Inter Specific Genetic Diversity Within the Ethnomedicinally Important Ipomoea L
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Research Article Int J Environ Sci Nat Res Volume 19 Issue 3 - May 2019 Copyright © All rights are reserved by Noor Muhammad DOI: 10.19080/IJESNR.2019.19.556012 Inter Specific Genetic Diversity within the Ethnomedicinally Important Ipomoea L. Species through Morphological and Biochemical Profiling Noor Muhammad1,2*, Nisar Uddin2, M Khalil Ullah Khan2 and Niaz Ali2 1College of Horticulture, Hebei Agricultural University, China 2Department of Botany, Hazara University Mansehra, KP Submission: April 15, 2019; Published: May 17, 2019 *Corresponding author: Noor Muhammad, College of Horticulture, Hebei Agricultural University, Baoding, China & Department of Botany, Hazara University Mansehra, KP, Pakistan Abstract The present study has attempted to highlight the medicinal importance as well as genetic relationships among of Ipomoea species growing in Swat Valley, KP, Pakistan. There was a general agreement among the informers regarding the medicinal uses of plants and Informants Consensus Factor (ICF) value ranged from 0.991 to 0.965. The study shown that most of the informants agreed in the application of I. hederacea to use as Ipomoea species viz., 14 genotypes of I. hedracea, 14 of I. purpurea morphologicalblood purifier (ICFcharacters 0.991) werethat exhibited counted forthe the highest assessment fidelity oflevel intra (100%). and inter The genetic intra and disagreement inter specific through dissimilarity traits similarityamong the index 28 genotypes and cluster of , were tested using morphometric and biochemical profiling. Twenty-fourI. Purpurea and I hedracea I. hedraceaanalysis. Total seed proteinI. purpurea profiling was carried out on 12% slab gel electrophoresis; 6 reproducible bands were detected in both with molecular weight ranging from 10KDa to 180KDa. Intra locus contribution towardIpomoea the genetic disagreement was 33.33% in on two-way and cluster 66.66% plotting in successfully. In the determined same way, theinter two species species locus into contribution separate clusters toward and genetic the diversity diversity found was 83.33%.between Interestingly, and within Ipomoea locus 2 species(L-2) was may monomorphic offer unique in avenues the collected for novel germplasms drug discovery. and may be shown as generic specific locus for species. SDS-PAGE profiling based Keywords: Ipomoea species; Ethno medicinal uses; Genetic diversity; SDS-PAGE; Cluster analysis Introduction sepals are lanceolate, longattenuate, and caudate. Leaves are The Ipomoea is an important genus belongs to family ovate and deeply 3-lobed with acuminate tips and cordate bases. convolovulaceae; with many species of Ipomoea have wide Stem twine with their apex in a dextral fashion [6-8]. therapeutic uses. Species are broadly dispersed throughout temperate and tropical regions and mostly found in tropical Morphological characterization has important role in the America and tropical Asia [1], used to treat blood disease, sterility exploration of genetic diversity in crop plants but effected by in women, urinary infection, constipation and gynecological environmental changes [9]. On the other hand, genetic diversity disorders [2]. The plant is also having laxative, psychedelic [3] evaluation through molecular methods such as biochemical anticarcinogenic, hepatoprotectivity, oxytocic and antioxidant assessment at protein level and DNA based techniques have a properties [4] as well as in rheumatism and fungal infection [5]. number of advantages over the classical morphology [10] but compared to biochemical assessment at protein level, molecular Ipomoea species are used for its medicinal contents and investigation of DNA level is highly robust but too expensive (Win psychoactive compounds mostly alkaloids. The genus Ipomoea et al., 2011). Among biochemical techniques, Sodium dodecyl comprises of food crops such as the tubers of sweet potatoes and sulphate polyacrylamide gel electrophoresis (SDS-PAGE) method the leaves of water spinachare are commercially important food stuffs. The most important species Ipomoea including I.hederacea SDS-PAGE is now broadly used biochemical practice to designate and I. purpurea are growing in the hilly areas of Swat, Pakistan. The is a simple, reliable, cheap and free of environmental fluxes [11]. the genetic structure of crop germplasm (Hameed et al., 2009). I. purpurea and I. hederacea are distinguishable by sepal and leaf Enormous attention has been focused on the use of SDS-PAGE characteristics. The I. purpurea sepals are ovate-lanceolate with over the last two decades for estimation of genetic diversity, an apex that is acute to abruptly acuminate while I. hederacea’s reliable judgment and identification of plant varieties. Seed Int J Environ Sci Nat Res 19(3): IJESNR.MS.ID.556012 (2019) 0064 International Journal of Environmental Sciences & Natural Resources storage protein markers have been successfully used to resolve percentage of informants demanding the application of a certain taxonomic relationships and characterize cultivated varieties plant species for the same major purpose, was counted for the in a number of crop plant species; An attempt for estimation of genetic relationships among the nine Ipomoea species using the Where, Np = number of informants that claim a use of a plant most frequently reported diseases as: FL (%) = (Np / N) × 100 molecular technique of sodium dodecyl sulphate polyacrylamide species to treat a particular disease; N = number of informants gel electrophoresis (SDS-PAGE) of seed proteins was carried out that use the plants as a medicine to treat any given disease [13]. by Pragati et al. [12]. Morphological characterization Pakistan is rich in naturally distributed species of Ipomea but Qualitative and quantitative descriptors were taken. there are no reports about the ethnomedicinal uses or genetic Qualitative traits were recorded on the general visualization relationship of Ipomoea species based on total seed protein (phenotypic observations). Nine qualitative traits i.e. Leaf Shape, Leaf Color, Leaf Pubescent, Flower color, Seed texture, Hilum uses and SDS-PAGE analyses of 28 genotypes of Pakistani Ipomea profiling available. In this article, we report on ethnomedicinal color, Seed coat color, Seed shape, Spots on the seed coat while species (I. hedracea and I. purpurea) growing in hilly areas of Swat. the quantitative traits were Petiole length, Leaf length, Leaf width, Both these species are locally adapted and are of widespread Seed length, Seed thickness, Seed width, Pod length, Seed per pod, use as ornamental and medicinal purposes. The objective of the Pod per plant, Flower length, Flower width, Seed weight, Branches study to document the ethnomedicinal uses and level of genetic per plant, Plant height, Biomass, Yield per plant. A total of 24 traits polymorphism and relationships among Pakistani Ipomoea were recorded for morphological description. species (I. hedracea and I. purpurea) growing in hilly areas of Swat using morphological and SDS-PAGE characterization. Protein Profiling To assess genetic diversity and genetic relationships, SDS- Material and Methods PAGE was conducted (Lameli, 1970). For seed storage protein Plant materials In the present study several exploratory trips were arranged profile, single seed of each genotypes was crushed into a powder. to different agro-ecological zones of Swat in 2017 – 2018. During 400μl of Protein Extraction Buffer PEB (0.5M Tris-HCL, 0.2%SDS, of seed powder. Then mixture in E-tube was Vortexed thoroughly expedition, of different zones i.e. Swegalai, Kohay, Ziarat, Jawand, 5M Urea, 1%B-mercaptoethanol under 8-pH) was added to 0.01g Gadi, Sharfabad, Rangila, Gharai, Khazana, Aboha, Barikot, to homogenize it. The Comassive Brilliant Blue (CBB) was added Shingardar Manyar, Nawagai, Shahdeari, Sarsinai, Khawazakhela, to the E-tube as tracking dye to see the movement of PEB-FP on Mangultan, Manglor, Chamtalai, Chaliyar, Baidara, Sherpalam, the separation PAG. The homogenated samples were centrifuged Kanju, Bandai, Derai and Hazara were selected for collection of at 13,000 rpm for 10 minutes under room temperature. The samples for Ethno medicinal uses, morphological characterization electrophoretic process was carried out using 12% polyacrylamide Specimens for each specie have been collected and processed and estimation of genetic diversity in seed storage protein profile. gel (composition of resolution gel: 3.0M Tris-HCl pH9.0, 0.4% SDS using standard herbarium techniques [13]. The data was collected and staking gel 0.4M Tris-HCl pH 7.0, 0.4% SDS). The electrode through semi structured interviews with inhabitants involved buffer containing 0.025M Tris, 129M Glycine and 0.125% SDS was in the plant’s management [13]. A total of 120 residents were poured in the Electrophoresis tank. Similarly, 15μl PEB-FP was 100V until the blue line passed through the bottom of gel plates. interviewed for information on uses of plants to cure various loaded in each well of 12% PAG. The electrophoresis was run at diseases of human being, parts used, of medicine have been The PAG were than stained and destained for data scoring of seed collected. Based on the information obtained from the informants storageData analysis protein profile. into 9 groups. The use value (UV) demonstrates the relative in the study area, all the reported diseases have been classified Morphological data was analyzed through cluster plotting importance of locally known plants [14]. It was calculated using using software’s SPSS and PC-ORD. the formula: UV = ∑ Ui/