New EST-SSR Markers for Individual Genotyping of Opium Poppy Cultivars (Papaver somniferum L.)

Abstract

High-quality simple sequence repeat (SSR) markers are invaluable tools for revealing genetic variability which could be utilized for many purposes, such as breeding new varieties or the identifying current ones, among other applications. Based on the analysis of 3.7 million EST sequences and 15 genomic sequences from bacterial artificial chromosome (BAC) libraries, 200 trinucleotide genic (EST)-SSR and three genomic (gSSR) markers were tested, where 17 of them fulfilled all criteria for quality markers. Moreover, the reproducibility of these new markers was verified by two genetics laboratories, with a mean error rate per allele and per locus equal to 0.17%. These markers were tested on 38 accessions of Papaver somniferum and nine accessions of another five species of the Papaver and Argemone genera. In total, 118 alleles were detected for all accessions (median = 7; three to ten alleles per locus) and 88 alleles (median = 5; three to nine alleles per locus) within P. somniferum alone. Multivariate methods and identity analysis revealed high resolution capabilities of the new markers, where all but three pair accessions (41 out of 47) had a unique profile and opium poppy was distinguished from other species.

Scientific Prospects for Cannabis-Microbiome Research to Ensure Quality and Safety of Products

“…In this regard, systems biology [1,2] is well-equipped to integrate findings and define the genetic variability and functional metabolites [3] in a model plant system. Model plant research, such as that of well-known Arabidopsis thaliana [4][5][6][7] and the extensively-studied medicinal plant, Papaver somniferum [8][9][10][11][12], highlight the significance of gathering information about regulation of plant fitness and homeostatic mechanisms of metabolites [13] orchestrated by the host genome and its affiliated microbial metagenome [14]. Such an approach notably reveals the importance of the plant's genome-microbiome interactions [15], which changes under stress or disease conditions [16,17] as plants selectively source their microbiomes to suit their needs [18,19].…”

OpiumPlex is a novel microsatellite system for profiling opium poppy (Papaver somniferum L.)

“…The total number of markers (27) represents a compromise between the effort to maximize discriminatory power, as we were aware of the relatively low genetic variability reported for opium poppy 16 , 18 , 19 , and obtain independently segregating loci. Unfortunately, there is a lack of information on the recombination rate in opium poppy in the literature, and it can substantially vary among species, chromosomes of the same species or even a chromosome itself 50 – 52 .…”

“…The opium poppy genome has not been published until recently 14 . Therefore, most of the STR markers were developed by the data mining of sequences from transcriptomic studies 15 – 18 or by screening sequences of unplaced genomic fragments retrieved via next-generation sequencing (NGS) 19 . Unfortunately, many of these markers suffer from one or more weaknesses, such as low polymorphism 20 , short lengths of repeated motifs or unknown localization on a chromosome.…”

Application of different molecular markers in biotechnology

“…(Asadi et al, 2020). Moreover, development of EST-SSR markers have been also studied in Indian mulberry (Thumilan et al, 2016), Lycium barbarum (Chen et al, 2017), Bletilla striata (Xu et al, 2018), Lilium (Biswas et al, 2018, Chinese Hawthorn (Ma et al, 2019) and opium poppy (Vašek et al, 2020). In addition to plants, these markers are also used for animal (Li et al, 2020;Silva Junior et al, 2020) and human (Pai et al, 2016).…”