Assessment of Genetic Diversity Landraces In Onobrychis Sativa Using SSR,RAPD, SRAP markers

Assessment of Genetic Diversity Landraces In Onobrychis Sativa Using SSR,RAPD, SRAP markers | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Assessment of Genetic Diversity Landraces In Onobrychis Sativa Using SSR,RAPD, SRAP markers Mohammad Sharifi Sedeh This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7173060/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In order to evaluate genetic diversity cultivated sainfoin accessions collected from different regions of the world, SSR markers from medicago genus were used. Polymorphism information content value ranged from 0.2 to 0.043 with an average of 0.33. Cluster analysis based on SSR markers grouped the sainfoin population in two main groups (Iranian and exotic cultivate sainfoin) with two sub cluster for each group. In another survey was performed to find out an efficiency of ecological factors on the genetic similarity was investigated among five population of Onobrychis viccifolia East Azarbaijan, Iran using RAPD and bulked DNA, The largest and shortest genetic were detected between Bonab and Heris (0.04904)and Amand and Khosroshahr(0.0572) on UPGMA dendrogram, two population of Sarab and Heris were nested within one cluster, and Amand and Khosroshahr in other cluster. Also Bonab was completely separated from all other population. Biotechnology and Bioengineering Sainfoin Markers Multiphase Key Points I survey Genetic Diversity in sainfoin to select best adjectives in sainfoin .after select best adjectives you can produce best lines for sainfoin modify . And use them as food for animal. Introduction Please include line numbering in your manuscript (Wikipedia.org). According to its seed specification, it has been divided into 270 categories. Resistance to the dryness is the most important characteristic of this plant, making it appropriate for the dry farming and pastures (Hume, et al., 1985). The larger content of tannins in common sainfoin leaves, prevents bloat and protein hydrolysis in the livestock rumen, and increases amino acid absorption in the intestine of ruminants (Carbanero et al. 2011). Fabaceae provender is of great importance Moreover, knowledge about the rate of polymorphism with differences between different examples is an important in order to reduce the rate of hereditary resources, kept in the gene bank and analyzing the purity of seeds in the cross- pollinated plants (Brummer, 1999). Studying genetic diversity via morphologic marker is accompanied with limitations such as low polymorphism, low heritability, and effectiveness due to environmental conditions (Werlemark et al. 1999). The Molecular markers reveal genetic diversity in DNA surface; do not show having the mentioned problems as in the morphologic indicators. Moreover, molecular markers are prominent tools in an evaluating relational selection of better plants as well as for analyzing the similarities and differences among various species (Awasthi et al .). In this regard, micro-satellites or simple sequential repeats (SSR) are the repeats of 106 pairs of DNA sequence that primarily was thought to be only exist in non-codifying regions of DNA sequence, while occurrence time of micro-satellites in codifying sequences or translated sequences were found by the passage of time, and EST-SSRs were developed (Choudhary et al., 2009). Micro-satellite markers usually shows the higher level of polymorphism, and by considering the frequency of alleles in each gene place, they can be distinguished a very similar species (Kuleung et al., 2008) Extracting SSR primer by genome libraries is an expensive and tedious method, requiring proper skills. By having access to a number of ESTs and other information about DNA sequence, developing SSR markers via extracting information has become effective and lower costs for many plant species (Eujayl et al., 2000; Varshney et al,. 2005). Although many EST databanks for modelled and non-modelled creatures are available in the recent years and new generation technologies for finding sequences have created a great evolution in the genetic researches, but its share for a plant like common sainfoin is rather little in the databanks. Thus, a simple method for developing a SSR primer in the species that are not studied is a great deal id using SSR primer for related species. Genomix has made an apparent comparison, and the order and contents of genes among closely related species are greatly protected. Harmony of markers shown in a comparative drawing indicates, marker of the same material or type can exist in another material or type (Kuleung et al., 2004). The Number of 27 SSR marker of Medicago sativa and Glycin max was tested for transferring in O. vicifolia species by Demdoum et al. (2011), and after identifying the markers with the potential to create polymorphs, 6 pairs of SSR markers were used for fingerprinting and estimating genetic similarities in 23 farming sainfoins and two wild types. Thus, masses of farming sainfoins of two wild types of sainfoils and 23 farming sainfoins based on geographical origins were separated (Demdoum et al., 2011). In another study by Avci et al. (2014), they identified the transferability of 95 markers for Medicago truncatula and phaseolus sativa to sainfoin, and 18 markers for analyzing the genetic diversity of sainfoin type. RAPD markers are used extensively in the studies regarding diversity, structure, and genetic similarities of the populations of many plant species (Williams et al. 1990 Nybomaind Bartish, 2000). These markers are suitable due studying genome, for studying the ordinary selection on the genetic structure of plant populations. Although this method has limitations, such as having a dominant nature and not identifying homozygosity against heterozygosity natures, hence, SRAP markers were used that are simple, and have high reliability, average overall coverage of the genome and potentials for reproduction in codifying regions in the genome. Materials and Methods The population of the considered farming sainfoin included 5 masses of Iranian and 17 foreign sainfoin masses, provided by direct collection and sampling from gene banks (IPK/ Germany and USDA/ USA) (Table 1). Table 1. Ecological characteristic of cultivated sainfoin population. S.No Population Height from Sea Surface(m) Geographical wide Weather 1 Banab 1290 37/3292 Warm 2 Amand 1400 38/1878 Warm 3 Khosrosha hr 1500 38/0011 Semi warm 4 Sarab 1680 37/9264 Cold 5 Haris 1900 38/2556 Cold A population of local Medicago sativa alfalfa was also used as the control sample. Two sets of seeds were cultivated in medium plastic pots in the research farm of Isfahan University of Technology in spring 2012. Two weeks after growing of seedlings, 10 samples were randomly selected from each population, and sampling was done from healthy and fresh leaves. Extraction of DNA from the leaf samples of sainfoin was done by Murry and Thompson method (1980). To determine the quality and quantity of DNA, the gel (0.7%) of agars in TBA 0.5x of ethidium bromide dye (10mg/ml) and Gel Doc device were used. Since the micro- satellites (SSR) with “ Onobrychis are not known, screening the identified SSRs with medicago provides the possibility of finding similar regions on onobrychis genome. In this study, 22 pairs of SSR and EST-SSR primer of medicago containing high polymorphism information, content were selected from different resources (Table 2). Table 2. Various pair of SSR and EST-SSR primer of medicago. S.no Code Country Geographical wide Geographical length Height 1 VICUNS8 Iran 53/00 32/00 1200 2 VICESfS9 Iran 53/00 32/00 1200 3 VICUNS18 Iran 53/00 32/00 1200 4 VICUNS39 Iran 53/00 32/00 1200 5 VICUNS42 Iran 53/00 32/00 1200 6 VICRUS112 Russia 55/75 37/61 144/17 7 VICRUS113 Russia 55/75 37/61 144/17 8 VICRUS122 Russia 55/75 37/61 144/17 9 VICGES104 Kyrgyzstan 48/01 67/1 342/9 10 VICGES108 Kyrgyzstan 48/01 67/1 342/9 11 VICMAS134 Hungary 47/26 19/15 378/2 12 VICENS107 England 51/5 0/11 12/19 13 VICENS118 England 51/5 0/11 12/19 14 VICASS127 Slovakia 48/66 19/68 1081/12 15 VICGES119 Germany 51/15 10/45 459/94 16 VICHOS130 Netherland 52/11 5/28 7/92 17 VICSWS123 Switzerland 46/81 8/21 1777/59 18 VICSWS124 Switzerland 46/81 8/21 - 19 VICROS114 Romania 45/93 24/96 596/18 20 VICROS120 Romania 45/93 24/96 596/18 21 VICSPS115 Spain 40/27 3/73 895/8 22 VICUNS106 Unknown - - - Polymerase chain reaction (PCR) for the samples in 15µL volume including 1.5 µL of buffer 10x, 1.5 µL of magnesium chloride, a unit of tag polymerase, 0.3 mmol of dNTP, 0.25 µmol. Of each starter, and 50 ng of genome DNA was prepared from each sample. Reaction was made in a thermal cycler device in a primary blotting cycle in 94°C for 4 minutes, and 35 cycles of secondary blotting in 94°C for 1 minutes, with connection of the primer to DNA string in 50-55°C for 45 seconds, and expansion of synthesized DNA in 72°C for 2 minutes, as well as a final expansion cycle in 72°C for 10 minutes. Polymerase chain product went under electrophoresis on acrylamide gel (12%) and dyeing was done by silver nitrate (Rohlf, 1998). A percentage of polymorph for each starter was obtained by division of polymorphic places and the number of alleles in the places. The polymorphic information content (PIC) was calculated for each band by the following relation (Botstemeta, 1980). Pij is the abundance of amino allele in marker “i”, population is “j”, and “n” is the number of alleles. In this study, 5 different populations of farming sainfoin that are growing wild in different regions in East Azerbaijan province were analyzed. Ten (10) samples of the plant were randomly selected from each population. Geographically, the individual plants in this population are away from each other, in 50 km distances. The studied populations are different with regards to the sea levels, geographical latitude, weather conditions, and soil salinity (Table 1). Two populations in Amand and Khosroshahr are grown in rather salty soil, and in rather close geographical distances. Two populations in Heris and Sarab are growing in cold climates and higher sea levels, in closer distances, while the population in Bonab is comparatively in further distances than the other 4 populations, and ecologically, due to growing in properly salty soil, it is different to the other populations. Regarding geographical latitudes, the populations in Sarab and Khosroshahr are closer to each other, and the populations in Heris and Amand are in almost equivalent geographical latitudes, while the latitude in Bonab has an enormous difference with others. In this test, DNA from young seedlings and seeds were extracted by Madden (2002) method, which is a faster and more economical method in comparison with other common methods in DNA extraction. The DNA of an individual in a population after extraction were mixed, in order to only prepare one DNA sample, i.e. mass DNA, as the representative of that population. Estimating the density of extracting DNA was done by electrophoresis of agar gel (0.8%) and spectrophotometry in 260 nm, to adjust the final density of DNA samples in 10ng/ml. Polymorphism of RAPD strings was tested for 5 studied populations by using 10 random starters. Hence, the contents of 10 ng of model DNA and primer with a density of 100 PM/µL in the final volume of 25µL were analyzed. The number of 17 different bulks of sainfoin plant were collected that belongs to the four species: O.viciifolia (13 bulks), O.viciifolia (1 bulk), O.viciifolia (1 bulk), and O.viciifolia (1 bulk) from different regions in the country, and after extraction of DNA from their fresh leaves and determining their quality and quantity, the extracted DNA was used in PCR reactions, and 88 strings were obtained from the total of 10 primer compositions, 73 of which were polymorphic. Eight pairs of SSR primer and one pair of EST- SSR primer in sainfoin out of 26 pairs of SSR and EST-SSR primers of medicago type (Table 2) indicated polymorphic forms (Table 3), which were used for analyzing genetic diversity in farming sainfoin populations. Table 3. Genetical distance matrix between five population of Onobrychis sativa . Population Amand Bonab Sarab Khosroshahr Haris Amand 0 - - - - Bonab 0.3851 0 - - - Sarab 0.1178 0.3851 0 - Khosroshahr 0.0572 0.3363 0.1178 0 - Haris 0.0870 0.4904 0.0870 0.1495 0 Nine(09) pairs of used micro-satellite primers could separate 44 alleles for evaluating 22 populations of sainfoins, and the number of alleles varied between 4 to 6, and the size of reproducing components were in the range of 100-400 pairs. Polymorphic information content was changing between 0.2 to 0.43 distances for studied micro- satellite places, having an average of 0.33. Only half of the 10 primers produced clear and gradable polymorphic strings. To ensure from repeatability of the produced strings, PCR was repeated for the studied samples with the 5 primers, for three times, and the patterns of strings were similar for the three repetitions, indicating the repeatability of the mentioned strings. The five primers produced 12 polymorphic strings in total, and the highest rate of polymorphic strings was related to no. 1 and no. 2 primers, each with 3 (0.25) polymorphic strings and the lowest rate was related to no. 2, no. 3, and no. 5 primers, each with 2 (16.7%) polymorphic strings. For the matrix of calculated genetic distance, the greatest genetic distance (0.4904) was between the populations of Bonab and Heris, and the lowest distance (0.0572) was between the populations of Amand and Khosroshahr, and 0.087 in the next stage, between populations of Heris and Sarab and also between Heris and Amand, based on the genetic distance of “Nei” on a UPGMA dendrogram. With the greatest genetic similarities, the populations of Amand and Khosroshahr were placed in a cluster, while the populations of Sarab and Heris were in another cluster, and the population of Bonab was placed in a separate cluster due to the least amount of genetic similarity as compared to the other mentioned populations. Discussion the genetic diversity of 48 individual plants from four cultivated populations of alfalfa (Medicago sativa L.) in Iran was assessed using sequence-linked amplified polymorphic markers (SRAP). Fourteen SRAP primer combinations produced 193 fragments, of which 95 were polymorphic. The number of polymorphic fragments identified in each primer combination ranged from 3 to 10 bands with an average of 6.78 bands. The average polymorphic information content (PIC) for all primer combinations was 0.343. Although AMOVA (analysis of molecular variance) results showed significant differences in genetic diversity among populations (P < 0.0001), genetic diversity, which was mainly due to intrapopulation variation, accounted for 93.17% of the total genetic diversity. Analysis of marker data using unweighted pair-group arithmetic mean (UPGMA) method clearly separated subtropical (Yazdi) and subcold (Hamedani and Nikshahri) populations as well as the improved Kodi population. It can be concluded that SRAP markers are useful for studying diversity and relationships between and within alfalfa populations. Overall, the results shows that the obtained markers of medicago type can be suitable resources for identifying the place of micro- satellites in sainfoin, micro-satellite markers considered to have capacity for analyzing relationships and genetic diversity in sainfoin plants. Moreover, the used SSR markers in the present study show the high polymorphism. Populations of an Iranian and foreign sainfoin plants were place in a separate population, indicating a high genetic diversity in the considered germplasm. Moreover, high genetic diversity between Iranian and foreign sainfoin populations is suitable for modification programs in order to reach maximum heterosis. On analyzing the genetic diversity, SRAP marker in sainfoin plant genetic diversity between the masses, the range of genetic distance between masses was found to be in between 124 and 349. Regarding the dispersion of bulks (masses), the lowest genetic distance was between the masses related to Russia (codes P and Q), and the greatest distance was in between the Damavand and Najafabad regions (codes G and M). The numerical value (33.1) obtained for gene movements between groups shows genetic moves in the studied populations, but the move is not to as such to cause homogeneity of in the populations. The range for information index for primer compositions was calculated to be in between 31 and 52. High overall heritability values for height, number of stems and percentage of stems indicate the major role of genetic effects in controlling these traits. Given that these traits are also considered components of forage yield, indirect selection through them can increase the return from selection for improved forage yield. China was superior to China in terms of forage yield. Iranian varieties had a significant advantage over foreign varieties in terms of yield and its most important components. On the other hand, foreign varieties were superior only in terms of leaf percentage and leaf to stem ratio, which are considered as criteria for palatability and forage quality. It is Results The present study results on the farming of wild populations of sainfoin shows the populations grow in a similar ecologic conditions (similar ecotype) and are in a close distance to each other have more genetic similarities. According to the present study findings using DNA, RAPD, we can identify ability of different ecotypes of the cultivated species. In cluster analysis of SRAP data, dendrograms drawn by the NTSYS VERSION 2 could not place the used genotypes in the separate groups. In other words, mixture of masses with each other indicates little inter-mass genetic diversity. Analysis of the main coordination also showed the first three components justified 62.95% of the changes. Thus, analysis of clusters by using SRAP data is a best method for evaluating inter-mass genetic diversity of sainfoin. Declarations Source of funding: None. Conflict of interest: The There is no financial conflict with the subject matter discussed in the manuscript. References Awasthi, A.K., Nagaraja, G.M., Naik, G.V., Kanginakudru, S., Thangavelu, K., and Nagaraju, J.(2004). Genetic diversity and relationships in mulberry (genus Morus ) as revealed by RAPD and ISSR marker assays. BMC Genetics, 5:1–9. Avci et al.(2014). Analysis of Onobrychis genetic diversity using SSR markers From Related Legume Species. The Journal of Animal & Plant Sciences , 24(2): 556-566. Botstein D, White RL, Skolnick M and Davis RW (1980) Construction of genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet 32:314-331. Brummer, E.C. (1999). Capturing hetrosis in forage crop cultivar development. Crop Science , 39: 943-954. Carbonero, C.H., Mueller-Harvey, I., Brown, T.A., and Smith, L. (2011). Sainfoin ( Onobrychis viciifolia ): a beneficial forage legume. Plant enetic Resources: Characterization Utilization,9:70–85. Choudhary, S., Sethy, N.K., Shokeen, B., and Bhatia, S. (2009). Development of chickpea EST-SSR markers and analysis of allelic variation across related species. Theoretical and Applied Genetics ,118:591- 608. Demdoum, S., Munoz, F., Delgado, I., Valderrabano, J., and Wunsch, A. (2011). EST-SSR cross- amplification and genetic similiarity in Onobrychis genus. Genetic Resources and Crop Evolution ,59: 253-260. Eujayl, I., Sledge, M.K., Wang, L., May, G.D., Chekhovsiy, K., Zwonitzer, J.C., and Mian, M.A.R. (2004). Medicago truncatula EST-SSR reveal cross- species markers for Medicago spp . Theoretical and Applied Genetics ,108:414-422. Hume, L. J., Withers, N. J., and Rhoades, D. A. (1985). Nitrogen fixation in sainfoin ( Onobrychis viciifolia ) 2. Effectiveness of the nitrogen-fixing system. New Zealand Journal of Agricultural Research , 28:337348. Kuleung, C., Baenziger, P.S., and Dweikat, I., 2004. Transferability of SSR markers among wheat, rye and triticale. Theoretical and Applied Genetics , 108:1147- 1150. Madden D. (2002). Investigating plant DNA. Technical guide.Version 2.3.The National Centre for Biotechnology Education.UK. Murray, M.G., and Thompson, W.F., (1980). Rapid isolation of high molecular weight plant DNA. Nucleic Acids Research , 8: 4321– 4325. Nybom H., Bartish, I (2000). Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants. Perspectives in plant ecology, evolution and systematics 3:93–114 Rohlf, F.J. (1998). NTSYS-pc numerical taxonomy and multivariate analysis system. Version 2.00. Exeter Software, Setauket, NY. Varshney RK, Graner A, Sorrells ME. (2005). Genic microsatellite markers in plants: features and applications. Trends Biotechnol , 23(1):48–55. Wikipedia.org accessed from https://en.wikipedia.org/wiki/Onobrychis_viciifolia Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A. and Tingey, S.V. (1990) DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18: 6331-6535. Werlemark G, Uggla M, Nybom H(1999). Morphological and RAPD markers show a highly skewed distribution in a pair of reciprocal crosses between hemisexual dogrose species, Rosa sect. Caninae. Theoretical and Applied Genetics ,98:557–563. Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7173060","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":488245995,"identity":"2ab957d4-44c3-4b99-a078-1be623945367","order_by":0,"name":"Mohammad Sharifi Sedeh","email":"data:image/png;base64,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","orcid":"","institution":"","correspondingAuthor":true,"prefix":"","firstName":"Mohammad","middleName":"Sharifi","lastName":"Sedeh","suffix":""}],"badges":[],"createdAt":"2025-07-21 04:12:59","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-7173060/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7173060/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":87440145,"identity":"ecdee5f9-a2eb-4c04-bc51-eba08100d34d","added_by":"auto","created_at":"2025-07-23 19:38:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":463544,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7173060/v1/54481c73-1bba-42a4-9087-4ebd3f9e0056.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eAssessment of Genetic Diversity Landraces In Onobrychis Sativa Using SSR,RAPD, SRAP markers\u003c/p\u003e","fulltext":[{"header":"Key Points","content":"\u003cp\u003eI survey Genetic Diversity in sainfoin to select best adjectives in sainfoin .after select best adjectives you can produce best lines for \u0026nbsp;sainfoin modify . And use them as food for animal.\u003c/p\u003e"},{"header":"Introduction","content":"\u003cp\u003e\u003cem\u003ePlease include line numbering in your manuscript\u003c/em\u003e (Wikipedia.org).\u0026nbsp;According\u0026nbsp;to\u0026nbsp;its\u003c/p\u003e\n\u003cp\u003eseed specification, it has been divided into 270 categories. Resistance to the dryness is the most important characteristic of this plant, making it appropriate for the dry farming and pastures (Hume, \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e1985). The larger content of tannins in common sainfoin leaves, prevents bloat and protein hydrolysis in the livestock rumen, and increases amino acid absorption in the intestine of ruminants (Carbanero \u003cem\u003eet al.\u0026nbsp;\u003c/em\u003e2011).\u003c/p\u003e\n\u003cp\u003eFabaceae provender is of great importance Moreover, knowledge about the rate of polymorphism with differences between different examples is an important in order to reduce the rate of hereditary resources, kept in the gene bank and analyzing the purity of seeds in the cross- pollinated plants (Brummer, 1999). Studying genetic diversity \u003cem\u003evia\u0026nbsp;\u003c/em\u003emorphologic marker is accompanied\u0026nbsp;\u0026nbsp;with\u0026nbsp;\u0026nbsp;limitations\u0026nbsp;\u0026nbsp;such\u0026nbsp;\u0026nbsp;as\u0026nbsp;\u0026nbsp;low\u003c/p\u003e\n\u003cp\u003epolymorphism, low heritability, and effectiveness due to environmental conditions (Werlemark \u003cem\u003eet al.\u0026nbsp;\u003c/em\u003e1999).\u003c/p\u003e\n\u003cp\u003eThe Molecular markers reveal genetic diversity in DNA surface; do not show having the mentioned problems as in the morphologic indicators. Moreover, molecular markers are prominent tools in an evaluating relational selection of better\u0026nbsp;plants as well as for analyzing the similarities and differences among various species (Awasthi \u003cem\u003eet al\u003c/em\u003e.). In this regard,\u0026nbsp;micro-satellites or\u0026nbsp;simple\u0026nbsp;sequential repeats (SSR) are the repeats of 106 pairs of DNA sequence that primarily was thought to be only exist in non-codifying regions of DNA sequence, while occurrence time of micro-satellites in codifying sequences or translated sequences were found by the passage of time, and EST-SSRs were developed (Choudhary \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2009). Micro-satellite markers usually shows the higher level of polymorphism, and by considering the frequency\u0026nbsp;of alleles in each gene place, they can be distinguished\u0026nbsp;a very\u0026nbsp;similar\u0026nbsp;species (Kuleung \u003cem\u003eet\u0026nbsp;al.,\u0026nbsp;\u003c/em\u003e2008)\u0026nbsp;Extracting\u0026nbsp;SSR\u0026nbsp;primer\u0026nbsp;by\u0026nbsp;genome\u0026nbsp;libraries\u0026nbsp;is\u003c/p\u003e\n\u003cp\u003ean\u0026nbsp;expensive\u0026nbsp;and\u0026nbsp;tedious\u0026nbsp;method,\u0026nbsp;requiring\u0026nbsp;proper skills. By having access to a number of ESTs and other\u0026nbsp;information\u0026nbsp;about\u0026nbsp;DNA\u0026nbsp;sequence,\u0026nbsp;developing SSR markers \u003cem\u003evia\u0026nbsp;\u003c/em\u003eextracting information has\u0026nbsp;become effective and lower costs for many plant species (Eujayl \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2000; Varshney \u003cem\u003eet al,.\u0026nbsp;\u003c/em\u003e2005). Although many EST databanks for modelled and non-modelled creatures are available in the recent years and new generation technologies for finding sequences have created a great evolution in the genetic researches, but its share for a plant like common sainfoin is rather little in the databanks. Thus,\u0026nbsp;a\u0026nbsp;simple\u0026nbsp;method\u0026nbsp;for\u0026nbsp;developing\u0026nbsp;a\u0026nbsp;SSR\u0026nbsp;primer in the species that are not studied is a great deal id using SSR primer for related species. Genomix has made an apparent comparison, and the order and contents\u0026nbsp;of\u0026nbsp;genes\u0026nbsp;among\u0026nbsp;closely\u0026nbsp;related\u0026nbsp;species\u0026nbsp;are greatly protected. Harmony of markers shown in a comparative\u0026nbsp;drawing\u0026nbsp;indicates,\u0026nbsp;marker\u0026nbsp;of\u0026nbsp;the\u0026nbsp;same material or type can exist in another material or type (Kuleung \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2004). The Number of 27 SSR marker of \u003cem\u003eMedicago sativa\u0026nbsp;\u003c/em\u003eand \u003cem\u003eGlycin max\u0026nbsp;\u003c/em\u003ewas tested for transferring in \u003cem\u003eO. vicifolia\u0026nbsp;\u003c/em\u003especies by Demdoum \u003cem\u003eet al.\u0026nbsp;\u003c/em\u003e(2011), and after identifying the markers\u0026nbsp;with\u0026nbsp;the\u0026nbsp;potential\u0026nbsp;to create\u0026nbsp;polymorphs,\u0026nbsp;6 pairs of SSR markers were used for fingerprinting and estimating genetic similarities in 23 farming sainfoins and two wild types. Thus, masses of farming\u0026nbsp;sainfoins\u0026nbsp;of\u0026nbsp;two\u0026nbsp;wild\u0026nbsp;types\u0026nbsp;of\u0026nbsp;sainfoils\u0026nbsp;and\u003c/p\u003e\n\u003cp\u003e23 farming sainfoins based on geographical origins were\u0026nbsp;separated\u0026nbsp;(Demdoum\u0026nbsp;\u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2011).\u0026nbsp;In\u0026nbsp;another study by Avci \u003cem\u003eet al.\u0026nbsp;\u003c/em\u003e(2014), they identified the transferability of 95 markers for \u003cem\u003eMedicago truncatula\u0026nbsp;\u003c/em\u003eand \u003cem\u003ephaseolus sativa\u0026nbsp;\u003c/em\u003eto sainfoin, and 18 markers for analyzing the genetic diversity of sainfoin type. RAPD markers are used extensively in the studies regarding diversity, structure, and genetic similarities of the populations of many plant species (Williams \u003cem\u003eet al.\u0026nbsp;\u003c/em\u003e1990 Nybomaind Bartish, 2000). These markers are suitable due studying genome, for studying the ordinary selection on the genetic structure of plant populations. Although this method has limitations, such as having a dominant nature and not identifying homozygosity against heterozygosity natures, hence, SRAP markers were used that are simple, and have high reliability, average overall coverage of the genome and potentials for reproduction in codifying regions in the genome.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eThe population of the considered farming sainfoin included 5 masses of Iranian and 17 foreign sainfoin masses, provided by direct collection and sampling from gene banks (IPK/ Germany and USDA/ USA) (Table 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1.\u0026nbsp;\u003c/strong\u003eEcological characteristic of cultivated sainfoin population.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 9.26724%;\"\u003e\n \u003cp\u003eS.No\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.8276%;\"\u003e\n \u003cp\u003ePopulation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.9828%;\"\u003e\n \u003cp\u003eHeight\u0026nbsp;from\u0026nbsp;Sea Surface(m)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.75%;\"\u003e\n \u003cp\u003eGeographical\u0026nbsp;wide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30.1724%;\"\u003e\n \u003cp\u003eWeather\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 9.26724%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.8276%;\"\u003e\n \u003cp\u003eBanab\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.9828%;\"\u003e\n \u003cp\u003e1290\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.75%;\"\u003e\n \u003cp\u003e37/3292\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30.1724%;\"\u003e\n \u003cp\u003eWarm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 9.26724%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.8276%;\"\u003e\n \u003cp\u003eAmand\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.9828%;\"\u003e\n \u003cp\u003e1400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.75%;\"\u003e\n \u003cp\u003e38/1878\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30.1724%;\"\u003e\n \u003cp\u003eWarm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 9.26724%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.8276%;\"\u003e\n \u003cp\u003eKhosrosha hr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.9828%;\"\u003e\n \u003cp\u003e1500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.75%;\"\u003e\n \u003cp\u003e38/0011\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30.1724%;\"\u003e\n \u003cp\u003eSemi warm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 9.26724%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.8276%;\"\u003e\n \u003cp\u003eSarab\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.9828%;\"\u003e\n \u003cp\u003e1680\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.75%;\"\u003e\n \u003cp\u003e37/9264\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30.1724%;\"\u003e\n \u003cp\u003eCold\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 9.26724%;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.8276%;\"\u003e\n \u003cp\u003eHaris\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.9828%;\"\u003e\n \u003cp\u003e1900\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.75%;\"\u003e\n \u003cp\u003e38/2556\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30.1724%;\"\u003e\n \u003cp\u003eCold\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eA population of local \u003cem\u003eMedicago sativa\u0026nbsp;\u003c/em\u003ealfalfa was also used as the control sample. Two sets of seeds were cultivated in medium plastic pots in the research farm of Isfahan University of Technology in spring 2012. Two weeks after growing of seedlings, 10 samples were randomly selected from each population, and sampling was done from healthy and fresh leaves. Extraction of DNA from the leaf samples of sainfoin was done by Murry and Thompson \u0026nbsp;method \u0026nbsp;(1980). \u0026nbsp;To \u0026nbsp;determine \u0026nbsp;the\u003c/p\u003e\n\u003cp\u003equality\u0026nbsp;and\u0026nbsp;quantity\u0026nbsp;of\u0026nbsp;DNA,\u0026nbsp;the\u0026nbsp;gel\u0026nbsp;(0.7%)\u0026nbsp;of\u0026nbsp;agars in TBA 0.5x of ethidium bromide dye (10mg/ml) and Gel Doc device were used. Since the micro- satellites (SSR) with \u0026ldquo;\u003cem\u003eOnobrychis\u0026nbsp;\u003c/em\u003eare not known, screening the identified SSRs with medicago provides the possibility of finding similar regions on \u003cem\u003eonobrychis\u0026nbsp;\u003c/em\u003egenome. In this study, 22 pairs of SSR and EST-SSR primer of medicago containing high polymorphism information, content were selected from different resources (Table 2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2.\u0026nbsp;\u003c/strong\u003eVarious pair of SSR and EST-SSR primer of medicago.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.71022%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS.no\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.0905%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCode\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4204%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCountry\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.943%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGeographical wide\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.938%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGeographical length\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.8978%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeight\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.71022%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.0905%;\"\u003e\n \u003cp\u003eVICUNS8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4204%;\"\u003e\n \u003cp\u003eIran\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.943%;\"\u003e\n \u003cp\u003e53/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.938%;\"\u003e\n \u003cp\u003e32/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.8978%;\"\u003e\n \u003cp\u003e1200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.71022%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.0905%;\"\u003e\n \u003cp\u003eVICESfS9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4204%;\"\u003e\n \u003cp\u003eIran\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.943%;\"\u003e\n \u003cp\u003e53/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.938%;\"\u003e\n \u003cp\u003e32/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.8978%;\"\u003e\n \u003cp\u003e1200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.71022%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.0905%;\"\u003e\n \u003cp\u003eVICUNS18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4204%;\"\u003e\n \u003cp\u003eIran\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.943%;\"\u003e\n \u003cp\u003e53/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.938%;\"\u003e\n \u003cp\u003e32/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.8978%;\"\u003e\n \u003cp\u003e1200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.71022%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.0905%;\"\u003e\n \u003cp\u003eVICUNS39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4204%;\"\u003e\n \u003cp\u003eIran\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.943%;\"\u003e\n \u003cp\u003e53/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.938%;\"\u003e\n \u003cp\u003e32/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.8978%;\"\u003e\n \u003cp\u003e1200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.71022%;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.0905%;\"\u003e\n \u003cp\u003eVICUNS42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4204%;\"\u003e\n \u003cp\u003eIran\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.943%;\"\u003e\n \u003cp\u003e53/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.938%;\"\u003e\n \u003cp\u003e32/00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12.8978%;\"\u003e\n \u003cp\u003e1200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICRUS112\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eRussia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e55/75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e37/61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e144/17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICRUS113\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eRussia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e55/75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e37/61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e144/17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICRUS122\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eRussia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e55/75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e37/61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e144/17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICGES104\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eKyrgyzstan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e48/01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e67/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e342/9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICGES108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eKyrgyzstan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e48/01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e67/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e342/9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICMAS134\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eHungary\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e47/26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e19/15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e378/2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICENS107\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eEngland\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e51/5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e0/11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e12/19\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICENS118\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eEngland\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e51/5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e0/11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e12/19\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICASS127\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eSlovakia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e48/66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e19/68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e1081/12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICGES119\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e51/15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e10/45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e459/94\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICHOS130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eNetherland\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e52/11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e5/28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e7/92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICSWS123\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eSwitzerland\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e46/81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e8/21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e1777/59\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICSWS124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eSwitzerland\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e46/81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e8/21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICROS114\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eRomania\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e45/93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e24/96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e596/18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICROS120\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eRomania\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e45/93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e24/96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e596/18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICSPS115\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eSpain\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e40/27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e3/73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e895/8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 7.8727%;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.598%;\"\u003e\n \u003cp\u003eVICUNS106\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.258%;\"\u003e\n \u003cp\u003eUnknown\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 15.0754%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.268%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.928%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003ePolymerase chain reaction (PCR) for the samples in 15\u0026micro;L volume including 1.5 \u0026micro;L of buffer 10x, 1.5 \u0026micro;L of magnesium chloride, a unit of tag polymerase, 0.3 mmol of dNTP, 0.25 \u0026micro;mol. Of each starter, and 50 ng of genome DNA was prepared from each sample. Reaction was made in a thermal cycler device in a primary blotting cycle in 94\u0026deg;C for 4 minutes, and 35 cycles of secondary blotting in 94\u0026deg;C for 1 minutes, with connection of the primer to DNA string in 50-55\u0026deg;C for 45 seconds, and expansion of synthesized DNA in 72\u0026deg;C for 2 minutes, as well as a final expansion cycle in 72\u0026deg;C for 10 minutes. Polymerase chain product went under electrophoresis on acrylamide gel (12%) and dyeing was done by silver nitrate (Rohlf, 1998). A percentage of polymorph for each starter was obtained by division of polymorphic places and the number of alleles in the places. The polymorphic information content (PIC) was calculated for each band by the following relation (Botstemeta, 1980).\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAOcAAAArCAYAAACD3p/vAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAAFiUAABYlAUlSJPAAABAaSURBVHhe7ZwJXE9Z/8c/v36tVJQMskTJUiQihLH8p2c8MzwMqbElBmN5xhiyZss64bGMjN3DWB6yPJZRD4ki8kSyNSFLaKG0aNdv+f7v794Tv5LkV8wvz32/XufVPd9z7v2de8/5nu/3fO+5SYgDIiIiWocO+ysiIqJliMopIqKliMopIqKliMopIqKliMopIqKliMopIqKliMopIqKliMopIqKliMopIqKliMopIqKliNv3RDRAjqTbN5CQJWd5zTGsWRe2No1gJBXy6Q9vYv2U7ph3JANoPgCnTuyGq40h4kNWoc/Q6Yh5pgQsnbFyxz5M/qIJpBLhvE8RUTlFNCAP+8a0xeCtd1lecxq6TkTgYX+0MpYjPmI7vLw2wLSbC2okXEDAybsYuScKowo3wXvjJbTs7AI8jsDew5eg03UqLp3wQ0tTptWfIKJy/skU3DuG8YtC0W/6PPS3r8mk2s+LOwcwsKc7QpKFfC3r3vDxmwBrPQXKGlFZD85i+bxfEJMr5G37TMaRf62GHaecEbvW42imJRb8MAjxe/+OTkN3o+PQASBZdUzxmYPeDnVQcHcbujYfjah2ExAT/AvszMtWzgdBSzF7UwTySAqpDmdmucYpFHIoS7RRR6oL9/l7MaxdNSbRAlTKKfLxyc9IoIMLPMjG3IhgbE9+IcmspKogp0sbviMLKVTDnHSNmtLkPTdZWVnIKfbQTGpRQ4c/j1NOiskWSmSFcuGACujEwt4k4WyHWf2hFPS8SE6Ucn0TteXOazBsFaUUMGEZFGQm0eUAH7LizlH9nioZmFqQhYV6qkUGnFxi/BlZtutHm8OekIKd/5o02u/jTvUtLcmyVTfaepk1Wo2004vJUlXOpVGbophUc8SA0EcnE+EBW+D1ZSu4LdiP++n5QHUj6FW5xZMUHYfMxqSBzaHL5eT597B54XQcjc0Rit+KFC36jYNnd2uWf42unmAFqTADt67d4DTJBB7LZ6J3rdfWMfFCCKIhgX1jKxhxGvUuDGrUQ/uBUzFjtD2f1zG2wOqwVKSmqqfnuLh7GlpIUpB09SjGDnDH9qgUvv5rlMjNSEFiUhKSkpKR/bKE6eVQ5mdyZaryJDzPfsmkmiMq50ck/ebv8PUagL94jMX+yEwmrcKYWmPi7Jno8ll1Ppt35xRmLNqBFAWffTtSK4zycuUHn7IUH1iWG4vI4CSgQQ/0dW7EpCqyEHn+IqfFjeHkYAsjJn0nEh2Y1qzNMgDn1b5Bu6E/Y/XEbkImLQJ+6/4D5nkzCEoFuzHZm26xClK+vnF5aRXeE1E5PwoKhK7zROfeI7Dkjg2OXYpGdPQ5LOrrBH1Wo6pi3mYolvv04a2n6j7v7PfBgv2xfK4s6ri6oSf3N++ljPc11cmODsPJbKC2Qws0tTBlUo6UizgUmABJvSawt23E2eDyQ6RkR29DBx0GD4Ujc2AST/8bkenCsYAZvpn7T1yL5vruYhCGtxUmJHXMenhz/arq22j4ezkyqeZ8EOWU5TxFVOhphJ0PR3h4OMLOnsZ/AgMRGBiEkNBwXIl7ymqqKMDD/4YjJOw8Xzc8PBRnzkbheSErfkUBYq6ex0n+OoE4dSYM4Rcvc25hxd2HD48UNs5fYdSItbh/YQu+6OgIR8e2sOYGXtV/E6AH53HTMca6hpBVZmHX9Kk4/IBz18vC2B4DPbuii2MzVC+mZQrERoYjh1P3jg4dUFctRlZ47yYuvACa2NnDoanwe3k5ldf/BuZNYVufZQqykVtsDOrCvL4N2jhyfdeqBWpXf1N1dE0tuX5V9a0jmtSuhMASW3tWKnlPo2nF0A7EzS2qSZFMGjrTiEmTaNJ4L7KrwS28m3SlaYt30708Ve0MOvPLdHLg5Kq6gAX1nexPt3P4S/HkJF2mLd6Dqaa+FfUdz11n0kRybW7M1dWhPr4hlM/qVS3SaOeI7nwgAnXa06rQp0xeNXl00o+amxf1oT45eq2hxFxW+F4k0jIXC4KBFXnvuk6vQ0FEMdsHE2ehqcNgf27UvKSgn71p9E9bKfXN6E1xlJm0a+rnfNu4NSetu8zkJch5cpoGNRLuwaS2O4WxmE9BXDDNnTufFi1ZSkuXLqWV67dRbLpQpiI/8RptWzGHfBcv4cuXLl1LwbHPWanmfLBorSJmL3VtLOVvtMPgtVTU1KjfxlMzfdUDMKRvlodxj1iFkja4m/J1YTmMInmlFZBnR9Lkto24Mj362+wTVBSgy76yhmw5mcuQ7aT2nKoQn5ZykiyT9vzY49WEjGoNaNrem6VEPd/Bo8PUQTVR13WmndFqA4EjbvdIMuSubVSrKXVzdSFrY1facO4WZXEL1zIpp3K+uLePehoK7Tfv7EtPmLww+QZtWTqWGhfdm3lT2nuXFXIUpsfTmWMryNVEiEAD1Wjc3hhWqjkfbM2pUEqgoys4bQpZAbJVzeaw69wHLZuo/BjOTT1+Cc94qRI6RZE3Yz0gX8YyhTjmOwZroh9Dt91gzPrhS7yq5vQtxnBrtkYNzMqx9lAiNz0Z9+/dw70KpvsPnqGA3YuIGro18O20BejtZC7k8xKweq4vQhILhHw5yU5OgW5DG7Rt3xrNLIuHfGzc58PXvQ3qmRbigX5XHP7jKMZ1s4dJJa0N7h4/ivN8c6Xo4vEFijxcvbqtMXrWMswb14Mfa0YmxtBT0xw9Myv07OuN33ZOYTEEExiqV9AUpqSVzssbe6mbjS4/k7Rz86OHbHaTx4eQh5MRJ5eQ/dfr6BkvldHGYcxy2nrRf9MKeWnhw0PUva5Qt/PYdZSs7uO8Fy9ov3cv4foVTn3orNC8CvKJWU5GfLA32ag9rzYjNlPquyzbh0bdcprUpi1/MLkaD0PXURdDYbzWaz2BItJKLpYK6OSCb8mUKze0cqQD95hYjewbG8mOv+86NPlALJNqzgeznMWQcFaUzW4v0hLw9LEqWKCD5q6dYCGISyXpajjuZqvqmqBRfWtovlPLCB0Hz8L2LVuwpYJp+44paKlxOz59rL6Yi2VjXFgOuLFrEZYExPKaqg1QQTa2zfwO332nnjww4G9TcKFAjqadv8fOQwvRyZxzoItBpb72UYeUysq9T6aklY665Wzdx4ciE5Mo6eENmv2VKRnoGlI9j2WUlFO0Iindct7aOYrq8muA2jTCL4QqxWBpDZVnORWFeZSZkUEZFU5Z9FJecTMnS75Ew52Evlelel086XIaK/wzULOcqqQjlZK0WNKjaq5TKOJaLCWkvm3bUT4Fzfco03JmXfuVWvK/USUsp2Au4yMDMGWEJzzH/ISIPGdM3XQScftmol4p4WgB4byszHzI3nilIlKSuGOz0cjMDGYVTl2w7VYqu6rm6NbtiHlz5sLeVOhfPYNq0P84Pto7Ue0QWhNRiMJC9VSA7FP/QKc2LVDfoiiq8efzgR+ZaqICmveagEPBwQgOPo0zZ4OxZNTnePMVrjrCeWa1uE7ln1UBcnNzIepp6VjYf4158+djfoXTJHSsW3bPlA8lpPqcW2jEDS9da3hNnQGH99jTfzfYH76+vth85CKy3rXbSAMkEh3o6JRIrEyrYBa00nlbQKh0Sndrcy/7k425Hi/vMHoNJcl4sRpKzs0vnxumqqdQKCohVVZ049MMCKl4mRJBY9vV4PttwKIQenOLeNkcnFCbP9d64EJ6LLxrqxjlfJVSNp+QWyuRSFRxIHas8+q4dLi6RS3R4Y7ZYTWnv2JY08/444d34vC82J7qBPi4tECfiQF4wSRvJxvHFrrB1MAABhVMxubDcbHoTY/ImyizsO7H/th89QVMe/tgzU+9YMyKykst6/aws7ND2+YNoF/muPm0+YDWXAaFTNjPWCjPfccPEQpzVBMbB7fOlBvqCccSa3y/fDpa1QKenw/Akn+GcA6TQGroDuyJiEdWdv4r2dvRQ6M2PTHMk1v3VjANG/I5PtNKH0gbkOPulhmYtf8ZTBu4wn/JRDTUwEvuMTUQMTExOLhkJOqwofA/CbOglUpO4iWa29eOqkv4xSPpmjWgISuOU2ap4dY0Ou47lmxNhLrcSpO6ey6h61msmONO2G80f4QL6ek1pN6DPMjDw42cG6nefxrRwOXhbJdRVSOdfvPq8cqtXR0mvPGtyiRe2kr/V6saQd+cJm6/rj3R9RJurf8VJn8vPhG31tC8OUb6BeAq54rGxcUhJuIU5ro7l9jgXIQJnIdMwZHLd/m6cXER+HXGYDRR2xzS7PPhWLDpd8TdOgm/2TMxc6YPtgVFI+5eLDZO7FRFv+zQgUzBvshQKvgdVVWaF7exduEChKTlofnA+Zgz3IHzV8pGlvcCqc8zkc88LEqKxCyvz9HKthlsbW3Rf8ZWPKuMJYREyq2W+CcNpUyBJynFPjcpJ6rAEbM26suwDwlTUpGPxLPrERQUGEgnti+iTk0Mhb6GHjm7z6YDnDwoNILUPvyvIuTQ7z7fkAl3LyZNvqfI3PLtqD30kw3BsAP5h7NdrEolyeUPaIGzBf9cHIb70ZPKcIuyb9GPPeowvTIk52l7KYMVlRtFPP3s5kjcFEqo0YB8Dj5kBa+pbMspKufHQp5JZ/bMI2cz4WOAtycpuQxcSKdup7ITtZ9Hp6dRE67teg3b0ZqzRdvF34HyIU235+63bi/aFVX8C47w5YIL2s5rBSVUUDkTwrfShP5tiz9jiTH18FhGofGZrFbZFD6NoHke3clM7RrGDTrQtD3nKFdtIq1s5RT/wdfHggrw8OZV3E+XwVC3jP1/nIubn68Daycn2Fho0T+behtJ5zDg66/w72t6GLQ8AHumub7TneVJPohuloMQbtsHR47sQz+715Gj6xvd4Tj+ADjlxLFN3qhfgXVL1uMoRN7OhH41g9dBSdUzzpPCtlN7NK5ZcpvemyhyEhBxIRbyakZg33LwH3NIatmic2sr6DFZ9o2N6NRmPP5AHUw+EIrVbi2EAk3hVVRERBNkj2jlt07c5C4hJ7cVFM/E70ZGR+e4EKdzJG3Rj47fLv7h59X1A3nrVBmW82OSdeUX3oOA1InWXkpgUs0RLaeIhihwzn8IPKcG4JFMHwOmrYGrFYFbbbLyMki/iRVrd+BR2kuYdRmBEwE70NmSlXFE/+qGdhMPVYrl/JgkHpqIBm6/Alb9EHh6N/7a9H3f8JaAV1ERkfck5/JWsjV7vQbTNNV08aQLJYxM1bScz2h9n5Z8u1sPW0WP8yq+k0x8nS7y3ijzo+EzzBtxGUxQAYwMq8HYhGUYqh1lVQVSFCA9OQp+w/vj77/HomGXgVi5cDwaGlX81Zjo1oq8Nzk3D2DS0t3IKKi4EtVr3xezpo5CQ7W4TOTaL9Fx8ik4jVzJubVTYanFu4QK06IwtmN77Mxshq//4o4FGxahPftfZxVFVE4RLSMT/l854IegJ7D1WIzQPT6w1OaP2znLeSVoB+4Y98LQHs2YsHIQlVNEe6Ak7Jz5A6atOoZUuRy6pnXRutUobDu7GG3/B3fAi8opIqKVAP8PYcPZ2fysEUQAAAAASUVORK5CYII=\"\u003e\u003c/p\u003e\n\u003cp\u003ePij is the abundance of amino allele in marker \u0026ldquo;i\u0026rdquo;, population is \u0026ldquo;j\u0026rdquo;, and \u0026ldquo;n\u0026rdquo; is the number of alleles. In this study, 5 different populations of farming sainfoin that are growing wild in different regions in East Azerbaijan\u0026nbsp;province\u0026nbsp;were\u0026nbsp;analyzed. Ten (10) samples of the plant were randomly selected from each population. Geographically, the individual plants in this population are away from each other, in 50 km distances. The studied populations are different with regards to the sea levels, geographical latitude, weather conditions, and soil salinity (Table 1). Two populations in Amand and Khosroshahr are grown in rather salty soil, and in rather close geographical distances. Two populations in Heris and Sarab are growing in cold\u0026nbsp;climates\u0026nbsp;and\u0026nbsp;higher\u0026nbsp;sea\u0026nbsp;levels,\u0026nbsp;in\u0026nbsp;closer\u003c/p\u003e\n\u003cp\u003edistances, while the population in Bonab is comparatively in further distances than the other 4 populations, and ecologically, due to growing in properly salty soil, it is different to the other populations. Regarding geographical latitudes, the populations in Sarab and Khosroshahr are closer to each other, and the populations in Heris and Amand are in almost equivalent geographical latitudes, while the latitude in Bonab has an enormous difference with others. In this test, DNA from young seedlings and seeds were extracted by Madden (2002) method, which is a faster and more economical method in comparison with other common methods in DNA extraction.\u003c/p\u003e\n\u003cp\u003eThe DNA of an individual in a population after extraction were mixed, in order to only prepare one DNA sample, i.e. mass DNA, as the representative of that population. Estimating the density of extracting DNA was done by electrophoresis of agar gel (0.8%) and spectrophotometry in 260 nm, to adjust the final density of DNA samples in 10ng/ml. Polymorphism of RAPD strings was tested for 5 studied populations by using 10 random starters. Hence, the contents of 10 ng of model DNA and primer with a density of 100 PM/\u0026micro;L in the final volume of 25\u0026micro;L were analyzed.\u003c/p\u003e\n\u003cp\u003eThe number of 17 different bulks of sainfoin plant were collected that belongs to the four species: \u003cem\u003eO.viciifolia\u0026nbsp;\u003c/em\u003e(13 bulks), \u003cem\u003eO.viciifolia\u0026nbsp;\u003c/em\u003e(1 bulk), \u003cem\u003eO.viciifolia\u0026nbsp;\u003c/em\u003e(1 bulk), and \u003cem\u003eO.viciifolia\u0026nbsp;\u003c/em\u003e(1 bulk) from different regions in the country, and after extraction of DNA from their fresh leaves and determining their quality and quantity, the extracted DNA was used in PCR reactions, and 88 strings were obtained from the total of 10 primer compositions, 73 of which were polymorphic.\u003c/p\u003e\n\u003cp\u003eEight pairs of SSR primer and one pair of EST- SSR primer in sainfoin out of 26 pairs of SSR and EST-SSR primers of medicago type (Table 2) indicated polymorphic forms (Table 3), which were used for analyzing genetic diversity in farming sainfoin populations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3.\u0026nbsp;\u003c/strong\u003eGenetical distance matrix between five population of \u003cem\u003eOnobrychis\u0026nbsp;sativa\u003c/em\u003e.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePopulation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 45px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAmand\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBonab\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 44px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSarab\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eKhosroshahr\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHaris\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003eAmand\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 45px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 44px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003eBonab\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 45px;\"\u003e\n \u003cp\u003e0.3851\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 44px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003eSarab\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 45px;\"\u003e\n \u003cp\u003e0.1178\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0.3851\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 44px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003eKhosroshahr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 45px;\"\u003e\n \u003cp\u003e0.0572\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0.3363\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 44px;\"\u003e\n \u003cp\u003e0.1178\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003eHaris\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 45px;\"\u003e\n \u003cp\u003e0.0870\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0.4904\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 44px;\"\u003e\n \u003cp\u003e0.0870\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e0.1495\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eNine(09) pairs of used micro-satellite primers could separate 44 alleles for evaluating 22 populations of sainfoins, and the number of alleles varied between 4 to 6, and the size of reproducing components were in the range of 100-400 pairs. Polymorphic information content was changing between 0.2 to 0.43 distances for studied micro- satellite places, having an average of 0.33. Only half of the 10 primers produced clear and gradable polymorphic strings. To ensure from repeatability of the produced strings, PCR was repeated for the studied samples with the 5 primers, for three times, and the patterns of strings were similar for the three repetitions, indicating the repeatability of the mentioned strings.\u003c/p\u003e\n\u003cp\u003eThe five primers produced 12 polymorphic strings in total, and the highest rate of polymorphic strings was related to no. 1 and no. 2 primers, each with 3 (0.25) polymorphic strings and the lowest rate was related to no. 2, no. 3, and no. 5 primers, each with 2 (16.7%) polymorphic strings. For the matrix of calculated genetic distance, the greatest genetic distance (0.4904) was between the populations of Bonab and Heris, and the lowest distance (0.0572) was between the populations of Amand and Khosroshahr, and 0.087 in the next stage, between populations of Heris and Sarab and also between Heris and Amand, based on the genetic distance of \u0026ldquo;Nei\u0026rdquo; on a UPGMA dendrogram. With the greatest genetic similarities, the populations of Amand and Khosroshahr were placed in a cluster, while the populations of Sarab and Heris were in another cluster, and the population of Bonab was placed in a separate cluster due to the least amount of genetic similarity as compared to the other mentioned populations.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003ethe genetic diversity of 48 individual plants from four cultivated populations of alfalfa (Medicago sativa L.) in Iran was assessed using sequence-linked amplified polymorphic markers (SRAP). Fourteen SRAP primer combinations produced 193 fragments, of which 95 were polymorphic. The number of polymorphic fragments identified in each primer combination ranged from 3 to 10 bands with an average of 6.78 bands. The average polymorphic information content (PIC) for all primer combinations was 0.343. Although AMOVA (analysis of molecular variance) results showed significant differences in genetic diversity among populations (P \u0026lt; 0.0001), genetic diversity, which was mainly due to intrapopulation variation, accounted for 93.17% of the total genetic diversity. \u0026nbsp;Analysis of marker data using unweighted pair-group arithmetic mean (UPGMA) method clearly separated subtropical (Yazdi) and subcold (Hamedani and Nikshahri) populations as well as the improved Kodi population. It can be concluded that SRAP markers are useful for studying diversity and relationships between and within alfalfa populations.\u003c/p\u003e\n\u003cp\u003eOverall, the results shows that the obtained markers of medicago type can be suitable\u0026nbsp;resources for identifying the place of micro- satellites in sainfoin, micro-satellite markers considered to have capacity for analyzing relationships and genetic diversity in sainfoin plants. Moreover, the used SSR markers in the present study show the high polymorphism. Populations of an Iranian and foreign sainfoin plants were place in a separate population, indicating a high genetic diversity in the\u0026nbsp;considered germplasm. Moreover, high genetic diversity between Iranian and foreign sainfoin populations is suitable for modification programs in order to reach maximum heterosis.\u003c/p\u003e\n\u003cp\u003eOn analyzing the genetic diversity, SRAP\u0026nbsp;marker\u0026nbsp;in\u0026nbsp;sainfoin\u0026nbsp;plant\u0026nbsp;genetic\u0026nbsp;diversity\u0026nbsp;between\u003c/p\u003e\n\u003cp\u003ethe masses, the range of genetic distance between masses was found to be in between 124 and 349. Regarding the dispersion of bulks (masses), the lowest genetic distance was between the masses related to Russia (codes P and Q), and the greatest distance was in between the Damavand and Najafabad regions (codes G and M).\u0026nbsp;The numerical value (33.1) obtained for gene movements\u0026nbsp;between\u0026nbsp;groups shows genetic moves in the studied populations, but the move is not to as such to cause homogeneity of in the populations. The range for information index for primer compositions was calculated to be in between 31 and 52.\u003c/p\u003e\n\u003cp\u003eHigh overall heritability values for height, number of stems and percentage of stems indicate the major role of genetic effects in controlling these traits. Given that these traits are also considered components of forage yield, indirect selection through them can increase the return from selection for improved forage yield. China was superior to China in terms of forage yield. Iranian varieties had a significant advantage over foreign varieties in terms of yield and its most important components. \u0026nbsp;On the other hand, foreign varieties were superior only in terms of leaf percentage and leaf to stem ratio, which are considered as criteria for palatability and forage quality. It is\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe present study results on the farming of wild populations of sainfoin shows the populations grow in a similar ecologic conditions (similar ecotype) and are in a close distance to each other have more genetic similarities. According to the present study findings using DNA, RAPD, we can identify ability of different ecotypes of the cultivated species. In cluster analysis of SRAP data, dendrograms drawn by the NTSYS VERSION 2 could not place the used genotypes in the separate groups. In other words, mixture of masses with each other indicates little inter-mass genetic diversity. Analysis of the main coordination also showed the first three components justified 62.95% of the changes. Thus, analysis of clusters by using SRAP data is a best method for evaluating inter-mass genetic diversity of sainfoin.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eSource of funding: None.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003ch2\u003eConflict of interest:\u003c/h2\u003e\u003cp\u003eThe There is no financial conflict with the subject matter discussed in the manuscript.\u003c/p\u003e\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAwasthi, A.K., Nagaraja, G.M., Naik, G.V., Kanginakudru, S., Thangavelu, K., and Nagaraju, J.(2004). Genetic diversity and relationships in mulberry (genus Morus ) as revealed by RAPD and ISSR marker assays. BMC Genetics, 5:1\u0026ndash;9.\u003c/li\u003e\n\u003cli\u003eAvci et al.(2014). Analysis of \u003cem\u003eOnobrychis \u003c/em\u003egenetic diversity using SSR markers From Related Legume Species. The \u003cem\u003eJournal\u003c/em\u003e\u003cem\u003eof\u003c/em\u003e\u003cem\u003eAnimal\u003c/em\u003e\u003cem\u003e\u0026amp;\u003c/em\u003e\u003cem\u003ePlant\u003c/em\u003e\u003cem\u003eSciences\u003c/em\u003e, 24(2): 556-566.\u003c/li\u003e\n\u003cli\u003eBotstein D, White RL, Skolnick M and Davis RW (1980) Construction of genetic linkage map in man using restriction fragment length polymorphisms. \u003cem\u003eAm J Hum Genet \u003c/em\u003e32:314-331.\u003c/li\u003e\n\u003cli\u003eBrummer, E.C. (1999). Capturing hetrosis in forage crop cultivar development. \u003cem\u003eCrop Science\u003c/em\u003e, 39: 943-954.\u003c/li\u003e\n\u003cli\u003eCarbonero, C.H., Mueller-Harvey, I., Brown, T.A., and Smith, L. (2011). Sainfoin ( \u003cem\u003eOnobrychis viciifolia \u003c/em\u003e): a beneficial forage legume. Plant enetic Resources: Characterization Utilization,9:70\u0026ndash;85.\u003c/li\u003e\n\u003cli\u003eChoudhary, S., Sethy, N.K., Shokeen, B., and Bhatia, S. (2009). Development of chickpea EST-SSR markers and analysis of allelic variation across related species. \u003cem\u003eTheoretical\u003c/em\u003e\u003cem\u003eand\u003c/em\u003e\u003cem\u003eApplied\u003c/em\u003e\u003cem\u003eGenetics\u003c/em\u003e,118:591- 608.\u003c/li\u003e\n\u003cli\u003eDemdoum, S., Munoz, F., Delgado, I., Valderrabano, J., and Wunsch, A. (2011). EST-SSR cross- amplification and genetic similiarity in Onobrychis genus. \u003cem\u003eGenetic Resources and Crop Evolution\u003c/em\u003e,59: 253-260.\u003c/li\u003e\n\u003cli\u003eEujayl, I., Sledge, M.K., Wang, L., May, G.D., Chekhovsiy, K., Zwonitzer, J.C., and Mian, M.A.R. (2004). Medicago truncatula EST-SSR reveal cross- species markers for \u003cem\u003eMedicago spp\u003c/em\u003e. \u003cem\u003eTheoretical and Applied Genetics\u003c/em\u003e,108:414-422.\u003c/li\u003e\n\u003cli\u003eHume, L. J., Withers, N. J., and Rhoades, D. A. (1985). Nitrogen fixation in sainfoin (\u003cem\u003eOnobrychis viciifolia\u003c/em\u003e) 2. Effectiveness of the nitrogen-fixing system. \u003cem\u003eNew Zealand Journal of Agricultural Research\u003c/em\u003e, 28:337348.\u003c/li\u003e\n\u003cli\u003eKuleung, C., Baenziger, P.S., and Dweikat, I., 2004. Transferability of SSR markers among wheat, rye and\u0026nbsp;triticale. \u003cem\u003eTheoretical and Applied Genetics\u003c/em\u003e, 108:1147- 1150.\u003c/li\u003e\n\u003cli\u003eMadden D. (2002). Investigating plant DNA. Technical guide.Version 2.3.The National Centre for Biotechnology Education.UK.\u003c/li\u003e\n\u003cli\u003eMurray, M.G., and Thompson, W.F., (1980). Rapid isolation of high molecular weight plant DNA. \u003cem\u003eNucleic Acids Research\u003c/em\u003e, 8: 4321\u0026ndash; 4325.\u003c/li\u003e\n\u003cli\u003eNybom H., Bartish, I (2000). Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants. \u003cem\u003ePerspectives in plant ecology, evolution and systematics \u003c/em\u003e3:93\u0026ndash;114 Rohlf, F.J. (1998). NTSYS-pc numerical taxonomy and multivariate analysis system. Version 2.00. Exeter Software, Setauket, NY.\u003c/li\u003e\n\u003cli\u003eVarshney RK, Graner A, Sorrells ME. (2005). Genic microsatellite markers in plants: features and applications. \u003cem\u003eTrends Biotechnol\u003c/em\u003e, 23(1):48\u0026ndash;55.\u003c/li\u003e\n\u003cli\u003eWikipedia.org accessed from https://en.wikipedia.org/wiki/Onobrychis_viciifolia Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A. and Tingey, S.V. (1990) DNA polymorphism amplified by arbitrary primers are useful as genetic markers. \u003cem\u003eNucleic\u003c/em\u003e\u003cem\u003eAcids\u003c/em\u003e\u003cem\u003eResearch,\u003c/em\u003e18: 6331-6535.\u003c/li\u003e\n\u003cli\u003eWerlemark G, Uggla M, Nybom H(1999). Morphological and RAPD markers show a highly skewed distribution in a pair of reciprocal crosses between hemisexual dogrose species, Rosa sect. Caninae. \u003cem\u003eTheoretical\u003c/em\u003e\u003cem\u003eand\u003c/em\u003e\u003cem\u003eApplied\u003c/em\u003e\u003cem\u003eGenetics\u003c/em\u003e,98:557\u0026ndash;563.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Sainfoin, Markers, Multiphase","lastPublishedDoi":"10.21203/rs.3.rs-7173060/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7173060/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn order to evaluate genetic diversity cultivated sainfoin accessions collected from different regions of the world, SSR markers from medicago genus were used. Polymorphism information content value ranged from 0.2 to 0.043 with an average of 0.33. Cluster analysis based on SSR markers grouped the sainfoin population in two main groups (Iranian and exotic cultivate sainfoin) with two sub cluster for each group. In another survey was performed to find out an efficiency of ecological factors on the genetic similarity was investigated among five population of Onobrychis viccifolia East Azarbaijan, Iran using RAPD and bulked DNA, The largest and shortest genetic were detected between Bonab and Heris (0.04904)and Amand and Khosroshahr(0.0572) on UPGMA dendrogram, two population of Sarab and Heris were nested within one cluster, and Amand and Khosroshahr in other cluster. Also Bonab was completely separated from all other population.\u003c/p\u003e","manuscriptTitle":"Assessment of Genetic Diversity Landraces In Onobrychis Sativa Using SSR,RAPD, SRAP markers","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-23 19:22:05","doi":"10.21203/rs.3.rs-7173060/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"072b8827-fbe7-4d78-90a0-9aad2e3d3889","owner":[],"postedDate":"July 23rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":51824977,"name":"Biotechnology and Bioengineering"}],"tags":[],"updatedAt":"2025-07-23T19:22:05+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-23 19:22:05","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7173060","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7173060","identity":"rs-7173060","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}