Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA | Mahabat KONURBAYEVA

Article | 2023 | Applied and Environmental Soil Science2023

Soil bacterial and fungal communities were investigated in relation to soil type and farm management practices after vegetation harvesting in autumn. Soils from fields cultivated with Phaseolus vulgaris (bean) and Pyrus comminus (pear) and nonarable, natural areas were studied. Microbial diversity was analysed using cultivation-dependent methods (isolation of pure cultures) and cultivation-independent methods (direct extraction of DNA from soil, followed by PCR amplification of the 16S rRNA and 18S rRNA genes). The use of cultivation-dependent methods revealed that there were no differences in the biodiversity of the soil bacterial . . .and fungal communities between fields cultivated with bean plants and pear trees. However, the use of cultivation-independent methods showed that there were clear soil and crop type-specific effects on the composition of the soil bacterial and fungal communities. The density of the bacterial population was two times higher in northern mountain-valley serozem (NMVS) soil samples than in light chestnut (LC) soil samples. In contrast, the densities of the fungal communities were almost equal in the studied soil types. The density of the actinomycetes community was almost two times higher in LC soil than in NMVS soil under bean plants. The Shannon index values showed that the bacterial biodiversity in the NMVS soil samples was greater than that in the LC soil samples. Soils under fallow appeared to have diverse bacterial communities that mainly consisted of local autochthonous microflora and a small amount of zymogenic microflora (since fresh plant residue does not enter the soil). The Shannon index results revealed two interesting facts: (1) the soil bacterial community was highly diverse in soils that supported bean plants and (2) the soil fungal biodiversity was high under pear trees in both soil types. Keyword: gradient gel-electrophoresis; bacterial communities; land-use; cropping systems; plant diversity; carbon; rhizosphere; management; proteobacteria; chrysanthemu More less

Saykal BOBUŞOVA | Tinatin DOOLOTKELDİEVA

Article | 2023 | Journal of Fungi9 ( 10 )

Background: The entomopathogenic fungus Metarhizium anisopliae sensu lato forms a species complex, comprising a tight cluster made up of four species, namely M. anisopliae sensu stricto, M. pinghaense, M. robertsii and M. brunneum. Unambiguous species delineation within this “PARB clade” that enables both the taxonomic assignment of new isolates and the identification of potentially new species is highly solicited. (2) Methods: Species-discriminating primer pairs targeting the ribosomal intergenic spacer (rIGS) sequence were designed and a diagnostic PCR protocol established. A partial rIGS sequence, referred to as rIGS-ID800, was i . . .ntroduced as a molecular taxonomic marker for PARB species delineation. (3) Results: PARB species from a validation strain set not implied in primer design were clearly discriminated using the diagnostic PCR protocol developed. Using rIGS-ID800 as a single sequence taxonomic marker gave rise to a higher resolution and statistically better supported delineation of PARB clade species. (4) Conclusions: Reliable species discrimination within the Metarhizium PARB clade is possible through both sequencing-independent diagnostic PCR and sequencing-dependent single marker comparison, both based on the rIGS marker. Keyword: diagnostic PCR; metarhizium anisopliae; metarhizium brunneum; metarhizium robertsii; metarhizum pinghaense; PARB clade; ribosomal intergenic spacer (rIGS); single-marker phylogeny; species delineatio More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA

Article | 2021 | Journal of Plant Pathology103 ( SUPPL 1 )

Erwinia amylovora, the causal agent of fire blight, was first reported in Kyrgyz orchards in 2008 and, within a few years, the disease has spread throughout most of the northern part of the country. As fire blight expanded and penetrated new areas, it also reached the unique ecosystem constituted by the Arslonbob wild walnut-fruit forests in South Kyrgyzstan. Here, we present the results of field surveys in the forests and in the nearby orchards and gardens of the Jalal-Abad region during the 2019 season. Further, we use genetic profiling of the CRISPR regions to compare the genotypes of the obtained isolates to those of previous ou . . .tbreaks in the north of the country and in nearby Kazakhstan. While fire blight was found in twelve of the twenty-six sites investigated in and around the Arslonbob walnut-fruit forest, it was mostly located on domesticated varieties of quince, apple or pear in gardens, orchards or nurseries. Plants in the forest and especially endangered wild species were not affected by the disease, but their proximity to the current infection herds is cause of major concern and calls for immediate protective action. Genetic analysis suggests that the main introduction route for fire blight to Central Asia was import of contaminated plant material from already affected countries in the Eurasian continent, which probably occurred on multiple independent instances. Nonetheless, indications of natural spread of the disease were identified between the regions of South Kazakhstan and North Kyrgyzstan More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA

Article | 2022 | Journal of Nanomaterials2022

The antibacterial activities of silver (Ag) and copper (Cu) monometallic and bimetallic nanoparticles (MNPs and BNPs) against the most economically important and common plant bacterial diseases in Kyrgyzstan were tested, as well as their phytotoxic effects on wheat and corn seeds. To achieve this, a range of Ag and Cu MNPs and BNPs were synthesized using different methods and were tested at different concentrations to compare their biological effects. The antibacterial activities of Ag and Cu MNPs and Ag-Cu BNPs against the plant pathogenic bacteria Erwinia amylovora, Pseudomonas syringae, and Pectobacterium carotovorum were evaluat . . .ed. Of the three pathogens, Ps. syringae was found to be the most sensitive to NPs. The antimicrobial activities of the tested NPs showed significant positive correlations with the concentration; hence, there was a dose-dependent effect. Due to their antibacterial properties, these Ag-Cu nanocomposites have high potential in plant protection applications More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA

Article | 2014 | MANAS Journal of Agriculture, Veterinary and Life Science (MJAVL)4 ( 1 )

For the first time in Kyrgyzstan Raltsonia solanacearum bacterium as a pathogen of bacterial wilt (quarantine for the country object) was obtained and identified by enzyme-linked immunosorbent assay (ELISA) and biochemical methods. Three potato (Solanum tuberosum) cultivars: Picasso, Sante and Nevskiy were used for isolation of pathogen, which were collected from different regions of Kyrgyzstan. Detection and identification of the pathogen by ELISA performed directly from diseased potato shoots and leaves, and from pure culture of Ralstonia solanacearum isolated from tubers of potato seed during storage. For ELISA was used Ralstonia . . . solanacerium PathoScreen R Kit DAS ELISA (Agdia product, USA).Isolated races of Ralstonia solanacearum by biochemical characteristics were classified as a 3-biotype. - Keywords: The Potato (Solanum tuberosum) Cultivars, Identification of Ralstonia Solanacearum , ELISA assay, Biochemical Tests More less

Emil ÖMÜRZAK | Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA

Proceedings Paper | 2018 | 2017 International Conference on Aerospace Technology, Communications and Energy Systems (ATCES 2017)302 ( UNSP 012075 )

By means of XRD and FESEM analysis, it is established that copper nanoparticles with sizes less than 10 nm are formed during the chemical reduction, which form aggregates mainly with spherical shape. Presence of gelatin during the chemical reduction of copper induced formation of smaller size distribution nanoparticles than that of nanoparticles synthesized without gelatin and it can be related to formation of protective layer. Synthesized Cu nano-powders have sufficiently high activity against the Erwinia amylovora bacterium, and the bacterial growth inhibition depends on the Cu nanoparticles concentration. At a concentration of 5 . . .mg / ml of Cu nanoparticles, the exciter growth inhibition zone reaches a maximum value within 72 hours and the lysis zone is 20 mm, and at a concentration of 1 mg / ml this value is 16 mm, which also indicates the significant antibacterial activity of this sample More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA | Ayperi MURATBEKOVA

Article | 2021 | Acta Parasitologica66 ( 4 )

Purpose Predatory fungi have been the subject of fundamental studies and their potential as biological control agents against parasitic plant nematodes has been assessed. The aim of the present study was to isolate and identify predatory fungi, performing in vitro and in vivo screening to select highly active strains to control parasitic nematodes. Methods Different nutrient media were used to isolate predatory fungi and determine their morphological and cultural properties. Identification was performed by classical and molecular biology methods. In vitro and in vivo screening was conducted to select highly active strains. Results T . . .welve isolates of Arthrobotrys oligospora (Orbiliomycetes) found in nature were investigated for their predaceous efficacy against garlic stem nematodes (Ditylenchus dipsaci). The effect of temperature and pH on the growth rate and trap formation of representative isolates was determined and isolates were characterized by light microscopy and molecular markers. BLAST was used to sequence the rDNA internal transcribed spacer of A. oligospora isolate KTMU-7. The optimum growth of A. oligospora strains was achieved at 20-25 degrees C on 1-2?orn meal agar (CMA) within the pH range of 5.6-8.6. The factors responsible for the trap formation of these fungal strains were identified. In vitro and in vivo experiments were performed to evaluate the nematicidal activity of local predatory fungal isolates against soil nematodes. Conclusions Preliminary studies proved A. oligospora to be a potentially effective biological control agent, immobilizing 85.7 /- 2.19?f garlic stem nematodes in soil from the rhizosphere of potato plants More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA

Article | 2012 | Фундаментальные исследования ( 9; Часть 2 )

С помощью классического метода микробиологии и анализа последовательностей 16SrRNA впервые было изучено количественное соотношение и биоразнообразие микробных комплексов низкотемпературных почв долины Сон-Куль. Структура видового состава бактерий отличалась в почвах под различным покровам растительности по мере возрастания высоты местности. Анализ последовательностей 16SrRNA показал, что в летний период в почвах высокогорной долины Сон-Куль доминирующими филогенетическими группами были Actinobacteria (55,0%), Gammaproteobacteria ( 33,0%) и Firmicutes phyla (22,0%). Видовой состав обнаруженных функциональных групп бактерий дает основ . . .ание полагать, что в почвах Сон-Кульской долины из-за постоянных низких температур в процесс минерализации органических растительных остатков вовлечен приемущественно автохтонная микрофлора и зимогенная микрофлора, интенсивно разлагающая остатки за крайне короткий летний период. Ключевые слова: экология микроорганизмов; биоразнообразие почвенных микроорганизмов; структура микробных сообществ; высокогорные и низкотемпературные экосистемы. The quantitative correlation and the soil microbial communities’ biodiversity of low-temperature soils of Son-Kul valley have been first studied by the classical microbiology method and the 16SrRNA sequence analysis. The structure of the bacterial species composition has been differed in soils under different vegetation with increasing altitude. The 16SrRNA sequence analysis had been showed, that the Actinobacteria (e.g.55,0 %), the Gammaproteobacteria (e.g. 33,0 %), and the Firmicutes phyla (e.g. 22,0 %) were the dominant phylogenetic groups during the summer in the alpine soils of Son-Kul valley.Of the 10 studied sites in 5 sites the representatives of Actinobacteria phyla have been dominated. In other sites, under the meadow vegetation the representatives of Gammaproteobacteria phyla have been dominated. The species composition of the detected bacterial functional groups has been suggested to be assumed, that under constantly low temperatures conditions of Son-Kul Valley an autochthonous microflora is predominantly involved in the mineralization process of organic crop residues. And the zymogenic microflora is rapidly decomposed the organic crop residues for the extremely short summer period. Keywords: ecology of microorganisms; soil microbial biodiversity; the structure of microbial communities; high-and low-temperature ecosystem More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA | Mahabat KONURBAYEVA

Article | 2021 | Air, Soil and Water Research14

Currently, in the territory of Kyrgyzstan, 50 storage facilities of obsolete pesticides exist; they store about 5000 tons of these hazardous chemicals. The storage conditions have become unusable for a long time. They pose a serious threat to the people living there, livestock, and the environment. The main purpose of this research was the use of selected bacteria with cytochrome P450 genes for the bioremediation of polluted soils around the burial sites in model soil experiments. In the first trial of biodegradation experiments, one contaminated soil was used without any changes in chemical contents, and in the second, the physical . . . and chemical contents of the soil were improved to maintain the bioremediation conditions. The soils in both variants were treated 3 times (ie, once a month) with suspensions of a single culture or a blend of active bacteria (1 x 10(8) cells/mL) selected from in vitro biodegradation experiments. Two control units without the addition of the bacteria culture were also run. The quantification of targeted persistent organic pollutants (POPs) before and after biodegradation was performed by capillary gas chromatography (GC) coupled to a mass spectrometer. In 6 months, obsolete pesticides such as dieldrin, alpha-endosulfan, beta-endosulfan, and 4-heptachlor-epox pure were able to degrade almost completely, up to 98?o 99.0?by the blend of bacteria and the single culture of bacteria. Endrin aldehyde showed more resistance as the blend of bacteria was able to degrade it to 59.77?To improve the aerobic degradation for elimination of pesticides from contaminated soils, it is necessary to create optimal agrotechnical and agrochemical conditions More less

Saykal BOBUŞOVA | Tinatin DOOLOTKELDİEVA

Article | 2023 | Journal of Plant Pathology ( Published online: 30 May 2023 )

Fire blight, caused by the bacterial pathogen Erwinia amylovora, is a severe bacterial disease of apple and pear that can quickly destroy whole plants. In the last decade, it was also detected in Central Asia, where wild pomaceous fruit plants represent the dominant species in mid-altitude forests and constitute a critical foundation for the entire ecosystem. Efficiently informing farmers, forestry services and private persons about the instances and dangers of fire blight, the correct way to recognize the symptoms, and the methods of disease control is thus of paramount importance in a vast and fragmented natural landscape like the . . . one characterizing countries like Kyrgyzstan, Kazakhstan and Tajikistan. For that purpose, we have developed an app for smartphones and mobile devices that can inform stakeholders about fire blight, simultaneously allowing a citizen science approach for mapping the spread of the disease in Central Asia. The app is available in the three national languages as well as in Russian, English, and German, and can easily be adapted to new countries, languages or even diseases. Keywords: malus; pyrus; citizen science; mobile device; androi More less

Tinatin DOOLOTKELDİEVA | Mahabat KONURBAYEVA | Saykal BOBUŞOVA

Article | 2018 | Environmental Science and Pollution Research25 ( 32 )

In Kyrgyzstan, many former storehouses and dump sites for obsolete pesticides exist. In 2009/2010, an inventory and assessment of these sites including risks of environmental hazard has been conducted by FAO and the World Bank. Monitoring revealed high concentration of pesticides listed as persistent organic pollutants (POPs). The purpose of this research was to study the microbial structural complexes of the pesticide-contaminated soils in these dumping zones, and to search for and select microorganism’s destructors with cytochrome P450 genes for pesticide degradation. Culture-dependent and culture-independent approaches were used . . .to determine the taxonomic composition of these bacterial communities. The universal primer set for the 16S ribosomal RNA (rRNA) gene and the specific primer set P450R were used to amplify the cytochrome P450 hydroxylase gene. In soils from Suzak A and B and soils from Balykchy dumping sites, the bacteria from the Actinobacteria phylum (Micrococcus genus) were dominant. These bacteria made up 32–47?f the indigenous local microflora; bacteria species from the Pseudomonas genus (Gammaproteobacteria phylum) made up 23?n Suzak, 12?n Balykchy soils. Bacillus species from the Firmicutes phylum were found only in Suzak soils. The 16S rRNA analyses and the specific primer set P450R have revealed bacteria with cytochrome genes which are directly involved in the degradation process of organic carbon compounds. Experiments were carried out to help select active degraders from the bacterial populations isolated and used to degrade Aldrin in laboratory. Active bacterial strains from the Pseudomonas fluorescens and Bacillus polymyxa population were selected which demonstrated high rates of degradation activity on Aldri More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA | Sezim COLDOŞBEKOVA

Article | 2019 | Journal of Invertebrate Pathology167

We report on the enzootic foci of the insect pathogenic fungus, Beauveria bassiana, found in high meadows in the middle mountain steppes of Kyrgyzstan, at elevations from 1000 m to 2200 m. The growth characteristics of various B. bassiana isolates on different media and as a function of temperature were studied. In addition, the ability of the fungal isolates to produce enzymes with amylase, protease and lipase activities was investigated. Dense biomass production on inexpensive solid media (oatmeal and bean oil meal) produced conidia used for insect bioassays targeting white grub larvae (Phyllophaga fullo, Coleoptera, Scarabaeidae) . . . and nymphal and adult populations of whiteflies (Trialeurodes vaporariorum, Hemiptera, Aleyrodidae). The efficacies of the tested B. bassiana strains for third instar white grub larvae varied, with only two strains showing high entomopathogenic activity. At 25 degrees C, mortality reached 73?or Bay.5-Gal and 74?or Bay.1-Lep at 55 d post-infection, but was lower, 27?nd 29?respectively, at 12 degrees C. These two strains produced significantly higher mortality in adult and whitefly nymphs, with 65-75?ortality 6 d post-infection. Based on morphological characters, including production of ellipsoidal conidia, and molecular characters (ITS, partial 18S (SSU rDNA) and EF1-alpha sequences), the isolates were identified as Beauveria bassiana belonging to Clade E from Asia. Our results add to data on the diversity of ecosystems inhabited by B. bassiana and provide a local resource for pest control efforts More less