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Assessment of Genetic Variation in Achillea Nobilis L. by Using ISSR

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Assessment of Genetic Variation in Achillea Nobilis L. by Using ISSR Assessment of Genetic Variation in Achillea nobilis L. by Using ISSR Markers Saleheh Saffar1, Adeleh Saffar2, Mohsen Karimi2 1-Tabriz University, Department of Biology- Iran 2- Gorgan University of Agricultural Sciences and Natural Resources, Department of Biology- Iran . Achillea (Yarrow) is one of the most important [email protected] of the Asteraceae family . Nineteen species of this genus are found in Iran, among which seven are endemic . Different species of Achillea such as A. nobilis is known for many years in the fold medicine. Pharmacological studies have shown these species have antimicrobial , antiallergic and anti-inflammatory activities. .Assessing the levels of genetic variation, and partitioning total genetic variation within and between populations in the cultivated accessions are major concerns in breeding programs, genecology and conservation genetics and it is essential to make valid iological interpretations about its breeding system and reproductive biology Summrary .In this work we study genetic variation in some population of Achillea nobilis L. by using ISSR markers. This study confirms the efficacy of ISSR markers for the identification of plant genotypes. This information should be helpful for breeding and genome mapping programs After collecting plants material from different floristic regions of Golestan Province: . DNA extraction from the leaves by liquid nitrogen. Assesmen of Exteracted DNA by Spectrophotometry & Electrophoresis. PCR reaction with six different ISSR primers. Electrophoresis & the ISSRs loci were scored. The ISSRs amplifications were repeated three times to insure the reproducibility of the banding patterns. The ISSRs loci were scored as 1 for present and 0 for absent. Genetic diversity within populations was estimated based on Nei,s gene index. Exprimental Genetic relationship between populations was measured on the basis of Nei,s genetic distance, F-statistic index and coancestry index. Total partitioning of genetic diversity of within and among populations using analysis of molecular variance (AMOVA) was applied to estimate the variance. A dendrogram show that genetic diversity population by cluster analysis based on UPGMA. In this study ISSR markers was used to detect intra- and inter- population genetic diversity of A. nobilis. 90 genotypes, belonging to Tuskestan six populations, were studied using six ISSR primers, to investigate Ghorogh the genetic structure between populations. Ramian Percentage of polymorphic loci, estimated using Nei's genetic Daland diversity index and Shannon's information index revealed low or moderate levels of genetic variations within each A. nobilis population. Golestan The ISSRs patterns were reproducible and clear for scoring. Applying 0.26 0.36 0.46 0.56 0.66 6 ISSRs primers produced a number of 86 polymorphic loci. Coefficient Cluster analysis grouped these five population into two clusters, Golestan populations and cluster including the other four populations.The analysis of molecular variance showed that most of the variation (65.23 %) occurred among individuals within all five populations, which expected for an outrossing populations. This highergenetic diversity within populations of an outcrossing A. nobilis also reflects a higher level of hetrozygosity. Results and Discussion and Results Results showed that ISSR is the useful marker for investigating the genetic diversity and assessing differentiation between A. nobilis populations belonging to different geographical regions of Iran . These results indicate that the conservation of existing natural population of A. nobilis is necessary by the development of different management practices such as: maintaining effective population sizes, construction of an in situ conservation area, ex situ conservation of seed and domestication of this wild plant species. [1]Farajpour et al. 2012. Assessment of genetic diversity in Achillea millefolium accessions from Iran using ISSR marker. Biochemical Systematics and Ecology. 43, 73–79. [2] Khadivi-Khub et al. 2014. Comprehensive genetic discrimination of Leonurus cardiaca populations by A FLP, ISSR, RAPD and IRAP molecular markers. Molecular Biology Reports. 41(6), 4007-4016. [3]poormohammad. 2013. Application of molecular markers in medicinal plant studies. Acta Universitatis Sapientiae Refrences Agriculture and invironment. 5, 80-90..
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