDNA;实心羽茎可以提出一定纯度的mtD-NA。因此该研究将拓宽珍稀濒危鸟类采样的范围,为珍稀濒危鸟类的分子生物学研究提供了更广阔的样品来源。
关键词(KeyWords)： 珍稀濒危鸟类;非伤害性取样;非损伤性取样;核DNA;线粒体DNA
基金项目(Foundation): 黑龙江省科技攻关项目(GC04B519)
作者(Author): 田秀华,刘铸,白素英
DOI: 10.13326/j.jea.2005.04.010
参考文献(References)：
- [1]Taberlet P,Waits L P,Luikart G.Noninvasive sampling:look be-fore you leap[J].Trends in Ecology and Evolution,1999,14:323-327.
- [2]李军林,加建斌,舒青,等.非损伤性取样在珍稀濒危鸟类———朱种群遗传学中的应用[J].榆林高等专科学校学报,2001,11(2):37-39.
- [3]Taberlet P,Griffin S,Goosens B.Reliable genotyping of sampleswith very low DNA quantities using PCR[J].Nucleic Acids Re-search,1996,24:3 189-3 194.
- [4]Morin P A,Chambers K E,Boesch C,et al.Quantitative poly-merase chain reaction analysis of DNA from noninvasive samples foraccurate microsatellite genotyping of wild chimpanzees(PantroglodytesVerus)[J].Molecular Ecology,2001,10:1 835-1 844.
- [5]Gagneux P,Boesch C,Woodruff D.Microsatellite scoring errorsassociated with non-invasive genotyping based on nuclear DNA am-plified from shed hair[J].Molecular Ecology,1997,6:861-868.
- [6]Kohn M H,York E C,Kamradt D A,et al.stimating populationsize by genotyping faeces[J].Proceedings of the Royal Society ofLondon,Series B:Biological Sciences,1999,266:657-663.
- [7]翟永功,马建岗,吴晓民,等.水貂Mustla Vison微量血DNA的提取与纯化[J].经济动物学报,1998,2(2):6-9.
- [8]Wang J J,Tian X H,Song J B,et al.Physiological testing of bloodfor great bustard[J].Journal of Forestry Research,1998,9(4):278-279.
- [9]任文社,陈宝江,黄玉亭,等.不同组合CoBFA对雏鸡部分血液指标的影响[J].河北农业大学学报,2001,24(1):49-50.
- [10]Marton B H,Begona M,Juan J N,et al.An overlooked DNAsource for non-invasive genetic analysis in birds[J].Journal ofAvain Biology,2005,36:84-88.
- [11]Segelbacher G.Noninvasive genetic analysis in birds:testing reli-ability of feather samples[J].Molecular Ecology Notes,2002,2:367-369.
- [12]Walsh P S,Metzger D A,Higuchi R.Chelex-100 as amediumfor simple extraction of DNA for PCR-based typing from forensicmaterial[J].Biotechniques,1991,10(4):506-513.