Bioactive Natural Product Laboratory, Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Jamia Hamdard, New Delhi, India.
Pharm Biol. 2013 Jun;51(6):771-7. doi: 10.3109/13880209.2013.765898. Epub 2013 Apr 11.
Stevia rebaudiana Bertoni (Asteraceae) is an important medicinal plant and is much used due to its zero calories sweetening property. Stevia leaves as well as its extracts and pure compounds are currently used in the preparation of several medicines, food products and neutraceuticals.
To study the genetic and metabolic variability in S. rebaudiana among accessions of different geographical regions of India using random amplified polymorphic DNA (RAPD) markers and high-performance thin layer chromatography (HPTLC) analysis.
The RAPD analysis of Stevia rebaudiana (11 accessions) was carried out using 20 random operon primers. Dendrogram was constructed for cluster analysis based on the unweighted pair group method with arithmetic means (UPGMA) using Winboot. The HPTLC analysis of all samples was carried out on silica using acetone:ethyl acetate:water (5:4:1, v/v/v) for fingerprinting and quantification of stevioside and rebaudioside A at 360 nm after spraying with anisaldehyde sulphuric acid.
Ten out of 20 primers screened were found most informative; amplification products of the genotypes yielded a total of 87 scorable bands (67 polymorphic), whereas genetic similarity (GS) coefficient (0.01-0.08) and polymorphism (67.24-92.40%) showed huge variability. Similarly, HPTLC analysis showed large variation among different samples with respect to their presence or absence of metabolite and their concentration.
Out of the 11 Stevia accessions, Delhi and Mohali varieties showed much relatedness with each other and were concluded to be the superior genotype in context to RAPD and HPTLC analysis. The information obtained here could be valuable for devising strategies for cultivating this medicinal plant.
甜菊（Stevia rebaudiana Bertoni）（菊科）是一种重要的药用植物，由于其零卡路里的甜味特性而被广泛使用。甜菊叶及其提取物和纯化合物目前用于制备几种药物、食品产品和营养保健品。
使用随机扩增多态性 DNA（RAPD）标记和高效薄层色谱（HPTLC）分析研究来自印度不同地理区域的甜菊品种间的遗传和代谢变异性。
对 11 个甜菊品种（品系）进行了 Stevia rebaudiana 的 RAPD 分析，使用了 20 个随机操作子引物。基于算术平均值的非加权对组方法（UPGMA）构建了基于聚类分析的聚类图。使用丙酮：乙酸乙酯：水（5：4：1，v/v/v）对所有样品进行 HPTLC 分析，在 360nm 处用茴香醛硫酸喷雾进行指纹图谱分析和甜菊糖苷和莱鲍迪甙 A 的定量。
筛选出的 20 个引物中有 10 个最具信息量；基因型的扩增产物共产生 87 个可评分条带（67 个多态性），而遗传相似性（GS）系数（0.01-0.08）和多态性（67.24-92.40%）显示出巨大的变异性。同样，HPTLC 分析显示，不同样品之间在代谢物的存在与否及其浓度方面存在很大差异。
在所研究的 11 个甜菊品种中，德里和 Mohali 品种彼此之间表现出很大的相关性，并且在 RAPD 和 HPTLC 分析方面被认为是较好的基因型。本研究获得的信息对于制定这种药用植物的种植策略可能具有重要价值。
