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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.
Elena O. Vidyagina; Nikolay N. Kharchenko; Konstantin A. Shestibratov. Efficient Cryopreservation of Populus tremula by In Vitro-Grown Axillary Buds and Genetic Stability of Recovered Plants. Plants 2021, 10, 77 .
AMA StyleElena O. Vidyagina, Nikolay N. Kharchenko, Konstantin A. Shestibratov. Efficient Cryopreservation of Populus tremula by In Vitro-Grown Axillary Buds and Genetic Stability of Recovered Plants. Plants. 2021; 10 (1):77.
Chicago/Turabian StyleElena O. Vidyagina; Nikolay N. Kharchenko; Konstantin A. Shestibratov. 2021. "Efficient Cryopreservation of Populus tremula by In Vitro-Grown Axillary Buds and Genetic Stability of Recovered Plants." Plants 10, no. 1: 77.
The breeding of forest trees is only a few decades old, and is a much more complicated, longer, and expensive endeavor than the breeding of agricultural crops. One breeding cycle for forest trees can take 20–30 years. Recent advances in genomics and molecular biology have revolutionized traditional plant breeding based on visual phenotype assessment: the development of different types of molecular markers has made genotype selection possible. Marker-assisted breeding can significantly accelerate the breeding process, but this method has not been shown to be effective for selection of complex traits on forest trees. This new method of genomic selection is based on the analysis of all effects of quantitative trait loci (QTLs) using a large number of molecular markers distributed throughout the genome, which makes it possible to assess the genomic estimated breeding value (GEBV) of an individual. This approach is expected to be much more efficient for forest tree improvement than traditional breeding. Here, we review the current state of the art in the application of genomic selection in forest tree breeding and discuss different methods of genotyping and phenotyping. We also compare the accuracies of genomic prediction models and highlight the importance of a prior cost-benefit analysis before implementing genomic selection. Perspectives for the further development of this approach in forest breeding are also discussed: expanding the range of species and the list of valuable traits, the application of high-throughput phenotyping methods, and the possibility of using epigenetic variance to improve of forest trees.
Vadim G. Lebedev; Tatyana N. Lebedeva; Aleksey I. Chernodubov; Konstantin A. Shestibratov. Genomic Selection for Forest Tree Improvement: Methods, Achievements and Perspectives. Forests 2020, 11, 1190 .
AMA StyleVadim G. Lebedev, Tatyana N. Lebedeva, Aleksey I. Chernodubov, Konstantin A. Shestibratov. Genomic Selection for Forest Tree Improvement: Methods, Achievements and Perspectives. Forests. 2020; 11 (11):1190.
Chicago/Turabian StyleVadim G. Lebedev; Tatyana N. Lebedeva; Aleksey I. Chernodubov; Konstantin A. Shestibratov. 2020. "Genomic Selection for Forest Tree Improvement: Methods, Achievements and Perspectives." Forests 11, no. 11: 1190.
Strawberry (Fragaria) and raspberry (Rubus) are very popular crops, and improving their nutritional quality and disease resistance are important tasks in their breeding programs that are becoming increasingly based on use of functional DNA markers. We identified 118 microsatellite (simple sequence repeat—SSR) loci in the nucleotide sequences of flavonoid biosynthesis and pathogenesis-related genes and developed 24 SSR markers representing some of these structural and regulatory genes. These markers were used to assess the genetic diversity of 48 Fragaria and Rubus specimens, including wild species and rare cultivars, which differ in berry color, ploidy, and origin. We have demonstrated that a high proportion of the developed markers are transferable within and between Fragaria and Rubus genera and are polymorphic. Transferability and polymorphism of the SSR markers depended on location of their polymerase chain reaction (PCR) primer annealing sites and microsatellite loci in genes, respectively. High polymorphism of the SSR markers in regulatory flavonoid biosynthesis genes suggests their allelic variability that can be potentially associated with differences in flavonoid accumulation and composition. This set of SSR markers may be a useful molecular tool in strawberry and raspberry breeding programs for improvement anthocyanin related traits.
Vadim G. Lebedev; Natalya M. Subbotina; Oleg P. Maluchenko; Tatyana N. Lebedeva; Konstantin V. Krutovsky; Konstantin A. Shestibratov. Transferability and Polymorphism of SSR Markers Located in Flavonoid Pathway Genes in Fragaria and Rubus Species. Genes 2019, 11, 11 .
AMA StyleVadim G. Lebedev, Natalya M. Subbotina, Oleg P. Maluchenko, Tatyana N. Lebedeva, Konstantin V. Krutovsky, Konstantin A. Shestibratov. Transferability and Polymorphism of SSR Markers Located in Flavonoid Pathway Genes in Fragaria and Rubus Species. Genes. 2019; 11 (1):11.
Chicago/Turabian StyleVadim G. Lebedev; Natalya M. Subbotina; Oleg P. Maluchenko; Tatyana N. Lebedeva; Konstantin V. Krutovsky; Konstantin A. Shestibratov. 2019. "Transferability and Polymorphism of SSR Markers Located in Flavonoid Pathway Genes in Fragaria and Rubus Species." Genes 11, no. 1: 11.
Weeds are a big problem in agriculture and forestry, and herbicides are the main tools to control them. Phosphinotricin (ammonium glufosinate, PPT) is one of the most effective non-selective herbicides, to which weeds hardly gain resistance, but the reasons for its effect and toxicity to plants are still unclear, and especially, it is little studied in trees, including transgenic ones. We studied the physiological responses of downy birch (Betula pubescens Ehrh.) containing the herbicide resistance bar gene or the cytosol glutamine synthetase GS1 gene (the target enzyme of the herbicide) to PPT-based Basta herbicide treatment in various doses under open-air conditions during two years. Birch saplings with the bar gene were resistant to a double field dose (10 L/ha), but the expression of the GS1 gene only slightly increased resistance compared to the control. Herbicide treatment increased the ammonium level in leaf tissue by 3–8 times, but this, apparently, was not the main cause of plant death. Among leaf pigments, chlorophyll B was the most resistant to PPT, and carotenoids were the most sensitive. Responses of birch trees with the GS1 gene (accumulation of ammonium, pigment content, and dehydration) during treatment with a low dose of herbicide were less pronounced than in control plants. One-year-old control and transgenic plants with the GS gene died after 2.5 L/ha treatment, and two-year-old plants lost foliage after such treatment but remained alive and developed buds four weeks after treatment. Herbicide treatment of plants with the bar gene did not cause significant deviations in height (first year) or the accumulation of aboveground biomass (second year). The obtained results improve our understanding of the effect of PPT on woody plants and can be used both to clarify mechanisms of herbicide action and in plantation forestry.
Vadim G. Lebedev; Konstantin V. Krutovsky; Konstantin A. Shestibratov. Effect of Phosphinothricin on Transgenic Downy Birch (Betula pubescens Ehrh.) Containing bar or GS1 Genes. Forests 2019, 10, 1067 .
AMA StyleVadim G. Lebedev, Konstantin V. Krutovsky, Konstantin A. Shestibratov. Effect of Phosphinothricin on Transgenic Downy Birch (Betula pubescens Ehrh.) Containing bar or GS1 Genes. Forests. 2019; 10 (12):1067.
Chicago/Turabian StyleVadim G. Lebedev; Konstantin V. Krutovsky; Konstantin A. Shestibratov. 2019. "Effect of Phosphinothricin on Transgenic Downy Birch (Betula pubescens Ehrh.) Containing bar or GS1 Genes." Forests 10, no. 12: 1067.
Raspberry is a valuable berry crop containing a large amount of antioxidants that correlates with the color of the berries. We evaluated the genetic diversity of differently colored raspberry cultivars by the microsatellite markers developed using the flavonoid biosynthesis structural and regulatory genes. Among nine tested markers, seven were polymorphic. In total, 26 alleles were found at seven loci in 19 red (Rubus idaeus L.) and two black (R. occidentalis L.) raspberry cultivars. The most polymorphic marker was RiMY01 located in the MYB10 transcription factor intron region. Its polymorphic information content (PIC) equalled 0.82. The RiG001 marker that previously failed to amplify in blackberry also failed in black raspberry. The raspberry cultivar clustering in the UPGMA dendrogram was unrelated to geographical and genetic origin, but significantly correlated with the color of berries. The black raspberry cultivars had a higher homozygosity and clustered separately from other cultivars, while at the same time they differed from each other. In addition, some of the raspberry cultivars with a yellow-orange color of berries formed a separate cluster. This suggests that there may be not a single genetic mechanism for the formation of yellow-orange berries. The data obtained can be used prospectively in future breeding programs to improve the nutritional qualities of raspberry fruits.
Vadim G. Lebedev; Natalya Subbotina; Oleg P. Maluchenko; Konstantin V. Krutovsky; Konstantin A. Shestibratov. Assessment of Genetic Diversity in Differently Colored Raspberry Cultivars Using SSR Markers Located in Flavonoid Biosynthesis Genes. Agronomy 2019, 9, 518 .
AMA StyleVadim G. Lebedev, Natalya Subbotina, Oleg P. Maluchenko, Konstantin V. Krutovsky, Konstantin A. Shestibratov. Assessment of Genetic Diversity in Differently Colored Raspberry Cultivars Using SSR Markers Located in Flavonoid Biosynthesis Genes. Agronomy. 2019; 9 (9):518.
Chicago/Turabian StyleVadim G. Lebedev; Natalya Subbotina; Oleg P. Maluchenko; Konstantin V. Krutovsky; Konstantin A. Shestibratov. 2019. "Assessment of Genetic Diversity in Differently Colored Raspberry Cultivars Using SSR Markers Located in Flavonoid Biosynthesis Genes." Agronomy 9, no. 9: 518.
The use of natural products that can serve as natural herbicides and insecticides is a promising direction because of their greater safety for humans and environment. Secondary metabolites of plants that are toxic to plants and insects—allelochemicals—can be used as such products. Woody plants can produce allelochemicals, but they are studied much less than herbaceous species. Meanwhile, there is a problem of interaction of woody species with neighboring plants in the process of introduction or invasion, co-cultivation with agricultural crops (agroforestry) or in plantation forestry (multiclonal or multispecies plantations). This review describes woody plants with the greatest allelopathic potential, allelochemicals derived from them, and the prospects for their use as biopesticides. In addition, the achievement of and the prospects for the use of biotechnology methods in relation to the allelopathy of woody plants are presented and discussed.
Vadim G. Lebedev; Konstantin V. Krutovsky; Konstantin A. Shestibratov. …Fell Upas Sits, the Hydra-Tree of Death †, or the Phytotoxicity of Trees. Molecules 2019, 24, 1636 .
AMA StyleVadim G. Lebedev, Konstantin V. Krutovsky, Konstantin A. Shestibratov. …Fell Upas Sits, the Hydra-Tree of Death †, or the Phytotoxicity of Trees. Molecules. 2019; 24 (8):1636.
Chicago/Turabian StyleVadim G. Lebedev; Konstantin V. Krutovsky; Konstantin A. Shestibratov. 2019. "…Fell Upas Sits, the Hydra-Tree of Death †, or the Phytotoxicity of Trees." Molecules 24, no. 8: 1636.
Successful acclimatization and ex vitro rooting are among the key factors reducing the cost of micropropagated plants. We compared the survival of seven Russian cultivars of raspberry (Rubus idaeus) after rooting in vitro and ex vitro. Rooted shoots adapted to nonsterile conditions much better than nonrooted ones, with survival rates of 81%–98% versus 43%–76%, respectively. We studied the effects of different combinations of plant-growth regulators and gelling agents added to a proliferation medium on ex vitro rooting of primocane-fruiting raspberry cultivar “Atlant”. Reducing the agar concentration from 8 to 6.5 g/L increased the multiplication rate, but caused shoot hyperhydricity. The highest survival rate (97.2%) was observed for shoots grown in a medium containing 0.2 and 0.1 mg/L IBA, and gelled with 5 g/L agar and 0.2 g/L Phytagel. The microshoot height at the multiplication stage did not correlate with the plant growth during acclimatization. The obtained results can be used in the commercial micropropagation of the raspberry.
Vadim Lebedev; Mikhail Arkaev; Mariya Dremova; Ivan Pozdniakov; Konstantin Shestibratov. Effects of Growth Regulators and Gelling Agents on Ex Vitro Rooting of Raspberry. Plants 2018, 8, 3 .
AMA StyleVadim Lebedev, Mikhail Arkaev, Mariya Dremova, Ivan Pozdniakov, Konstantin Shestibratov. Effects of Growth Regulators and Gelling Agents on Ex Vitro Rooting of Raspberry. Plants. 2018; 8 (1):3.
Chicago/Turabian StyleVadim Lebedev; Mikhail Arkaev; Mariya Dremova; Ivan Pozdniakov; Konstantin Shestibratov. 2018. "Effects of Growth Regulators and Gelling Agents on Ex Vitro Rooting of Raspberry." Plants 8, no. 1: 3.
The growing importance of forest plantations increases the demand for phytopathogen resistant forest trees. This study describes an effective method for early detection and identification of the main fungal phytopathogens in planting material of silver birch (Betula pendula) and downy birch (B. pubescens), based on the estimation of the size of the internal transcribed spacers (ITS1 and ITS2) in the 18S-5.8S-28S rDNA gene cluster, which are species-specific for most micromycetes. The electrophoretic assay of the ITS1 and ITS2 loci has allowed us to identify predominant phytopathogenic fungal species in downy and silver birch in planta. This new molecular genetic method can be used to screen birch and other forest trees for different fungal pathogens to evaluate disease resistance. This information can be useful in breeding new genotypes of forest trees, including transgenic clones with modified wood composition.
Konstantin A. Shestibratov; Oleg Yu. Baranov; Natalya M. Subbotina; Vadim G. Lebedev; Stanislav V. Panteleev; Konstantin V. Krutovsky; Vladimir E. Padutov. Early Detection and Identification of the Main Fungal Pathogens for Resistance Evaluation of New Genotypes of Forest Trees. Forests 2018, 9, 732 .
AMA StyleKonstantin A. Shestibratov, Oleg Yu. Baranov, Natalya M. Subbotina, Vadim G. Lebedev, Stanislav V. Panteleev, Konstantin V. Krutovsky, Vladimir E. Padutov. Early Detection and Identification of the Main Fungal Pathogens for Resistance Evaluation of New Genotypes of Forest Trees. Forests. 2018; 9 (12):732.
Chicago/Turabian StyleKonstantin A. Shestibratov; Oleg Yu. Baranov; Natalya M. Subbotina; Vadim G. Lebedev; Stanislav V. Panteleev; Konstantin V. Krutovsky; Vladimir E. Padutov. 2018. "Early Detection and Identification of the Main Fungal Pathogens for Resistance Evaluation of New Genotypes of Forest Trees." Forests 9, no. 12: 732.
An alternative way to increase plant productivity through the use of nitrogen fertilizers is to improve the efficiency of nitrogen utilization via genetic engineering. The effects of overexpression of pine glutamine synthetase (GS) gene and nitrogen availability on growth and leaf pigment levels of two Betula species were studied. Untransformed and transgenic plants of downy birch (B. pubescens) and silver birch (B. pendula) were grown under open-air conditions at three nitrogen regimes (0, 1, or 10 mM) for one growing season. The transfer of the GS1a gene led to a significant increase in the height of only two transgenic lines of nine B. pubescens, but three of five B. pendula transgenic lines were higher than the controls. In general, nitrogen supply reduced the positive effect of the GS gene on the growth of transgenic birch plants. No differences in leaf pigment levels between control and transgenic plants were found. Nitrogen fertilization increased leaf chlorophyll content in untransformed plants but its effect on most of the transgenic lines was insignificant. The results suggest that birch plants carrying the GS gene use nitrogen more efficiently, especially when growing in nitrogen deficient soil. Transgenic lines were less responsive to nitrogen supply in comparison to wild-type plants.
Vadim G. Lebedev; Nina P. Kovalenko; Konstantin A. Shestibratov. Influence of Nitrogen Availability on Growth of Two Transgenic Birch Species Carrying the Pine GS1a Gene. Plants 2017, 6, 4 .
AMA StyleVadim G. Lebedev, Nina P. Kovalenko, Konstantin A. Shestibratov. Influence of Nitrogen Availability on Growth of Two Transgenic Birch Species Carrying the Pine GS1a Gene. Plants. 2017; 6 (1):4.
Chicago/Turabian StyleVadim G. Lebedev; Nina P. Kovalenko; Konstantin A. Shestibratov. 2017. "Influence of Nitrogen Availability on Growth of Two Transgenic Birch Species Carrying the Pine GS1a Gene." Plants 6, no. 1: 4.
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.
Vadim G. Lebedev; Elena Vidyagina; Alla A. Larionova; Konstantin A. Shestibratov. Decomposition of Leaves, Stems and Roots of Transgenic Aspen with the Xyloglucanase (sp-Xeg) Gene under Laboratory Microcosm Conditions. Environments 2016, 4, 4 .
AMA StyleVadim G. Lebedev, Elena Vidyagina, Alla A. Larionova, Konstantin A. Shestibratov. Decomposition of Leaves, Stems and Roots of Transgenic Aspen with the Xyloglucanase (sp-Xeg) Gene under Laboratory Microcosm Conditions. Environments. 2016; 4 (1):4.
Chicago/Turabian StyleVadim G. Lebedev; Elena Vidyagina; Alla A. Larionova; Konstantin A. Shestibratov. 2016. "Decomposition of Leaves, Stems and Roots of Transgenic Aspen with the Xyloglucanase (sp-Xeg) Gene under Laboratory Microcosm Conditions." Environments 4, no. 1: 4.