AGRIS - 

Data provenance by Country

2020

Belousova, M.E. | Grishechkina, S.D. | Ermolova, V.P. | Antonets, K.S.

One of the key microorganisms used as the basis of biological preparations production is the gram-positive spore-forming bacterium Bacillus thuringiensis (Bt). The goal of the work was to identify the molecular determinants of the insecticidal properties of the industrial strain B. thuringiensis var. darmstadiensis 56 as well as to test its activity in the field. Field trials of the effectiveness of the biological preparation based on BtH10 56 against the Colorado potato beetle was carried out on potatoes (Solanum tuberosum L.) of the Vineta and Rocco varieties in 2018 and 2019 (Voronezh Province) in the area of 1 ha. To evaluate the entomocidal activity, we used a liquid form of the preparation. The application rate of the preparation was 20 l/ha. The counts were carried out in 5, 10 and 14 days after treatment. The biological effectiveness of the preparation was determined by analyzing a decrease in the number of pests according to the Abbott formula. According to the test results, the high efficiency of the developed preparation against the Colorado potato beetle was established. This efficiency varied from 83.8 to 87.8% and did not differ from the chemical standards. Using Illumina and Oxford Nanopore technology, we obtained the whole genome sequence of the ВtH10 56 strain. After assembly and annotation of the genome, a search for toxins was conducted. The CryProcessor and BtToxin_scanner programs were used to search and classify genes encoding the Bt insecticidal toxins. As a result, a gene belonging to the cry1E group, cry1Ea7, was found. It was found that the genome of the strain does not contain genes encoding Vip, Sip and Cyt. toxins, however, it harbors several genes encoding synthetases of non-ribosomally synthesized peptides (nrp) that may explain its multifunctional properties. Thus, considering the data obtained the liquid form of the biological preparation based on BtH10 56, can be recommended for use in the industry and organic farming./Один из ключевых микроорганизмов, используемых в качестве основы для создания биологических препаратов, - грамположительная спорообразующая бактерия Bacillus thuringiensis (Bt). Цель работы - выявление молекулярных детерминант инсектицидных свойств штамма-продуцента B. thuringiensis var. darmstadiensis 56 (BtH10 56), а также испытание его активности в полевых условиях. Полевые испытания эффективности указанного биопрепарата против колорадского жука проводили на картофеле (Solanum tuberosum L.) сортов Винета и Рокко в 2018 и 2019 гг. в Воронежской обл. на площади 1 га. Для оценки энтомоцидной активности использовали жидкую форму препарата в норме- 20 л/га. В качестве химического эталона использовали инсектицидные препараты Цепеллин, КЭ и Колорадо, ВРК в дозах соответственно 100 г/л и 0,1 л/га. Учеты проводили перед и после обработки (на 5-е, 10-е и 14-е сут). По результатам испытаний была установлена высокая эффективность разработанного препарата против колорадского жука, которая на 10-е сут составляла 83,8-87,8% и не отличалась от химического эталона. С использованием технологии Illumina и мономолекулярного нанопорового секвенирования была получена полная последовательность генома штамма-продуцента ВtH10 56. Для поиска и классификации генов инсектицидных токсинов Вt использовали программы CryProcessor и BtToxin_scanner, в результате чего был идентифицирован ген cry1Ea7, относящийся к группе сry1E, для которой описана активность в отношении Lepidoptera. Исследованный штамм не содержал генов, кодирующих токсины групп Vip, Sip и Cyt, однако нес ряд генов, кодирующих синтетазы нерибосомно синтезируемых пептидов (nrp), что объясняет его полифункциональные свойства. Полученные данные позволяют рекомендовать биологический препарат на основе BtH10 56 для использования в промышленности и органическом земледелии.

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Central Scientific Agricultural Library - Russian Federation

2020

Kalmykova, G.V. | Cheshkova, A.F. | Akulova, N.I., Siberia Federal Scientific Center of Agrobiotechnology, Novosibirsk Region

The development of the nutriculture medium (NM) composition and the optimal concentrations of its constituent components to increase the bacteriocin like activity of the strain Bacillus thuringiensis ssp. Dakota are presented. The study was carried out using a multifactor experiment with further processing of statistical data in order to optimize the basic NM and maximize the activity of the target product. Sources of nitrogen (peptone and yeast extract) and carbon (glycerin and glucose) were optimization factors. The control growth medium was medium "A" used for the cultivation of bacteria of the g. Bacillus. The degree of impact of the studied factors on the effective feature was determined using the model of multiple linear regression of the 1 order, whereby the optimal ratio of the components was calculated on the basis of the quadratic model. The ability of the B. thuringiensis ssp. dakota strain to produce a bacteriocin-like substance (BLIS) was revealed. The dependence of BLIS synthesis on the cultivation medium was established: on the carbohydrate-free medium, the antimicrobial activity of BLIS was 1.5 times lower than that on the medium containing glucose and glycerin. To obtain maximum BLIS activity, the carbohydrate medium was optimized using a multifactor experiment performed by the method of orthogonal Latin rectangles. Mathematical models of linear regression of the 1 and 2 orders were constructed depending on the concentration of nutrient medium components. The optimal concentration of the components was determined on the basis of a second-order regression model that takes into account the effects of the interaction of factors and the nonlinearity of the process. As a result of optimization of the NM, the optimum number of the components of the culture medium was determined: peptone – 9 g/l; yeast extract – 2.6; glycerol – 5.6 g/l. The antimicrobial activity of BLIS on this medium was higher by 60% than on the initial medium. The synthesis of BLIS, unlike the synthesis of delta-endotoxin, is not regulated by catabolite repression of carbon. | Summary (Ru, En) | 5 tables | 18 ref. | Представлена разработка состава питательной среды (ПС) и оптимальные концентрации входящих в ее состав компонентов для повышения бактерициноподобной активности штамма Bacillus thuringiensis ssp. dakota. Для оптимизации базовой ПС с целью максимального повышения активности целевого продукта проведены исследования при помощи многофакторного эксперимента с дальнейшей статистической обработкой данных. Факторами оптимизации являлись источники азота (пептон и дрожжевой экстракт) и углерода (глицерин и глюкоза). Контрольной средой выращивания была среда "А", используемая для культивирования бактерий р. Bacillus. Степень влияния изучаемых факторов на результативный признак определяли с помощью модели множественной линейной регрессии 1-го порядка, оптимальные соотношения компонентов рассчитывали на основе квадратичной модели. Выявлена способность штамма B. thuringiensis ssp. dakota продуцировать бактериоциноподобное вещество (BLIS). Установлена зависимость синтеза BLIS от среды культивирования: на безуглеводной среде антимикробная активность BLIS в 1,5 раза ниже активности на среде, содержащей глюкозу и глицерин. Для получения максимальной активности BLIS среду с углеводами оптимизировали с помощью многофакторного эксперимента, выполненного методом ортогональных латинских прямоугольников. Построены математические модели линейной регрессии 1-го и 2-го порядка в зависимости от концентрации компонентов питательных сред. Оптимальную концентрацию компонентов определяли на основе регрессионной модели 2-го порядка, учитывающей эффекты взаимодействия факторов и нелинейность процесса. В результате оптимизации определено оптимальное количество компонентов среды культивирования: пептон – 9 г/л; дрожжевой экстракт – 2,6; глицерин – 5,6 г/л. Антимикробная активность BLIS на этой среде была на 60% выше, чем на исходной среде. Синтез BLIS, в отличие от синтеза дельтаэндотоксина, не регулируется катаболитной регрессией углерода.

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Central Scientific Agricultural Library - Russian Federation

2019

Andreeva, I.V. | Shatalova, E.I. | Kalmykova, G.V. | Akulova, N.I. | Ul'yanova, E.G., Russian Academy of Sciences, Novosibirsk region (Russian Federation). Siberian Federal Scientific Center for Agrobiotechnology

The results of assessing the effect of Bacillus thuringiensis ssp. aizawai strain on five species of lepidopteran pests (Lepidoptera) belonging to different families are presented. The mortality of larvae infected with B. thuringiensis strain varied significantly depending on the species of insect. In four species: the cabbage white butterfly (Pieris brassicae L.), the cabbage moth (Mamestra brassicae L.), the greater wax moth (Galleria melonella L.) and the black-veined white (Aporia crataegi L.), death of individuals was noted on the second day of the experiment. However, mortality rate was significantly different between species and increased over time. The diamondback moth (Plutella xylostella L.) was more susceptible to the pathogen. Significant mortality of the larvae of this phytophage was observed already on the first day after infection; while on the second day of the experiment all individuals died. Lethal time LT90 of more resistant species (black-veined white) and more sensitive ones (diamondback moth) differed sevenfold. Semi-lethal concentrations of B. thuringiensis strain for the tested insect species were determined: on the third day of the experiment, lethal concentration LC50 for the cabbage moth larvae, the greater wax moth and black-veined white ranged from 1.7 to 4.5*10E-8spores/ml. For the cabbage white butterfly, it was much lower (1.8*107 spores/ml). For diamondback moth caterpillars, LC50 was 4.25*10E-7 spores/ml already one day after infection. It was noted that the specificity of B. thuringiensis strain is associated with various mechanisms of insect protection from pathogens, and is due to individual characteristics of the species. In particular, the pH of excrement of intact caterpillars of the cabbage white butterfly, a species sensitive to B. thuringiensis, was 2 times higher than that of caterpillars of the greater wax moth – 8.9 and 4.3, respectively, which is one of the reasons for the susceptibility (or resistance) of the species to bacterial entomopathogen. | Представлены результаты оценки специфичности действия штамма Bacillus thuringiensis ssp. aizawai по отношению к пяти видам чешуекрылых вредителей (Lepidoptera), относящихся к разным семействам. Смертность личинок, зараженных штаммом B. thuringiensis, значительно варьировала в зависимости от вида насекомого. У четырех видов – капустной белянки (Pieris brassicae L.), капустной совки (Mamestra brassicae L.), большой вощинной огневки (Galleria melonella L.) и боярышницы (Aporia crataegi L.) – гибель особей отмечена на вторые сутки опыта. При этом она существенно различалась между видами и увеличивалась со временем. Капустная моль (Plutella xylostella L.) была более восприимчива к патогену. Существенную смертность личинок этого фитофага наблюдали уже через одни сутки после заражения, на вторые сутки эксперимента погибали все особи. ЛТ90 у устойчивого (боярышница) и чувствительного (капустная моль) видов насекомых различалось в 7 раз. Определены полулетальные концентрации изучаемого штамма B. thuringiensis для тестируемых видов насекомых: на третьи сутки опыта ЛК50 для личинок капустной совки, большой вощинной огневки и боярышницы составляла от 1,7 до 4,5*10Е-8 спор/мл. У капустной белянки она была на порядок ниже – 1,8*10Е-7 спор/мл. Для гусениц капустной моли ЛК50 уже через сутки после заражения составляла 4,25*10Е-7 спор/мл. Отмечено, что специфичность действия штамма B. thuringiensis связана с различными механизмами системы защиты насекомых от патогенов и обусловлена индивидуальными особенностями вида. В частности, рН экскрементов интактных гусениц чувствительного к B. thuringiensis вида – капустной белянки – был в 2 раза выше, чем у гусениц вощинной огневки – 8,9 и 4,3 соответственно, что является одной из причин восприимчивости (или устойчивости) вида к бактериальному энтомопатогену.

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Central Scientific Agricultural Library - Russian Federation

2018

Gorobej, I.M. | Kalmykova, G.V. | Davydova, N.V | Andreeva, I.V., Russian Academy of Sciences, Novosibirsk (Russian Federation). Siberian Federal Scientific Center for Agrobiotechnology

The article presents the results of the study of growth-stimulating and antifungal activity of strains Bacillus thuringiensis ssp. fukuokaensis ssp. morrisoni, ssp. toumanoffi, ssp. dakota and ssp. amagiensis. The positive effect of all studied strains on morphometric parameters of spring rape and spring wheat seedlings was revealed. Treatment of spring rape seeds with spore-crystal mixtures of B. thuringiensis contributed to a significant increase in the length of roots by 1.9-3.4 times and sprouts – by 1.3-1.9 times, depending on the strain. Stimulation of plant growth was also observed in the inoculation of spring wheat seeds with suspension strains of B. thuringiensis. However, a significant positive effect was revealed only in the variant with the use of the strain of B. thuringiensisssp. fukuokaensis. There was a downward trend in the number of spring wheat seedlings affected with root rot when the seeds were treated with the strains of B. thuringiensisssp. morrisoni, ssp. dakota, ssp. amagiensis. In the experiments with pure cultures, it was found that all tested strains of B. thuringiensis showed a high degree of antagonistic activity to two types of phytopathogens. The inhibitory activity of the strains of B. thuringiensis on the 10th day of the experiment with regard to Fusarium oxysporum ranged from 68 to 83%, Alternaria alternate – from 71 to 89%. The analysis showed that all studied strains of B. thuringiensis stimulated the growth of spring rape and spring wheat and showed antifungal activity. The obtained data are of interest for further research in order to create multifunctional biological preparations for plant protection. | Представленырезультатыизученияростостимулирующейиантифунгальнойактивностиштаммов Bacillus thuringiensis ssp. fukuokaensis. ssp. morrisoni, ssp. toumanoffi, ssp. dakotaи ssp. amagiensis. Выявлено положительное влияние всех изученных штаммов на морфометрические показатели проростков ярового рапса и яровой пшеницы. Обработка семян ярового рапса спорово-кристаллическими смесями B. thuringiensis способствовала достоверному увеличению длины корней в 1,9–3,4 раза, проростков в 1,3–1,9 раза в зависимости от штамма. Стимуляция роста растений отмечена и при инокуляции семян яровой пшеницы суспензиями штаммовB. thuringiensis, однако достоверное положительное действие выявлено только в варианте с использованием штамма B. thuringiensisssp. fukuokaensis. Выявлена тенденция снижения количества пораженных корневой гнилью проростков яровой пшеницы при обработке семян штаммами B. thuringiensisssp. morrisoni, ssp. dakota, ssp. amagiensis. В экспериментах с использованием чистых культур установлено, что все тестируемые штаммы B. thuringiensisпроявили высокую степень антагонистической активности к двум видам фитопатогенов. Ингибирующая активность штаммов B. thuringiensis на 10-е сутки эксперимента в отношении Fusariumoxysporumварьировала от 68 до 83%, Alternariaalternate– от 71 до 89%. Проведенный анализ показал, что все исследуемые штаммы B. thuringiensis стимулировали рост растений ярового рапса и яровой пшеницы и проявляли антифунгальную активность. Полученные данные представляют интерес для дальнейших исследований с целью создания полифункциональных биологических препаратов для защиты растений.

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Central Scientific Agricultural Library - Russian Federation

2013

АНДРЕЕВА И.С. | МАЗУРКОВА Н.А. | МОКРУШИНА О.С. | ПУЧКОВА Л.И. | ЗАКАБУНИН А.И.

Исследована возможность подавления размножения вируса гриппа человека A/Aichi/2/68 (H3N2) и вируса гриппа птиц A/chicken/Kurgan/05/2005 (H5N1) на культуре ткани MDСK водорастворимыми метаболитами штаммов Bacillus thuringiensis. Проведенные исследования показали, что наряду с отсутствующей или незначительной токсичностью исследуемые опытные образцы препаратов показали высокую активность против вирусов гриппа, что делает перспективным дальнейшую их разработку в целях борьбы с вирусами гриппа человека и птиц. | The possibility of inhibiting the replication of human influenza virus A/Aichi/2/68 (H3N2) and avian influenza virus A/chicken/Kurgan/05/2005 (H5N1) on MDCK tissue culture by water-soluble metabolites of Bacillus thuringiensis strains was studied. The performed investigations showed that along lacking or insignificant toxicity the studied pilot samples of the preparations demonstrated high activity against influenza viruses, which makes promising their further development for control of human and avian influenza viruses.

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Marina Sokolova Publishings - Russian Federation

2013

АНДРЕЕВА И.С. | МАЗУРКОВА Н.А. | МОКРУШИНА О.С. | ПУЧКОВА Л.И. | ЗАКАБУНИН А.И.

Исследована возможность подавления размножения вируса гриппа человека A/Aichi/2/68 (H3N2) и вируса гриппа птиц A/chicken/Kurgan/05/2005 (H5N1) на культуре ткани MDСK водорастворимыми метаболитами штаммов Bacillus thuringiensis. Проведенные исследования показали, что наряду с отсутствующей или незначительной токсичностью исследуемые опытные образцы препаратов показали высокую активность против вирусов гриппа, что делает перспективным дальнейшую их разработку в целях борьбы с вирусами гриппа человека и птиц. | The possibility of inhibiting the replication of human influenza virus A/Aichi/2/68 (H3N2) and avian influenza virus A/chicken/Kurgan/05/2005 (H5N1) on MDCK tissue culture by water-soluble metabolites of Bacillus thuringiensis strains was studied. The performed investigations showed that along lacking or insignificant toxicity the studied pilot samples of the preparations demonstrated high activity against influenza viruses, which makes promising their further development for control of human and avian influenza viruses.

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Marina Sokolova Publishings - Russian Federation

2010

Kirillova, N.E. | Levitin, E.I. | Vojtsekhovskaya, Ya.A. | Zalunin, I.A. | Chestukhina, G.G., State Research and Development Inst. of Genetics and Breeding of Industrial Microorganisms, Moscow

A system for expression allowing to obtain effective production of the recombinant protein Cry9A in Bacillus cell has been suggested. The obtained protein is similar to the native one in immunological properties, sensitivity to proteolysis and biological activity. It was shown that the recombinant Cry9A forms crystal bodies in the sporulating cells of Bacillus thuringiemn Thus, we are the first to obtain the evidence that Cry-proteins that naturally form crystal bodies jointly with other endotoxins are capable of producing crystals on their own. The represented system including vector replicative carriers, expression cassettes holders and protocol of obtaining and cultivation of the strain-producer J allows to easily manipulate of the gene for the delta-endotoxinl Cry9A in gene-engineering experiments

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Central Scientific Agricultural Library - Russian Federation

2010

Smirnov, O.V. | Grishechkina, S.D., All-Russia Research and Development Inst. of Agricultural Microbiology, St. Petersburg

In vitro experiments there was studied an antifungal effect of Bacillus thuringiensis-based biopreparations on phytopathogenic fungi. Bacikol suppressed the growth of Botrytis cinerea colonies even in the minimal concentration of 0.1%, and in the concentration of 2% it completely blocked the growth of pathogen already 3 days after the experiment started. Bitoxybacillin and bactoculicide also exhibited antifungal activity towards B. cinerea but in higher concentrations. Testing different components of liquid bacikol showed its low effectiveness against B. cinerea even in 4% concentration, whereas spore-crystalline complex exhibited antifungal activity already in the concentration of 0.1%. Bacikol had an antifungal effect on other species of phytopathogenic fungi as well, in particular, Bipolaris sorokiniana and Fusarium oxysporum. The estimation of the enzymic activity of bacikol producing strain has shown that it has protease and chitinase activity being inferior to that of the reference strain Serratia marcescens producing chitanase. It is concluded that using Bacillus thuringiensis as a basis for biofungicides is perspective for creating biopreparations with polyfunctional activity

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Central Scientific Agricultural Library - Russian Federation

2009

Ryazanova, A.G. | Eremenko, E.I. | Tsygankova, O.I. | Tsygankova, E.A., Stavropol Research and Development Inst. of Plague Control

Studied were biological properties of atypical strains (AS) Bacillus anthracis and improved were methods for their identification and differentiation from the group of closely related Bacillus cereus by analyzing 1101 strains of microorganisms isolated in 33 regions of the RF and 6 near border countries and identified as B. anthracis. The occurrence of isolating AS of the anthrax agent in natural conditions and the frequency of misidentification of bacillus strain like B. anthracis were evaluated. The phenotypic test was improved to determine the sensibility of strain to penicillin. Variant of multiplex PCR was developed to differentiate B. anthracis strain with any set of plasmids from closely related bacilli strains. Feasibility of multiple loci analysis of 6 chromosomal and 2 plasmid regions of B. anthracis genome having a variable number of tandem repeats (MLVA) for differentiation of anthrax strains from other bacilli of genus Bacillus was demonstrated. MLVA allows at the same time and very exactly to determine genotype, to differ some atypical B. anthracis strains from typical ones and to differentiate any anthrax strains from other bacilli. It is concluded that the isolation occurrence of B. anthracis AS is equal to that of bacilli misidentified as B. anthracis. According to the final identification of 1101 strains, when 969 strains (88.01%) were B. anthracis, and 132 strains (11.99%) did not correspond to the features of the anthrax agent by the basic tests, laboratory diagnosis of anthrax. In order to optimize the processes of identification of B. anthracis typical and atypical strains and differentiation between closely related bacilli, it is rational to use the disk-diffusion method with commercially available penicillin disks, multiprimer PCR and MLVA method on 6 chromosomal and 2 plasmid loci

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Central Scientific Agricultural Library - Russian Federation

2008

Ryabchinskaya, T.A. | Kharchienko, G.L., All-Russia Research and Development Inst. of Plant Protection, Saint-Petersburg

The entomocoenosis of an orchard is able to keep the relative biological balance between its complexes of phytophages and entomophages. The biodiversity of the last ones is wider, polyphages and wide oligophages are dominate among them, therefore the mechanisms of regulation are rather soft. These processes can be activated due to ecologically safe plant protection using. For example, the parasites of apple leaf miner moth keep down the host population even at subthreshold number (10-20 millions per 100 leaves). Some microbiological preparations have shown their effect in plant protection especially against caterpillars. The preparations containing Bacillus thuringiensis have pronounced aftereffect at relatively weak start impact under conditions of high pest number (50-60%) and they can decrease the final number of pests on 95-99%. The method of adaptive immunity induction is popular in plant protection and it is realized by biofungicide and polyfunctional preparation treatments (Immunotsitofit, Akat-25K, Planriz and others). The treatments against apple scab and current American true mildew excitants have been effective in early spring and the acquired immunity has continued for 1.5-2 months. At the ecological system of plant protection the treatments can be hold only at the base of both pest and entomophages monitoring. The chemical treatments are to be let at the pest threshold number higher in 3-4 times and epiphytes danger. The ecological methods are used at less pest number. The use of the methods can decrease treatment costs in 1.5-1.9 times in compare with the chemical ones and it decrease the pesticide load on the phytocoenosis on 20-90%. The quality of the products increases on 15-20%

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Central Scientific Agricultural Library - Russian Federation