Genetic Diversity Among Saudi Peganum harmala and Rhazya stricta Populations Using Chemical and ISSR Markers.

BACKGROUND: The vernacular name 'Harmal' is used for two plant species in Saudi Arabia, i.e. Peganum harmala L. and Rhazya stricta Decne. Both are important medicinal plants which offer interesting pharmacological properties. OBJECTIVE: This study aimed to evaluate the genetic diversity among different populations of harmal based on chemical variations of alkaloids and molecular polymorphism. METHODS: Total alkaloids were extracted from plants of three populations of each species and estimated by using spectrophotometer and the chemical compounds were analyzed by Gas chromatography mass spectrometry (GC-MS). Molecular polymorphism was estimated by using the Inter Simple Sequence Repeat (ISSR) fingerprints. RESULTS: The results showed that the alkaloids content of R. stricta was higher than P. harmala populations. The GC-MS analysis revealed the presence of (65-53) compounds in R. stricta and P. harmala, and the percentage of polymorphism was found to be 93.2%. Sixteen ISSR primers produced 170 scorable bands with an average of 9.6 bands per primer and 75%-100% polymorphism. The cluster analysis using the unweighted pair-group method of the arithmetic average (UPGMA) method based on combined data of GC-MS and ISSR markers divided the six harmal genotypes into two major groups. CONCLUSION: The existence of variations in chemical and genetic markers is useful for the selection of potential genotypes for medicinal use, and for breeding lines for medicinal substances production to spare wild plants from uncontrolled harvesting for folk medicine.

Classification and differentiation of the genus Peganum indigenous to China based on chloroplast trnL-F and psbA-trnH sequences and seed coat morphology.

The genus Peganum (Zygophyllaceae) consists of six species and one subspecies; three of which are distributed in China, P. harmala Linn, P. nigellastrum Bunge and P. multisectum (Maxim.) Bobr. A probable new or intermediate species, Peganum sp., has been suggested in the wild in northwest China. Traditional classification in genus Peganum has focused on hairs on the plant surface. In this study, seed coat characteristics of Peganum species were investigated using light and scanning electron microscopy, demonstrating clear differences in morphology between species. In addition, DNA sequence data from two sequences (chloroplast: trnL-F, psbA-trnH) were used to differentiate Peganum sp. and study polygenetic relationships. Diversity in DNA sequences among Peganum species was found, with inter-specific sequence divergence ranging from 0.6% to 5.6% in psbA-trnH, and 0.0% to 1.8% in trnL-F. The variations within species were low: from 0.0% to 0.4% in psbA-trnH and 0.0% to 0.4% in trnL-F. Therefore, Peganum species can now be easily identified as separate entities based on variations in DNA sequences. Phylogenetic trees were constructed from the combined data set for the two gene fragments, and the results indicate that Peganum sp. is monophyletic and sister to P. harmala and P. nigellastrum, while P. multisectum is also monophyletic. DNA data further confirmed that P. multisectum is an independent species and that a new species, Peganum sp., exists within the genus Peganum growing wild in China.