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根据长芒苋和苋属其他物种的一个2 bp ITS序列差异设计特异引物和探针,建立了长芒苋的Taq Man实时荧光PCR鉴定方法。应用该方法测试了82个样品,包括23个不同来源的长芒苋样品、57个其他10种苋属样品,以及2个与长芒苋种子形态近似的鸡冠花、藜样品。试验结果表明,所有供试的长芒苋样品均表现为阳性扩增,非长芒苋和空白对照均没有出现阳性扩增。不同浓度长芒苋DNA的检测结果显示,该实时荧光PCR方法的检测灵敏度达2.2 fg/μL。该检测方法可能快速、准确鉴定长芒苋及其种子。
Abstract：A rapid method for the identification of the pigweed Amaranthus palmeri S.Watson was developed based on TaqMan Real-time PCR. Primers and Taqman probes were developed based on a 2-bp polymorphisms of ITS which could distinguish A. palmeri from other species of the genus. A total of 82 samples include 23 A. palmeri samples of various sources and 57 samples representing 10 Amaranthus species,as well as 2 species of Celosia cristata and Chenopodium album,which seeds are similar to A.palmeri,were tested with the probe and primers for TaqMan RT-PCR. Results showed that all the A.palmeri samples are positive and others are negative,meanwhile the sensitivity reached to 2.2 fg/μL. The reliable and sensitive method could make a contribution to fast and accurate identification of A. palmeri,especially for its seeds.
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基本信息:
DOI：10.19662/j.cnki.issn1005-2755.2020.06.002
中图分类号:S451
引用信息:
[1]李友军,薛华杰,张宁等.长芒苋实时荧光PCR鉴定方法[J].植物检疫,2020,34(06):25-30.DOI:10.19662/j.cnki.issn1005-2755.2020.06.002.
基金信息:
国家重点研发计划（2017YFC1200601）