نوع مقاله : مقاله پژوهشی
نویسندگان
گروه کشاورزی، دانشگاه پیام نور، تهران، ایران
چکیده
کلیدواژهها
موضوعات
عنوان مقاله [English]
نویسندگان [English]
The Iranian poppy (Papaver bracteatum) is a medicinal plant known for its ability to produce the alkaloid tebaine and grows in specific regions of Iran. This study aimed to identify the optimal protein extraction method and investigate the genetic diversity of 18 populations of Iranian poppy collected from various parts of Iran, along with a control sample of Papaver somniferum, using seed protein electrophoresis patterns. The results indicated that the protein extraction method utilizing phenol yielded the best results. Additionally, after staining with Coomassie Brilliant Blue G250, the band patterns displayed higher clarity and provided greater precision in tracking protein bands. A total of 68 bands were observed within a molecular weight range of 13-120 kDa when comparing electrophoresis patterns. The least and the greatest number of bands were recorded in populations P2 (Lar Dam, Alborz) and P17 (Pol Zanguleh region, Alborz), respectively. Genetic similarity among the studied populations varied significantly, ranging from 0.4 to 0.8. The greatest distance was noted between populations P15 and P7 from two different regions: Yush-Baladeh village in Alborz and Pir-Imran village in Kurdistan. Seed protein electrophoresis effectively distinguished between species P.bracteatum and P.somniferum. Cluster analysis using UPGMA method along with Dice similarity coefficient revealed five distinct groups among the studied populations. The findings suggest that geographic location has minimal impact on population similarity and their clustering; therefore, seed protein electrophoresis can be utilized for isolating and identifying different populations of this species in conservation programs.
کلیدواژهها [English]
Extended Abstract
Introduction
Iranian poppy (Papaver bracteatum) is a valuable medicinal plant that grows in limited regions of Iran. This species contains a high concentration of thebaine, an important alkaloid with significant pharmaceutical properties that can be readily converted into codeine. Moreover, thebaine can be transformed into Bentley compounds and various narcotic antagonists. As a precursor alkaloid with notable therapeutic value, thebaine plays a key role in the synthesis of numerous morphine-derived drugs, including codeine (a cough suppressant), morphine (a narcotic analgesic), noscapine (an antitumor agent), sanguinarine (an antimicrobial compound), and papaverine (a vasodilator).
Seed storage proteins are widely used as biochemical markers due to their physiological stability and ease of handling. These proteins are typically polymorphic, heritable, and represent the direct products of gene expression, with minimal influence from environmental factors. Analysis of seed storage proteins using SDS-PAGE electrophoresis is a common and reliable method for evaluating genetic variation within and between plant species.
In the present study, seed protein biochemical markers were employed to assess the genetic diversity of wild populations of Iranian poppy collected from different regions of Iran, highlighting the species’ importance for pharmaceutical applications.
Materials and Methods
This study aimed to (1) determine the most efficient method for protein extraction and staining, and (2) investigate the genetic diversity of 18 wild populations of Papaver bracteatum collected from various regions of Iran, along with one sample of P. somniferum used as a control, based on seed protein electrophoresis patterns.
Four protein extraction methods—Tris’s buffer, phosphate buffer, trichloroacetic acid (TCA), and phenol—were evaluated. Additionally, three staining techniques were applied: silver nitrate (AgNO₃), Coomassie Brilliant Blue G-250, and Coomassie Brilliant Blue R-250.
Polymorphic protein bands were scored as presence (1) or absence (0) using Phoretix Pro software. Molecular data were analyzed using Photoshop CS11 for image processing, GenAlEx for genetic analysis, and NTSYS v2.0.2 for cluster analysis.
Results and Discussion
Comparison of the extraction methods revealed that the phenol method produced the most reliable and reproducible results. Among the staining techniques, Coomassie Brilliant Blue G-250 provided higher resolution and improved visualization of protein bands.
A total of 68 protein bands, ranging from 13 to 120 kDa, were detected. The lowest and highest numbers of bands were observed in populations P2 and P17, respectively. Variation in banding patterns reflects genetic diversity among populations, whereas the presence of shared bands indicates genetic relatedness and possibly a common evolutionary origin. Differences in band intensity suggest that multiple protein polypeptides with similar molecular weights may overlap within the same band.
Genetic similarity among the studied populations ranged from 0.4 to 0.8. The greatest genetic distance was observed between populations P15 (Yushbeldeh village, Alborz region) and P7 (Piromran village, Kurdistan region). Seed protein electrophoresis clearly distinguished P. bracteatum from P. somniferum, as expected given their taxonomic differences.
The highest similarity was detected between populations grouped in the first and second clusters. Principal component analysis (PCA) results were consistent with cluster analysis findings, confirming the reliability of the grouping pattern. Cluster analysis using the UPGMA method and Dice similarity coefficient divided the populations into five distinct groups.
Because seed storage proteins are more stable in mature seeds than proteins in other plant tissues, SDS-PAGE effectively revealed polymorphisms based on variations in protein intensity among germplasms. These findings indicate that seed storage protein profiles provide a reliable tool for identifying and differentiating populations of P. bracteatum.
Conclusion
The results suggest that geographic origin has only a minor influence on population similarity and grouping. In contrast, seed protein electrophoresis patterns effectively differentiate and identify populations of Papaver bracteatum, demonstrating their usefulness as reliable biochemical markers for genetic diversity studies.
Author Contributions
Dr. Soheila afkar a conceived and planned the experiments. Ms. akbari carried out the experiments. Dr. Soheila afkar analyzed data, wrote the first manuscript and revised according to reviewers’ comments.
Data Availability Statement
Data available on request from the authors.
Acknowledgements
The authors would like to thank the research council of the University of Payame Noor, Iran for the financial support of this research.
Ethical considerations
The authors avoided data fabrication, falsification, plagiarism, and misconduct.
Conflict of interest
The author declares no conflict of interest.
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