A plant that can cure the deadliest diseases known to human. A plant that could replace chemicals used for insomnia and depression. A plant that could reduce the destruction of forests, thus preserving the habitat of many endangered animals. A plant that has wide commercial use, in textiles, paper production, and much more. But the cultivation of this plant is illegal, for some irrelevant reason. This plant is Cannabis.
Cannabis is a plant that has been used for centuries. Seeds of cannabis found in Siberia, which dates back to the 30th century BC. The Chinese used cannabis for medical purposes. And so go through the centuries, you will come across countless examples of using cannabis by a man. Even the US President Washington has cultivated cannabis in his backyard. And in my country, cannabis (hemp) was cultivated in large quantities
'' At one time in our region hemp growing to 50,000 ha, of which 80% were in Vojvodina. It is expected that in the 5-10 years under hemp will be several tens of thousands of hectares. This is a logical trail, since the growth of areas under this plant is obvious. In 2015, there were 60 ha, while in 2016, as many as 250 hectares of hemp were planted'', explains Maja Timotijevic, President of the Association "Hemp".
In the literature, research is available in which DNA sequences used for the differentiation of genus Cannabis from its closest relatives, plant from genus Humulus. For these studies were used molecular markers (RAPD,markers),however, they were not always able to distinguish two Cannabis variants. And for this reason, this scientific work has been done. And for this reason this study was done in order to discovered the AFLP polymorphic locus that will be able to differentiate the psychoactive plant from its closest relative, industrial hemp.
The first stage was the procurement of marijuana seeds (HortaPharm, Amsterdam), as well as seeds of industrial hemp. In addition, they also took the hemp from abandoned fields.
Before AFLP analysis, thc was isolated from the plants and its quantification was made. Psychoactive individuals contained about 7 % of the thc, and the hemp contains 0.03 % thca
The DNA is isolated from young leaves of 2 weeks old plants, and from dried blooms 60-160 days after harvest. The amount of DNA was determined spectrometrically. For AFLP analysis, 30 ng genomic DNA was used as well as two endonucleases MseI I EcoRI.
First, they compared AFLP profiles obtained with the help of three different pairs of selective primers on DNA. The results of the statistical tests have shown that although there may be 12% non-overlap between the obtained fragments, there is no significant difference.
Using 10 primers, the authors generated 1206 fragments of different lengths. 52 fragments appeared only once, and 178 were present in all individuals. The remaining 976 fragments (88%) were polymorphic. Among the remaining fragments, as many as 47 were stable for the differentiation of psychoactive and industrial cannabis (19 were unique for hemp and 28 for psychoactive). In addition, 4 fragments were present in all three industrial hemp, but they were absent from the psychoactive plant, while as many as 7 fragments present in the psychoactive were not present in the industrial hemp.
ANOVA with 4 cannabis populations has shown that 27.2% of genetic variability losing on the difference between marijuana and hemp, 20.9% on the difference between populations, and as much as 51.9% of the variability within the population. PCA clearly separated marijuana from hemp.
