Anatomical Variation and Molecular Characterization of Catharanthus roseus Using RAPD Analysis
Abstract
Catharanthus roseus is one of the most interesting groups of ornamental plants in the world with high medicinal value and member of the family Apocynaceae. The goal of this study was to investigate the anatomical and molecular characterization of five types (White yellow, White red, Pink yellow, Pink red, and Red yellow) of C. roseus plant. The anatomical study was made by cutting transverse sections of the materials and stained with double stained techniques and observed data with the help of high-powered compound microscope. Molecular analysis was done by using RAPD molecular markers with four primers. Anatomical analysis indicated that the epidermis was recorded as a single layer. Bi-collateral vascular bundle was present and frequently arranged in radial symmetry in the stem. The results of RAPD markers revealed a total of sixty-nine (69) amplified bands, forty-six (46) of them were monomorphic and twenty-three (23) of them were polymorphic from using four primers. The result of dendrogram separated the five types of C. roseus into two major clusters (C1 and C2). Cluster C1 is represented by a single type Red yellow indicating that it is distinct from other four types, and Cluster C2 is represented by the other four types of C. roseus. The dendrogram and linkage distance revealed that the highest similarity was 70% between S1 (White yellow types) and S2 (White red types). However, the lowest similarity was 41.7% between S3 (Pink yellow types) and S5 (Red yellow types). This study could provide a key platform for further crop improvement at molecular level for genetic variability and cross breeding.
Introduction
Catharanthus roseus (L.) is a perennial tropical plant. C. roseus also referred to as Nayantara” or“Sadabahar”. It is also referred to as vinca, bright eyes, old-maid, periwinkle, pink periwinkle, rose periwinkle, and madagascar periwinkle. The word Catharanthus derives from the Greek language meaning "pure flower." While, roseus means red, roseor rosy. Although C. roseus is native to Madagascar, it has been grown as an ornamental plant for centuries in the tropics and occasionally in the subtropics. As are sult, it has naturalized in many places. Madagascar, an island in the Indian Ocean, is home to C. roseus. C. roseus is simultaneously an ornamental and amedicinal plant species. It is a member of the Apocynaceae family, which has 411 genera and 4650species, many of which have ornamental and therapeutic uses (Simpson, 2006).
It is one of the most interesting groups of or namental plants in the world, considerable variation of different colored flowers can be observed including purple, red, pink, or white corolla (Plaizier, 1981). According to Kumar et al., (2013) and Nejat et al., (2015), there are five variations of flower’s color, namely white-yellow, white-red, pink-red, pink-white, and red white.
C. roseus is an important medicinal plant. It has been used in folkloric remedies for the treatment of many different disease including diabetes (Ahmed et al.,2010) malaria (Gathirwa et al., 2007) insect bites diarrhea (Sukumar and Osmani, 1981), skin, eye and throat inflammations, indigestion, toothache, fever and lung congestion (Nejat et al., 2015) menstrual disorder (Kumar et al., 2013 ) enhances kidney and liver functions (Adekomi, 2010) vinblastine alkaloidsare used in the treatment of various types of lymphoma and leukemia (Lucas et al., 2010).Antimicrobial, antitumor, Anti-proliferative activityof extracts from the plant has been proved (Patil andGhosh, 2010; Vega-´ Avila, 2012).
It contains flavonoids, saponins, tannins, and several anticancer alkaloids namely vinblastine (VLB),vincristine (VCR) and leurosin (Jaleel et al., 2008;Pandiangan, 2012; Nejat et al., 2015). The leaves ofthis plant are used as alkaloid-producing the rapeutic plant components (Renault et al., widely 1999).Several studies revealed that the leaves of this medicinal plant can treat various diseases.
The leaves are infused and used to treat me norrhagia.To treat wasp stings, the leaf juice is externally applied. Alkaloids are abundant throughout the entire plant, with the root bark containing the highest concentrations, especially during flowering. All partsof the plant are also said to have hypoglycemic and antioxidant properties. This plant has long been use das a remedy for hypertension, diabetes mellitus, malaria, constipation, and diuretics.
The assessment of the genetic diversity of Catharanthus species and cultivars was accomplished by secondary metabolites, according to the literature. Inter simple sequence repeat (ISSR), amplified fragment-length polymorphism (AFLP), random amplified polymorphic DNA (RAPD), and other techniques. (Kalpana et al., 2004; Arif et al., 2010;Leal et al., 2010; Chaudhary et al., 2012; El Domyatiet al., 2012). They provide quick results by revealing genetic variations without regard to stage, physiological factors, or surroundings. It isanticipated that biochemical genetic markers wouldbe crucial in characterizing the genotypes of therapeutic plants (Tharachand et al., 2012). RAPD iswidely used to study the genetic diversity of many plants (Shaw et al., 2009; El- Domyati et al., 2012; Lalet al., 2011), as this method is straightforward, quick, and affordable in comparison to other kinds of DNA based approaches. Kim et al., (2007) Shaw et al.,(2009) Vardhan et al., (2012) and Prasad (2014)discovered low- moderate to high genetic variation among Catharanthus species and cultivars.
The aim of this study to investigate the internodes, leaf midrib, and petiole of Catharanthus roseus to determine their gross anatomical characteristics and anatomical quantitative and qualitative traits as well as to ascertain the genetic diversity and relationships between the various types of C. roseus.123 Sima and AfsanaInt. J. Biosci.
Source : Anatomical variation and molecular characterization of Catharanthus roseus through RAPD analysis
