Tracing Oregano Diversity: Genetic Variability Revealed by RAPD Markers | InformativeBD

Hafiza Faiza, Nadir Ali Rind,  Muhammad Rafiq, and  Muhammad Umar Dahot, from the institute of Pakistan. wrote a Research Article about, Tracing Oregano Diversity: Genetic Variability Revealed by RAPD Markers. Entitled, Study of genetic variability of local Oregano (Origanum vulgare) through RAPD markers. This research paper published by the Journal of Biodiversity and Environmental Sciences | JBES. an open access scholarly research journal on Biodiversity. under the affiliation of the International Network For Natural Sciences| INNSpub. an open access multidisciplinary research journal publisher.

Abstract 

In the present study Origanum vulgare seeds were exposed to chemical mutagens at different concentration of Ethyl methane sulphonate and Sodium azide. Mutants and control (without mutagen treatment) of Origanum vulgare seeds were than culture in-vitro on MS media (supplemented with hormones) for regeneration and callus induction. DNA was isolated from all treated and control Origanum vulgare and polymorphism of isolated DNA was studied using six RAPD markers. In this study six RAPD primers were used as markers to study the genetic differences in mutagenized Oregano plants and callus cultures. The DNA samples of plants and callus cultures showed common bands. As common amplified bands in RAPD separated in agarose gels also showed variable intensity of their bands. This type of polymorphism may be due to different copy numbers of corresponding DNA loci. RAPD showed variations in mutagenized Oregano plants and callus cultures. The callus cultures can be used for several studies like for mass propagation, production of secondary compounds and gene transfer studies. RAPD has frequently used as molecular marker for genetic variability studies in plants due to its simplicity, rapidity and lack of prior genetic information about the plant.

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Introduction

Tissue culture is the method through which genetic exploitation studies can be done fruitfully (Parrott et al. 1994). Molecular marker practices, for example random amplified polymorphic DNA (RAPD) analysis, have been used to evaluate somaclonal changes (Al-Zahim et al., 1999; Hashmi et al., (1997), Rind et al., (2016) Rout and Das, (2002) and Zucchi et al., (2002) in addition to assess the effects of growth regulators (Mangolin et al., (2002) and chemical mutagens (Teparkum and Veilleux, 1998) in various discrete loci inside a genome (Samal et al., (2003). As primers customized by even a single nucleotide create diverse banding profiles, the RAPD technique can produce polymorphisms among very closely-related genotypes (Deng et al., (1995). The RAPD assay is well-organized for selection of nucleotide sequence polymorphism between individuals as each primer (on average) will straight to the amplification of numerous discrete loci inside a genomeSamal et al., (2003).

Origanum onites and Origanum vulgare taxa can be known by the most flawlessly using RAPD markers. RAPD has also been used to differentiate extraordinary plant material under the name Mediterranean oregano for class control (Marieschi et al.(2009). Currently, many special objectives have been effectively recognized by the concerned of molecular markers, on different oregano species for example RAPD in Origanum majorana (Klocke et al., (2002). and in Origanum subspecies (Katsiotis et al., (2009), and simple sequence repeat (SSR) in Origanum vulgare (Novak et al., (2008). Though, it is not yet finely available about the connection among genetic diversity and chemical compounds of essential oils of Origanum onites. RAPD is still one of the most successfully and frequently used molecular techniques because of its simplicity, low cost and high speed. Thus, genetic diversity among diverse genotypes of many crops can be easily and rapidly evaluate by victoriously using RAPD markers (Rafalski and Tingey, (1993), Ragot and Hoisington, (1993), Williams et al., (1990). Universally, RAPD can present valuable data for the assessment of population, genetic structure as well as genetic diversity within and among populations, population sections and degree of inbreeding and individual relation (Ragot and Hoisington, (1993). In case of herbaceous plants, RAPD markers have been hardly applying for genetic diversity studies, e.g. in Origanum majorana L. (Klocke et al., (2002), Cunila galioides Benth (Fracaro et al., (2005), Curcuma zedoaria (Christm) Rosc (Islam et al., (2007), Syamkumar and Sasikumar, (2007), Matricaria chamomilla L. (Solouki et al., (2008), Satureja hortensis L. Solouki et al., (2008) and Foeniculum vulgare Mill Zahid et al., (2009). RAPD-PCR is a quick attempt in which no complex technology and no previous sequence information is necessary, it also allows high-quality appearance levels when the DNA to be probed in its low quantities. Marieschi et al., (2009) have used RAPD-PCR method to evaluate its competence as correspondingly, quick and steady assay to probe the attendance of existing contamination and speed pharmacognostic examination of large batches of samples.

In recent study, six different arbitrary primers were used as RAPD markers to assess the genetic diversity among mutagen treated plants and callus cultures. Origanum vulgare germplasm collected from the local market of Hyderabad, Pakistan.

The seeds were treated with various concentrations of EMS and Na2N grown in the field and under in vitro conditions, mutant plantlets and four to six month old calli were used to study the genetic variability created through chemical mutagens in plants and callus cultures through the use of Random amplified polymorphic DNA.

Reference

Al-Zahim M, Ford-Lloyd B, Newbury H. 1999. Detection of somaclonal variation in garlic (Allium sativum L.) using RAPD and cytological analysis. Plant Cell Reports1 8, 473-477.

Bogani P, Simoni A, Lio’ P, Germinario A, Buiatti M. 2001. Molecular variation in plant cell populations evolving in vitro in different physiological contexts. Genome 44, 549-558.

Fracaro F, Zacaria J, Echeverrigaray S. 2005. RAPD based genetic relationships between populations of three chemotypes of Cunila galioides Benth. Biochemical systematics and ecology 33, 409-417.

Hashmi G, Huettel R, Meyer R, Krusberg L, Hammerschlag F. 1997. RAPD analysis of somaclonal variants derived from embryo callus cultures of peach. Plant cell reports 16, 624-627.

Islam M, Meister A, Schubert V, Kloppstech K, Esch E. 2007. Genetic diversity and cytogenetic analyses in Curcuma zedoaria (Christm.) Roscoe from Bangladesh. Genetic Resources and Crop Evolution 54, 149-156.

Katsiotis A, Nikoloudakis N, Linos A, Drossou A, Constantinidis T. 2009. Phylogenetic relationships in Origanum spp. based on rDNA sequences and intra-genetic variation of Greek O. vulgare subsp. hirtum revealed by RAPD. Scientia horticulturae 121, 103-108.

Khan IA, Dahot MU, Seema N, Bibi S, Khatri A. 2008. Genetic variability in plantlets derived from callus culture in sugarcane. Pakistan Journal of Botany 40, 547-564.

Klocke E, Langbehn J, Grewe C, Pank F. 2002. DNA Fingerprinting by RAPD on Origanum majorana L. Journal of Herbs, Spices & Medicinal Plants9, 171-176.

Kozyrenko M, Artyukova E, Lauve L, Zhuravlev YN, Reunova G. 2001. Genetic variability of the Panax ginseng callus lines. Russian Journal of Biotechnology C/C of Biotekhnologiia 1, 29-34.

Mangolin CA, Ottoboni LM, Maria de Fátima P. 2002. RAPD markers to evaluate callus tissue of Cereus peruvianus Mill. (Cactaceae) maintained in different growth regulator combinations. Biochemical Genetics 40, 351-358.

Marieschi M, Torelli A, Poli F, Sacchetti G, Bruni R. 2009. RAPD-based method for the quality control of Mediterranean oregano and its contribution to pharmacognostic techniques. Journal of agricultural and food chemistry 57, 1835-1840.

Martin K. 2002. Rapid propagation of Holostemma adakodien Schult., a rare medicinal plant, through axillary bud multiplication and indirect organogenesis. Plant Cell Reports 21, 112-117.

Morell M, Peakall R, Appels R, Preston L, Lloyd H. 1995. DNA profiling techniques for plant variety identification. Australian Journal of Experimental Agriculture 35, 807-819.

Murray MG, Thompson WF. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic acids research 8, 4321-4326.

Nei M, Li W-H. 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proceedings of the National Academy of Sciences 76, 5269-5273.

Novak J, Lukas B, Bolzer K, Grausgruber‐Gröger S, Degenhardt J. 2008. Identification and characterization of simple sequence repeat markers from a glandular Origanum vulgare expressed sequence tag. Molecular Ecology Resources 8, 599-601.

Parrott W, All J, Adang M, Bailey M, Boerma H, Stewart C. 1994. Recovery and evaluation of soybean plants transgenic for a Bacillus thuringiensis var. Kurstaki insecticidal gene. In Vitro Cellular & Developmental Biology-Plant 30, 144-149.

Rafalski JA, Tingey SV. 1993. Genetic diagnostics in plant breeding: RAPDs, microsatellites and machines. Trends in Genetics9, 275-280.

Ragot M, Hoisington D. 1993. Molecular markers for plant breeding: comparisons of RFLP and RAPD genotyping costs. Theoretical and Applied Genetics 86, 975-984.

Rind NA, Aksoy O, Dahot, MU, Dikilitas S, Rafiq M, Tutunoglu B. 2016. Evaluation of Genetic Diversity amaong Melia azedarach L (Meliaceae) with RAPD markers. Fresenius Environmental Bulletin 25, 2374-2382.

Rout G, Das G. 2002. An assessment of genetic integrity of micropropagated plants of Plumbago zeylanica by RAPD markers. Biologia plantarum 45, 27-32.

Salvi ND, George L, Eapen S. 2001. Plant regeneration from leaf base callus of turmeric and random amplified polymorphic DNA analysis of regenerated plants. Plant cell, tissue and organ culture 66, 113-119.

Samal S, Rout G, Nayak S, Nanda R, Lenka P, Das P. 2003. Primer screening and optimization for RAPD analysis of cashew. Biologia plantarum 46, 301-304.

Solouki M, Mehdikhani H, Zeinali H, Emamjomeh A. 2008. Study of genetic diversity in Chamomile (Matricaria chamomilla) based on morphological traits and molecular markers. Scientia Horticulturae 117, 281-287.

Sridhar T, Naidu C. 2011. An efficient callus induction and plant regeneration of Solanum nigrum (L.)-An important antiulcer medicinal plant. Journal of Phytology 3.

Stegniĭ V, Chudinova I, Salina E. 2000. RAPD analysis of the flax (Linum usitatissimum L.) varieties and hybrids of various productivity. Genetika 36, 1370-1373.

Syamkumar S, Sasikumar B. 2007. Molecular marker based genetic diversity analysis of Curcuma species from India. Scientia Horticulturae 112, 235-241.

Teparkum S, Veilleux RE. 1998. Indifference of potato anther culture to colchicine and genetic similarity among anther-derived monoploid regenerants determined by RAPD analysis. Plant cell, Tissue and Organ Culture 53, 49-58.

Tyagi RK, Agrawal A, Mahalakshmi C, Hussain Z, Tyagi H. 2007. Low-cost media for in vitro conservation of turmeric (Curcuma longa L.) and genetic stability assessment using RAPD markers. In Vitro Cellular & Developmental Biology-Plant 43, 51-58.

Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic acids research 18, 6531-6535.

Zahid N, Abbasi N, Hafiz I, Ahmad Z. 2009. Genetic diversity of indigenous fennel (Foeniculum vulgare Mill). Germplasm in Pakistan assessed. By RAPD markers. Pak. J. Bot 41, 1759-1767.

Zucchi MI, Arizono H, Morais VA, Fungaro MHP, Vieira MLC. 2002. Genetic instability of sugarcane plants derived from meristem cultures. Genetics and Molecular Biology 25, 91-96.

Article source : Study of genetic variability of local Oregano (Origanum vulgare) through RAPD markers