季节和天气因素对麋鹿非损伤性遗传取样的影响  

The Season and Weather Impacts on Noninvasive Genetic Sampling of the David's Deer(Elaphurus davidianus)

作　　者：王琰[1] 钱晨[1] 丁晶晶[1,2] 陈亚[1] 刘佳秀玉 丁玉华[3] 朱立峰[1] 常青[1] 

机构地区：[1]南京师范大学生命科学学院,江苏南京210023 [2]江苏省林业科学研究院,江苏南京211153 [3]江苏大丰麋鹿国家级自然保护区,江苏大丰224136

出　　处：《南京师大学报（自然科学版）》2014年第1期123-127,共5页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：国家自然科学基金(41271057);江苏省林业三新工程(lysx[2011]09)项目

摘　　要：非损伤性遗传取样在濒危动物遗传多样性的研究中具有重要作用.在运用非损伤性遗传取样对麋鹿（ Elaphurus davidianus）进行研究时发现，冬季采集粪样提取的核酸质量优于夏季.利用QIAamp DNA Stool Mini Kit试剂盒，冬季样品获得的DNA OD 260/280在1.60~1.90之间的比例为61.57%，而夏季的仅为43.51%.基于PCR扩增的性别鉴定和微卫星分型表明，冬季样品的成功率分别为84.98%和96.25%，夏季的分别为93.13%和95.25%.天气因素会对样品质量造成极显著的影响，非雨雪天气采集样品的OD 260/280在1.60~1.90之间的比例为61.57%，大雪后的为23.68%，大雨后的为23.88%.雨雪可能会冲刷掉部分粪样表面的肠壁组织、粘膜或者血液，并且降解其中的DNA.对麋鹿的非损伤性遗传取样建议选择在晴朗的冬天进行.In recent years,noninvasive genetic sampling plays an important role in genetic researches of many endangered animals. When we used this method to analyze the genetic diversity of David＆#39;s Deer, Elaphurus davidianus, one of the world endangered species,we found the DNA quality from fecal samples which collected in winter was better than that in summer. The proportion of OD 260/280 ratio which between 1. 60 and 1. 90 was about 61. 57% for the winter samples, and about 43. 51% for the summer samples,according to the standard procedure of the QIAamp DNA Stool Mini Kit. The success rate of PCR amplification for gender identification and microsatellite genotyping was 84. 98% and 96. 25% for winter samples,and 93. 13% and 95. 25% for summer samples respectively. There was no significant difference between the different seasons. The weather such as rain and snow influenced the DNA quality significantly. The proportion of OD 260/280 ratio which between 1. 60 and 1. 90 was about 61. 57% for the sunny winter samples,about 23. 68% and 23. 88% for the samples after raining and snowing respectively. The rain would wash out the intestinal tissue,mucosa or blood on the surface of fecal samples, and made the degradation of the DNA. The results suggest that the noninvasive genetic sampling for David＆#39;s Deer should be done in sunny winter,as it can improve the DNA quality effectively.

关 键 词：非损伤性遗传取样 麋鹿 OD 260/280 DNA质量 

分 类 号：Q953[生物学—动物学]