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Morphological diversity among the studied poppy (Papaver) accessions. Representative images showing (a) the distinct floral morphologies, (b) the diversity in capsule shape and size and (c) the different seed colour types recorded for the accessions. Accession names are indicated.
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Opium poppy ( Papaver somniferum ) is one of the world’s oldest medicinal plants and a versatile model system to study secondary metabolism. However, our knowledge of its genetic diversity is limited, restricting utilization of the available germplasm for research and crop improvement. We used genotyping-by-sequencing to investigate the extent of g...
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... The DNA barcode system based on the SNP markers derived from chloroplast DNA [27] also has limits for intraspecific discrimination within P. somniferum L. genotypes due to the high conservativeness of the chloroplast genome. The genotyping-by-sequencing analysis [28] approach is probably the most efficient but also the most technically, time-, and financially demanding. Functional applications were developed for human genetic forensics and some farm and wild animal species. ...
Papaver somniferum L. is cultivated for its edible seeds and for the production of alkaloids. A serious problem in seed trade and processing is the intentional mixing of excellent food-quality seeds with non-food-grade-quality seeds. Tracking the correct or illegitimate handling of seeds requires an efficient method for discrimination and individualization of poppy varieties. As in human and animal forensics, DNA variable regions containing short tandem repeats (STRs) located either in non-coding DNA or in gene sequences (EST-STRs) are preferred markers for discrimination between genotypes. Primers designed for 10 poppy EST-STR loci not analyzed so far were tested for their discriminatory ability on a set of 23 related P. somniferum L. genotypes. Thirty-three EST-STR alleles were identified together. Their polymorphic information content (PIC) values were in the range of 0.175‒0.649. The PI value varied in the range of 0.140‒0.669, and the cumulative PI was 1.2 × 10−5. PIsibs values varied between 0.436 and 0.820 and the cumulative value was lower (5.0 × 10−3). All analyzed genotypes were distinguished mutually, each with its own unique EST-STR profile. These newly developed EST-STR markers more effectively discriminated P. somniferum L. genotypes, even those genotypes whose DNA profiles were previously identical.
... Opium poppy (Papaver somniferum L., diploid no.=22), is an annual, self-pollinated plant species with a history dating back to the earliest Neolithic ages (Hong et al., 2022). Native to the Western Mediterranean region of Europe, it is now grown in various countries including India, Russia, Egypt, Yugoslavia, Poland, Germany, Netherlands, China, Japan, Argentina, Spain, Bulgaria, Hungary, and Portugal for its legal therapeutic application. ...
... Examples of such molecular markers include Conserved DNA and gene family-based markers (CDMs), Transposable elements-based markers (TEMs), RNA-based markers like EST-SSRs, cDNA-amplified fragment length polymorphism (cDNA-AFLP), sequence-related amplified polymorphism (SRAP), Single nucleotide polymorphism (SNPs), and promoter based markers such as Start Codon targeted polymorphism (SCoT), CAAT box derived polymorphism (CBDP), among others (Poczai et al., 2013). In the context of opium poppy, various PCR-based molecular markers, including arbitrarily amplified, gene-targeted, and sequence-specific markers, have been extensively utilized to evaluate genetic assortment in numerous studies (Dittbrenner et al., 2008;Saunders et al., 2001;Hari et al., 2009;Acharya, Sharma, 2009;Selale et al., 2013;Masarova et al., 2015;Guclu et al.,2014;Parmaksiz, Ozcan, 2011;Celik et al., 2016;Sharma et al., 2020;Vašek et al., 2020;György et al., 2022;Hong et al., 2022). However, selecting the most appropriate marker techniques for genetic differentiation and relatedness studies is not always straightforward and depends on factors like available resources, species biology, and research objectives. ...
... Assessment of genetic assortment in licit CPS poppy is a crucial step towards genetic improvement and researchers across the globe have already utilized various arbitrarily amplified molecular markers to achieve this goal. However, recent advancements in genomic research in opium poppy initiated a trend toward the utilization of sequence-targeted functional markers such as genomic SSRs, EST-SSRs, and SNPs for genetic diversity, and trait mapping (Hong et al., 2022;György et al., 2022;Vašek et al., 2020;Celik et al., 2016;Selale et al., 2013) studies. Conversely, the applicability of molecular descriptors or markers depends on several factors like the availability of genetic resources, nature of polymorphism, inheritance, cost, reproducibility, informativeness, reliability, level of resolution, methodology, etc. Nowadays, gene-targeted and functional markers are being used more frequently for varietal identification, purity testing, and quality assessment in various field crops along with genetic diversity studies as these are inexpensive, relatively easy to handle, quick, highly reproducible and highly informative, however, utilization of these markers in case of medicinal plants is very limited and not well defined. ...
Targeted fingerprinting marker techniques have emerged as promising tools in plant sciences, particularly in plant breeding. To initiate an effective breeding program for a licit but industrially potential plant, the opium poppy, a comparative assessment of these markers was conducted. In this study, we compared the Start Codon Targeted polymorphism (SCoT), CAAT box-derived polymorphism (CBDP), and Expressed sequence tag-simple sequence repeats (EST-SSRs) markers for their competence and informativeness in revealing genetic identity, relatedness, and diversity among ten released varieties of the Indian opium poppy. Among the markers studied, EST-SSR markers demonstrated higher discriminating power, attributed to their ability to unveil comparatively higher polymorphic information content, marker index, and effective marker index. While all three marker systems effectively discriminated the poppy varieties, EST-SSR markers showed greater genetic diversity and a more informative ancestral history-based dendrogram topology. Significant correlation coefficients were observed between EST-SSR and CBDP markers, particularly in the combined marker dataset. Furthermore, the co-phenetic correlation was significant for three marker systems, with EST-SSR markers showing maximum correlation. Notably, the dendrograms constructed using EST-SSR markers and combined marker data displayed a significant and positive correlation with phenotypic traits assessed for the ten poppy varieties. Overall, the findings highlight the superiority of EST-SSR markers in deciphering genetic relationships among Indian opium poppy varieties and suggest their potential as valuable tools in breeding and genetic diversity studies.
... For example, the active ingredient in aspirin was first isolated from the bark of the willow tree, which had been used for centuries to relieve pain and reduce fever. Similarly, morphine, still widely used today as a painkiller, was first isolated from the opium poppy, a traditional medicine that had been used for thousands of years (Desborough & Keeling, 2017;Hong et al., 2022). Other traditional medicines used for centuries and have since been studied scientifically include quinine, used to treat malaria; digitoxin, used to treat heart failure; and pilocarpine, used to treat dry mouth and other conditions (Pilocarpine, 2016). ...
The safest and most effective sources of medications are natural and traditional medicines derived from plants and herbs. In Western India, various parts of the Dalbergia sissoo plant, which belongs to the Fabaceae family, have been traditionally used to treat different types of cancer by the local tribes. However, this claim has not been scientifically proven yet. Thus, the purpose of this study was to examine the antioxidant (2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity) and anticancer effects of different plant extracts from Dalbergia sissoo bark, root, and branch on six different cancer cell lines (K562, PC3, A431, A549, NCIH 460, and HEK 293 T) using in vitro cell viability and cytotoxicity assays. The study also involved in silico docking, MD simulation, and ADME studies of previously reported bioactive compounds from the same parts of the plant to confirm their bioactivity. The DPPH radical scavenging experiment findings showed that the methanol: water extract of the bark had a more significant antioxidant activity IC50 (45.63 ± 1.24 mg/mL). Furthermore, the extract prevented the growth of the A431, A549, and NCIH 460 cancer cell lines with the lowest IC50 values of 15.37, 29.09, and 17.02 g/mL, respectively, demonstrating remarkable anticancer potential. Molecular docking and dynamic simulation studies revealed that Prunetin, Tectorigenin, and Prunetin 4'-O-Galactoside show efficient binding to the EGFR binding domain. This study suggests that tested hits may have antioxidant and anticancer agents and can be considered for future applications in the pharma sector.
... The Papaver somniferum/setigerum seeds reported in the archaeological record are, so far, not identified to subspecies/status level (i.e. at the wild/domesticated level) due to the lack of discriminating criteria [33,34], since all three taxa are very close in both size and shape. However, they are genetically separated (see [21]). P. setigerum is native to the Western Mediterranean area, and for this reason, it is assumed that the plant must have been first collected and cultivated in this area [11,14]. ...
... The whole process possibly involved repeated exchanges and inter-breeding of genetic stock or cultivated varieties between the Alps and the Mediterranean and maybe even other areas during a very long period of time. This is also shown by the considerable genetic variability of P. somniferum as shown by Hong et al. [21]. Yet unresolved is the fact that P. setigerum and P. somniferum show considerable genetic differences (most setigerum is tetraploid, whereas somniferum is diploid), therefore more research is needed in order to understand the story of the domestication of opium poppy. ...
Domesticated opium poppy Papaver somniferum L. subsp. somniferum probably originated in the Western Mediterranean from its possible wild progenitor, Papaver somniferum L. subsp. setigerum and spread to other European regions. Seeds of opium poppy have been identified in different European regions since the Early Neolithic (from the 6th millennium cal. BC onwards) period. However, until recently, the absence of morphological identification criteria has prevented the discrimination between wild and domestic morphotypes. New morphometric approaches to distinguish modern subspecies have been proven to be applicable to waterlogged archaeological remains, opening the possibility of understanding the process of domestication of the plant in both time and space. This paper applies seed outline analyses, namely elliptic Fourier transforms, combined with size and number of cells to archaeological waterlogged Papaver seeds throughout the Neolithic period in the NW Mediterranean and the surroundings of the Alps. Furthermore, one example from the Late Bronze Age (LBA) was added to see what kind of differences appeared during the >1000 years between the end of the Neolithic and the LBA. The aim of the study is to classify the archaeological seeds as domestic or wild morphotypes and observe morphometric changes in connection to geographical and chronological patterns that can explain the spread and domestication process(es) of this important crop. A total of 295 archaeological seeds coming from 10 waterlogged sites dating between 5300-2300 cal. BC (Neolithic), and one LBA site dating to 1070 cal. BC were analysed. The results indicate the presence of seeds, similar to the wild morphotype, in the Mediterranean sites and larger seeds, similar to the domestic morphotype, in the regions surrounding the Alps. The number of cells mainly increased during the Late Neolithic (3300 to 2300 cal. BC) and, finally, in the Late Bronze Age (ca. 1050-800 cal. BC), larger, morphologically domesticated seeds are clearly predominant. A change in the shape of the seeds is only clearly visible in the LBA material. Altogether our results suggest that opium poppy seeds show no sign of domestication in the early periods of the Neolithic, despite the fact that the plant was very probably already cultivated at that time in the western Mediterranean region.
... The National Gene Bank for Medicinal and Aromatic Plants (CIMAP) in Lucknow contains a large opium poppy germplasm collection that includes accessions from all around India and Thailand, as well as wild relatives, mutants, released varieties, and breeding lines. Germplasm collections essentially provide the basis of genetic and phenotypic variability that can be utilized to accelerate the breeding programs (Hong et al., 2022;Karthika and Govintharaj, 2022). Although enormous genetic diversity in opium poppy has been reported from several countries, including India, Turkey, the Czech Republic, Germany, Slovakia, and Australia (Börner, 2006;Brezinova. ...
... et al., 2009;Saunders et al., 2001;Celik et al., 2016;Srivastava et al., 2020), the genetic potential yet remains underutilised in poppy. This is mainly due to the lack of detailed studies on the genetic diversity of the poppy germplasm (Hong et al., 2022). Therefore, the present study was conducted to evaluate the degree of genetic variability in the 40 germplasm of opium poppy using 13 morphological and biochemical traits. ...
The goal of this study was to characterize forty genotypes of Papaver somniferum L. and assess the genetic variability using data from morphological as well as biochemical studies. For the characterization, the morphological characters, viz., plant height, primary branch, peduncle length, days to maturity, number of selfed-capsules, weight of selfed-capsule, number of open-pollinated capsule, weight of open-pollinated capsules were complimented with five biochemical alkaloids morphine, codeine, thebaine, papaverine, and narcotine. The results show that morphine and codeine had a highly significant positive correlation (0.68 **). Similarly, a highly positive significant correlation (0.63 **) was established between morphine and narcotine. Further, all 40 genotypes were subjected to cluster analysis, where eight primary clusters were obtained using morphological analysis, whereas nine clusters were formed based on biochemical analysis. On the basis of morphological analysis, 32 genotypes were assigned to a one, large cluster which was further organized into 12 sub-clusters. However, clustering based on biochemical analysis yielded one large cluster with 25 genotypes, which was further subdivided into eight subclusters. The findings from the present study can be successfully employed for developing superior lines of the opium poppy.
... 2020 procedures for DNA extraction from Opium Poppy (Papaver somniferum L.) and Poppy Seed-containing products [462]; comparative cp genome analyses to study the evolutionary pattern in Papaveracea [463]; MALDI-MS method for detection of opiates in Papaver somniferum [464]; new EST-SSR markers for individual genotyping Opium Poppy Cultivars (Papaver somniferum L.) [465]; development of a quantitative real-time PCR (qPCR) method for the quantification of opium poppy DNA, evaluation of three commercial DNA extraction kits for their ability to isolate DNA from poppy seeds, and evaluation of nineteen opium poppy short tandem repeat (STR) markers for their use in a forensic identification panel [466]; 2021 characterization of a novel variable number tandem repeat markers of forensically important poppy species based on the genetic analysis of 164 samples collected in South Korea [467]; genomic analysis of P. somniferum [468]; genomic characterization of California poppy using the draft genome sequence [469] genomic profiling of WRKY genes involved in Benzylisoquinoline Alkaloid biosynthesis in California poppy [470]; 2022 molecular identification and phylogenetic analysis of Papaver based on ITS2 barcoding to detect and identify P. somniferum as well as common adulterants of the same genus [471]; genotyping-by-sequencing to investigate the genetic diversity and population structure in a collection of poppy germplasm consisting of 91 accessions originating in 30 countries of Europe, North Africa, America, and Asia [472]; IsoSeq on opium poppy to improve the gene annotation [473]; eleven simple sequence repeats (SSR) markers and two single nucleotide polymorphism (SNP) markers were mined to distinguish opium poppy from other six Papaver species [474]. ...
... Genotyping-by-sequencing (GBS) is one of most widely used RRS methods in which the barcode system is improved to allow the discovery of genome-wide single nucleotide polymorphisms (SNPs) with a lower error rate and low cost [26]. Because of these advantages, the high-throughput SNPs detected by GBS are widely used for genetic diversity analysis in many plant species [24,[27][28][29][30][31][32][33][34]. ...
Zabelia tyaihyonii (Nakai) Hisauti and H. Hara is a perennial shrub endemic to Republic of Korea that grows naturally in only a very limited region of the dolomite areas of Gangwon-do and Chungcheongbuk-do Provinces in the Republic of Korea. Given its geographical characteristics, it is more vulnerable than more widely distributed species. Despite the need for comprehensive information to support conservation, population genetic information for this species is very scarce. In this study, we analyzed the genetic diversity and population structure of 94 individuals from six populations of Z. tyaihyonii using a genotyping-by-sequencing (GBS) approach to provide important information for proper conservation and management. Our results, based on 3088 single nucleotide polymorphisms (SNPs), showed a mean expected heterozygosity (He) of 0.233, no sign of within-population inbreeding (GIS that was close to or even below zero in all populations), and a high level of genetic differentiation (FST = 0.170). Analysis of molecular variance (AMOVA) indicated that the principal molecular variance existed within populations (84.5%) rather than among populations (17.0%). We suggested that six management units were proposed for conservation considering Bayesian structure analysis and phylogenetic analysis, and given the various current situations faced by Z. tyaihyonii, it is believed that not only the in situ conservation but also the ex situ conservation should be considered.
... Recently, next-generation sequencing (NGS) has identified sufficient single nucleotide polymorphisms (SNPs) from population genetic studies to facilitate the study of genetic diversity in Malus [43][44][45] . Molecular diagnosis of genetic diversity within the nuclear genome using GBS has the potential to examine hybrid signals, introgressions, and the population-species boundary [46][47][48] . This is the first study to explore the possibility of hybridization and evolutionary relationship of Malus across the Korean Peninsula. ...
This study was to understand the genetic structure and diversity of the Korean Malus species. We used genotyping-by-sequencing (GBS) technology to analyze samples of 112 individuals belonging to 18 populations of wild Malus spp. Using GBS, we identified thousands of single nucleotide polymorphisms in the species analyzed. M. baccata and M. toringo, two dominant mainland species of the Korean Peninsula, were distinguishable based on their genetic structure. However, M. toringo collected from Jeju Island exhibited a different genetic profile than that from the mainland. We identified M. cf. micromalus as a hybrid resulting from the Jeju Island M. toringo (pollen donor) and the mainland M. baccata, (pollen recipient). Putative M. mandshurica distributed on the Korean Peninsula showed a high structural and genetic similarity with M. baccata, indicating that it might be an ecotype. Overall, this study contributes to the understanding of the population history and genetic structure of Malus in the Korean Peninsula.
... Hong et al. utilized genotyping-by-sequencing (GBS) of whole germplasm data of various Papaver species from five countries and demonstrated that their SNP profiles do group together to represent their biogeographical structure, even with limited data and no full genome available. This study also showed a grouping pattern between alkaloid profiles and the nuclear genome, but the results need to be validated through field studies, as alkaloid profiles are affected by the environment; the samples used in this study were grown under laboratory conditions from seeds [32]. ...
The opium poppy, Papaver somniferum L., is a forensically important plant due to the medicinal and illegal uses for the milky latex stored in the pods. This latex contains the alkaloids morphine, codeine, and thebaine that are used for their analgesic properties and/or for synthesizing other opioids. However, these compounds are highly addictive and have caused a national opioid epidemic. Two other Papaver species, P. setigerum DC. and P. bracteatum Lindl., are also of forensic interest because they pose both forensic and legal issues. They are largely uncontrolled under the Controlled Substances Act, making these species a common defense strategy. Current morphological and chemical identification methods have been moderately successful but have drawbacks. There is also a lack of sequencing data available. Therefore, exploiting the genome using chloroplast DNA barcoding markers could help to accurately identify these species of interest when plant material is taken. This study screened and assessed the genetic variation both between species and within populations of P. somniferum in nine cpDNA barcode regions (ndhF-rpl32, petA-psbJ, rpl32-trnL, rps16-trnQ, trnE-trnT, trnH-psbA, trnL-trnF, rpl16 intron, and psbE-petL). Published reference genomes from the NCBI GenBank database were aligned and compared for an initial in silico screening. Additionally, ten P. somniferum seed samples from various vendors were sequenced and compared across samples and to published reference data at the various barcode regions of interest. This study showed that the regions trnH-psbA and petA-psbJ have promise for utility in individualization for both inter- and intra-species individualization of P. somniferum.
... One striking property of opium poppy cultivars is their remarkable variation in alkaloid content and composition. For instance, some plants produce large amounts of morphine and codeine while others synthesise high levels of thebaine or oripavine [72]. This is due to the extensive artificial selection the species has been subjected to which has resulted in genetic variation between cultivars, landraces and lines. ...
The individual tissues and cell types of plants each have characteristic properties that contribute to the function of the plant as a whole. These are reflected by unique patterns of gene expression, protein and metabolite content, which enable cell-type-specific patterns of growth, development and physiology. Gene regulatory networks act within the cell types to govern the production and activity of these components. For the broader organism to grow and reproduce successfully, cell-type-specific activity must also function within the context of surrounding cell types, which is achieved by coordination of signalling pathways. We can investigate how gene regulatory networks are constructed and function using integrative 'omics technologies. Historically such experiments in plant biological research have been performed at the bulk tissue level, to organ resolution at best. In this review, we describe recent advances in cell- and tissue-specific 'omics technologies that allow investigation at much improved resolution. We discuss the advantages of these approaches for fundamental and translational plant biology, illustrated through the examples of specialised metabolism in medicinal plants and seed germination. We also discuss the challenges that must be overcome for such approaches to be adopted widely by the community.
