Inter-retrotransposon Amplified Polymorphism Research Articles

Elucidate Genetic Diversity and Population Structure of Bread Wheat (Triticum Aestivum L.) Cultivars Using IRAP and REMAP Markers

Analysis of genetic diversity and population structure in bread wheat is an essential step in their conservation, utilization, and breeding. Retrotransposons are ubiquitous and abundant a throughout the plant genomes, therefore extensively used as ideal molecular markers for genetic variability, DNA fingerprinting and genetic mapping studies in plant species. In the current research, we used two retrotransposon-based marker systems, inter-retrotransposon amplified polymorphisms (IRAPs), and the retrotransposon-microsatellite amplified polymorphisms (REMAPs) markers to evaluate the genetic diversity and survey activity of long terminal repeat retrotransposon (LTR-retrotransposon) elements in a collection of 49 bread wheat (Triticum aestivum L.) cultivars that mainly bred in Iran. In general, 90 and 126 loci were amplified using 9 IRAP and 20 REMAP primers, respectively. Both techniques produced a satisfactory number of bands for cultivar analysis; however, the technique IRAP, particularly single primer Nikita generated a large number of bands, indicating the wide activity of Nikita family under various environmental conditions of bread wheat. The percentage of polymorphic loci (PPL) in the studied collection for IRAP and REMAP markers was 81.78 and 86.40%, respectively. A model-based Bayesian method, Principal coordinate analysis (PCoA) and cluster analysis using Minimum Evolution (ME) algorithm hinted of the existence of two groups. This grouping was in agreement with the growing season and conformed by the high within-group bootstrap value. These results demonstrated that these markers developed using transpositionally active retrotransposons (RTNs) are efficient and reliable markers in determining level of genetic diversity and population structure in bread wheat in breeding programs.

Assessment of Genetic Relationships among Cultivated and Wild Pistachios (Pistacia vera L.) using Molecular Markers

Abstract Iran is one of the main diversity centers and origins of pistachios in the world. Pistachio cultivation spread first within the ancient Persian Empire and then moved gradually westward. Knowledge of the genetic relationships among wild and cultivated varieties of pistachio is important for the efficient utilization of the available germplasm resources. Three molecular marker strategies, namely, inter-simple sequence repeat (ISSR), inter-retrotransposon amplified polymorphism (IRAP), and retrotransposon microsatellite amplified polymorphism (REMAP), were used to study the genetic relationships among 35 pistachio accessions including 15 wild-type genotypes of Pistacia vera and 20 important cultivars from Iran. According to the results, high levels of polymorphism were observed for all three marker systems. REMAP and IRAP techniques had the higher mean values of genetic relationships parameters than ISSR technique. The results from this study showed that the 5′LTR2, Sukkula, Sukkula + UBC855, and 5′LTR2 + UBC811 primers were the most informative and could be used to evaluate the genetic relationships of pistachios accessions. Cluster analysis using the unweighted pair group method with arithmetic mean (UPGMA) properly separated the accessions and divided them into four main groups. The presence of most cultivated genotypes in a group indicates genetic erosion of cultivated pistachio in Iran. Wild-type genotypes of P. vera are located in different clusters indicating the high diversity of the genotypes. The results provide useful genetic information about wild pistachios in northeastern of Iran and indicate that the use of wild pistachios in breeding programs could be useful for generating new genotypes with interesting characters.

Cytological variations and long terminal repeat (LTR) retrotransposon diversities among diploids and B-chromosome aneuploids in Lilium amabile Palibin.

B chromosomes are supernumerary chromosomes found in numerous plant species, including in the genus Lilium. Lilium amabile, an endemic Korean Lilium species, carries B chromosomes which are highly variable in terms of numbers and shape among the accessions collected throughout the Korea. Class 1 retrotransposons are highly abundant in the genome of Lilium species, but their biological functions are still obscure. Lilium species were known to hold high diversities derived from retrotransposons. In this study, genetic diversities among the L. amabile accessions were analyzed to better understand relationships between genetic variations and cytological diversities. Chromosomes were prepared from 95 L. amabile accessions for cytological identification. Genetic variations were analyzed by inter-retrotransposon amplified polymorphism (IRAP), and genetic differentiation was evaluated via Tajima's D neutrality and FST analyses. Population structure and phylogenetic analyses were also carried out. The L. amabile accessions were classified into 11 cytotypes by the chromosome constitutions. Genetic diversity measured by IRAP analysis revealed high genetic diversity among the accessions. In the joint analysis of cytological variation with genetical variation, IRAP diversity was not related to the cytological diversities of diploid and aneuploids among L. amabile accessions, and genetic differentiation was not obvious. Moreover, the geographical distribution of L. amabile was not related to either IRAP diversity or cytological diversity. The B chromosome-carrying aneuploids occurred randomly among diploids throughout Korea, and IRAP diversification predated L. amabile dispersion in Korea without genetic differentiation.

Analyzing Sunn pest resistance in bread wheat genotypes using phenotypic characteristics and molecular markers.

Sunn pest is one of the most destructive insects in the western and central parts of Asia causing severe reductions of wheat yield and flour quality. Therefore, an effort was undertaken to find effective resistance by analyzing genetic variation among 25 wheat genotypes artificially infested in field-cages as well as using start codon targeted (SCoT) polymorphism and inter- retrotransposon amplified polymorphism (IRAP) markers. High variation was revealed amongst genotypes with Sunn pest resistance characteristics including Bayat, Bezostaya, Sayson, Line93, Line120, Rashagol, Golsepi and AarasGolsoor, which were classified as resistant to moderately resistant. SCoTs and IRAPs were determined as efficient markers for studying genetic diversity. The non-parametric Kruskal-Wallis test was conducted to evaluate the effect of specific SCoT and IRAP amplicons on Sunn pest resistance characteristics for wheat genotypes. The stepwise regression analysis exhibited seven informative SCoTs and IRAPs explaining the highest resistance characteristics variation ranging from 25.7-50.1 to 17.6-40.1, respectively. The relationship between resistance of genotypes and informative SCoT and IRAP amplicons was found based on canonical discriminant analysis showing the capacity of informative markers for functional marker selection method and screening the wheat germplasms for Sunn pest resistance characteristics.

Genetic characterization of Moniliophthora perniciosa from Ecuador and in vitro sensitivity to compost tea

Moniliophthora perniciosa is the causal agent of the witches’ broom disease (WBD) in cacao but the morphological and molecular diversity of M. perniciosa in Ecuador—the top fine flavor cacao producer worldwide—are poorly understood and the pathogen’s sensitivity to compost teas is unknown. A total of 90 isolates of M. perniciosa were obtained from symptomatic samples of cocoa branches and pods collected from four cocoa-producing regions of Ecuador. Growth of each isolate was assed and fitted to a Gompertz model, and the molecular variability was evaluated by sequencing the ITS1, 5 .8s, and ITS2 rDNA regions as well as by inter-retrotransposon amplified polymorphism (IRAP), retrotransposon microsatellite amplified polymorphism (REMAP), and simple sequence repeats (SSR) analyses. Sensitivity of M. perniciosa to a compost tea was evaluated at concentrations of 0.5, 1, 1.5, 2, 2.75, and 3.5% in PDA. Results showed morphological and growth rate homogeneity across isolates. Mycelial growth fitted to Gompertz model gave similar parameter estimates for isolates from the different sampling sites. No molecular variability was observed based on ITS and SSR analysis but IRAP and REMAP tests showed high polymorphism. The total genetic diversity based on IRAP and REMAP tests was 0.28 and the genetic diversity within population was 0.24, with Nei’s diversity, Shannon’s Information, and percent polymorphism values of 0.315, 0.48, and 95.24, respectively for the whole sampled population; isolates from Los Rios province being the most polymorphic. Compost tea sensitivity analysis performed on 85 of the isolates showed a significant inhibitory effect at concentrations of 2.75% or above but concentrations below 1.5% significantly enhanced the growth of most isolates of the pathogen. Results confirmed the high genetic diversity of M. perniciosa and provided insights into growth rate and compost tea sensitivity of the pathogen in Ecuador.

Development and use of retrotransposons-based markers (IRAP/REMAP) to assess genetic divergence among table grape cultivars

AbstractFor more than four decades after the introduction of cv. Italia (Vitis vinifera L.) in Brazil, several somatic mutations in the genome of cv. Italia and its somatic mutants gave rise to phenotypes which generated at least five new cultivars of fine table grapes. Since no molecular marker proved to be effective in discriminating cv. Italia (V. vinifera L.) and its coloured mutants (Rubi, Benitaka, Brasil, Black Star), primers for the long terminal repeat (LTR) sequences were developed to analyse Inter Retrotransposon Amplified Polymorphism (IRAP) and Retrotransposon-Microsatellite Amplified Polymorphism (REMAP), and investigate how the coloured cultivars derived from clonal propagations of somatic mutations are genetically structured. Primers for LTR sequences of IRAP and REMAP markers were edited from grape sequence databases available at a GenBank. Twenty-four primers, denominated DKS001–DKS024, were edited. Three hundred and forty-nine DNA segments were amplified by individual DKS primers and DKS/ISSR (Inter Simple Sequence Repeats) primer combinations, at an average of 13.96 amplicons per primer pair. High genetic divergence between the five cultivars was inferred from polymorphism in retrotransposons IRAP and REMAP. The analysis of polymorphism of IRAP and REMAP retrotransposons was crucial to show that clonal propagation of somatic mutations may lead towards the formation of genetically divergent cultivars by the formation of genetically structured vineyards and show the mixture of genomes within each cultivar.

LTR retrotransposons and highly informative ISSRs in combination are potential markers for genetic fidelity testing of tissue culture-raised plants in sugarcane

Testing the genetic fidelity of micropropagated clones with DNA markers could curtail the losses caused by somaclonal variation in sugarcane tissue culture industry. Evaluation of 32 inter-retrotransposon amplified polymorphism (IRAP) markers and 100 inter-simple sequence repeats (ISSRs) in 47 sugarcane accessions identified 20 (8 IRAPs and 12 ISSRs) polymorphic markers representing 98 loci. IRAP system was superior to ISSR in terms of marker index (2.68 as against 1.76), resolving power (2.85 as against 1.98), and polymorphic loci per assay (6 compared to 4.1), except mean polymorphic information content (0.31 and 0.34). Further evaluation of the 20 polymorphic markers in testing the genetic fidelity of micropropagated sugarcane clones identified a variant clone by three primers of ISSR and one IRAP marker (UBC810, UBC813, UBC840, and LTR6149 + 3′LTR). The unique amplicons from the somaclonal variant were validated by sequencing. Based on the number of bands amplified and proportion of polymorphic loci, a set of six ISSR and four IRAP markers has been recommended to test clonal fidelity for sugarcane tissue culture industry.

Transferability of Barley Retrotransposons (Sukkula and Nikita) to Investigate Genetic Structure of Pimpinella anisum L.

Transferability of barley retrotransposons (Nikita and Sukkula) were examined in Pimpinella anisum L. seeds by using a retrotransposon-based molecular marker: IRAP (inter-retrotransposon amplified polymorphism). Furthermore, transposons’ sequences identified in medically important plants were obtained form NCBI, and multiple alignment analyses were performed to investigate the evolutionary relationships. These two retrotransposons were identified in Pimpinella anisum L., showing homomorphic band profiles. In addition, limited similar sequences were detected as a result of clustal analyses. Till date, no study about retrotransposons evaluation using IRAP as molecular marker has been published in aniseed. Our results are expected to contribute a new perspective for genome architect of medically important plants in addition to aniseed.

Evaluation of Water Stress Tolerance of Soybean Using Physiological Parameters and Retrotransposon-Based Markers

Drought is one of the most important environmental stresses that severely reduce plant growth and crop productivity. This study was carried out to investigate the difference in the response of six soybean cultivars (Giza 21, 22, 35, 82, 83 and 111) under water stress and the genetic difference between these cultivars using retroelements technique. The results showed that drought stress caused reduction in morphological criteria, photosynthetic pigments, starch, phospholipids, glycolipids, pectin, cellulose and lignin in shoots of all soybean cultivars except Giza 22 and Giza 83. On the other hand, there was a considerable increase in root length, soluble sugars, proline, glycine betaine, total lipids and hemicellulose contents in the shoots of the soybean cultivars in response to water stress. The soybean cultivars Giza 22 and 83 were more drought tolerant than the other cultivars while Giza 21 and Giza 111 were the most sensitive. Inter-primer binding sites (iPBS) and inter-retrotransposon amplified polymorphism (IRAP) techniques were used to fingerprint the six soybean cultivars using a set of eight primers. The techniques successfully tagged each cultivar with specific bands and detected molecular genetic markers related to drought tolerance in soybean.

IRAP and REMAP based genetic diversity among varieties of Lallemantia iberica.

This study describes the genetic relationships among 34 varieties of Lallemantia iberica using inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP). Samples were collected from Agriculture Research Center of Urmia city (northwest Iran). Ten IRAP and REMAP primers generated 76 scorable electrophoretic bands with 78.94% polymorphism. The pair-wise Jacquard genetic similarity varied from 0.48 to 0.94 for IRAP and REMAP data combined. Average PIC values for IRAP and REMAP markers were 0.38. The retro-elements marker system produced 76 alleles in range of 100- 3000 bp. The cophenetic correlation coefficient between Jaccard's similarity matrix and the plotted dendrogram was 0.66. A dendrogram constructed based on COMPLETE LINKAGE. Cluster analysis of IRAP and REMAP data using the NTSYSpc 2.02 resulted in five clusters. The present study represents high genetic distance at genotype level suggesting that IRAP and REMAP markers are useful for Lallemantia iberica genetic diversity analysis.

Potential role of transposable elements to differentiate tuna and xoconostle Opuntia varieties

Opuntia, an important horticultural crop in Mexico, is cultivated mainly for its two fruitsvariants: sweet fruits or prickly pears (tunas) and acidic fruits (xoconostles). The interretrotransposonamplified polymorphism (IRAP) technique was applied to evaluate geneticdiversity of Opuntia varieties and to differentiate xoconostle fruits from tunas. Five IRAPprimers previously described for other plant species and classified into three retrotransposonfamilies, namely Copia, Gypsy and TRIM, were analysed in 43 Opuntia varieties (eightxoconostles and 35 tunas). The five individual IRAP primers generated a total of 264fragments, where 64.8 % of them were polymorphics. The retrotransposon of the Gypsyfamily (60 fragments) was more represented than Copia (average of 52 fragments) or TRIM(48 fragments) families. Moreover, the percentage of polymorphic fragments was higher (61.9%) in xoconostles than in tunas (56.5 %). A larger number of total amplified fragments (262)was found among tunas, compared to those amplified from xoconostle varieties (257fragments). In contrast, a lower number of polymorphic bands were counted among tunas(148) than among xoconostle varieties (159). Unlike the UPGMA analysis, where three of thexoconostle-producing varieties were grouped with other tunas, the PCoA analysis allowed abetter separation of all xoconostle varieties. These results suggest a potential role of thetransposable elements in genetic divergence within the Opuntia genus.

Investigation of IRAP transposon-based molecular markers for analysis of genetic diversity in pea germplasm

Retrotransposons diversity has been extensively studied in monocots, but it is not well documented in dicot species. Transposition activity of transposons creates DNA polymorphism and their abundant presence in genomes is making transposons a promising marker system for varietal identification and fingerprinting. In this study, four transposon-based markers (two DNA- and two RNA-transposons) were employed to evaluate the effectiveness of Inter-Retrotransposon Amplified Polymorphism (IRAP) transposon system in assessing genetic diversity in pea germplasm accessions. A total of 28 alleles were detected across the 35 pea accessions with number of alleles per locus ranged from 5 (Mutator) to 9 (Cyclops). RNA transposons produced a higher number of polymorphic alleles (Ogre: 8, Cyclops: 9) than DNA transposon markers (Mutator: 5, MITE: 6). Overall mean PIC value and D values for these transposon markers were 0.810 and 0.817 respectively. Genetic similarity values ranged from 0.143 to 0.823 with a mean similarity value of 0.403. Cluster analysis classified pea genotypes into six major groups that were somewhat consistent with their geographical origins. The molecular analyses differentiated all the 35 accessions and generated higher PIC and D values that can be useful for MAS-based breeding programs in pea.

LTR retrotransposons from the Citrus x clementina genome: characterization and application

Long terminal repeat retrotransposons (LTR-RTs) are a large portion of most plant genomes, and can be used as a powerful molecular marker system. The first citrus reference genome (Citrus x clementina) has been publicly available since 2011; however, previous studies in citrus have not utilized the whole genome for LTR-RT marker development. In this study, 3959 full-length LTR-RTs were identified in the C. x clementina genome using structure-based (LTR_FINDER) and homology-based (RepeatMasker) methods. LTR-RTs were first classified by protein domain into Gypsy and Copia superfamilies, and then clustered into 1074 families based on LTR sequence similarity. Three hundred fifty Copia families were grouped into four lineages: Retrofit, Tork, Sire, and Oryco. One hundred seventy-eight Gypsy families were sorted into six lineages: Athila, Tat, Renia, CRM, Galadriel, and Del. Most LTR-RTs (3218 or 81.3%) were anchored to the nine Clementine mandarin linkage groups, accounting for 9.74% of chromosomes currently assembled. Accessions of 25 Rutaceae species were genotyped using 17 inter-retrotransposon amplified polymorphism (IRAP) markers developed from conserved LTR regions. Sequence-specific amplified polymorphism (SSAP) makers were used to distinguish ‘Valencia’ and ‘Pineapple’ sweet oranges (C. x sinensis), and 24 sweet orange clones. LTR-RT markers developed from the Clementine genome can be transferred within the Rutaceae family demonstrating that they are an excellent tool for citrus and Rutaceae genetic analysis.

Molecular insights into genetic diversity and population dynamics of five medicinal Eulophia species: a threatened orchid taxa of Africa.

Genetic diversity existing amongst five Eulophia orchid species were assessed using start codon targeted polymorphism (SCoT) and inter-retrotransposon amplified polymorphism (IRAP) markers. A total of 12 SCoT and 5 IRAP markers revealed an average of 63% genetic variability [SCoT = 63.87; IRAP = 64.95%] amongst the five Eulophia species investigated. The genetic similarities were assessed using both UPGMA and Bayesian approaches which indicated identical clustering patterns at a genetic similarity level of 50%. Analysis of molecular variance (AMOVA) revealed the presence of a significant degree of genetic variability, mostly compartmentalized within the species level. Amongst the five assessed Eulophia species, E. parviflora was the most genetically diverse representative whereas E. welwitschii was found to be least diverse based on a comparative assessment of various population genetic parameters like Nei's gene diversity (h) and Shannon's information index (I) with an overall gene flow value greater than 1. In order to evaluate the comparative marker efficiency, SCoT and IRAP marker data were subjected to various benchmark analyses like marker index, resolving power, polymorphic index content, multiplex ratio and effective multiplex ratio which revealed the robustness of both the marker techniques in assessment of genetic diversity. The present report provides the first molecular insights into the aspects of inter and intra specific genetic variability in medicinally as well as horticulturally important Eulophia species along with addressing their conservation concerns. In a nutshell, the present approach is simple, rapid and cost effective and can be extended for analysis of genetic diversity of other related plant species.

Detection of HERV-K6 and HERV-K11 transpositions in the human genome.

Mobile genetic elements classed as transposons comprise an estimated 45% of the human genome, and 8% of these elements are human endogenous retroviruses (HERVs). Endogenous retroviruses are retrotransposons, containing 5' and 3' long terminal repeat sequences and encoding envelope, group-specific antigen and DNA polymerase proteins. The aim of the present study was to analyse genome integration polymorphisms of HERV type K member 6 (HERV-K6) and HERV-K11 by using the retrotransposon based molecular marker technique, inter-retrotransposon amplified polymorphism (IRAP). For this purpose, blood samples of 18 healthy individuals within the age range of 10-79 years (10 females and 8 males) were collected, genomic DNAs were isolated and IRAP-polymerase chain reaction (PCR) was performed. IRAP-PCR analyses demonstrated that there were 0-70% polymorphism rates for HERV-K6, and 0-38% polymorphism rates for HERV-K11 among all the samples. Furthermore, the polymorphism rates were 0-70% among females and 11-60% among males for HERV-K6, and 0-38% among females and 0-25% among males for HERV-K11. Age-associated polymorphism was also investigated, but no age-associated polymorphism was observed among the samples. Therefore, HERV-K6 and HERV-K11 polymorphisms may arise on an individual-specific basis. Various previous studies have investigated the associations between the expression of HERVs and cancer or other major diseases. However, few reports have analysed HERV-K movements among individuals. This is the first report to investigate HERV-K6 and HERV-K11 retrotransposon polymorphisms between the genders and different age groups.

Short-term mutagenicity test by using IRAP molecular marker in rice grown under herbicide treatment

ABSTRACTRice is an economically important plant as well as a model organism. The rice genome consists of 35% retrotransposons. Although most of the retrotransposons are inactivated through evolutionary processes, they can be activated under various biotic and abiotic stress conditions. The main objective of this study was to explore the effects of herbicides on retrotransposon activities and the usage of retrotransposons in short-term mutagenicity tests. In this study, bentazone and an MCPA (2-methyl-4-chlorophenoxyacetic acid)-containing herbicide was used. Plant samples were grouped into three classes: control (untreated), 1% and 2% herbicide treatment. Retrotransposon activities were investigated by using the inter-retrotransposon amplified polymorphism (IRAP) marker technique. IRAP analyses were performed for Houba (Tos5/Osr13) retrotransposon. Polymorphism ratios were calculated with the Jaccard similarity index, and the significance of polymorphism was evaluated by one-way analysis of variance (ANOVA). We observed that the polymorphism ratios ranged from 8%–90% for Houba among plant samples. ANOVA showed that these variable ratios were statistically significant. Bentazone and the MCPA-containing herbicide increased the retrotransposon activities, and they might be responsible for DNA mutations. This study indicated valuable data for establishing retrotransposon-based short-term mutagenicity test in rice with suitable retrotransposons such as Houba.

Identification of SSR and retrotransposon-based molecular markers linked to morphological characters in oily sunflower ( $$\textit{Helianthus annuus}$$ Helianthus annuus L.) under natural and water-limited states

Sunflower is an important source of edible oil. Drought is known as an important factor limiting the growth and productivity of field crops in most parts of the world. Agricultural biotechnology mainly aims at developing crops with higher tolerance to the challenging environmental conditions, such as drought. This study examined a number of morphological characters, along with relative water content (RWC) in 100 inbred sunflower lines. A 10 × 10 simple lattice design with two replications was employed to measure the mentioned parameters under natural and water-limited states during two successive years. In molecular trial, 30 simple sequence repeat (SSR) primer pairs, as well as 14 inter-retrotransposon amplified polymorphism (IRAP) and 14 retrotransposon-microsatellite amplified polymorphism (REMAP) primer combinations were used for DNA fingerprinting of the lines. Most of the examined characters had lower average values under water-limited than natural states. Maximum and minimum reductions were observed in the cases of yield and oil percentage, respectively. The broad-sense heritabilities for all the examined characters were 0.20-0.73 and 0.10-0.34 under natural and water-limited states, respectively. In the studied samples, 8.97% of the 435 possible locus pairs of the SSRs represented significant linkage disequilibrium (LD) levels. In the association analysis using SSR markers, 22 and 21 markers were identified (P ≤ 0.05) for the studied characters under natural and water-limited states, respectively. The corresponding values were 50 and 37 using retrotransposon-based molecular markers. Some detected markers were communal between the characters under water-limited and natural states. This was in line with the phenotypic correlations detected between the characters. Communal markers facilitate the simultaneous selection of several characters and can thus improve the efficacy of selection based on markers in the plant-breeding activities.

Retrotransposons in Betula nana, and interspecific relationships in the Betuloideae, based on inter-retrotransposon amplified polymorphism (IRAP) markers.

The Betulaceae family comprises two subfamilies, Betuloideae and Corylaceae. The subfamily Betuloideae contains two genera, Alnus Mill. and Betula L. Twenty putative long terminal repeat (LTR) retrotransposons were mined from 171 scaffolds containing 5,208,995bp of dwarf birch (Betula nana) genome sequences. Five retrotransposons were finally selected after filtering the retrotransposon canonical features and nucleotide similarities between left and right LTR sequences. Of the five retroelements, three elements were found to be Ty1/Copia retrotransposons; identity of the other two elements could not be ascertained due to sequence undetermined 'N' bases in the sequence database. Inter-retrotranposon amplified polymorphism (IRAP) analysis, based on the LTR sequences of the mined LTR-retrotransposons, produced 179 discernible IRAP bands among the Alnus and Betula genera. Sequence analysis revealed no size homoplasy among the homologous IRAP bands. Phylogenetic and principle coordinate analysis, based on the band sharing among the taxa, showed the species in two different genera were clearly separated. The subgenera in each genus of Alnus and Betula were also distinguishable from the IRAP profiles. In the genus Betula, the species in subgenus Betula showed mixed clustering between species. This is incongruent with the phylogeographical distribution of the species.

Retraction Note to: Potential Start Codon Targeted (SCoT) and Interretrotransposon Amplified Polymorphism (IRAP) Markers for Evaluation of Genetic Diversity and Conservation of Wild Pistacia Species Population.

"This article has been retracted by the Publisher in agreement with the Editor-in-Chief, because it contains portions of writings on the same topic already published and without sufficient attribution to these earlier works being given. The principal authors of the paper acknowledged that text from background sources was mistakenly used in this article without proper reference to the original source. Upon investigation carried out according to the Committee on Publication Ethics guidelines, it has been found that the authors have duplicated or rephrased parts from other articles of which the main sources.

RETRACTED ARTICLE: Comparison of traditional and new generation DNA markers declares high genetic diversity and differentiated population structure of wild almond species

Wild almond species as sources of genetic variation may have crucial importance in breeding. A total of 389 accessions of 18 species have been analysed using inter-retrotransposon amplified polymorphism (IRAP), retrotransposon-microsatellite amplified polymorphism (REMAP), sequence-specific amplification polymorphism (S-SAP), amplified fragment length polymorphism (AFLP), inter simple sequence repeat (ISSR) and simple sequence repeats (SSR). Retrotransposon markers indicated the presence and movement of some Ty3-gypsy and Ty1-copia-elements in almond genome. Since transposable elements are associated with large-scale genome alterations, REMAP produced more reliable phylogenetic inferences than AFLP where homoplasy may affect clustering. In addition, high resolution melting (HRM) analysis was developed to detect SNPs. HRM analysis revealed 1:189 bp frequency of SNPs in exon positions, and the transition-to-transversion proportion was 1.84:1. The low transition bias suggests low methylation levels in almond genome. The polymorphic information content (PIC) was the highest for SSR markers, while SNPs had an average PIC of 0.59, which is close to the values of the rest of the markers. Huge genetic diversity, fragmented population structure and footprints of human selection was confirmed by merging information from all marker strategies. Considering time, cost and performance HRM can be a marker of choice in future studies of Prunus diversity.