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大熊猫粪便野外暴露时间对微卫星实验的影响
Effect of Exposure Time of Ailuropoda melanoleuca Feces on Microsatellites Analysis
王承东1, 杨波1, 黄杰2,3, 黄炎1, 成彦曦1, 李仁贵1, 黄山1, 李德生1, 张和民1*
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作者单位:1. 中国保护大熊猫研究中心, 四川卧龙 623006;
2. 四川省濒危野生动物保护生物学重点实验室, 四川大学生命科学学院, 成都 610064;
3. 河南商丘师范学院生命科学学院, 河南商丘 476000
中文关键字:大熊猫;粪便;暴露时间;微卫星
英文关键字:Ailuropoda melanoleuca; feces; exposure time; microsatellite
中文摘要:非损伤性取样,如从粪便中获取DNA用于分子生物学研究已经在珍稀野生动物中得到了广泛应用。DNA质量与后续研究很大程度上依赖于粪便暴露时间的长短。本研究通过分析不同新鲜程度粪便样品的DNA质量和微卫星基因分型效果探讨大熊猫粪便野外暴露时间对分子实验的影响。研究表明,暴露时间越短,粪便DNA降解程度越低,所提取的DNA质量也较高。同时粪便DNA的降解程度与季节和暴露环境也有一定的关系:春、秋季14 d以内、夏季7 d以内、冬季21 d以内的粪便样品提取的DNA质量较好,微卫星能成功分型;另外,野外卧穴口的粪便样品DNA保存时间相对较长。本研究对野外大熊猫粪便采集工作具有指导意义。
英文摘要:Noninvasive sampling methods, such as acquiring host genomic DNA from fecal samples for molecular biology research, are increasingly used in endangered wild animals. However, it is difficult to obtain fresh feces in the field. By focusing on the effects of feces after different exposure times in the field on molecular studies for giant pandas (Ailuropoda melanoleuca), we found that shorter exposure time of feces would lower the level of DNA degradation, and increase the quality of the feces DNA. The degradation level of feces DNA was also related to seasonal variation and field environment. The valid time was no longer than 14 days in spring and autumn, less than 7 days in summer and no longer than 21 days in winter. Moreover, dry environment such as the entrance of the cave may be the best place for preserving feces samples in the field. The results of this study provided guidance for the feces collection of wild giant pandas in the field.
2016,35(4): 481-487 收稿日期:2016-02-16
DOI:10.11984/j.issn.1000-7083.20160033
分类号:Q958.1;Q953
基金项目:大熊猫国合资金项目(AD1413):大熊猫饲养、繁育及管理;重点区域野生大熊猫疫病本底调查与监测
作者简介:王承东,博士,高级工程师,主要从事大熊猫饲养管理、兽医及放归研究
*通讯作者:张和民,E-mail:wolong-zhm@126.com
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