PLANT GENETICS
Investigation of the effect of the cytoplasm on the combining ability (CA) of lines with cytoplasmic male sterility (CMS) is of considerable interest in terms of understanding the genetic functions of the cytoplasm and for practical purposes to create hybrids with improved economically valuable traits. In order to investigate the effect of different types of sterile cytoplasm (A3, A4, 9E) on CA in sorghum, we studied the manifestation of a number of biological and agronomic traits in 54 F1 hybrid combinations obtained using iso-nuclear CMS lines with the nuclear genome of the line Zheltozernoye 10, differing only in the types of sterile cytoplasm (A3, A4 and 9E). Eighteen varieties and lines of grain sorghum developed at the Russian Research and Project-technological Institute of Sorghum and Maize were used as paternal parents. The CA was determined by the topcross method. F1 hybrids and their parents were grown in 2015–2017 in conditions of insufficient (2015–2016: HTC (hydro-thermal coefficient) = 0.32–0.66), or good water availability conditions (2017: HTC = 1.00). On average, for three years of testing, a positive effect of the 9E cytoplasm on the general combining ability (GCA) (0.63) and negative effects of the A3 and A4 cytoplasms (–0.32 and –0.31) for the inflorescence length were noted. In dry seasons, significant positive effects of the 9E cytoplasm on GCA for the length of the largest leaf, and positive effects of the A3 cytoplasm on GCA for the plant height, and negative effects of the A4 cytoplasm on GCA for these traits were observed. No differences were observed during the wet season. The type of CMS did not affect the GCA for the width of the largest leaf and grain yield. The dispersion of specific combining ability (SCA) in the dry seasons was significant for the following traits: leaf length, plant height, panicle length and width, and grain yield, the 9E cytoplasm had the highest SCA dispersion, whereas the A4 cytoplasm had the smallest one. The data obtained indicate that different types of sterile cytoplasm of sorghum make a different contribution to CA under conditions of drought stress.
Bread wheat (Triticum aestivum L.), the varieties of which are widely used for the grain production, is difficultly crossable with related species of Triticeae Dum. This factor limits the chance of introduction of alien genetic material into the wheat gene pool and the possibility of new varieties breeding with good adaptation to adverse environmental factors. The crossability between wheat and related species is controlled by Kr1-Kr4 genes (Crossability with Rye, Hordeum and Aegilops spp.) and the SKr gene (Suppressor of crossability). SKr and Kr1 have the largest influence on the trait. In the case of the recessive alleles, these genes do not function and the quantity of hybrid seeds after pollination with alien species can achieve more than 50 %. SKr is located on 5BS between the GBR0233 and Xgwm234 markers, closely linked with the markers Xcfb341, TGlc2 and gene12. Kr1 was mapped on 5BL, proximally to the Ph1 gene, between the EST-SSR markers Xw5145 and Xw9340. The markers of SKr were used to control the transfer of its recessive allele into other wheat genotypes, which made it possible to obtain highly crossable forms. However, the advantages of using the SKr and Kr1 markers in marker-assisted selection and in the screening of ex situ collections are not sufficiently studied. The published Kr1 sequence for varieties with different crossability offers great prospects, because it will be possible to create allele-specific markers. In this review, the following issues are considered: genetic resources created by wheat and rye hybridization, the geographical distribution of easy-to-cross forms of wheat, genetic control of the wheat and rye compatibility, advances of the use of molecular markers in the mapping of Kr-genes and their transmission control.
Awns are bristle‐like structures, typically extending from the tip end of the lemmas in the florets of cereal species, including such economically important crops as wheat (Triticum aestivum L., T. durum Desf.), barley (Hordeum vulgare L.), rice (Oryza sativa L.), and rye (Secale cereale L.). The presence of long awns adhered at tip end of glumes is a characteristic feature of “Persian wheat” T. carthlicum Nevski spike. Glume outgrowth of T. carthlicum Nevski spike passes into a long awn, equal in length to the lemma awn. Awned glumes can be formed in T. aestivum and T. aethiopicum wheats, however, such forms are rare. Features of the awned glume development and the genetic determinants of this trait have been little studied. In this paper, we described the features of the development and inheritance of the tetra-awness (awned glume) trait of the bread wheat T. aestivum line CD 1167-8, using classical genetic analysis, molecular genetic mapping, and scanning electron microscopy. It was shown that the trait is inherited as a recessive monogenic. The gene for the awned glume trait of CD 1167-8 was mapped in the long arm of chromosome 5A, using the Illumina Infinium 15K Wheat Array (TraitGenetics GmbH), containing 15,000 SNPs associated with wheat genes. Results of allelism test and molecular-genetic mapping suggest that the gene for awned glumes in bread wheat is a recessive allele of the B1 awn suppressor. This new allele was designated the b1.ag (b1. awned glume). Analysis of the CD 1167-8 inflorescence development, using scanning electron microscopy, showed that awns had grown from the top of the lemmas and glumes simultaneously, and no differences in patterns of their development were found.
Clonal propagation of conifers using somatic embryogenesis is essential for the selection of tree species, and for the implementation of afforestation and reforestation. In combination with cryopreservation, somatic embryogenesis creates the basis for the development of economically valuable lines of clones and elite genotypes. The industrial use of such genetically verified clone lines in forestry can significantly increase forest productivity compared to any conventional methods for improving tree crops that are available. Larch is considered as one of the main conifer candidates for large-scale reforestation, not only due to the vastness of its habitat, but also due to the unique quality of its wood, rapid growth and high ecological plasticity. However, the vast majority of larch species are characterized by uneven yields and extremely low seed quality. In this regard, obtaining planting material for reforestation from larch seeds on seed plantations is not advisable, but can be successfully implemented in afforestation programs using somatic embryogenesis technologies. Research on the somatic embryogenesis of larch has been conducted for over 30 years, which allowed considerable experience in this field to be accumulated. To date, the conditions for the initiation and maintenance of embryogenic cultures, as well as for the formation and development of somatic embryos have been determined. Significant progress has been made in the study of both the factors affecting these processes and the molecular mechanisms that underlie the various stages of embryogenesis. Nevertheless, despite the successes achieved, knowledge available today on the somatic embryogenesis of representatives of the genus Larix is still not enough to develop technologies for producing valuable plant-breeding material in vitro. This review analyzes the current state of research on the problem of somatic embryogenesis of representatives of the genus Larix. Particular attention is paid to the choice of explants for somatic embryogenesis, the composition of the media for cultivation, the dependence of the potential of somatic embryogenesis on the duration of cultivation, and the genetic control of somatic embryogenesis.
Of all the subspecies of Zea mays L. cultivated in the world, sweet maize is the most important for the global economy. The leading seed-growing companies and research institutions around the world are engaged in breeding this crop. To meet the increasing demands of the industry to grain quality, it is important to select appropriate local varieties and lines for hybridization. Local (usually heterogeneous) varieties are a valuable source material for creating self-pollinated lines that contribute to a significant broadening of the genetic base of parental forms used in breeding. The advantages of sweet maize varieties and the interest of the food industry in them make it possible to consider accessions from the maize collection of the N.I. Vavilov Institute (VIR) as a potentially valuable source material for breeding. The present research concentrated on 19 local sweet maize varieties with different grain colors from the VIR collection, that is, 9 varieties with the blue color of ripe grain, 4 with white (colorless) grain, 3 with yellow, and 3 with red. The research included an analysis of zein electrophoretic patterns (protein markers); a study of their biotype composition and the nature of genetic polymorphism, as well as the creation of a protein pattern database for each accession. For a series of accessions with the same varietal name, but different catalog numbers, the degree of their identity was determined from their biotype composition in order to exclude duplication. Zein electrophoresis was carried out in vertical plates of 10 % polyacrylamide gel according to the standard ISTA technique developed with the participation of the Biochemistry and Molecular Biology Department of VIR. Zein patterns were used for the first time to electrophoretically study sweet maize varieties with different grain colors. Unique zein patterns were established for all the accessions studied, which makes possible their identification by specific marker components. The results of this work characterize zein electrophoresis as a useful tool for the identification and registration of duplicate accessions in the VIR collection of sweet maize varieties.
Low-temperature stress is one of the main factors limiting the distribution and reducing the yield of many subtropical crops, including the tea crop. Efficient breeding to develop frost-tolerant cultivars requires a reliable set of genetic markers for identifying resistance donors, and that is why it is necessary to reveal the specific genetic response in frost-tolerant genotypes in comparison with frost- susceptible ones. In this work, we performed a comparative analysis of the expression of 18 tea genes (ICE1, CBF1, DHN1, DHN2, DHN3, NAC17, NAC26, NAC30, bHLH7, bHLH43, P5CS, WRKY2, LOX1, LOX6, LOX7, SnRK1.1, SnRK1.2, SnRK1.3) under cold and frost conditions in two tea genotypes, tolerant and susceptible. Low-temperature stress was induced by placing the potted plants in cold chambers and lowering the temperature to 0…+2 °С for 7 days (cold stress), followed by a decrease in temperature to –4…–6 °С for 5 days (frost stress). Relative electrical conductivity of leaf was measured in response to the stress treatments, and a significant difference in the frost tolerance of the two tea genotypes was confirmed. Cold exposure did not lead to a change in the electrical conductivity of leaf tissue. On the other hand, frost treatment resulted in increased REC in both genotypes and to a greater extent in the susceptible genotype. Increased expression of all the genes was shown during cold and frost. The genes that were strongly expressed in the tolerant tea genotype were revealed: ICE1, CBF1, DHN2, NAC17, NAC26, bHLH43, WRKY2, P5CS, LOX6, SnRK1.1, SnRK1.3. These genes can be proposed as markers for the selection of frost-tolerance donors in tea germplasm collections. Additionally, it was shown that the tolerant genotype is characterized by an earlier response to stress at the stage of cold acclimation. The study of the expression of the identified genes in different organs of tea plants and in different exposures to low temperature is relevant for further investigations.
PLANT GENE POOL AND BREEDING
New cultivars adapted to major durum wheat growing environments are essential for the cultivation of this crop. The development of new cultivars has required the availability of diverse genetic material and their extensive field trials. In this work, a collection of tetraploid wheat consisting of 85 accessions was tested in the field conditions of Almaty region during 2018 and 2019. The accessions were ranged according to nine agronomic traits studied, and accessions with the highest yield performance for Almaty region of Kazakhstan were revealed. The ANOVA suggested that the performance of agronomic traits were influenced both by Environment and Genotype. Also, the collection was analyzed using seven SSR (simple sequence repeats) markers. From 3 to 6 alleles per locus were revealed, with an average of 4.6, while the effective number of alleles was 2.8. Nei’s genetic diversity was in the range of 0.45–0.69. The results showed high values of polymorphism index content (PIC) in the range of 0.46–0.70, with an average of 0.62, suggesting that 6 out of 7 SSRs were highly informative (PIC > 0.5). Phylogenetic analysis of the collection has allowed the separation of accessions into six clusters. The local accessions were presented in all six clusters with the majority of them grouped in the first three clusters designated as A, B, and C, respectively. The relations between SSR markers and agronomic traits in the collection were studied. The results can be efficiently used for the enhancement of local breeding projects for the improvement of yield productivity in durum wheat.
Radish and small radish (Raphanus sativus L.) are popular and widely cultivated root vegetables in the world, which occupy an important place in human nutrition. Edaphic stressors have a significant impact on their productivity and quality. The main factor determining the phytotoxicity of acidic soils is the increased concentration of mobile aluminum ions in the soil solution. The accumulation of aluminum in root tissues disrupts the processes of cell division, initiation and growth of the lateral roots, the supply of plants with minerals and water. The study of intraspecific variation in aluminum resistance of R. sativus is an important stage for the breeding of these crops. The purpose of this work was to study the genetic diversity of R. sativus crops including 109 accessions of small radish and radish of various ecological and geographical origin, belonging to 23 types, 14 varieties of European, Chinese and Japanese subspecies on aluminum tolerance. In the absence of a rapid assessment methodology specialized for the species studied, a method is used to assess the aluminum resistance of cereals using an eriochrome cyanine R dye, which is based on the recovery or absence of restoration of mitotic activity of the seedlings roots subjected to shock exposure to aluminum. The effect of various concentrations on the vital activity of plants was revealed: a 66-mM concentration of AlCl3 · 6Н2О had a weak toxic effect on R. sativus accessions slowing down root growth; 83 mM contributed to a large differentiation of the small radish accessions and to a lesser extent for radish; 99 mM inhibited further root growth in 13.0 % of small radish accessions and in 7.3 % of radish and had a highly damaging effect. AlCl3 · 6Н2О at a concentration of 99 mM allowed us to identify the most tolerant small radish and radish accessions that originate from countries with a wide distribution of acidic soils. In a result, it was possible to determine the intraspecific variability of small radish and radish plants in the early stages of vegetation and to identify genotypes that are contrasting in their resistance to aluminum. We recommend the AlCl3 · 6Н2О concentration of 83 mM for screening the aluminum resistance of small radish and 99 mM for radish. The modified method that we developed is proposed as a rapid diagnosis of aluminum tolerance for the screening of a wide range of R. sativus genotypes and a subsequent study of contrasting forms during a longer cultivation of plants in hydroponic culture (including elemental analysis of roots and shoots, contrasting in resistance of accessions) as well as reactions of plants in soil conditions.
Narrow-leaved lupine (Lupinus angustifolius L.), a valuable leguminous crop adapted to a wide range of climatic conditions, has a very short history of domestication. For many centuries it was used mainly as a green manure, since the success and prospects of the multi-purpose use of the species depend on its breeding improvement, in particular, on a particular concentration of alkaloids in seeds and green mass. The first varieties of scientific breeding were created only in the 1930s after the appearance of low-alkaloid mutants. Despite wide prospects for use in various areas of the national economy, unstable productivity and susceptibility to diseases hinder the production of this crop. Obviously, breeders deal only with a small part of the gene pool of the species and limited genetic resources, using mainly low-alkaloid (sweet) genotypes to create new varieties. The genetic potential of the species can be used more efficiently. At the same time, it is rational to create highly alkaloid (bitter) varieties for green manure, while food and feed varieties should not lose their adaptive potential, in particular, resistance to pathogens, due to the elimination of alkaloids. In this regard, it seems to be a productive idea to create ‘bitter/sweet’ varieties combining a high content of alkaloids in the vegetative organs and low in seeds, which can be achieved by regulating the synthesis/transport of alkaloids in the plant. The paper discusses the current state of use of the species as a green manure, fodder, food plant. Information is given on the quantity and qualitative composition of narrow-leaved lupine alkaloids, their applied value, in particular, fungicidal, antibacterial, insecticidal, the use of lupine alkaloids as active principles of drugs. Along with promising breeding considerations, the possibility of using technologies for processing raw high-alkaloid materials with the accompanying extraction of valuable ingredients for pharmaceuticals is discussed. Information is briefly presented about the genomic resources of the species and the prospects for their use in marker-assistant selection and genome editing.
MOLECULAR AND CELL BIOLOGY
The free-living flatworm Macrostomum mirumnovem is a neopolyploid species whose genome underwent a recent Whole Genome Duplication (WGD). In the result of chromosome fusions of the ancient haploid chromosome set, large metacentric chromosomes were formed. In addition to three pairs of small metacentrics, the current karyotype of M. mirumnovem contains two pairs of large metacentric chromosomes, MMI1 and MMI2. The generation of microdissected DNA libraries enriched for DNA repeats followed by DNA probe preparation and fluorescent in situ hybridization (FISH) were performed. The DNA probes obtained marked chromosome regions enriched for different DNA repeats in the M. mirumnovem chromosomes. The size and localization of these regions varied in different copies of large chromosomes. They varied even in homologous chromosomes, suggesting their divergence due to genome re-diploidization after a WGD. Besides the newly formed chromosome regions enriched for DNA repeats, B chromosomes were found in the karyotypes of the studied specimens of M. mirumnovem. These B chromosomes varied in size and morphology. FISH with microdissected DNA probes revealed that some Bs had a distinct DNA content. FISH could paint differently B chromosomes in different worms and even in the same sample. B chromosomes could carry a bright specific fluorescent signal or could show no fluorescent signal at all. In latter cases, the specific FISH signal could be absent even in the pericentromeric region of the B chromosome. Possible mechanisms of B chromosome formation and their further evolution are discussed. The results obtained indicate an important role that repetitive DNAs play in genome re-diploidization initiating a rapid differentiation of large chromosome copies. Taking together, karyotype peculiarities (a high level of intraspecific karyotypic diversity associated with chromosome number variation, structural chromosomal rearrangements, and the formation of new regions enriched for DNA repeats) and some phenotypic features of M. mirumnovem (small body size, short lifecycle, easy maintenance in the laboratory) make this species a perspective model in the studies of genomic and karyotypic evolution in species passed through a recent WGD event.
The paper describes some biological features of the radioprotective effect of double-stranded RNA preparation. It was found that yeast RNA preparation has a prolonged radioprotective effect after irradiation by a lethal dose of 9.4 Gy. 100 % of animals survive on the 70th day of observation when irradiated 1 hour or 4 days after 7 mg RNA preparation injection, 60 % animals survive when irradiated on day 8 or 12. Time parameters of repair of double-stranded breaks induced by gamma rays were estimated. It was found that the injection of the RNA preparation at the time of maximum number of double-stranded breaks, 1 hour after irradiation, reduces the efficacy of radioprotective action compared with the injection 1 hour before irradiation and 4 hours after irradiation. A comparison of the radioprotective effect of the standard radioprotector B-190 and the RNA preparation was made in one experiment. It has been established that the total RNA preparation is more efficacious than B-190. Survival on the 40th day after irradiation was 78 % for the group of mice treated with the RNA preparation and 67 % for those treated with B-190. In the course of analytical studies of the total yeast RNA preparation, it was found that the preparation is a mixture of single-stranded and double-stranded RNA. It was shown that only double-stranded RNA has radioprotective properties. Injection of 160 μg double-stranded RNA protects 100 % of the experimental animals from an absolutely lethal dose of gamma radiation, 9.4 Gy. It was established that the radioprotective effect of double-stranded RNA does not depend on sequence, but depends on its double-stranded form and the presence of “open” ends of the molecule. It is supposed that the radioprotective effect of double-stranded RNA is associated with the participation of RNA molecules in the correct repair of radiation-damaged chromatin in blood stem cells. The hematopoietic pluripotent cells that have survived migrate to the periphery, reach the spleen and actively proliferate. The newly formed cell population restores the hematopoietic and immune systems, which determines the survival of lethally irradiated animals.
Myeloid dendritic cells (DCs) play an important role in the immune response; therefore, the search for compounds that can effectively activate DCs is a needful goal. This study was aimed to investigate the effect of synthetic CpG oligodeoxynucleotides (CpG-ODN) on the maturation and allostimulatory activity of myeloid DCs in comparison with other PAMP and DAMP molecules. For the research, we synthesized known CpG-ODN class C (SD-101 and D-SL03) containing thiophosphate internucleotide groups, and their original phosphate-modified analogues (SD-101M and D-SL03M) with mesylphosphoramide internucleotide groups (M = μ-modification). The effects of CpG-ODN and other activators were evaluated on DCs generated from blood monocytes in the presence of GM-CSF and IFN-α (IFN-DC) or IL-4 (IL4-DC). Evaluation of the intracellular TLR-9 expression showed that both types of DCs (IFN-DC and IL4-DC) contained on average 52 and 80 % of TLR-9-positive cells, respectively. The CpG-ODNs studied enhanced the allostimulatory activity of IFN-DCs, and the effect of μ-modified CpG-ODNs was higher than that of CpG-ODNs with thiophosphate groups. The stimulating effect of CpG-ODN at a dose of 1.0 μg/ml was comparable (for D-SL03, D-SL03M, SD-101) with or exceeded (for SD-101M) the effect of LPS at a dose of 10 μg/ml. At the same time, IFN-DCs were characterized by greater sensitivity to the action of CpG-ODNs than IL4-DCs. The enhancement of DC allostimulatory activity in the presence of CpG-ODNs was associated with the induction of final DC maturation, which was confirmed by a significant decrease in the number of CD14+DC, an increase in mature CD83+DC and a trend towards an increase in CD86+DC. Interestingly, the characteristic ability of LPS to enhance the expression of the co-stimulatory molecule OX40L on DCs was revealed only for the μ-analogue SD-101M. In addition, CpG-ODNs (SD-101 and SD-101M) had a stimulatory effect on IFN-γ production comparable to the action of LPS. The data obtained indicate a stimulating effect of CpG-ODN on the maturation and allostimulatory activity of human myeloid DCs, which is more pronounced for μ-modified analogs.
MICROBIAL GENETICS
This article presents a general overview of the prevalence, genetic diversity and detection methods of picobirnaviruses (PBVs), which are small, non-enveloped icosahedral viruses with a segmented double-stranded RNA genome consisting of two segments taxonomically related to the genus Picobirnavirus of the family Picobirnaviridae. This review of scientific papers published in 1988–2019 provides data on the PBV distribution in the nature and a broad host range. PBV infection is characterized as opportunistic, the lack of understanding of the etiological role of PBVs in diarrhea is emphasized, since these viruses are detected both in symptomatic and asymptomatic cases. The concept of PBV infection as a chronic disease caused by a long-lasting persistence of the virus in the host is considered. Such factors as stress syndrome, physiological conditions, immune status and host age at the time of primary PBV infection influence the virus detection rate in humans and animals. The possible zoonotic nature of human PBV infection is noted due to the capacity for interspecies PBV transmission acquired during evolution as a result of the reassortment of the genome segments of different viruses infecting the same host. Data providing evidence that PBVs belong to eukaryotes and a challenging hypothesis stating that PBVs are bacterial viruses are presented. The need to intensify work on PBV detection because of their wide distribution, despite the complexity due to the lack of the cultivation system, is emphasized. Two strategies of RT-PCR as main PBV detection methods are considered. The genomes of individual representatives of the genus isolated from different hosts are characterized. Emphasis is placed on the feasibility of developing primers with broader specificity for expanding the range of identifiable representatives of the genus PBV due to a huge variety of their genotypes. The importance of effective monitoring of PBV prevalence for studying the zoonotic and anthroponotic potential using metagenomics analysis is highlighted, and so is the possibility of using PBV as a marker for environmental monitoring.
Aphids are a diverse family of crop pests. Aphids formed a complex relationship with intracellular bacteria. Depending on the region of study, the species composition of both aphids and their facultative endosymbionts varies. The aim of the work was to determine the occurrence and genetic diversity of Wolbachia, Spiroplasma and Rickettsia symbionts in aphids collected in 2018–2019 in Moscow. For these purposes, 578 aphids from 32 collection sites were tested by PCR using specific primers. At least 21 species of aphids from 14 genera and four families were identified by barcoding method, of which 11 species were infected with endosymbionts. Rickettsia was found in six species, Wolbachia in two species, Spiroplasma in one species. The presence of Rickettsia in Impatientinum asiaticum, Myzus cerasi, Hyalopterus pruni, Eucallipterus tiliae, Chaitophorus tremulae and Wolbachia in Aphis pomi and C. tremulae has been described for the first time. A double infection with Rickettsia and Spiroplasma was detected in a half of pea aphid (Acyrthosiphon pisum) individuals. For the first time was found that six species of aphids are infected with Rickettsia that are genetically different from previously known. It was first discovered that A. pomi is infected with two Wolbachia strains, one of which belongs to supergroup B and is genetically close to Wolbachia from C. tremulae. The second Wolbachia strain from A. pomi belongs to the supergroup M, recently described in aphid species. Spiroplasma, which we observed in A. pisum, is genetically close to male killing Spiroplasma from aphids, ladybirds and moths. Both maternal inheritance and horizontal transmission are the pathways for the distribution of facultative endosymbiotic bacteria in aphids.