بررسی تنوع مورفولوژیکی و بیوشیمیایی در برخی از گونه‌های گیاه دارویی بنفشه

به‌منظور بررسی تنوع مورفولوژیکی و بیوشمیایی و تعیین شباهت‌ها و فواصل خویشاوندی، تعداد 22 اکوتیپ گیاه دارویی بنفشه (viola spp.) که شامل 5 گونه بودند، از مناطق مختلف استان‌های گیلان، مازندران و چهارمحال و بختیاری جمع‌آوری شدند. صفات مورفولوژیکی شامل طول ریشه، ارتفاع ساقه گل‌دهنده، تعداد برگ، طول و عرض برگ، طول و عرض کاسبرگ‌ و طول و عرض گلبرگ و صفات بیوشیمیایی شامل کلروفیل a، کلروفیل b، کاروتنوئیدها، آنتوسیانین و موسیلاژ اندازه‌گیری شدند. گونه‌های مطالعه‌شده ازنظر صفات طول ریشه، تعداد برگ و طول گلبرگ اختلاف معنی‌داری داشتند. با توجه به مقایسه میانگین صفات مورفولوژیکی، دو گونه v. sieheana و v.reichenbachiana قرابت بیشتری را نشان دادند. ازنظر میزان آنتوسیانین و کاروتنوئیدها تفاوت معنی‌داری بین گونه‌های مورد مطالعه مشاهده شد. از مقایسه میانگین صفات بیوشیمیایی نیز شباهت دو گونه v.odarata و v. alba منتج شد. درنهایت به‌منظور تعیین شباهت‌ها و فواصل خویشاوندی و با استفاده از روش تجزیه خوشه‌ای، 5 گونه بنفشه در 3 خوشه جداگانه قرار گرفتند. خوشه اول شامل گونه‌های v.odarata و v. alba خوشه دوم شامل گونه v. ignobilis بود. خوشه سوم نیز گونه‌های v. sieheana و  v.reichenbachianaرا شامل شد.

study of morphological and biochemical diversity in some violet species (viola spp.)

to investigate the morphological and biochemical diversity and determine their relationships, 22 ecotypes of the violet medicinal plant (viola spp.), comprising five species, were collected from different regions of gilan, mazandaran, and chaharmahal and bakhtiari provinces. morphological traits, including root length, flowering stem height, number of leaves, leaf length and width, sepal length and width, and petal length and width, were measured. biochemical traits, including chlorophyll a, chlorophyll b, carotenoids, anthocyanin, and mucilage, were also evaluated. significant differences were observed among the studied species in terms of root length, number of leaves, and petal length. based on the mean comparison of morphological traits, v. sieheana and v. reichenbachiana exhibited greater affinity. regarding anthocyanin and carotenoid content, a significant difference was noted among the species. the mean comparison of biochemical traits revealed a similarity between v. odorata and v. alba. finally, cluster analysis grouped the studied species into three distinct clusters: the first cluster included v. odorata and v. alba, the second cluster comprised v. ignobilis, and the third cluster included v. sieheana and v. reichenbachiana.introductionthe use of morphological, biochemical, and physiological traits remains one of the traditional methods for classifying and evaluating plant diversity and is still widely applied in various studies. the distribution of a species across different geographical and altitudinal regions induces diversity in its morphological and physiological characteristics. this diversity, driven by environmental factors, can result in population differentiation in many species. the medicinal plant violet (viola spp.) has been used for centuries to treat inflammation, eczema, skin itching, rheumatism, chronic coughs, acute bronchitis, sore throat, and other ailments. its flowers and leaves are rich in essential oils, salicylic acid, saponins, polyphenols, alkaloids, and flavonoids. one of the most significant active compounds in this genus is a group of mini-peptides known as cyclotides, including cycloviolacin. this plant is a valuable genetic resource for pharmaceutical, phytomedical, and breeding research. the plant extract contains a diverse array of secondary metabolites, enabling the production of medicines, combating pathogens and pests, and resisting biotic and abiotic stresses. to fully utilize the potential of this medicinal plant, it is essential to conduct comprehensive research on the morphological, biochemical, and molecular traits of various species and ecotypes of violets. the observed differences in morphological traits, biochemical contents, secondary metabolites, and molecular diversity among violet species emphasize the importance of studying these aspects in detail. this research aims to investigate the morphological and biochemical diversity of some species of the medicinal plant violet. understanding the genetic diversity of this plant will significantly enhance efforts to identify the status of its genetic resources and provide valuable information for future studies on its applications. materials & methodsin this study, to determine the affinities and distances among species, morphological and biochemical traits of violet species were analyzed across 22 ecotypes representing 5 different species, including viola odorata l. (7 ecotypes), v. alba besser (6 ecotypes), v. ignobilis rupr. (3 ecotypes), v. sieheana w. becker (3 ecotypes), and v. reichenbachiana jord. ex boreau (3 ecotypes). five plant samples from each ecotype were collected from their natural habitats, with precise locations recorded using the universal transverse mercator (utm) coordinate system (table 1). simultaneously, herbarium samples were prepared for identification, verification, and herbarium code assignment and were sent to the herbarium of sfahan for registration and storage. at the sampling sites, morphological traits were measured with a precision of 1 millimeter. these traits included root length, flowering stem height, number of leaves, leaf length and width, sepal length and width, and petal length and width. statistical analyses were performed using sas statistical software. for the extraction of metabolites and biochemical compounds, complete plant samples from each ecotype were collected. fresh petals and leaves were immediately frozen in liquid nitrogen and transferred to a -80°c freezer until extraction. the measured biochemical traits included photosynthetic pigments, carotenoids, mucilage, and anthocyanin content. finally, statistical analyses, including variance analysis, mean trait comparison, and cluster analysis, were conducted using the data obtained from the measurements of morphological and biochemical traits. the affinities among violet species were examined based on the results of these analyses. research findingsthe study compared the morphological traits of five violet species, revealing that viola sieheana had the longest average root length, while v. odorata had the shortest. no significant differences were observed between v. sieheana and v. reichenbachiana for all examined traits. analysis of variance showed a significant difference at the 5% level among species for root length. positive and significant correlations were identified between root length and leaf length, while negative correlations were observed with petal width at the 5% level. v. odorata exhibited the longest average stem length, whereas v. ignobilis had the shortest. no significant differences were detected among the species for stem length. positive correlations at the 1% level were noted between stem height and sepal length. the highest number of leaves was recorded in v. sieheana and v. reichenbachiana, while the lowest was found in v. odorata, with significant differences at the 1% level. negative correlations at the 1% level were identified between the number of leaves and petal width. no significant differences were observed for leaf length and width or sepal length and width. regarding petal length, v. sieheana had the longest petals, followed by v. odorata, with significant differences at the 5% level. in terms of biochemical traits, v. sieheana exhibited the highest chlorophyll a content, whereas v. odorata showed the highest chlorophyll b content. however, no significant differences were observed among the species for these traits. positive correlations at the 5% level were found between chlorophyll a, chlorophyll b, and carotenoid content. v. ignobilis had the highest carotenoid and anthocyanin contents. cluster analysis grouped the ecotypes into two main clusters. the first cluster included 14 ecotypes comprising v. odorata, v. alba, and v. ignobilis. the second cluster consisted of 8 ecotypes, including v. reichenbachiana and v. sieheana. discussion of results & conclusionmorphological evaluation plays a vital role in managing collections, confirming sample identities, identifying errors in identification, and determining phenological relationships. although violet plants grow wild in some forests of iran, particularly in the northern regions, limited research has been conducted on their morphological and biochemical characteristics. the distribution of species across different geographical and altitudinal regions contributes to diversity in their morphological and biochemical traits. in this study of five violet species, significant differences were observed in root length, number of leaves, and petal length. among the studied species, viola odorata and v. alba exhibited a wide distribution across various altitudes, whereas the other species were primarily found in mountainous and high-altitude regions. morphological trait measurements revealed that the longest leaf lengths were recorded in low-altitude areas, while the longest root lengths were observed in high-altitude areas. this suggests that in low-altitude regions, where soil development constraints are minimal and nutrients are readily available, roots penetrate less deeply and grow more superficially. as a result, the aerial parts, such as the number of leaves, leaf dimensions, and stem length, show improved growth. in the biochemical and phytochemical trait analysis, significant differences were found in carotenoid and anthocyanin content among species, while no significant differences were observed for mucilage content. the findings indicate that biochemical synthesis in violets tends to increase at higher altitudes. by combining morphological and biochemical data, the study clustered v. odorata and v. alba together, while v. reichenbachiana and v. sieheana formed another group, with v. ignobilis displaying intermediate traits. these results highlight the importance of integrating morphological and biochemical evaluations for understanding species diversity. however, further studies focusing on molecular and genetic diversity assessments are recommended to achieve more precise and comprehensive results.