Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 75(3): 101-108, 2022 Firenze University Press www.fupress.com/caryologia ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.36253/caryologia-1533 Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Citation: Syamand Ahmed Qadir, Chnar Hama Noori Meerza, Aryan Mahmood Faraj, Kawa Khwarahm Hamafaraj, Sherzad Rasul Abdalla Tobakari, sahar hussein hamarashid (2022). Comparative study and genetic diversity in Malva using srap molecu- lar markers. Caryologia 75(3): 101-108. doi: 10.36253/caryologia-1533 Received: January 01, 2022 Accepted: November 23, 2022 Published: April 5, 2023 Copyright: © 2022 Syamand Ahmed Qadir, Chnar Hama Noori Meer- za, Aryan Mahmood Faraj, Kawa Khwarahm Hamafaraj, Sherzad Rasul Abdalla Tobakari, sahar hussein hamarashid. This is an open access, peer-reviewed article published by Firenze University Press (http://www. fupress.com/caryologia) and distrib- uted under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, pro- vided the original author and source are credited. Data Availability Statement: All rel- evant data are within the paper and its Supporting Information files. Competing Interests: The Author(s) declare(s) no conflict of interest. Comparative study and genetic diversity in Malva using srap molecular markers Syamand Ahmed Qadir1, Chnar Hama Noori Meerza2, Aryan Mahmood Faraj3, Kawa Khwarahm Hamafaraj4, Sherzad Rasul Abdal- la Tobakari5, Sahar Hussein Hamarashid6,* 1 Medical laboratory techniques department/Halabja Technical Institute, Research center/ Sulaimani Polytechnic University, Sulaymaniyah, Iraq 2 Food Science and Quality Control Department, Bakrajo Technical Institute, Sulaimani Polytechnic University, Sulaymaniyah, Iraq 3 Medical Laboratory Science Department/ Technical College of Applied Science/Sulaima- ni Polytechnic University, Sulaymaniyah, Iraq 4 Nursing department/Halabja Technical Institute/ Sulaimani Polytechnic University, Sulaymaniyah, Iraq 5 Medical laboratory techniques department/Halabja Technical Institute, Research center/ Sulaimani Polytechnic University, Sulaymaniyah, Iraq 6 Agricultural Project Management Department/ Technical College of Applied Science/ Sulaimani Polytechnic University, Sulaymaniyah, Iraq *Corresponding author. E-mail: sahar.rashid@spu.edu.iq Abstract. The Malva genus has 25-40 species and it can be considered as an annual and/or biannual herb. Malva species are indicated with potential therapeutic as cic- atrizing and analgesic by the Ministry of Health. The aim of this study was to analyze SRAP (Sequence-related amplified polymorphism) markers in a total of 70 accessions of Malva species, which included five species Malva neglecta Wallr., Malva pusilla Sm., Malva sylvestris L., Malva verticillata L., Malva nicaeensis All.. A total of 89 (Number of total loci) (NTL) DNA bands were produced through polymerase chain reaction amplifications (PCR) amplification of five Malva species. These bands were produced with the combinations of 5 selective primers. The total number of amplified fragments ranged from 10 to 27. The predicted unbiased gene diversity (UHe) varied between 0.077 (Malva sylvestris) and 0.382 (Malva pusilla). The genetic similarities between three species are estimated from 0.70 to 0.91. Neighbor-Joining tree results showed two major clusters. According to the SRAP (Sequence-related amplified polymorphism) markers analysis, Malva pusilla and Malva aegyptia had the lowest similarity. Our results provided great molecular identification of all assayed genotypes, which have shown that there is large quantity of genetic diversity among the Malva accessions. Objectives of the study were; a) to estimate genetic diversity; b) to evaluate population relationships using NJ approaches. Current results have implications in breeding and conservation programs. Keywords: Sequence-related amplified polymorphism, Genetic Diversity, Medicinal Plants Malva,Taxonomy. 102 Syamand Ahmed Qadir et al. INTRODUCTION The use of medicinal plants can be influenced by the economic condition, the high cost of medicines and the difficult access to public consultations. In addition to that, there is difficulty of access by residents in rural are- as to health care units located in urban areas. Moreover, the increase the trend for considering traditional knowl- edge that supports using natural resources as an alterna- tive to synthetic drugs (Battisti et al., 2013). Given the significance of genetic diversity in conservation strate- gies, it is of utmost importance to disentangle genetic diversity in plant species, particularly threatened and rare species (Esfandani-Bozchaloyi et al. 2018a, 2018b, 2018c, 2018d). The indiscriminate use of plants due to the lack of phytochemical, pharmacological and mainly toxicologi- cal knowledge is of great concern for public health. The correct identification of medicinal plant species is nec- essary, especially when they are processed in order to avoid misuse of medicinal plants (Romitelli & Martins, 2013). The Malva genus presents different species with therapeutic potential and inadequate consumption can occur due to the incorrect identification of the plant in the market. Malvaceae or the mallow family is the family of flowering plants containing over 200 genera with close to 2300 species (la Duke and Dobley 1995). Many researches have been published on the ecology, taxono- my, genetic, cytology, chemotaxonomy, physiology, seed germination and economic uses of family Malvaceae such as (El-Rjoob and Omari 2009) in ecology; in tax- onomy in chemotaxonomy (Blunden et al., 2001) and in genetic researches (Baum et al., 2004) studied the pollen. Malva L. (mallow) is the genus within the Malva- ceae Juss. family, which includes 25–40 species and sev- eral hybrids (Ray 1995). This genus contains herbaceous annual, biennial, and perennial species that are native to regions of Africa, Asia, and Europe (Shaheen et al., 2009). In medicine, mallow species are used in the treat- ment of respiratory, urinary, and digestive problems as they have high bactericidal, antiulcerogenic, anti-inflam- matory, hepatoprotective, and antidiabetic activities (Pandey et al, 2012). The Malva genus is morphologically very diverse, but some species are hardly distinguish- able based on morphological features (Escobar et al., 2009). Several studies have been conducted to clarify the taxonomic affiliation of Malva species using differ- ent features, such as molecular data (nuclear ribosomal DNA (rDNA), internal transcribed spacer (ITS) region, intron–exon splice junction (ISJ), and inter simple sequence repeat polymerase chain reaction (ISSR) mark- ers), differentiation of seed and seed coat structure (El Naggar, 2001), morphology of pollen grains (El Naggar, 2004), epidermal structures and stem hairs (Akçin and Özbucak, 2006), and plant morphological traits (Michael et al., 2009). The variability in mallow species is due, at least in part, to hybridization. Natural crossings between M. pusilla Sm. and M. neglecta, M. alcea L., and M. mos- chata L. as well as M. sylvestris and M. neglecta were found in Europe. Ray (1995) stated that hybridization or polyploidy is probably a factor in the evolution of these species, but this aspect has not been investigat- ed so far. The taxonomy and systematics of the Malva genus are still unclear and very complicated. Taxo- nomic doubts have appeared because of the high level of homoplasty in morphological traits that are usually used as diagnostic features (Chen et al. 2021 ; BI et al. 2021). Sequence-related amplified polymorphism (SRAP) is PCR –based marker system. It is one of the efficient and simple marker systems to study gene mapping and gene tagging in plant species (Li and Quiros 2001), and SRAP are potential markers to assess plant systematics and genetic diversity studies (Robarts and Wolfe 2014). Previously, Wu et al. (2010) assessed genetic diversity and population structure in Pogostemon cablin  with the aid of SRAP markers. SRAP markers were successfully implemented in Lamiaceae, Geraniacea, Caryophyl- lacea and Rosaceae family to study natural populations and variations within the family (Peng et al., 2021; Ma et al.,2021). Objectives of the study were; a) to estimate genetic diversity; b) to evaluate population relationships using NJ approaches. Current results have implications in breeding and conservation programs. The present study is the first report on genetic diversity and phyloge- netic relationships between and within Malva species in Iraq using SRAP markers. MATERIALS AND METHODS Plants collection Five wild Malva species (Malva neglecta Wallr., Mal- va pusilla Sm., Malva sylvestris L., Malva vericillata L., Malva nicaeensis All..) in Halabja, Sulaimanieh, Kalar, Chamchamal and Basreh Provinces of Iraq were selected and sampled during July-August 2015-2020. Morpho- metric and SRAP analyses on 70 plant accessions were carried out. Five to twelve samples from each population belonging to three different species were selected based on other eco-geographic characteristics. Detailed infor- mation about locations of samples and geographical dis- tribution of species are mentioned. 103Comparative study and genetic diversity in Malva using srap molecular markers Morphological studies Five to twelve samples from each species were used for Morphometry. In total 36 morphological (13 quali- tative, 23 quantitative) characters were studied. Data obtained were standardized (Mean= 0, variance = 1) and used to estimate Euclidean distance for clustering and ordination analyses (Podani 2000). Morphological characters studied are: corolla shape, bract shape, calyx shape, calyx length, calyx width, calyx apex, calyx mar- gins, bract length, corolla length, corolla width, corolla apex, leaf length and leaf width, leaf apex, leaf margins, leaf shape, leaf gland and bract margins. Sequence-related amplified polymorphism method Fresh leaves were used randomly from one to twelve plants. These were dried with silica gel powder. Genomic DNA was extracted while following previous protocol. SRAP assay was performed as described previously (Li and Quiros 2001). Five SRAP in different primer combi- nations were used (Table 1). The overall reaction volume consisted of 25 μl. This PCR reaction was carried out in Techne thermocycler (Germany). The following cycles and programs were observed. The initial denaturation step was performed for 5 minutes at 94°C. The initial denaturation step was followed by 40 cycles for 1 min- ute at 94°C; 1 minute at 52-57°C, and 2 minutes at 72°C. The reaction was completed by a final extension step of 7-10 min at 72°C. Staining was performed with the aid of ethidium bromide. DNA bands/fragments were com- pared against a 100 bp molecular size ladder (Fermentas, Germany). Data analyses ANOVA (Analysis of variance) was conducted to assess morphological differences among species. Prin- cipal component analysis (PCA) was implemented to identif y variable morphological characters in Malva species. Multivariate statistical analyses i.e., PC analy- sis, were performed in PAST software version 2.17 (Hammer et al. 2001). Molecular analyses Sequence-related amplified polymorphism (SRAP) bands were recorded. Presence and absence of bands were scored present (1) and absent (0), respectively. Total loci (NTL) and the number of polymorphism loci (NPL) for each primer were calculated. Further- more, the polymorphic ratio was assessed based on NPL/NTL values. Polymorphism information content was calculated as previously suggested by Roldan-Ruiz et al. (2000). Parameter like Nei’s gene diversity (H), Shannon information index (I), number of effective alleles, and percentage of polymorphism (P% = number of polymorphic loci/number of total loci) were deter- mined. Nei’s genetic distance among populations was used for Neighbor Joining (NJ) clustering and Neigh- bor-Net networking. Mantel test checked the correla- tion between geographical and genetic distances of the studied populations (Podani 2000). These analyses were done by PAST ver. 2.17 (Hammer et al. 2012), DARwin ver. 5 (2012) and SplitsTree4 V4.13.1 (2013) software. To assess the population structure of the pistachio geno- types, a heuristic method based on Bayesian clustering algorithms were utilized. The clustering method based on the Bayesian-model implemented in the software program STRUCTURE () was used on the same data set to better detect population substructures. This clus- tering method is based on an algorithm that assigns genotypes to homogeneous groups, given a number of clusters (K) and assuming Hardy-Weinberg and link- age equilibrium within clusters, the software estimates allele frequencies in each cluster and population mem- berships for every individual (Pritchard et al. 2000). The number of potential subpopulations varied from two to ten, and their contribution to the genotypes of the accessions was calculated based on 50,000 iteration burn-ins and 100,000 iteration sampling periods. The most probable number (K) of subpopulations was iden- tified following Evanno et al. (2005). In K-Means clus- tering, two summary statistics, pseudo-F, and Bayesian Information Criterion (BIC), provide the best fit for k. Pairwise genetic similarity between species was evalu- ated to reveal genetic affinity between species (Jaccard, 1908). Unbiased expected heterozygosity and Shannon information index were calculated in GenAlEx 6.4 soft- ware (Peakall and Smouse, 2006). Table 1. SRAP primer information and results. Primer name NTLa NPLb Pc PICd RPe Em3-Me4 27 27 100.00% 0.55 33.24 Em3-Me1 16 10 75.00% 0.11 55.55 Em4-Me1 17 17 100.00% 0.39 11.23 Em5-Me1 10 10 100.00% 0.50 38.55 Em5-Me2 19 13 66.00% 0.32 44.65 Mean 19 15 83.10% 0.44 39.23 Total 89 80 104 Syamand Ahmed Qadir et al. RESULTS Morphometry The ANOVA findings showed substantial differenc- es (p<0.01) between the species in terms of quantitative morphological characteristics. Principal component anal- ysis results explained 60% cumulative variation. The first PCA axis explained 40% of the total variation. The high- est correlation (> 0.7) was shown by morphological char- acters such as corolla apex, seed length; number of seg- ment stem leaves; calyx length, calyx width; bract length and leaf shape. The morphological characters of five Mal- va species are shown in PCA plot (Figure 1). Each species formed separate groups based on morphological charac- ters. The morphometric analysis showed clear difference among Malva species and separated each groups. Species identification and genetic diversity Five (5) suitable primer combinations (PCs), out of 25 PCs were screened in this research. Figure 2 illus- trates the banding pattern of Em2-Me4 and Em4-Me1 primer by the SRAP marker profile. Eighty (80) ampli- fied polymorphic bands (number of polymorphic loci) were produced. These bands (fragments) had differ- ent range i.e. 100bp to 3000 bp. Maximum and mini- mum numbers of polymorphic bands were 27 and 10 for Em3-Me4 and Em5-Me1, respectively. Each primer produced 15 polymorphic bands on average. The PIC ranged from 0.11 (Em3-Me1) to 0.55 (Em1-Me4) for the 5 SRAP primers, with an average of 0.44 per primer. RP of the primers ranged from 11.23 (Em4-Me1) to 55.55 (Em3-Me1) with an average of 39.23 per primer (Table 2). The calculated genetic parameters of Malva species are shown (Table 2). The unbiased heterozygosity (H) varied between 0.077 (Malva sylvestris) and 0.382 (Mal- va pusilla) with a mean of 0.23. Shannon’s information index (I) was maximum in Malva nicaeensis (0.395), where as we recorded minimum Shannon’s information index in Malva sylvestris (0.13). The observed number of alleles (Na) ranged from 1.11 in Malva aegyptia to 1.580 in Malva nicaeensis. The significant number of alleles (Ne) ranged from 1.077 (Malva nicaeensis) to 1.432 (Malva pusilla). Figure 1. Morphological characters analysis of the Malva species by PCoA plot. Table 2. Genetic diversity parameters in the different Malva popu- lations, species, and cultivars; Abbreviations. Population %P N Na Ne I He UHe Malva pusilla Sm. 53.00% 15.000 1.500 1.432 0.388 0.310 0.382 Malva sylvestris L. 22.11% 10.000 1.333 1.177 0.130 0.033 0.077 Malva vericillata L. 33.33% 12.000 1.300 1.388 0.271 0.167 0.288 Malva nicaeensis All. 49.00% 17.000 1.580 1.077 0.395 0.156 0.277 Malva aegyptia L. 30.33% 13.000 1.110 1.366 0.299 0.238 0.144 105Comparative study and genetic diversity in Malva using srap molecular markers Analysis of Molecular Variance results in signifi- cant genetic difference (p = 0.01) among Malva species. The majority of genetic variation occurred among spe- cies. AMOVA findings revealed that 66% of the total variation was between species and comparatively less genetic variation was recorded at the species level (Table 3). Genetic difference between Malva species was high- lighted by genetic statistics (Nei’s GST), as evident by sig- nificant p values i.e. Nei’s GST (0.578, p = 0.01) and D_est values (0.829, p = 0.01) . NJ tree and UPGMA clustering produced similar results therefore only NJ tree is presented and discussed (Figure 3). This result show that molecular characters studied can delimit Malva species in two different major clusters or groups. In general, two major clusters were formed in NJ tree (Fig. 3), 20 individual of Malva nicaeen- sis and Malva aegyptia formed a single cluster. Cluster II contained two sub-clusters, and most of individual Malva pusilla; Malva sylvestris and Malva vericillata formed clus- ter II. There were 50 individuals in this cluster. We detected strong correlation between geographi- cal and genetic distances (r = 0.45, p=0.0002) and gene flow (Nm) score of 0.48 was reported among species. Detailed information about genetic distances and genetic identity (Nei’s) are described (Table not included). The findings suggested that there was the highest degree of genetic similarity (0.91) between Malva vericillata and Malva nicaeensis. On the contrary to this, Malva pusil- la and Malva aegyptia (0.70) had lowest genetic resem- blance. The Evanno test ΔK =5 (Figure not included), showed the genetic details of the Malva species. Accord- ing to STRUCTURE analysis, the Malva species are genetically differentiated due to different allelic struc- tures (Figure not included). Limited gene flow results were supported by K-Means and STRUCTURE analyses too. We could not identify substantial gene flow among the Malva species. This result is in agreement with grouping we obtained with NJ tree (Figure 3), as these populations were placed close to each other. As evi- denced by STRUCTURE plot based on admixture mod- el, these shared alleles comprise very limited part of the genomes in these populations and all these results are in agreement in showing high degree of genetic stratifica- tion within Malva populations. Figure 2. Electrophoresis gel of studied ecotypes from DNA frag- ments produced by SRAP profile; 1, 7: Malva neglecta 2,8: Malva parviflora 3,9: Malva pusilla. 4,10: Malva sylvestris 5,11: Malva veri- cillata 6,12: Malva nicaeensis = Ladder 100 bp. Table 3. Analysis of molecular variance (AMOVA) of the studied species. Source df SS MS Est. Var. % ΦPT Among Pops 70 2701.394 77.782 10.166 66% 66% Within Pops 10 111.449 390.19 27.833 34% Total 80 2875.807 37.060 100% df: degree of freedom; SS: sum of squared observations; MS: mean of squared observations; EV: estimated variance; ΦPT: proportion of the total genetic variance among individuals within an accession, (P < 0.001). Figure 3. Neighbor-Joining tree of populations in Malva species based on SRAP molecular markers. 106 Syamand Ahmed Qadir et al. DISCUSSION In the present study, we used morphological and molecular (SR AP) data to evaluate species relation- ships in Malva. Morphological analyses of Malva spe- cies showed that quantitative indicators (ANOVA test results) and qualitative characteristics are well differ- entiated from each other. PCA analysis suggests that morphological characters such as corolla shape, bract shape, calyx shape, calyx length, calyx width, calyx apex, calyx margins, bract length have the potentials to identify and delimitate Malva species. Principal compo- nent analysis results suggests the utilization of morpho- logical characters to identify and delimitate Malva spe- cies. Morphological characters including corolla length, corolla width, corolla apex, leaf length and leaf width, leaf apex, leaf margins, leaf shape, leaf gland and bract margins play key role in plant systematics and taxono- my. Our work also highlighted the significance of mor- phological characters and molecular data to identify and study species genetic diversity. In general, genetic relationships obtained from SRAP data coincides with morphometric results. This is in accordance with the parameters of AMOVA and genetic diversity results. SRAP molecular markers detected clear genetic differ- ence among species. These results indicate that SRAP have potentials to study plant systematics and taxonomy in Malva members. Given the negative impact of biodiversity threats and overexploitation of Malva plant species in Iran, it is necessary to conduct genetic diversity studies on Malva species. Genetic diversity based studies pave our understanding to develop conservation strategies (Jia et al. 2020; Shi et al., 2021; Zheng et al., 2021; Zhu et al., 2021). Genetic diversity studies are conducted through appropriate selection of primers and indexes includ- ing Polymorphic information content (PIC) and mark- er index (MI) are important indexes to fathom genetic variation in species (Wang et al., 2021; Yin et al., 2021; Zhao et al., 2021). Common logic suggests that different makers have different abilities to assess genetic diversity, and usually, genetic diversity is linked with polymor- phism (Sivaprakash et al. 2004). In the present work, 5 Malva species were charac- terized with 5 SRAP markers. The results confirm the efficiency of microsatellite markers for fingerprinting purposes. Our results demonstrated that the PIC ranged from 0.11 (Em3-Me1) to 0.55 (Em3-Me4) for the 5 SRAP primers, with an average of 0.44 per primer. RP of the primers ranged from 11.23 (Em4-Me1) to 55.55 (Em3- Me1) with an average of 39.23 per primer. Diversity study in Malva species Malvaceous germplasm has been variously investi- gated by different molecular marker techniques but the earlier studies either focused on the comparison of the Malvaceae with other families in the order Malvales or to explore the genetic relationships and diversity within and among population and limited number of species in the same genus. Very little attention has been given to the analysis at interspecific and intergeneric levels. La Duke and Dobley (1995) has the only worth mentioning work in this regard. Their results showed that, the genet- ic relationships and diversity within and between 12 malvaceous species belonging to five genera are investi- gated by using the Amplified fragment length polymor- phism (AFLP). Shaheen et al., (2009) with used AFLP (Amplified fragment length polymorphism) marker to explore phe- netic relationships and diversity within and between 13 Malvaceae species belonging to 5 different genera. Their primary objective of the study was to evaluate the taxo- nomic potential, usefulness and applicability of AFLP marker system to reconstruct genetic relationships at interspecific and intergeneric level in Malvaceae. Two primer pairs produced a total of 73 bands, of which 70 were polymorphic. According to Celka et al (2010) two categories of DNA markers were used to determine genetic relation- ships among eight Malva taxa. A maximum parsimony analysis validated the division of the genus Malva into the sections Bismalva and Malva. The species classified into those sections formed separate clusters. M. mos- chata was a distinctive species in the section Bismalva, as confirmed by previous genetic research based on ITS and cpDNA sequence analyses. The applied markers revealed a very high level of genetic identity between M. alcea and M. excisa and enabled molecular identification of M. alcea var. fastigiata. Jedrzejczyk and Rewers (2020) applied flow cytom- etry and inter simple sequence repeat polymerase chain reaction (ISSR-PCR) for fast and accurate species identi- fication. Genome size estimation by flow cytometry was proposed as the first-choice method for quick accession screening. Out of the 12 tested accessions, it was possi- ble to identify six genotypes based on genome size esti- mation, whereas all species and varieties were identified using ISSR markers. Flow cytometric analyses revealed that Malva species possessed very small (1.45–2.77 pg/2C), small (2.81–3.80 pg/2C), and intermediate (11.06 pg/2C) genomes, but the majority of accessions pos- sessed very small genomes. The relationships between the investigated accessions showed the presence of two clus- 107Comparative study and genetic diversity in Malva using srap molecular markers ters representing malvoid and lavateroid group of spe- cies. Their results showed that Flow cytometry and ISSR molecular markers can be effectively used in the identifi- cation and genetic characterization of Malva species. CONCLUSIONS The present study investigated the molecular varia- tion of five species. Molecular and morphometric analy- sis confirmed morphological and genetical difference between Malva species. This was first attempt to assess genetic diversity through Sequence-related amplified polymorphism and morphometrics analysis in Iraq. Current study reported two major clusters. These two major groups were separated on the basis of genetic and morphological characters. The genetic similarities between three species was estimated from 0.70 to 0.91. Current study also reported correlation between genetic and geographical distances. This clearly indicated isola- tion mechanism envloved in the ecology of Malva spe- cies . Present results indicated the potential of sequence- related amplified polymorphism to assess genetic diver- sity and genetic affinitiy among Malva species. Current results have implications in biodiversity and conserva- tion programs. Besides this, present results could pave the way for selecting suitable ecotypes for forage and pasture purposes in Iraq. REFERENCES Akçin, Ö.E.; Özbucak, T.B. (2006). 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Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Volume 75, Issue 3 - 2022 Firenze University Press Chromosome Mapping of Repetitive DNAs in the Picasso Triggerfish (Rhinecanthus aculeatus (Linnaeus, 1758)) in Family Balistidae by Classical and Molecular Cytogenetic Techniques Kamika Sribenja1, Alongklod Tanomtong1, Nuntaporn Getlekha2,* Chromosome number of some Satureja species from Turkey Esra Kavcı1, Esra Martin1, Halil Erhan Eroğlu2,*, Fatih Serdar Yıldırım3 L-Ascorbic acid modulates the cytotoxic and genotoxic effects of salinity in barley meristem cells by regulating mitotic activity and chromosomal aberrations Selma Tabur1,*, Nai̇me Büyükkaya Bayraktar2, Serkan Özmen1 Characterization of the chromosomes of sotol (Dasylirion cedrosanum Trel.) using cytogenetic banding techniques Kristel Ramírez-Matadamas1, Elva Irene Cortés-Gutiérrez2, Sergio Moreno-Limón2, Catalina García-Vielma1,* Contributions of species Rineloricaria pentamaculata (Loricariidae:Loricariinae) in a karyoevolutionary context A Cius¹, CA Lorscheider2, LM Barbosa¹, AC Prizon¹, CH Zawadzki3, LA Borin-Carvalho¹, FE Porto4, ALB Portela-Castro1,4 Cadmium induced genotoxicity and antioxidative defense system in lentil (Lens culinaris Medik.) genotype Durre Shahwar1,2,*, Zeba Khan3, Mohammad Yunus Khalil Ansari1 Biogenic synthesis of noble metal nanoparticles using Melissa officinalis L. and Salvia officinalis L. extracts and evaluation of their biosafety potential Denisa Manolescu1,2, Georgiana Uță1,2,*, Anca Șuțan3, Cătălin Ducu1, Alin Din1, Sorin Moga1, Denis Negrea1, Andrei Biță4, Ludovic Bejenaru4, Cornelia Bejenaru5, Speranța Avram2 Polyploid cytotypes and formation of unreduced male gametes in wild and cultivated fennel (Foeniculum vulgare Mill.) 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