Yang Yao-Hua, Zhao Zhi-Li, Gaawe Dorje, Mi Ma, Meng Qian-Wan, Ni Liang-Hong
Department of Pharmacognosy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Tibetan Traditional Medical College, Lhasa 850000, China.
Zhongguo Zhong Yao Za Zhi. 2013 Nov;38(21):3773-5.
To identify the common Tibetan herb Chuan-Bu.
Local herbalists were visited to observe which plants were being used as Chuan-Bu. Samples of the indigenous plants were collected at the same time. Leaf materials were collected from field surveys. Total genomic DNA was extracted from silica gel-dried leaf samples. The PCR products were purified and directly sequenced.
As the origin of Chuan-Bu in Tibet autonomous region was authenticated, two species were determined, i. e. Euphorbia stracheyiand E. wallichii. Also, based on our earlier research, the origin of Chuan-Bu in Gansu province, is from E. kansuensis. The sequences of ITS1 for E. stracheyi and E. wallichii were 261 bp in size, and 221 bp in ITS2, respectively. The size of the 5.8S coding region was 164 bp for all species examined in the genus. Especially, there was a heterozygous locus in ITS1 (C/G; position 72) for E. stracheyi. The nucleotide divergence between sequences of the 6 species in pairwise comparisons was calculated and the result showed that the variable site could be detected in each pairwise comparison of sequences. Also, there were 8 point mutations in the 5.8S coding region.
nrDNA ITS sequences can be used as the molecular markers to identify the Tibetan herb Chuan-Bu and such Traditional Chinese Medicines from the same genus Euphorbia as E. lathyris, E. humifusa and E. pekinensis.
鉴定常见藏药川布。
走访当地草药医生，观察哪些植物被用作川布。同时采集当地植物样本。通过实地调查采集叶片材料。从硅胶干燥的叶片样本中提取总基因组DNA。对PCR产物进行纯化并直接测序。
随着西藏自治区川布来源的确定，鉴定出两种植物，即西藏大戟和多裂大戟。此外，根据我们早期的研究，甘肃省川布的来源是甘肃大戟。西藏大戟和多裂大戟的ITS1序列大小分别为261 bp，ITS2为221 bp。在该属所有检测物种中，5.8S编码区大小为164 bp。特别是，西藏大戟的ITS1中存在一个杂合位点（C/G；位置72）。计算了6个物种序列两两比较之间的核苷酸差异，结果表明在每个序列两两比较中都能检测到可变位点。此外，5.8S编码区有8个点突变。
nrDNA ITS序列可作为分子标记用于鉴定藏药川布以及大戟属的其他中药，如续随子、地锦草和大戟。
