Evidence of the Ability of Microsatellite Method to Distinguish Cannabis Strains with High Cannabinoid Content.

Introduction: Cannabis is a plant with high potential for use in several sectors of the industry; however, it is also a controversial crop due to its tetrahydrocannabinol (THC) content. Moreover, the plant has a rather unclarified classification. Traditionally, two types of Cannabis have been distinguished, hemp as a source of fiber and low THC content, and marijuana with high THC levels, which is used as a drug. With the increasing use of CBD strains and wide range of commercially used THC strains, it is becoming paramount to be able to develop an easy and reliable method for Cannabis strain differentiation. The use of simple sequence repeat markers, or microsatellites, seems to be an applicable choice. Materials and Methods: In this study, 52 strains of Cannabis with variable cannabinoid content were collected from growers from different geographical regions and analyzed using 17 different microsatellite markers. For more precise differentiation, five strains were selected and a higher number of individuals of each were analyzed. Results: Fragment analysis and cluster analysis showed that when one to three individual plants per strain were analyzed, the method was able to classify these samples into distinguishable groups with similar gene structure. They also revealed that when a larger sample set was used (10 individual plants per strain), highly specific strain clusters could be fully discriminated. Conclusion: Our study involved the highest number of cannabinoid-rich strains up to now and showed that the microsatellite method can be used to reliably differentiate Cannabis strains and show their relationships.

Development of microsatellite markers for the perennial plant Tofieldia calyculata.

PREMISE: Polymorphic microsatellite markers were developed to study genetic diversity and genetic structure of populations of the locally endangered species Tofieldia calyculata (Tofieldiaceae). METHODS AND RESULTS: Nineteen polymorphic microsatellite loci were developed using DNA-enriched libraries sequenced by Ilumina technology and were then used to genotype 101 individuals from five populations from Austria, Slovakia, Poland, and the Czech Republic. Of the markers tested, 68% were polymorphic in four of the five investigated populations, with one marker polymorphic in all populations. The number of alleles per locus in the populations ranged from one to 11. Levels of observed and expected heterozygosity ranged from 0.00 to 0.75 and from 0.00 to 0.84, respectively. Moreover, six of the 19 loci amplified when tested in the congeneric species T. pusilla. CONCLUSIONS: The 19 newly developed microsatellite markers can be used to describe genetic diversity and population structure of populations of T. calyculata.

Microsatellite markers for study of the invasive species Rumex alpinus (Polygonaceae).

PREMISE OF THE STUDY: Polymorphic microsatellite markers were developed to study genetic diversity, population structure, and dispersal strategies of the highly invasive species Rumex alpinus (Polygonaceae). METHODS AND RESULTS: Fifteen polymorphic microsatellite loci were developed using a 454 sequencing approach and used to genotype 72 individuals from six populations in Austria and France. All markers were polymorphic in at least one investigated population, and the number of alleles ranged from one to four alleles per locus. Observed and expected heterozygosity ranged from 0.06 to 1.0 and from 0.18 to 0.72, respectively. CONCLUSIONS: These 15 markers will be useful for characterizing dispersal strategies and gene flow assessment between R. alpinus populations and other, often weedy Rumex species.

Hungarosoma bokori Verhoeff, 1928 (Diplopoda: Chordeumatida): new insights into its taxonomy, systematics, molecular genetics, biogeography and ecology.

Hungarosoma bokori Verhoeff, 1928 is a millipede species which was originally classified solely on the basis of a female specimen. Subsequently, a long history of field searching for and surmising about the systematic position of this small, enigmatic species followed. In April 2013, 85 years after its first description, a series of nine specimens were sampled in the type locality, the Abaliget Cave, in southern Hungary. An adult male was collected for the first time, along with females and juveniles. Descriptions of the gonopods and the female vulvae, both important for considerations of the systematic position of the species, are presented for the first time. Revision and re-designation of the type material was made.The cryptic life of the species is connected with its activity in winter, and its known fragmented distribution corresponds with its presence in undisturbed microhabitats having a specific microclimate, often in the soil at cave entrances.Molecular methods showed a positive detection of the intracellular prokaryotic parasite Wolbachia in H. bokori, reflecting its highly probable parthenogenetic character in the main part of its known area of occurrence. This is the first demonstration of Wolbachia in a millipede.The legitimacy of the family Hungarosomatidae Ceuca, 1974, as a separate taxon was analysed using morphological and molecular approaches. Results of both methods confirmed the existence of a distinct phyletic line. DNA barcoding has shown its closest position to Attemsiidae Verhoeff, 1899, or Neoatractosomatidae Verhoeff, 1901. Based on records from Austria, the Czech Republic, Hungary and Slovakia, the residual circum-pannonic distribution that the whole genus (family) probably represents is proposed.

The complete mitochondrial genome of the facultative entomopathogenic nematode Oscheius chongmingensis (Rhabditida: Rhabditidae).

We determined the complete mitochondrial genome of the facultative entomopathogenic nematode Oscheius chongmingensis. The mitogenome length was 15,413 bp and similar to other Rhabditids contains genes for 2 rRNAs, 22 tRNAs, and 12 proteins (ATPase subunit 8 is missing). Predicted tRNAs indicated the secondary structure typical for chromadorean nematodes. Gene order is similar to that observed in the genus Caenorhabditis. The control AT-rich region is considerably large (2061 bp, 84% of AT), positioned in between tRNA(Ala) and tRNA(Pro) and has several microsatellite-like (AT)n elements.