Tinatin DOOLOTKELDİEVA

Chapter of Books | 2018 | Agricultural Research Updates22 ( Chapter 2 )

Erwinia amylovora as a fire blight pathogen and Venturia inaequalis as a scab pathogen were isolated from the blossoms, exudates, infected fruits, leaves and bent branches of diseased apple, pear and hawthorn trees, selected in the Chy, Osh and Jalal Abad regions. Biochemical and pathogenicity tests, alongside PCR analyses were conducted to identify the local isolates of Erwinia amylovora and Venturia inaequalis. The alternative antagonistic microorganisms which combat bacterium E. amylovora and fungus Venturia inaequalis were tested within in vitro and in vivo conditions. The results revealed the ability of Streptomyces antagonisti . . .c bacteria to decrease fire blight severity on pear and apple trees during the first stage of the fire blight disease in leaf tissues. Streptomyces strain C1-4 suppressed E. amylovora disease symptoms in the leaf tissues and excised apple and pear shoots. The incidence of fire blight on leaves was reduced by about 70 with two applications of bacterial antagonists. Streptomyces sp strains were tested for apple scab control in vitro and field conditions. Two applications of Streptomyces strain C1-4 within 35 days completely stopped the scab disease in seedling leaves. Within 40 days, the seedlings were recovered; the new leaves have blossomed on branches. Seedlings grew new shoots and leaves around 50 days after the second treatment. Orchard experiment results provide great hope that a biological product based on Streptomyces could work as an effective agent to suppress the development of the pathogen in the early spring, when leaves start to show scab disease symptoms. Further studies at different locations in Kyrgyzstan, using large scale application, would allow for stronger recommendations to be made, including studies and recommendations on their ability to prevent diseases and to use them as main components in an integrated pest management program More less

Tinatin DOOLOTKELDİEVA

Chapter of Books | 2012 | Diversity of biodegradative gene populations in aquatic sediments examined by gene-targeted metagenomics ( Chapter 3 )

Previous studies in our lab and elsewhere showed that different microbial gene groups are obtained from geographically distant sites. Alkane monooxygenases activate terminal carbon-carbon bonds in the first step of alkane degradation with the addition of an oxygen to the compound, resulting in an alcohol product. The aim of this study was to test the hypothesis that unique microbial populations and alkane monooxygenase (alkB and Cytochrome P450) genes can be recovered from geographically distinct sites, with varying degrees of anthropogenic impact. Sediment was collected from six river and stream sites with various levels of human i . . .mpact located in Bishkek, Kyrgyzstan as well as Dushanbe and Khorog, Tajikistan, in Central Asia. The sites included rural and mountain streams, as well as rivers near or in cities. All sediments were enriched in media containing hexadecane as a carbon source and subcultured into fresh media with the alkane two times. DNA was extracted from each enrichment culture at various points throughout the enrichment for 16S rRNA and alkB clone library construction. The enrichment cultures were also analyzed by terminal restriction fragment length polymorphism (TRFLP) analysis of both 16S rRNA and alkB genes. The alkB and P450 genes, as well as 16S rRNA, were targeted with 454 pyrosequencing to obtain an estimate of microbial and alkane hydroxylase diversity at the sample locations and enrichments. The data show that distinct populations of alkB and P450 genes can be enriched from different locations and countries in Central Asia confirming that there is a wide diversity of alkane degrading genes in the environment More less

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

Article | 2009 | Microbiology Insights ( 2 )

During this work we detected 5 species of pathogenic fungi that damage seedlings and saplings of the Tiang-Shang spruce fir, which cause a perceptible damage to the coniferous nurseries of Northern Kyrgyzstan and require activities to protect the woods. The biological activity of Streptomyces metabolites for raising the resistance of seedlings to phytopathogenic fungi was tested in laboratory and field conditions during three seasons. In order to evaluate the biological activity of Streptomyces preparations as biological agents, we have soaked the seeds in the suspension of Streptomyces metabolites with the concentrations of 50, 100 . . ., 500 mg per liter. In other variant, the seeds were at first processed in the culture of pathogenic fungi as infectious drowning of sprouts (Fusarium, Alternaria), crown rot of sprouts (Sclerotinia graminearum Elen.) and grey dew of needles (Hypodermella sulsigena Tub.), then they were treated in the suspension of Streptomyces. Also we have used a spraying treatment with the suspension of Streptomyces metabolites of the one-year-old seedlings of Tiang-Shang spruce fir already staggered and dying from infectious drowning. The best results against the Alternaria culture were demonstrated by the use of preparation S. griseogromogenes 2ч-8 at the concentration of 500 mgs per liter, which provided the safety of sprouts to 80.2%. Preparations of S. rubrogriseus ТК2-5, S. wistariopsis СП3-13 and S. griseogromogenes 2ч-8 were effective in protection of sprouts from Fusarium sp. cultures. They provided safety of sprouts to 80% on average 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 | Saykal BOBUŞOVA

Article | 2017 | Advances in Microbiology7 ( 6 )

Among the most economically important fruit crops in South and North Kyrgyzstan affected by scab disease are apples (Malus domestica). Natural isolates of Venturia inaequalis were isolated from diseased parts of apple trees, selected from different regions, through sedimentation of air microflora on the crowns of fruit trees. Two biocontrol agents as Trichoderma viride and Streptomyces sр. were tested for apple scab control in vitro and field conditions. Two applications of Trichoderma viride within 35 days completely stopped the scab disease in seedling leaves. Within 40 days, the seedlings were recovered; the new leaves have bloss . . .omed on branches. The Streptomyces sp. antagonists were inferior in effectiveness compared to Trichoderma viride; nevertheless, they prevented progression of the disease in the leaves. Seedlings grew new shoots and leaves around 50 days after the second treatment. Orchard experiment results provide great hope that a biological product based on Trichoderma viride could work as an effective agent to suppress the development of the pathogen in the early spring, when leaves start to show scab disease symptom More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA

Article | 2014 | Global Advanced Research Journal of Microbiology3 ( 10 )

A total of 255 species of wild medicinal plants were analyzed in Northern and Northeastern Kyrgyzstan; from 150 plant species, 278 endophytic fungal isolates were identified. Endophytic fungi were frequently recorded on members of large plant families: Asteraceae, Ranunculaceae, Rutaceae, Fabaceae,Lamiaceae, Boraginaceae, Moraceae, Iridaceae, Euphorbiaceae and Brassicaceae. In the foothill ecosystems, rich biodiversity and numerous species of endophytic fungi genera - Fusarium, Acremonium, Cladosporium, Penicillium, Curvularia, Monilia, Rhizoctonia, Papularia, Botrytis and Stemphylium - were registered. The comparison of the Shannon . . . diversity index (HS) for the endophytic fungi community showed a higher diversity of light-colored species (HS = 3.47) in foothill ecosystems than in alpine systems (HS = 2.75), and a lower diversity in the middle mountains (HS = 2.46). Endophytes were identified from all organs of plants. The most favorite habitats for fungi in plant body were root tissues, followed by the stems and leaves, while the flower clusters and seeds were rarely colonized by endophytes. - Keywords: wild medicinal plants, biodiversity of endophytic fungi, distribution on vertical zonation, tissues colonize More less

Tinatin DOOLOTKELDİEVA | Saykal BOBUŞOVA

Article | 2016 | Advances in Microbiology6 ( 2 )

Ten isolates of Erwinia carotovora ssp. carotovora (Ecc) were isolated from infected potato tubers of Picasso, Sante, and Nevskiy varieties collected from different regions in Kyrgyzstan. Isolates were identified as Erwinia carotovora ssp. carotovora (Ecc) by standard bacteriological techniques and pathogenicity tests on tubers and also by PCR analyses. Tests on the pathogenicity of E. carotovora ssp. carotovora (Ecc) strains to host plants by artificial inoculation have shown a high sensibility of the Picasso variety. As a result, five isolates were chosen, three isolates (EcPo1, EcPo2, and Eco3) were highly pathogenic, while two i . . .solates (Eco4 and Eco5) were weakly pathogenic. The antagonistic bacteria, Streptomyces diastatochromogenes strain sk-6, and Streptomyces graminearuss strain sk-2, have a highly significant effect on soft rot bacteria isolates (Ecc), more than the other tested antagonistic organisms in vitro screening biotests. The Streptomyces diastatochromogenes sk-6 was selected for the control assay of storage potatoes against the most common soft rot bacterial strain in Kyrgyzstan, Erwinia carotovora sp. carotovora EcPo2. The pretreatment of potato tubers with antagonistic bacteria successfully prevented the initial infection multiplication of soft rot bacteria and reduced soft rot disease of potatoes in storage. These results justify selection of the dose 106 cells/ml of bacteria Streptomyces diastatochromogenes sk-6 for use in powdering the infected or non-infected potato tubers to suppress the development soft rot during storage. Streptomyces diastatochromogenes sk-6 as a biological disinfectant could destroy surface and internal infections, protect the tubers from the growth of phytopathogenic bacteria in the early period of their reproduction, and improve the overwintering of winter crops. - KEYWORDS: Potato Tubers, Soft Rot Disease, Erwinia carotovora ssp. carotovora, Biological Control, Streptomyces diastatochromogenes 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

Article | 2016 | Advances in Microbiology6 ( 11 )

Erwinia amylovora species were isolated from the blossoms, exudates, infected fruits, leaves and bent branches of diseased apple, pear and hawthorn trees, selected in the Chy, Osh and Jalal Abad regions. Biochemical and pathogenicity tests, alongside PCR analyses, were conducted to identify the local isolates of Erwinia amylovora. The alternative antagonistic microorganisms which combat bacterium E. amylovora were tested within in vitro and in vivo conditions. The results revealed the ability of Streptomyces antagonistic bacteria to decrease fire blight severity on pear and apple trees during the first stage of the fire blight dise . . .ase in leaf tissues. Streptomyces strain C1-4 suppressed E. amylovora disease symptoms in the leaf tissues and excised apple and pear shoots. The incidence of fire blight on leaves was reduced by about 70% with two applications of bacterial antagonists. Further studies at different locations in Kyrgyzstan, using large scale application, would allow for stronger recommendations to be made, including studies and recommendations on their ability to prevent disease and to use them as main components in an integrated pest management program.- Keyword: Biological Control Agents, Fire Blight, Erwinia amylovora, Antagonistic Bacteri More less

Tinatin DOOLOTKELDİEVA

Chapter of Books | 2013 | Multifunctional Agriculture, Ecology and Food Security: International Perspectives ( Chapter 14 )

The main agricultural activities in the Kyrgyz Republic are include the production of cereal grains, vegetables, potatoes and commercial crops such as cotton, tobacco, sugar beets and livestock. The sustainability of the agriculture sector was severely affected by the large scale mono-cropping approach of the Soviet era. Intensive agriculture has caused environmental pollution due to the excessive use of mineral fertilizers and pesticides. To combat this menace we have isolated and introduced non-chemical crop protection agents, the antifungal micro-organisms Streptomyces spp., producing antibiotics and bio-stimulating compounds. Ma . . .ny isolates were screened for their ability to suppress soil-borne plant pathogenic fungi and to also promote growth of plants. Strains that are able to survive under field conditions were selected. The 16 S RNA genes of bio-control related strains were studied by PCR amplification. Strains of Streptomyces fumanus, gn-2were obtained by submerged cultivation in a bioreactor. Streptomyces fumanus, gn-2 was evaluated under field conditions against fungal pathogens and also for the growth-promoting effect on wheat, beans, barley, potatoes, tomatoes, peppers, strawberries and other crops. The pre-seed treatment provided 90 percent more safety to seedlings than the control species and the resistance of crops to fungi pathogens-Alternaria, Aspergillus, Acremonium, Botrytis, Fusarium and Penicillium species. © 2013 Nova Science Publishers, Inc. All rights reserved More less