Authors:

Paola Diaz

Date Created:

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About Paper:

Many plants can cause poisoning or death of animals following ingestion. Currently, identification of the plant is performed by visual identification of plant material in stomach/rumen contents. In instances where plant parts are too small and digested to be identified, another method is needed to identify the causative plant. The goal of this study is to develop a quantitative polymerase chain reaction (qPCR)-based test to identify Taxus (yew) DNA in stomach/rumen contents.Taxus DNA can be identified using qPCR. Taxus and non-Taxus samples were collected around the Purdue campus. DNA from each plant was extracted utilizing Qiagen's DNeasy Plant Pro Kit. qPCR primers were designed to amplify sequences specific to Taxus. Amplified DNA was verified by direct sequencing of amplicons. Sensitivity was determined by extracting decreasing concentrations of Taxus, or by decreasing the concentration of Taxus in spiked stomach/rumen contents. Specificity was determined by introducing controls such as non-Taxus plant species, non-spiked stomach/rumen contents, or with nuclease free water. Results: Cycle threshold (Ct) values were positive for samples containing Taxus DNA. Samples from other trees were also positive. DNA from poison hemlock, an herbaceous plant, was negative. Nuclease-free water was negative. Sequencing of amplicons from reactions of each plant correctly identified the plant in each case, including the samples containing Taxus at each concentration. This study indicates that PCR with sequencing can be utilized as a specific and sensitive test to identify Taxus DNA in stomach/rumen contents.

Source:

Purdue University / 2023

Topics:

No topics listed

Co-authors:

Paola Diaz