Nicotiana glauca (tree tobacco) is a naturally transgenic plant, containing sequences acquired from Agrobacterium rhizogenes by horizontal gene transfer. Besides, N. glauca contains a wide profile of alkaloids of medical interest. We report a high-depth sequencing and de novo assembly of N. glauca full genome and analysis of genome elements with bacterial origin. The draft genome assembly is 3.2 Gb, with N50 size of 31.1 kbp. Comparative analysis confirmed the presence of single, previously described gT insertion. No evidence was acquired to support idea of multiple T-DNA insertions in the N. glauca genome. Our data is the first comprehensive de novo assembly of tree tobacco and provide valuable information for researches in pharmacological and in phylogenetic fields.

Naturally transgenic plants represent the result of Agrobacterium-mediated gene transfer. T-DNA of soil bacteria Agrobacterium integrated into plant’s genome is called cellular T-DNA (cT-DNA) [1]. Today, more than 50 species of naturally transgenic plants, or natural GMO (nGMO) are known [2, 3]. The function of cT-DNA in plants remains unknown. It is assumed that the fixation of transgenes could give plants different selective advantages depending on which genes had been integrated into the plant [4]. In order to clarify this issue, it is necessary to study more naturally transgenic plants. Until recently, the list of nGM plants contained less than 2 dozen species, but a search through genomic and transriptomic sequencing data made it possible to more than double this list [2]. In this work, we used the same approach, looking for cT-DNA genes in whole genome sequencing data that have appeared in the NCBI WGS since 2021. We found 14 new species of naturally transgenic plants, among which the most extended cT-DNAs were found in Triadica sebifera, Lonicera japonica, and Lonicera maackii. The cT-DNAs in these species are organized as imperfect inverted repeats. In the genomes of the species Paulownia fortunei, Apocynum venetum, Elaeagnus angustifolia, Erythroxylum havanense, E. densum, E. daphnites, E. cataractarum, Ceriops decandra, Camellia oleifera, Silene uniflora, short cT-DNAs containing only opine synthesis genes were found. We also estimated the approximate age of the cT-DNAs. The first described examples date back to the Late Paleogene, and the process continues to the present. Thus, we can conclude that natural GMOs are a widespread phenomenon, many aspects of which remain unclear, requiring additional research on the topic. The article was made with support of the Ministry of Science and Higher Education of the Russian Federation in accordance with agreement No. 075-15-2022-322 dated 22.04.2022 on providing a grant in the form of subsidies from the federal budget of the Russian Federation. The grant was provided for state support for the creation and development of a world-class scientific center, “Agrotechnologies for the Future”.

Agrobacterium (Rhizobium)-mediated transformation leads to the formation of crown galls or hairy roots on infected plants. These effects develop due to the activity of T-DNA genes, gathered on a big plasmid, acquired from agrobacteria during horizontal gene transfer. However, a lot of plant species are known to contain such sequences, called cellular T-DNAs (cT-DNAs), and maintain normal phenotypes. Some of the genes remain intact, which leads to the conclusion of their functional role in plants. In this study, we present a comprehensive analysis of the cT-DNAs in the Nicotiana noctiflora Hook. genome, including gene expression and opine identification. Deep sequencing of the Nicotiana noctiflora genome revealed the presence of two different cT-DNAs, NnT-DNA1 and NnT-DNA2, which contain the intact genes iaaM, iaaH, acs, orf13, orf13a, and orf14. According to the expression analysis results, all these genes are most active in roots in comparison with other organs, which is consistent with data on cT-DNA gene expression in other plant species. We also used genetic engineering approaches and HPTLC and HPLC-MS methods to investigate the product of the acs gene (agrocinopine synthase), which turned out to be similar to agrocinopine A. Overall, this study expands our knowledge of cT-DNAs in plants and brings us closer to understanding their possible functions. Further research of cT-DNAs in different species and their functional implications could contribute to advancements in plant genetics and potentially unveil novel traits with practical applications in agriculture and other fields.

Агротрансформация видов Nicotiana glauca и Nicotiana sylvestris

Background: A growing body of research supports the role of the microbial communities residing in the digestive system in the host’s cognitive functioning. Most of these studies have been focused on the gut microbiome and its association with clinical phenotypes in middle-aged and older adults. There is an insufficiency of population-based research exploring the association of normative cognitive functioning with the microbiome particularly with the oral microbiota. Methods: In this study, using metagenomics and metabolomics, we characterized the salivary microbiome diversity in a sample of 51 males of Hispanic and African American origin aged 12–18 years and explored the associations between the microbiome and the youths’ cognitive performance captured with the Kaufman Assessment Battery for Children II (KABC-II). Results: Several bacterial species of the oral microbiota and related metabolic pathways were associated with cognitive function. In particular, we found negative associations between indicators of general intelligence and the relative abundance of Bacteroidetes and Lachnospiraceae and positive associations with Bifidobacteriaceae and Prevotella histicola sp. Among metabolic pathways, the super pathways related to bacterial cell division and GABA metabolism were linked to cognitive function. Conclusions: The results of our work are consistent with the literature reporting on the association between microbiota and cognitive function and support further population work to elucidate the potential for a healthy oral microbiome to improve cognitive health.

Background. Sunflower seed production is based on utilization of the heterosis effect, manifesting itself in improving both yield and plant height in hybrids. Short-stemmed lines need to be used to develop commercial hybrids with an optimum height. Molecular bases of the trait manifestation in the dwarf lines from VIR’s sunflower genetic collection have not yet been studied. Materials and methods. The material included 27 short-stemmed and 10 tall sunflower lines from VIR’s genetic collection, as well as the F₁ and F₂ hybrid generations derived from crossing the tall (VIR 340) and dwarf (VIR 171) genotypes. The parental lines and hybrids were phenotyped for plant height, leaf number, and internode length. Genotyping for the Rht1 locus (HaDella1 candidate gene), encoding the negative regulator of the gibberellin response, the DELLA protein, was performed using the developed CAPS marker. Results. The average plant height in the VIR 340 line over a three-year study was 162 cm, the number of leaves 29, and the internode length 6 cm. The VIR 171 line demonstrated the plant height of 66 cm, leaf number of 24, and internode length of 2.8 cm. The F₁ hybrids were uniform, with the height of 180–190 cm, that indicated the dominance of the long stem trait. Analyzing the segregation in the F₂ hybrid generation led to an assumption admitting the digenic control of the dwarf phenotype in the VIR 171 line. The CAPS marker G-D-1/ Bmt I was developed to identify a missense mutation T>C in the first exon of the HaDella1 gene, which results in the substitution of leucine with proline in the DELLA motif. Using the marker, the mutant Rht1 allele was identified in the VIR 171 and VIR 434 dwarf lines, similar in their origin and phenotype. The results of validation in the F₂ hybrid population (VIR 340 × VIR 171) confirmed the efficiency of the G-D-1 / Bmt I marker for selecting dwarf genotypes homozygous for the Rht1 mutant allele.