Abstract

Purpose and objectives. To reveal peculiarities of the inheritance of the traits of cannabinoid compound presence and content upon self-pollination in monoecious hemp, to compare the pair correlation coefficients between the contents of major cannabinoid compounds in initial genotypes and self-pollinated lines, and to evaluate the effectiveness of self-pollination in hemp breeding.Material and methods. The study was carried out at the Institute of Bast Crops of the National Academy of Agricultural Sciences (Hlukhiv, Sumy Oblast, Ukraine) in 2009–2019. Inbred lines of industrial hemp varieties Hlukhivski 58, Hlesiia, Mykolaichyk, and Hlukhivski 46 belonging to the Central European eco-geographical type and variety Zolotoniski 15 belonging to the Southern eco-geographical type were taken as test objects. Self-pollination of plants (with and without cannabinoids) was carried out under individual agrotextile bags in a greenhouse. The offspring were grown in a nursery. Evaluation and analysis of cannabinoid compounds were conducted by thin-layer chromatography. In all the genotypes under study, the tetrahydrocannabinol (THC) content did not exceed 0.08% that is standard allowed by the current legislation of Ukraine. Data were statistically processed with calculating arithmetic mean, sampling mean error, pairwise correlation coefficients, and curvilinear regression.Results and discussion. Provided directional selection of initial plants that do not contain cannabidiol (CBD), THC or cannabinol (CBN), self-pollination reduced their contents to complete absence. Stabilization (homozygation) of the lines occurred in I2–I6 and was specific to a particular variety. Inbred lines of these generations are recommended to involve as parents in crossing. The ability to segregate CBD-, THC-, and CBN-free families as early as in I1. is a characteristic feature of the hemp varieties under investigation. There were strong positive correlations between the cannabinoid contents, in particular there was a strong positive correlation was found between the CBD and THC contents (r = 0.72–0.79 and 0.71–0.90, respectively), a strong or medium positive correlation between the CBD and CBN contents (0.68–0.80 and 0.67–0.82, respectively), a strong positive correlation between the THC and CBN contents (0.71–0.83 and 0.80–0.85, respectively) in I1–I3 Hlukhivski 58 and I1–I3 Zolotoniski 15. This makes selection for reduced contents of all cannabinoid compounds and easier, but at the same time significantly complicates breeding for increased CBD content with concurrent reduced THC content or increased contents of non-psychotropic cannabinoids. The correlations between the contents of cannabinoid compounds in inbred genotypes are weaker than those in the original breeding genotypes, with the coefficients ranging significantly, which allows using closely related reproduction in breeding aimed at reducing THC content or increasing non-psychotropic cannabinoid contents.Conclusions. Self-pollination is an effective method of determination of the hemp population stability in terms of the cannabinoid compound presence and contents and at the same time is a method of creating breeding genotypes with stable traits of cannabinoid absence or presence

Highlights

  • The presence of cannabinoid compounds is one of the main biological features of cannabis (Cannabis sativa L.), which is constantly monitored at all stages of breeding and seed production: from creation of starting material to production of certified seeds and cultivation on an industrial scale

  • Cannabinoids are found in excretory tissue cells. This attests to the fact that genes for the synthesis of these compounds can be expressed in all plant cells; it is glandular trichomes that specialize in the synthesis of high amounts of cannabinoids, in other tissues the contents of these substances are much lower [6]

  • Plants without cannabinoids were selfpollinated, in I1 and I2 the cannabinoid contents increased: the CBD content was found to be 0.21, THC – 0.15 and CBN – 0.13 points in I1, and 0.18, 0.24, 0.17 points in I2, respectively. This indicates that genes determining the expression of cannabinoid content genes were converted into a homozygous state, and that mutant plants with cannabinoids segregated as a consequence of the genotypic environment effect on persistent changes in the hereditary apparatus upon the extreme variant of closely related reproduction

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Summary

Introduction

The presence of cannabinoid compounds is one of the main biological features of cannabis (Cannabis sativa L.), which is constantly monitored at all stages of breeding and seed production: from creation of starting material to production of certified seeds and cultivation on an industrial scale. Cannabinoids are specific substances in cannabis that belong to the class of aromatic compounds and are synthesized and accumulated mainly in glandular trichomes (hairs) [1,2,3,4,5]. Cannabinoids are found in excretory tissue cells This attests to the fact that genes for the synthesis of these compounds can be expressed in all plant cells; it is glandular trichomes that specialize in the synthesis of high amounts of cannabinoids, in other tissues the contents of these substances are much lower [6]. To avoid damage and cell death, an adaptation, which consists in the fact that cannabinoids are accumulated and stored in the glandular trichome cavities (specialized secretory tissues), evolved. Cannabinoids play a protective role in the plant, while reduction in cannabinoid levels and in the number of glands in technical (industrial) hemp do not change these physiological functions; the synthesis of these substances in small quantities by other cells of the plant is sufficient [6]

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