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利用NCBI核酸数据库中ITS(internal transcribed spacer)序列,结合产地分析和形态分析对新发现的一种黑果枸杞混伪品进行溯源研究。通过提取样品DNA,PCR扩增及双向测序,获得样品的ITS序列。所得序列用Conting Express软件进行拼接,截取ITS序列全长;利用NCBI核酸数据库中已登录的ITS序列进行相似度分析对其进行DNA条码溯源;根据其产地分析DNA条形码溯源结果;再根据形态特征进一步验证结果,3种方法联用获得溯源结果。新发现的黑果枸杞混伪品来源于小檗科小檗属植物喀什小檗Berberis kaschgarica。与黑果枸杞的主要区别:喀什小檗果实表面平滑,质地轻脆,黑果枸杞表面皱缩,质地坚实而硬;喀什小檗外果皮细胞壁连珠状增厚,石细胞壁平直,层纹不明显,黑果枸杞外果皮细胞壁不增厚,石细胞壁波浪状,层纹明显;喀什小檗的ITS序列长度为606 bp,黑果枸杞的长度为654 bp,二者序列对齐后的相似度为53.32%。
Using the sequence of ITS (internal transcribed spacer) in NCBI nucleic acid database, combined with the origin analysis and morphological analysis, a new type of black fruit Lycium barbarum was found. The ITS sequence of the sample was obtained by extracting the sample DNA, PCR amplification and bidirectional sequencing. The obtained sequence was spliced by Conting Express software and intercepted the full length of ITS sequences. The similarity of ITS sequences registered in the NCBI nucleic acid database was used to carry out DNA barcode tracing. The DNA barcode tracing results were analyzed according to its origin. Verification results, three ways to get traceability results. The newly discovered black fruit of Lycium barbarum from the Berberidaceae Berberis Berberis kaschgarica Berberis. The main difference between blackberry and Chinese wolfberry: Kashi berry fruit surface smooth, crisp texture, black fruit wolfberry surface shrinkage, solid and hard texture; Kashi berberis outer pericarp wall thickening even cell wall, stony cell wall straight, stratum not Obviously, the outer wall of Lycium barbarum did not thicken, the wavy and streaked stone cell wall was obvious. The length of ITS sequence of barberry in Kashgar was 606 bp, and the length of black fruit Lycium barbarum was 654 bp. The alignment of the two sequences was 53.32%.