葛根超微饮片ITS序列指纹图谱的研究

目的探求葛根超微饮片的品种鉴别方法。方法以来自不同产地的4个野葛样品和10个粉葛样品制成的超微饮片为实验材料，用分子克隆方法获得并测定其ITS序列。结果标记它们的ITS1，5．8S，ITS2的全长序列，构建了葛根的ITS序列指纹图谱，通过分析比较，发现葛根属中野葛之间，野葛和粉葛之间有显著差异；粉葛之间的差异较小。野葛序列总的同一性为80．74％，长度差异可达20bp，但可分为2个同一性为99．7％以上的组，提示来源可能为2个种。野葛和粉葛序列总的同一性为62％，最大长度差异可达180bp。粉葛之间序列的同一性为98％以上，长度差异仅有4bp。结论以ITS序列测定与分析作为葛根超微饮片品种鉴定和质量控制标准是完全可行的。

ITS sequence fingerprint of ultramicro Radix Puerariae

Objective To establish the identification and evaluation of ultramicro Radix Puerariae.Methods Radix Puerariae from 4 kinds of Pueraia(Willd.) Ohwi and 10 kinds of P.thomsonii Benth.which came from different regions were used as materials,their total DNA were extracted and cloned,and the ITS sequence was determined by molecular biological technique.Results Some remarkable differences among the group of Pueraia lobata(Willd.) Ohwi,and between the 2 groups of Pueraia lobata(Willd.) Ohwi and P.thomsonii Benth were found.But the difference among the group of P.thomsonii Benth.was inconspicuous.The identity of the ITS sequence of the group of Pueraia lobata(Willd.) Ohwi was about 80.74%,and the length varied from 0 to 20 bp,whereas there were two groups among these ITS sequence whose identities were over than 99.7%,which showed they probably came from 2 sources.The percentage of identity between the 2 groups of Pueraia lobata(Willd.) Ohwi and P.thomsonii Benth.was only 62%,and their maximum length difference reached 180 bp.But the identity among the group of P.thomsonii Benth.was more than 98%,and their length varied by only 4 bp.Conclusion It is quite feasible to use the ITS sequence fingerprint as a criterion for the identification of ultramicro Radix Puerariae,which also supplies a reference for the evluation of quality.