Procedures for DNA Extraction from Opium Poppy (Papaver somniferum L.) and Poppy Seed-Containing Products

Abstract

Several commonly used extraction procedures and commercial kits were compared for extraction of DNA from opium poppy (Papaver somniferum L.) seeds, ground seeds, pollen grains, poppy seed filling from a bakery product, and poppy oil. The newly developed extraction protocol was much simpler, reduced the cost and time required for DNA extraction from the native and ground seeds, and pollen grains. The quality of extracted DNA by newly developed protocol was better or comparable to the most efficient ones. After being extended by a simple purification step on a silica membrane column, the newly developed protocol was also very effective in extracting of poppy DNA from poppy seed filling. DNA extracted from this poppy matrix was amplifiable by PCR analysis. DNA extracted from cold-pressed poppy oil and suitable for amplifications was obtained only by methods developed previously for olive oil. Extracted poppy DNA from all tested matrices was analysed by PCR using primers flanking a microsatellite locus (156 bp) and two different fragments of the reference tubulin gene (553 bp and 96 bp). The long fragment of the reference gene was amplified in DNA extracted from native seeds, ground seeds, and pollen grains. Poppy DNA extracted from the filling of bakery product was confirmed only by amplification of short fragments (96 bp and 156 bp). DNA extracted from cold-pressed poppy oil was determined also only by amplification of these two short fragments.

cor1 Gene: A Suitable Marker for Identification of Opium Poppy (Papaver somniferum L.)

“…The extraction of nucleic acids is the first and essential step in the molecular-based analysis of food. Many factors affect the success of DNA isolation; for example, the plant matrix and the related variety of metabolites contained in the plant cell, as well as other undesirable substances (lipids, proteins, and others) [37][38][39]. There is no universal protocol for DNA extraction yet; the most appropriate extraction method should be chosen on a matrix basis.…”

Interpol Review of Drug Analysis 2019-2022

“…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 ].…”

Target and Non-Target Approaches for Food Authenticity and Traceability

“…These techniques are highly specific and sensitive and are frequently considered as the most suitable tools for the identification of species. Various research papers on the use of DNA-based approaches are also included in this Special Issue, from the comparison of different DNA extraction methods [ 28 ] to the use of multiplex polymerase chain reaction (PCR) [ 23 ], real-time PCR [ 19 , 22 , 24 , 29 , 30 ], or more advanced techniques such as Digital PCR [ 31 ]. Kim et al [ 23 ] proposed the use of a simple qualitative assay based on the use of multiplex PCR to identify three deer species, namely red deer ( Cervus elaphus ), roe deer ( Capreolus capreolus ), and water deer ( Hydropotes inermis ).…”