Universal DNA-Based Methods for Assessing the Diet of Grazing Livestock and Wildlife from Feces
Universal DNA-Based Methods for Assessing the Diet of Grazing Livestock and Wildlife from Feces
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DOI：
10.1021/jf803680c
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发表时间：
2009-07-08
影响因子：
6.1
通讯作者：
Pompanon, Francois
中科院分区：
文献类型：
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作者：
Pegard, Anthony;Miquel, Christian;Pompanon, Francois
由于控制牲畜饲料的需求，两种表征粪便中存在的植物DNA的方法被开发出来。从粪便样品中提取DNA后，用植物通用引物对其进行聚合酶链式反应，扩增出一小段叶绿体trnL内含子。第一种方法产生作为PCR产物的电泳性迁移模式的签名。第二种方法是通过焦磷酸测序从聚合酶链式反应产物中测序数百个DNA片段。这些方法通过对精料和草料饲养的羔羊的粪便进行盲法分析而得到验证。签名方法允许区分这两种日粮，并确认其中一种日粮中存在精料。焦磷酸测序方法允许在饮食中识别多达25个分类群。这些方法是对已经使用的化学方法的补充。它们可以应用于饮食控制和食物偏好的研究。
Because of the demand for controlling livestock diets, two methods that characterize the DNA of plants present in feces were developed. After DNA extraction from fecal samples, a short fragment of the chloroplastic trnL intron was amplified by PCR using a universal primer pair for plants. The first method generates a signature that is the electrophoretic migration pattern of the PCR product. The second method consists of sequencing several hundred DNA fragments from the PCR product through pyrosequencing. These methods were validated with a blind analysis of feces from concentrate- and pasture-fed lambs. The signature method allowed differentiation of the two diets and confirmed the presence of concentrate in one of them. The pyrosequencing method allowed the identification of up to 25 taxa in a diet. These methods are complementary to the chemical methods already used. They could be applied to the control of diets and the study of food preferences.