Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 74(4): 77-83, 2021 Firenze University Press www.fupress.com/caryologia ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.36253/caryologia-1163 Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Citation: Fahimeh Koohdar, Neda Aram, Masoud Sheidai (2021) Biosystemat- ics, fingerprinting and DNA barcoding study of the genus Lallemantia based on SCoT and REMAP markers. Caryo- logia 74(4): 77-83. doi: 10.36253/caryolo- gia-1163 Received: December 14, 2021 Accepted: December 17, 2021 Published: March 08, 2022 Copyright: © 2021 Fahimeh Koohdar, Neda Aram, Masoud Sheidai. This is an open access, peer-reviewed article published by Firenze University Press (http://www.fupress.com/caryologia) and distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distri- bution, and reproduction in any medi- um, provided 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. Biosystematics, fingerprinting and DNA barcoding study of the genus Lallemantia based on SCoT and REMAP markers Fahimeh Koohdar*, Neda Aram, Masoud Sheidai Department of Plant Sciences and Biotechnology, Faculty of Life Sciences and Biotechnol- ogy, Shahid Beheshti University, Tehran, Iran *Corresponding author. E-mail: f_koohdar@yahoo.com Abstract. Lallemantia is a medicinally important plant in the world. Due to interspe- cific hybridization and horizontal gene transfer, spices relationship and delimitation on the genus Lallemantia is difficult based on different molecular markers. Therefore, selecting the appropriate marker can be important. Fingerprinting techniques continue to be used for genomic profiling for the characterization of germplasm and the estab- lishment of the identity of varieties/hybrids/parental sources of aromatic and medicinal plants. For this, we need to produce detailed information on genetic diversity avail- able in Lallemantia as well as investigate spices relationship and delimitation. There- fore, the present study was performed on Lallemantia species in Iran. We used the start codon targeted and retrotransposon-microsatellite amplified polymorphism molecular marker for our genetic investigation with the following aims: 1- To reveal the species delimitation and species relationship in Lallemantia, and 2- To investigate discrimi- nating power of the start codon targeted and retrotransposon-microsatellite amplified polymorphism markers by Gst and NM analysis. The results obtained revealed that the start codon targeted marker is the best to show the relationships between species while the retrotransposon-microsatellite amplified polymorphism marker is the best for spe- cies delimitation. we found the loci with the high value of Gst (1.00) in start codon targeted and retrotransposon-microsatellite amplified polymorphism markers that can be used in barcoding and fingerprinting of L. royleana. Keyword: Lallemantia, fingerprinting, ScoT, REMA, Iran. INTRODUCTION The genus Lallemantia (Lamiaceae) is composed of 5 species (Lallemantia royleana (Benth.) Benth., L. canescens (L.) Fisch. & C.A.Mey.,L. baldschuanica Gontsch., L. iberica (M.Bieb.) Fisch. &C.A. Mey. and L. peltata(L.) Fisch. & C. A. Mey.) that are widely distributed in Afghanistan, China, India, Kazakh- stan, kyrgyzstan, Iran, Russia, Tajikistan, Turkmenistan, Uzbekistan and Europe (Sheidai et al. 2018). There are all five species in Iran (Rechinger 1982). Lallemantia species are herbaceous with simple leaves, interrupted inflorescence, aristate-toothed bracteoles, and oblong, trigonous, smooth, and mucilaginous nutlets (Harley et al. 2004). These species are well known 78 Fahimeh Koohdar, Neda Aram, Masoud Sheidai as a source of food and medicine plant. For example, L.ibericais used as an oil seed plant in Iran and USSR (Rivera-Nunez and Obonde-Gastro 1992, Dinç et al. 2009), L. royleana Seeds have considerable anti - bacte- rial properties and is a suitable remedy for skin diseas- es and gastrointestinal diseases and also L. peltata that grows in limited area in Iran is known as medicinal plant that contains volatile and essential oil (Mahmood et al. 2013). The molecular systematic study of plants is per- formed with different purposes like: species delimita- tion, population divergence, species relationships, date of divergence determination, etc (Broadhurst et al., 2004; Millar et al., 2011). Various molecular markers have been used to perform the above tasks such as, amplified frag- ments length polymorphism (AFLP), simple sequence repeats (SSRs), inter-simple sequence repeats (ISSRs), start codon targeted (SCoTs), retrotransposon-micro- satellite amplified polymorphism (REMAPs) etc. (e.g., Sheidai et al. 2012, 2013, 2014, Minaeifar et al. 2015, Sab- oori et al. 2019). DNA barcoding is a sequence of DNA that can help in rapid and accurate recognition of species. It has been used in the identification of medicinal plants and has been able to detect actual and original products from its fake type (Heubl et al. 2010, Sheidai et al. 2018). Nucle- ar and chloroplast DNA have been examined for their suitability as barcodes through DNA fingerprinting and DNA sequencing-based approaches. In principle, both approaches can be used to differentiate between indi- viduals, species, and populations and to detect the pres- ence of adulterants. Notwithstanding the increasing use of DNA sequence-based approaches, fingerprinting techniques continue to be used for genomic profiling for characterization of germplasm and establishment of the identity of varieties/hybrids/parental sources of aromatic and medicinal plants (Sheidai et al., 2019). Among DNA markers, there are ubiquitous retro elements in the plant genome like IRAP and REMAP (retrotransposon-microsatellite amplif ied polymor- phism). The REMAP is produced by amplifying the frag- ments between a retrotransposon insertion site and a microsatellite site and employed in fingerprinting, link- age analysis, mapping, analysis of genome evaluation and genetic diversity. REMAP describe the profile of a population, discriminate between species or genotypes and analyze population diversity (Kumar et al. 2010). Start Codon Targeted (SCoT) polymorphisms (Col- lard and Mackill 2009) are dominant and reproducible markers based on the short conserved region flanking the ATG start codon in plant genes and use a single 18-mer primer in the polymerase chain reaction (PCR) assays and high annealing temperature (50 °C). These markers could have potential in genotyping and to reveal polymorphisms that might be directly related to gene function. SCoT markers have been used to assess genetic diversity and structure, in bulked segregant anal- ysis, and for quantitative trait loci (QTL) mapping and DNA fingerprinting (Collard and Mackill 2009, Luo et al. 2010). Due to the morphological similarity of Lallemantia species and sell them in the market as seed, the present study was performed with the following aims: 1- To reveal the species delimitation and species relationship in Lallemantia by SCoT and REMAP markers, and 2- To investigate discriminating power of the SCoT and REM- AP markers by Gst and NM analysis for barcoding and fingerprinting of medicinal spices in Lallemantia. MATERIAL AND METHODS Plant materials Extensive field investigations and collections were under taken during 2013–2015. Forty-two specimens of five species, Lallemantia royleana, L. canescens, L. bald- schuanica Gontsch., L. iberica and L. peltata were ran- domly collected from different geographic populations for molecular study. SCoT and REMAP assay Fresh leaves were put to dry in silica gel powder. Cetyltrimethyl-ammonium bromide -activated charcoal protocol (CTAB) was applied to extract the genomic DNA. The extraction was done by activating charcoal and poly vinyl pyrrolidone (PVP) for binding of poly- phenolics during extraction; for mild extraction and pre- cipitation conditions, the high-molecular weight DNA isolation was boosted without the interference of impuri- ties. The extracted DNA was examined in terms of qual- ity by running on 0.8% agarose (Sheidai et al. 2013). Three REMAP primer combinations, derived from one single IRAP primer (NIKITA) with 3 ISSR primers ((CA)7GT, (GA)9T, (GA)9C) were tested on plants sam- ples. Using a 25 µL volume containing 20 ng genomic DNA and 3 U of Taq DNA polymerase (Bioron, Germa- ny); 50 mMKCl; 10 mMTris-HCl buffer at pH;8 1.5mM MgCl2; 0.2 mM of each dNTP (Bioron, Germany); 0.2 µM of each primer, polymerase chain reaction (PCR) was implemented. The following program was used for amplification of nuclear region in a PCR reaction: 5 min initial denatura- 79Biosystematics and fingerprinting study of the genus Lallemantia based on SCoT and REMAP markers tion step 94°C, followed by 40 cycles of 1 min at 94°C; 1 min at 53.5°C and 2 min at 72°C. The reaction was completed by a final extension step of 7 min at 72°C. The amplification products were observed by running on 1% agarose gel, followed by the ethidium bromide stain- ing. The fragment size was estimated by using a 100 bp molecular size ladder (Fermentas, Germany). Four primers (SCOT1, SCOT2, SCOT36, and SCOT41) based on Collard and Mackill (2009) for monocotyledons plants were selected (Collard and Mackill 2009). These primer sequences are: SCoT1: CAACAATGGCTACCACCA, SCoT2: CAACAATG- GCTACCACCC , SCoT36: GCA ACA ATGGCTAC- CACC and SCoT41: CAATGGCTACCACTGACA. PCR reaction mixture with total volume of 25 μl contained 10 mMTris-HCl buffer (pH = 8), 50 mMKCl, 1.5 mM MgCl2, 0.2 mM of dNTP (Bioron, Germany), 0.2 μM of primer, 20 ng genomic DNA and 1U of Taq DNA poly- merase (Bioron, Germany). The amplification reactions were performed in Tech- ne thermocycler (Germany) with the following program: 5 min at 94 ºC, 40 cycles of 1 min at 94 ºC, 1 min at 49–58 ºC (SCOT1 50 ºC, SCOT2 49 ºC, SCOT36 50 ºC, SCOT41 58 ºC) and 1 min at 72 ºC and a final cycle of 7 min at 72 ºC. The amplification products were visualized by running on 2% agarose gel, stained with syber green (Powerload, Kosar Co. Iran). The fragment size was esti- mated by using a 100 bp molecular size ladder (Fermen- tas, Germany). Data analyses The SCoT and REMAP bands obtained were treated as binary characters and coded accordingly (presence = 1, absence = 0). The number of private bands versus common bands and genetic diversity parameters like: The percentage of allelic polymorphism, allele diversity (Weising, 2005), Nei’ gene diversity (He), and Shannon information index (I) (Weising 2005), were determined. We used GenAlex 6.4 for these analyses (Peakall and Smouse 2006). Discriminating power of REMAP and SCoT markers investigated by Gst and NM analysis as implemented in POPGENE32. Grouping of the species was done by different clus- tering and ordination methods such as unweighted paired group using average (UPGMA), Multidimen- sional scaling (MDS), and Principal components analysis (PCA) (Podani 2000). PAST version 2.17 (Hammer et al., 2012) was used for multivariate analysis. RESULTS SCoT results Almost all the SCoT primers produced bands were used and finally a data matrix of 70 × 42 was formed for further analysis. Based on band pattern in Lallemantia genus, the highest number of privet bands were observed in L. iberica and L. baldschuanica had the lowest value (Fig. 1). Genetic variation parameters were investigated in 5 species of Lallemantia genus. Highest level of Shannon index (0.336), expected heterozygosity (0.218), and per- centage of polymorphism (70.15) in L. iberica and lowest level of Shannon index (0.105), expected heterozygosity (0.072) and percentage of poly morphism (17.91) were observed in L. baldschuanic species (Table 1). The AMOVA test showed significant genetic differ- ences among Lallemantia species (P = 0.001). The results show that the species of this genus have been genetically distinguished from each other using SCoT marker. Different ordination and clustering methods like PCA, MDS and UPGMA produced similar results; therefore, only UPGMA plot is presented here. UPGMA Figure 1. Band patterns of ScoT marker in Lallemantia spices. 1: L. royleana, 2: L. iberica, 3: L. canescens, 4: L. peltata, 5: L. baldschuanica. 80 Fahimeh Koohdar, Neda Aram, Masoud Sheidai plot of SCoT markers(Fig. 2) grouped the specimens of L. platata, L. canescens and L. baldshuanica together in a single cluster, separated from each other but in L. iberica and L. royleana the specimens were divided in two sepa- rate clades. In this plot, L. royleana and L. baldshuanica as well as L. peltata were placed close to each other while L. iberica and L. canescens was placed far from them. REMAP results Almost all the REMAP primers produced bands were used and finally a data matrix of 55 (number of bands) × 42 (number of samples) was formed for further analysis. Based on band pattern in Lallemantia genus, the highest number of privet bands were observed in L. iberica and L. baldschuanica had the lowest value (Fig. 3). Genetic variation parameters and band patterns were investigated in 5 species of Lallemantia genus. Highest level of Shannon index (0.111), expected het- erozygosity (0.073), and percentage of polymorphism (21.88%) in L. baldschuanic and lowest level of Shannon index (0.017), expected heterozygosity (0.011) and per- centage of poly morphism (0.012) were observed in L. royleana species (Table 2). AMOVA showed significant genetic differences among Lallemantia species (P = 0.001). The AMOVA test showed 45% inter-species diversity and 55% intra-species diversity. The results show that the species of this genus Table 1. Genetic diversity parameters in Lallemantia species based on SCoT marker. Species N Na Ne I He uHe P L. royleana 10.000 0.925 1.204 0.201 0.128 0.135 44.78% L. iberica 10.000 1.522 1.354 0.336 0.218 0.230 70.15% L. peltata 5.000 0.761 1.191 0.163 0.110 0.122 29.85% L. canescens 7.000 0.821 1.207 0.183 0.121 0.130 35.82% L. bahdchuanic 5.000 0.493 1.128 0.105 0.072 0.080 17.91% Abbreviations: N = No of plants studied; Na = No. of alleles; Ne = Effective No. of alleles; He = Gene diversity; uHe = Unbiased gene diversity; P = Polymorphism percentage. Figure 2. UPGMA plot of SCoT marker in Lallemantia. R: L. royleana, IB: L.iberica, C: L. canescens, P: L. peltata, B: L. baldschuanica. 81Biosystematics and fingerprinting study of the genus Lallemantia based on SCoT and REMAP markers have been genetically distinguished from each other using SCoT marker. Different ordination and clustering methods like PCA, MDS and UPGMA produced similar results; therefore, only UPGMA plot is presented here. UPGMA plot of molecular markers (Fig. 4) grouped the speci- mens of all species together in a single cluster, separated from the other species. This means that REMAP molec- ular markers are of taxonomic value and can delimit the Lallemantia species. In this plot, L. royleana and L. baldschuanica as well as L. peltata and L. canescens were placed close to each other while L. iberica was placed far from them. Barcoding and fingerprinting Discriminating power analysis of molecular markers is important for fingerprinting and barcoding. for this purpose, Gst and Nm parameters was measured. The value of Gst in Seven loci in SCoT and 13loci in REMA marker in Lallemantia was 1 .00 while the mean Nm value was 0.00 which indicated that these markers can be used in L. royleana differentiation of other species. Comparison of SCoT and REMAP markers In this study, SCoT marker was able to separate only three species of five species, while REMAP marker was able to demonstrate the boundary between any five species (Figs 1 and 2). According to AMOVA result, SCoT marker was more successful in showing intraspecific diversity. DISCUSSION The present study revealed that species delimitation based on REMAP markers was more successful than SCoT marker. however, this marker was more successful in showing intraspecific diversity. These results are con- sisting with previous findings (Al-Qurainy et al. 2015, Saboori et al. 2019). L. baldschuanica and L. royleana as well as L. cane- scens and L. iberica are similar to each other based on morphological and micro morphological (nutlet and pollen structure) studies (Talebi and Rezakhanlou 2010; Kamrani et al. 2018). Lamiaceae family has many phylogenetically unre- solved genera and therefore many species are of not determined relationship due to the conf lict between molecular data and potential inter-specific hybridization as well as horizontal gene transfer. Sheidai et al. 2018, revealed that the relationships between Lallemantia species based on cp- DNA, ITS and ISSR molecular markers differed from morphological and micromorphological as well as to each other due to inter-specific hybridization and horizontal gene transfer. Table 2. Genetic diversity parameters in Lallemantia species based on REMAP marker. Species N Na Ne I He uHe P L. royleana 5.000 0.594 1.017 0.017 0.011 0.012 3.13% L. iberica 5.000 0.656 1.075 0.075 0.048 0.054 15.63% L. canescens 5.000 0.656 1.036 0.053 0.030 0.033 15.63% L. Peltata 5.000 0.594 1.109 0.090 0.061 0.068 15.63% L. baldschuanic 5.000 0.719 1.123 0.111 0.073 0.081 21.88% Abbreviations: N = No of plants studied; Na = No. of alleles; Ne = Effective No. of alleles; He = Gene diversity; uHe = Unbiased gene diversity; P = Polymorphism percentage. Figure 3. Band patterns of REMAP marker in Lallemantia Spices. 1: L. royleana, 2: L.iberica, 3: L. canescens, 4: L. peltata, 5: L. baldschuanica. 82 Fahimeh Koohdar, Neda Aram, Masoud Sheidai Our result suggested conf lict between SCoT and REMAP marker in relationship of Lallemantia species but in both markers, L. baldschuanica and L. royleana were placed to each other. The SCoT result was more similar to the previous studies (Talebi and Rezakhanlou 2010; Kamrani et al. 2018), so this marker can be sug- gested to study the inter-species relationships in Lalle- mantia. In the present study, we found the loci with the high value of Gst (1.00) in SCoT and REMAP markers that can be used in barcoding and fingerprinting of L. royleana. REFERENCES Al-Qurainy, F., Khan, S., Nadeem, M., Tarroum, M. (2015) SCoT marker for the assessment of genetic diversity insaudiarabian date palm cultivars. Pak. J. Bot. 47: 637-643. Broadhurst, L., Byrne, M., Craven, L., Lepschi, B. (2004) Genetic congruence with new species boundaries in the Melaleuca uncinata complex (Myrtaceae). Aust. J. Bot. 52, 729–737. Collard, B.C.Y., D.J. Mackill., 2009. Start codon targeted (SCoT) polymorphism: a simple, novel DNA marker technique for generating gene-targeted markers in plants. Plant. Mol. Biol. Report. 27: 86-93. Dinç, M., Pinar, N.M., Dogu, S.L., Yildirimli, S.I. (2009) Micromorphological studies of Lallemantia L. (Lami- aceae) species growing in Turkey. ACTA. BIOL. CRACOV. SER. BOT. 51: 45–54. Hammer, Ø., Harper, D.A.T., Ryan, P.D. (2012) PAST: Paleontological Statistics software package for educa- tion and data analysis. Palaeontol. Elec. 4: 9 Harley, R.M., Atkins, S., Budantsev, A.L., Cantino, P.D., Conn, B.J., Grayer, R., Harley, M.M., De Kok, R., Krestovskaja, T., Morales, R., Paton, A.J., Ryding, O., Upson, T. (2004) The families and genera of vascu- Figure 4. UPGMA plot of REMAP markers in Lallemantia. R: L. royleana, IB: L.iberica, C: L. canescens, P: L. peltata, B: L. baldschuanica 83Biosystematics and fingerprinting study of the genus Lallemantia based on SCoT and REMAP markers lar plants.In: Kadereit, JW (ed)Lamiaceae (Lamiales). Springer, Berlin, pp 167-282. Heubl, G., 2010. New aspects of DNA-based authentica- tion of Chinese medicinal plants by molecular bio- logical techniques. Planta. Med. 76, 1963–1974. htt- ps://doi.org/10.1055/s-0030-1250519. Kamrani, A., Riahi, M. (2017) Using molecular data to test the monophyly of Lallemantia in the subtribe Nepetinae (Mentheae, Lamiaceae). Plant. Biosyst. 152: 857-862. Kumar, M.,Qiang, X.X., Deng, B. (2010) Utility of RAPD, ISSR, IRAP and REMAP markers for the genetic analysis of Citrus Spp. Sci. Hortic. 124: 254-261. Luo, C., X.H. He, H. Chen, S.J. Ou, M.P. Gao, J.S. Brown, C.T. Tondo and R.J. Schnell. (2011) Genetic diversity of mango cultivars estimated using SCoT and ISSR markers. Biochem. Syst. Ecol. 39: 676-684. Mahmood, S., Hayat, M.Q., Sadiq, A., Ishtiaq, S.h., Malik, S., Ashra, M. (2013) Antibacterial activity of LallemantiaroyleanaBenth. indigenous to Pakistan. Afr. J.Microbiol. Res. 7: 4006-4009. Minaeifar, A.A., Sheidai, M., Attar, F., Noormohammadi, Z., Ghasemzadeh-Baraki, B. (2015) Genetic and mor- phological diversity in Cousiniatabrisiana (Asterace- ae) populationsBiologia. 70: 328-338. Millar, M. A., Byrne, M., O’Sullivan, W.O. (2011) Defin- ing entities in the Acacia saligna (Fabaceae) species complex using a population genetics approach. Aust. J. Bot. 59: 137-148. Peakall, R., Smouse, P.E., 2006. GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Mol. Ecol. Notes. 6: 288-295. Podani, J. (2000) Introduction to the Exploration of Mul- tivariate Data. Backhuyes, Leiden. Sheidai, M., Seif, E., Nouroozi, M., Noormohammadi, Z. (2012) Cytogenetic and molecular diversity of Cir- siumarvense (Asteraceae) populations in Iran. Journ. Jap. Bot. 87:193-205. Sheidai, M., Zanganeh, S., Haji-Ramezanali, R., Nouroozi, M., Noormohammadi, Z., Ghsemzadeh- Baraki, S. (2013) Genetic diversity and population structure in four Cirsium (Asteraceae) species. Bio- logia.68: 384-397. Sheidai, M., Ziaee, S., Farahani, F., Talebi, S.M., Noor- mohammadi, Z., Hasheminejad Ahangarani Fara- hani, Y. (2014) Infra-specific genetic and morpho- logical diversityinLinum album (Linaceae). Bio- logia. 69: 32e39 Sheidai, M., Koohdar, F., Moradiyanpoode, Z. (2018) Molecular phylogeny ofLallemantia L. (lamiaceae): incongruence between phylogenetic trees and the occurrence of hgt. Genetika. 50 (3): 907-918. Sheidai, M.,Tabaripour, R., Talebi, S.M., Noormohamma- di, Z.,Koohdar, F. (2019) Adulteration in medicinally important plant species of Ziziphora in Iran market: DNA barcoding approach .Ind. Crops. Prod. 130: 627–633. Rivera Nunez, D., Obon, D.E., Gastro, C. (1992) The eth- nobotany of Lamiaceaeof old world. In: Harley RM, Reynolds T (eds.) Advances in Lamiaceae. Science. Royal Botanical Gardens, Kew, pp 455-473. Rechinger, K. H., 1982. Labiatae. In: Rechinger KH (eds.) Flora Iranica, Austria, Graz, Wiena: Academische- Druck-und Verlasantalt, pp 25-44. Saboori S, Noormohammadi Z, Sheidai M, Marashi S.M. (2019) SCoT molecular markers and genetic finger- printing of date palm (Phoenix dactylifera L.) culti- vars. Genet. Resour. Crop. Evol. 67 (1): 73-82. Talebi, S.M., Rezakhanlou, R. (2010) Micromorphologi- cal study in Lallemantia Fisch. et Met. (Lamiaceae) in Iran. Weising, K., Nybom, H., Wolff, K., Kahl, G. (2005) DNA Fingerprinting in Plants, second ed. Boca Rayton, USA. Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Volume 74, Issue 4 - 2021 Firenze University Press Cytogenetic analyses in three species of Moenkhausia Eigenmann, 1903 (Characiformes, Characidae) from Upper Paraná River (Misiones, Argentina) Kevin I. Sánchez1,*, Fabio H. Takagui2, Alberto S. Fenocchio3 Genetic variations and interspesific relationships in Lonicera L. (Caprifoliaceae), using SCoT molecular markers Fengzhen Chen1, Dongmei Li2,* , Mohsen Farshadfar3 The new chromosomal data and karyotypic variations in genus Salvia L. 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