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The red raspberry is one of the world’s most popular berries. The main direction of its breeding has switched to nutritional quality, and the evaluation of raspberry germplasm for antioxidant content and activity is very important. As berries, raspberry leaves contain valuable bioactive compounds, but the optimal time for their collection is unknown. We evaluated 25 new breeding lines and standard raspberry cultivars for their polyphenolic content and antioxidant capacity. The antioxidant activity of berries correlated better with the content of total phenolics (0.88 and 0.92) and flavonoids (0.76 and 0.88) than with anthocyanins (0.37 and 0.66). Two breeding lines were significantly superior to the standard cultivars and can be used in further breeding. Leaves collected in three phenological phases of the raspberry contained more phenolics (5.4-fold) and flavonoids (4.1-fold) and showed higher antioxidant activities (2.4-fold in FRAP assay, 2.2-fold in ABTS) than berries. The optimal time for harvesting raspberry leaves is the fruit ripening stage, with exceptions for some cultivars. Genetic diversity analysis using microsatellite (SSR) markers from flavonoid biosynthesis genes divided the genotypes into five clusters, generally in agreement with their kinships. The relationship between genetic data based on metabolism-specific SSR markers and the chemical diversity of cultivars was first assessed. The biochemical and genetic results show a strong correlation (0.78). This study is useful for further the improvement of raspberry and other berry crops.

Axillary buds of in vitro microshoots were successfully frozen at –196 °C by the one-step freezing method using the protective vitrification solution 2 (PVS2). Microshoots were taken from 11 transgenic lines and three wild type lines. Influence of different explant pretreatments were analyzed from the point of their influence towards recovery after cryopreservation. It was found out that the use of axillary buds as explants after removal of the apical one increases recovery on average by 8%. The cultivation on growth medium of higher density insignificantly raises the regenerants survival rate. Pretreatment of the osmotic fluid (OF) shows the greatest influence on the survival rate. It leads to the increase in survival rate by 20%. The cryopreservation technology providing regenerants average survival rate of 83% was developed. It was based on the experimental results obtained with explant pretreatment. Incubation time in liquid nitrogen did not affect the explants survival rate after thawing. After six months cryostorage of samples their genetic variability was analyzed. Six variable simple sequence repeat (SSR) loci were used to analyze genotype variability after the freezing-thawing procedure. The microsatellite analysis showed the genetic status identity of plants after cryopreservation and of the original genotypes. The presence of the recombinant gene in the transgenic lines after cryostorage were confirmed so as the interclonal variation in the growth rate under greenhouse conditions. The developed technique is recommended for long-term storage of various breeding and genetically modified lines of aspen plants, as it provides a high percentage of explants survival with no changes in genotype.

Abstract:The genetic transformation of trees by wood modification genes for the improvement of forest plantations results in shifts in plant litter quality. These alterations in plant chemistry lead to changes in decomposition rates, thus affecting the carbon and nitrogen cycling in ecosystems and nutrient availability for plants. To assess the environmental impacts of transgenic trees, we studied the decomposition of plant litter from aspen plants (Populus tremula L.) transformed with the xyloglucanase gene from Penicillium canescens. Mass, carbon and nitrogen losses in the leaves, stems and roots of greenhouse-grown plants were evaluated during incubation in laboratory microcosms. After 12 months of the decomposition experiment, leaves, stems, and roots lost on average 51%, 46%, and 37% of initial mass, respectively. Decomposition of the transgenic stems was not different from wild-type aspen, but we observed significant differences for the leaves (only at the end of the experiment) and the roots (at the early stage). These differences may be related to the nitrogen content and the C/N ratio in the initial samples. Since the litter decomposability determines the availability of nutrients, such alterations should be taken into consideration when cultivating transgenic trees.

Gooseberry is one of the most promising and underrated small fruit crops. There is a need to improve and genotype the existing cultivars, and this work requires the use of new efficient DNA marking techniques. Marker-assisted selection (MAS) is a modern approach for work with genetic resources. However, the genetic determinants of key qualitative traits are poorly studied. Therefore, we developed SSR markers located in flavonoid biosynthesis genes that can affect the resistance of plants to biotic and abiotic stresses to study the genetic diversity of gooseberry cultivars grown in the Russian Federation and varying in genetic and geographical origin. We have genotyped 22 gooseberry cultivars using a set of 19 of our original microsatellite markers and two neutral previously published ones. A total of 53 alleles were found. Nine of the 21 analyzed loci were polymorphic. The most polymorphic loci of flavonoid biosynthesis were found in the DFR gene (RucDFR1-2, RucDFR1-3, RucDFR2-1), their mean PIC (polymorphism information content) being 0.65, comparable to the PIC values of neutral markers. Our set of gene-targeted SSR markers showed that most of all the studied gooseberry cultivars differed in origin, based on which they were divided into three groups: European cultivars, Russian cultivars, and interspecies hybrids. Thus, the polymorphic markers can be used for cultivar identification and certification as well as for the marker-assisted selection of gooseberry plants having different origins and phenotypic traits.

Background Recombinant carbohydrases genes are used to produce transgenic woody plants with improved phenotypic traits. However, cultivation of such plants in open field is challenging due to a number of problems. Therefore, additional research is needed to alleviate them. Results Results of successful cultivation of the transgenic aspens (Populus tremula) carrying the recombinant xyloglucanase gene (sp-Xeg) from Penicillium canescens in semi-natural conditions are reported in this paper for the first time. Change of carbohydrate composition of wood was observed in transgenic aspens carrying the sp-Xeg gene. The transformed transgenic line Xeg-2-1b demonstrated accelerated growth and increased content of cellulose in wood of trees growing in both greenhouse and outside in comparison with the control untransformed line Pt. The accelerated growth was observed also in the transgenic line Xeg-1-1c. Thicker cell-wall and longer xylem fiber were also observed in both these transgenic lines. Undescribed earlier considerable reduction in the wood decomposition rate of the transgenic aspen stems was also revealed for the transformed transgenic lines. The decomposition rate was approximately twice as lower for the transgenic line Xeg-2-3b in comparison with the control untransformed line Pt. Conclusion A direct dependence of the phenotypic and biochemical traits on the expression of the recombinant gene sp-Xeg was demonstrated. The higher was the level of the sp-Xeg gene expression, the more pronounced were changes in the phenotypic and biochemical traits. All lines showed phenotypic changes in the leave traits. Our results showed that the plants carrying the recombinant sp-Xeg gene do not demonstrate a decrease in growth parameters in semi-natural conditions. In some transgenic lines, a change in the carbohydrate composition of the wood, an increase in the cell wall thickness, and a decrease in the rate of decomposition of wood were observed.

Background: Recombinant carbohydrases genes are used to produce transgenic woody plants with improved phenotypic traits. However, cultivation of such plants in open field is challenging due to by a number of problems. Therefore, additional research is needed to alleviate them. Results: Results of successful cultivation of the transgenic aspens ( Populus tremula ) carrying the recombinant xyloglucanase gene ( sp-Xeg ) from Penicillium canescens in semi-natural conditions are reported in this paper for the first time. Change of carbohydrate composition of wood was observed in transgenic aspens carrying the sp-Xeg gene. The transformed transgenic line Xeg-2-1b demonstrated accelerated growth and increased content of cellulose in wood of trees growing in both greenhouse and outside in comparison with the control untransformed line Pt. The accelerated growth was observed also in the transgenic line Xeg-1-1c. Thicker cell-wall and longer xylem fiber were also observed in both these transgenic lines. Undescribed earlier considerable reduction in the wood decomposition rate of the transgenic aspen stems was also revealed for the transformed transgenic lines. The decomposition rate was approximately twice as lower for the transgenic line Xeg-2-3b in comparison with the control untransformed line Pt. Conclusion: A direct dependence of the phenotypic and biochemical traits on the expression of the recombinant gene sp-Xeg was demonstrated. The higher was the level of the sp-Xeg gene expression, the more markedly were changes in the phenotypic and biochemical traits. All lines showed phenotypic changes in the leave traits. Our results showed that the plants carrying the recombinant sp-Xeg gene do not demonstrate a decrease in growth parameters in semi-natural conditions. In some transgenic lines, a change in the carbohydrate composition of the wood, an increase in the cell wall thickness and a decrease in the rate of decomposition of wood were observed.