Allelopathic effects of <Emphasis Type="Italic">Thymus kotschyanus</Emphasis> on seed germination and initial growth of <Emphasis Type="Italic">Bromus tomentellus</Emphasis> and <Emphasis Type="Italic">Trifolium repens</Emphasis>

The allelopathic influence of aqueous extracts of Thymus kotschyanus on Bromus tomentellus and Trifolium repens germination (%), germination speed, and seedling growth (length, fresh and dry weight) was examined. It was noted that aqueous extracts had a considerable inhibitory effect on target plant germination, and the effect at 50%, 75%, and 100% concentration was found to be significantly higher than that at lower concentrations (5% and 25%) and control treatment (distilled water). Seedling length in addition to fresh and dry weights was also reduced significantly over control. The inhibitory effect was increased as the extract concentration was increased. B. tomentellus showed a higher sensitivity against T. kotschyanus in allelopathic effects compared to T. repens, which indicates that B. tomentellus planted in rangelands with leaf litter of T. kotschyanus will be adversely affected in terms of its germination, growth, and ultimately low forage production.     

Expression analysis of <Emphasis Type="Italic">RUS1</Emphasis> and construction of <Emphasis Type="Italic">RUS1</Emphasis> plant expressing vector

RUS1 was one of the disease resistance gene analogs obtained from Setaria italica Beauv. Semi-quantitative RT-PCR analysis result showed that RUS1 gene could be induced by Uromyces setariae-italicae and had relation to the resistance response of Setaria italica Beauv. against Uromyces setariae-italicae infection. Promoter sequence of RUS1 was obtained by the method of Genome Walking, and its length was 675 bp. RUS1 promoter and pCAMBIA1300 vector were fused to construct RUS1::GUS vector. GUS histochemical staining result showed that promoter could activate gene expression. RUS1 gene (including the promoter sequence) was obtained through PCR amplification and then fused with pCAMBIA1300 vector to construct pCAMBIA1300:RUS1 plant expressing vector. The research laid a foundation for gene functional identification of RUS1.     

Use of molecular markers of potato golden nematode resistance genes <Emphasis Type="Italic">H1</Emphasis> and <Emphasis Type="Italic">GRO1</Emphasis>

By means of DNA markers of potato golden nematode resistance genes H1 and Gro1, 109 Russian- and foreign-bred potato varieties are assessed. A sufficiently high level of coincidence of the presence of marker alleles with phenotypic resistance of potato varieties is revealed. However, not all resistant forms have specific amplification products. The use of DNA markers of only two genes H1 and Gro1 will not make it possible to completely replace mass laboratory testing, but it will make it possible to select potato forms resistant to this parasite by a simple and reliable method in a shorter time and thereby to considerably reduce the number of genotypes in the sample for further breeding.     

Founder effect and genogeography of <Emphasis Type="Italic">CV</Emphasis>- and <Emphasis Type="Italic">BL</Emphasis>-mutations in black-and-white cattle

The results are obtained by simultaneous diagnostics of mutant CV- and BL-alleles. Five groups of animals–carriers and noncarriers of mutations are identified. In Russia the mutant CV-alleles are met more frequently because of the intensive use of their carriers. While elimination of BL-allele was started earlier due to developed methods, the diagnostics of CV-mutations has been undertaken later. Propagation of missense-mutations in the world and the Russian Federation has been occurring through the breeding bulls and cows–carriers. Initially spermobanks and, afterwards, cows were reserves for the diagnosed BLand CV-mutations.     

Activity of the photosynthetic apparatus and antioxidant enzymes in leaves of transgenic <Emphasis Type="Italic">Solanum lycopersicum</Emphasis> and <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> plants,with <Emphasis Type="Italic">FeSOD1</Emphasis> gene

Introduction of the FeSOD gene enhanced the stability of the photosynthetic apparatus of plants to the action of oxidative stress caused by UV irradiation. The expression of Arabidopsis thaliana FeSOD gene, targeting the enzyme in chloroplasts due to a signal sequence, leaded to significant changes in ultrastructure of cell subcompartments of tobacco and tomato leaves. The activity of superoxide dismutase in leaves of transgenic tomato plants exceeded the value of activity of this enzyme of control plants. Transgenic tobacco plants showed increasing in SOD activity compared with control non-transgenic tobacco. The activity of AP in the leaves of transgenic tobacco and tomato plants was similar with that of control non-transgenic plants, but activity of one accession of transgenic tomato, which is also characterized by high values of SOD activity, exceeded the value of control plant. Differences in ultrastructural organization of chloroplasts in the cells of transgenic and control tobacco and tomato plants have been manifested in a strong enlargement in the size of plastoglobuli. These distinctions were evident especially in the cells of the leaf parenchyma of transgenic tomato as well as transgenic tobacco. Also, a quantity of starch grains in the plastids of guard cells was increased. Chloroplasts in the cells of leaf parenchyma in transgenic plants contained less a starch grains than in wild-type plants.     

Photoperiod sensitivity and molecular marking of genes <Emphasis Type="Italic">Ppd</Emphasis> and <Emphasis Type="Italic">Vrn</Emphasis> in connection with breeding alternative-habit wheat varieties

The response of wheat varieties and breeding lines to vernalization and photoperiod in pot experiments is studied and tested by means of allele-specific markers for genes Ppd and Vrn. These markers effectively predict the degree of photoperiod sensitivity of plants to vernalization, which attests to the expediency of using them in breeding practice.     

Increase of resistance of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> plants to phytopathogenic fungi expressing hevein-like peptides from weed plant <Emphasis Type="Italic">Stellaria media</Emphasis>

It is shown that constitutive hyperexpression of new hevein-like peptides from the weed plant chickweed (Stellaria media)in mouse-ear cress (Arabidopsis thaliana) plants leads to a substantial increase of their resistance to phytopathogenic fungi Botrytis cinerea and Bipolaris sorokiniana. Thus, common chickweed peptides can play a definite role in protecting this weed plant and be useful as a new genetic tool for producing plants resistant to fungal diseases.     

Introduction of grain cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> subsp. <Emphasis Type="Italic">cylindrical</Emphasis> (L.) Verdc.) in the Lower Volga

Introduction of cowpea (Vigna unguiculata subsp. cylindrical (L.) Verdc.) in the Lower Volga region is the possibility of using this culture for agricultural production, fodder production, and expanded range of food products. Selection for earliness in the model population of cowpea should be accompanied by the identification of genotypes with high intensity of biomass accumulation in the shoots-flowering interphase period.     

Identification of <Emphasis Type="Italic">Habrobracon hebetor</Emphasis> Populations Using RAPD Markers

It was found that food preferences and conditions of breeding of Habrobracon hebetor laboratory populations vary considerably. In this regard, it is necessary to identify populations within the studied species using DNA markers: an effective and reliable means for assessing the genetic differences between samplings of this insect species. A molecular genetic analysis of two different geographic populations of the Habrobracon hebetor entomophage (from Krasnodar, Russia, and Chimkent, Kazakhstan) was carried out using RAPD markers; 21 RAPD primers were tested for specificity to H. hebetor DNA. Five RAPD primers (OPA05, OPA10, OPB01, OPB04, and UBC519) were identified that have high specificity and the ability to differentiate H. hebetor populations. DNA markers that are specific for the Krasnodar and Chimkent entomophage populations and that can clearly identify them were revealed: for the Krasnodar population, RAPD markers with a molecular weight of 550 bp (UBC 519); 500 and 700 bp (OPA05); 1100, 1200, and 1300 bp (OPA10); 220 and 800 bp (OPB04); and 880 bp (OPB01); for the Chimkent population, 400, 600 and 1200 bp (UBC519); 600 and 950 bp (OPA10); and 800 bp (OPB01). It is concluded that these RAPD primers can be used for identification and differentiation of other H. hebetor populations.     

Formation of the hypersensitivity response due to the expression of <Emphasis Type="Italic">FeSOD1</Emphasis> gene in tomato when it is inoculated with <Emphasis Type="Italic">Phytophthora infestans</Emphasis>

The activity of antioxidant enzymes and ultrastructural changes in tissues inoculated with P. infestans isolate have been studied in the previously developed independent transgenic lines of tomato with FeSOD1 gene and control plants. It is shown that the activity of superoxide dismutase is significantly higher in transgenic plants than that in control plants (nontransgenic plants). Chlorosis and obvious changes in tissue turgor were observed when the control tomato plants were inoculated, which indicates irreversible damages and unimpeded progression of infection. At the same time, the transgenic lines were characterized by the formation of clearly limited zones of damaged cells that rapidly arrested the infection. In addition, the damages differed from those in nontransgenic plants: the cells along the edges of the infection site were smaller and had heavy invaginations of the cell wall. The contacts between the cells were disrupted in this zone, but they were preserved in undisturbed zones of the tissue. Thus, the expression of the FeSOD1 transgene promotes the emergence of the resistance to P. infestans in tomato transgenic plants.     

Identification of disease resistances in wheat-<Emphasis Type="Italic">Leymus multicaulis</Emphasis> derivatives and characterization of <Emphasis Type="Italic">L. multicaulis</Emphasis> chromatin using microsatellite DNA markers

To identify resistance to Fusarium head blight (FHB), cereal yellow dwarf virus (CYDV), stem rust (Sr), and powdery mildew (Pm) in 24 common wheat (Triticum aestivum)-Leymus multicaulis addition/translocation lines that were developed cytogenetically and to verify the authenticity of these lines using microsatellite (SSR) DNA markers. Resistance to FHB was identified in the wheat-L. multicaulis addition lines, Line 9 and Line 26, which both contained L. multicaulis-specific fragments as shown by SSR markers. The translocation line, Trans 1, and the addition lines, Line 5 and Line 29, have resistance to stem rust (IT 0). Resistance to CYDV was evaluated based on virus titers measured by enzyme linked immunosorbent assay (ELISA). The addition line, Line 23, showed low virus titer (0.15), indicating resistance to CYDV. The segregation distribution of CYDV resistance in 98 F₂ plants of Line 23/CS showed a significant deviation from 3:1. Inoculation with a set of 14 differential Blumeria graminis f. sp. tritici (Bgt) isolates did not detect powdery mildew resistance in translocation line Trans 1, addition line Line 9 and the amphiploid of wheat-L. multicaulis. However, Line 26 exhibited the resistance response pattern of Kavkaz, which carries Pm8, indicating that Line 26 most likely has the powdery mildew resistance gene Pm8 inherited from its parent lines Feng Kang 7 or Feng Kang 10. Twelve SSR markers, distributed on different homeologous chromosome groups of wheat, which distinguished L. multicaulis addition/translocation chromosomes, were used to verify the presence of L. multicaulis chromatin in the putative wheat-L. multicaulis addition/translocation lines. Of the 24 addition/translocation lines investigated using the 12 polymorphic SSR markers, 18 wheat-L. multicaulis derivatives showed the expected L. multicaulis-specific fragments, indicating that all of these 18 addition/translocation lines would most likely have the introgressed L. multicaulis chromosome(s). Chromosomal rearrangements also were detected in some of the wheat-L. multicaulis introgression lines.     

Likely threat of the spread of race UG99 of <Emphasis Type="Italic">Puccinia graminis</Emphasis> f. sp. <Emphasis Type="Italic">tritici</Emphasis> on wheat in southeastern Russia

In 2013 and 2014, a Ug99 race virulent to the resistance gene Sr31 was revealed on spring bread wheat fields in Saratov oblast and its inoculum was collected under conditions of stem rust epiphytoties (with the fungus Puccinia graminis f. sp. tritici acting as an agent). In March to April 2015, when a set of soft wheat lines was artificially inoculated in a climatic chamber, the virulence of the collected inoculum to gene Sr31 was confirmed, and four monopustular clones were isolated. Based on the isolated monopustular clones of Puccinia graminis f. sp. tritici, an assessment for resistance of four lines of spring soft wheat to novel Sr-gene combinations was made.     

Expression of special genes inhibited by powdery mildew (<Emphasis Type="Italic">Blumeria graminis</Emphasis> f. sp. <Emphasis Type="Italic">tritici</Emphasis>) in wheat germplasm N9436

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important fungal diseases in common wheat (Triticum aestivum L.) worldwide. Wheat germplasm N9436 is resistant to powdery mildew. In the present study, a backward subtracted cDNA library was constructed with cDNA from N9436 leaves inoculated by Blumeria graminis as the driver and cDNA from uninoculated N9436 leaves as the tester. A total of 120 positive clones were randomly chosen from the SSH-cDNA library and were amplified with sp6 and t7 primers to examine the insert size. After screening the repeated and redundant sequences, 59 expressed sequence tags (EST) were acquired. Nucleic acid and protein homology search were performed using the basic local alignment search tool (BLAST) program with the default settings at NCBI website (http://www.ncbi.nlm.nih.gov). BlastX results in nr-protein database revealed that 23 ESTs were highly homologous with known proteins involved in primary metabolism, energy metabolism, transport, signal transduction, and disease resistance and defenses. BlastNr results showed that 47 and 10 ESTs had high identities with known Unigene and function-unknown ESTs, respectively, and two ESTs matched none in the nr-database. Twenty-one ESTs were both in the nucleic acid and protein databases, including seven ESTs associated with powdery mildew resistance. Among them, one was responsible for signal transduction and six for systemic acquired resistance (SAR) system.     

Diversity of Dagestan barleys for the duration of the period between shooting and earing stages and alleles in the <Emphasis Type="Italic">Ppd-H1</Emphasis> and <Emphasis Type="Italic">Ppd-H2</Emphasis> loci

The duration of the shooting–earing period of 265 barley samples from Dagestan was studied. During the 3 years of study at the Dagestan Experimental Station of VIR (Derbent), fast-ripening samples k-15008 and k-15013 were identified. Evaluation of spring forms in the northwestern region of the country made it possible to identify sample k-15027, which had a high rate of development over 2 years. It was found that Dagestan barleys are strongly influenced by growing conditions; that is, they have a high rate of response. Vernalization temperatures, short photoperiod, and high temperatures during the growing season contribute to fast barley ripening. Using molecular markers, the allelic diversity of genes Ppd-H1 and Ppd-H2, which are involved in the control of the duration of the shooting–earing period, was investigated. Most samples of local forms of barley carry dominant allele Ppd-H2, which causes early earing under short photoperiod. Translocation of the studied barley group to unusual conditions of northwest Russia leads to significant delay in plant development.     

Proximate and phytochemical screening of different parts of <Emphasis Type="Italic">Moringa oleifera</Emphasis>

Lack of proper nutrition is one of the most serious problem across the globe, affecting majorly African countries, where many children died not only due to inadequate food but also as a result of insufficient nutrient in their diet. This research was aimed to study the proximate composition and screening of chemical components in different part of Moringa oleifera Lam. Proximate analysis was adapted from the standard methods developed by Association of Official Analytical Chemists (AOAC). We have determined percentage of moisture content, ash content, crude protein, crude fibre, crude fat and carbohydrate which were ranges from 8.1–4%, 5.59–9.28%, 11.70–26.57%, 21.03–44.27, 11.82–20.19 and 8.16–31.12% respectively. Different bioactive compounds of polyphenol, flavonoids, tannins and saponin were determined. Different parts of plants accumulate different levels of polyphenol, flavonoids, tannins and saponin. Our study suggest that Moringa oleifera can served as a good supplement to our nutrition, due to high content of fibre, protein, fat. However intake of Moringa plant might reduce the risk of many oxidative related problems due to the present of some polyphenols and flavonoid in almost all part of the plant used.     

Breed characteristics associated with <Emphasis Type="Italic">LEP</Emphasis> gene polymorphisms in Holstein cattle

Genotyping of the leptin gene locus (LEP) (SNP: R25C, Y7F, and A80V) has been performed in the animals from two groups in the Holstein cattle herd. Groups 1 and 2 included the fourth to seventh lactation cows and the heifers, respectively. The objective of the surveys was to study the Holstein cattle polymorphisms of the leptin gene associated with the productive longevity according to the related literature data. It has been ascertained that the LEP gene (SNP: R25C, Y7F, and A80V) polymorphism in the Holstein animals determines the certain breed characteristics that can be used for introduction to marker assisted selection. (Particularly, a low (0.04–0.15) frequency of occurrence of the F allele is typical for the LEP-Y7F SNP; as for SNP LEP-A80V, a low frequency of occurrence (0–0.14) is typical for the homozygous animals with the VV genotype when compared to a rather high frequency of occurrence (0.36–0.45) in the heterozygous animals).     

Spread of <Emphasis Type="Italic">Listeria</Emphasis> in fauna of the central region of Russia

The results of studying the spread of Listeria in nature, among environmental objects, wild ungulates, and hydrobionts are given. Both pathogenic and nonpathogenic Listeria species circulate in the territories of the Tver and Kaluga oblasts.     

Detection of indigenous endophytic bacteria in <Emphasis Type="Italic">Eucalyptus urophylla in vitro</Emphasis> conditions

The presence of indigenous endophytic bacteria in aseptically grown seedlings of Eucalyptus urophylla germinated from surface sterilized seeds was investigated using dilution plating, microscopy, and PCR detection. No culturable endophytic bacteria could be detected in suspensions of ground plant tissue incubated on solid or in liquid cultivation media. However, a large number of endophytic bacterial cells, mostly rod-shaped and measured 2–3 μm×0.5–0.8 μm, were observed in in vitro cultured seedlings of E. urophylla using both light and electron microscopy. Using the universal bacterial 16S rDNA primers, a predicted 190-bp fragment was amplified from total DNA isolated from the seedlings of E. urophylla. We concluded that the endophytic bacteria originated from the seed were present in seedlings of E. urophylla. However, the bacterial cells observed appeared to be nonculturable.     

Genetic diversity of genus <Emphasis Type="Italic">Malus</Emphasis> Mill. for scab resistance genes

The results of molecular genetic analysis of wild species and varieties of genus Malus Mill. for loci of monogenic scab resistance are presented. Dominant allele of gene Rvi6 was identified in varieties of M. robusta 43199, M. hupehensis, and M. floribunda. Locus Rvi4 was detected in genotypes of M. robusta v. persicifolia, M. asiatica 2343, M. sieversii 13280, M. niedzwetzkyana 13279, M. purpurea 2392, M. orientalis 41623, M. orientalis 29476, M. orientalis 29 460, M. spectabilis v. albi plena, and M. spectabilis v. rubra plena. Gene Rvi5 in the test collection of wild species is absent.