Interactions between the Translation Machinery and Microtubules

Microtubules are components of eukaryotic cytoskeleton that are involved in the transport of various components from the nucleus to the cell periphery and back. They also act as a platform for assembly of complex molecular ensembles. Ribonucleoprotein (RNP) complexes, such as ribosomes and mRNPs, are transported over significant distances (e.g. to neuronal processes) along microtubules. The association of RNPs with microtubules and their transport along these structures are essential for compartmentalization of protein biosynthesis in cells. Microtubules greatly facilitate assembly of stress RNP granules formed by accumulation of translation machinery components during cell stress response. Microtubules are necessary for the cytoplasm-to-nucleus transport of proteins, including ribosomal proteins. At the same time, ribosomal proteins and RNA-binding proteins can influence cell mobility and cytoplasm organization by regulating microtubule dynamics. The molecular mechanisms underlying the association between the translation machinery components and microtubules have not been studied systematically; the results of such studies are mostly fragmentary. In this review, we attempt to fill this gap by summarizing and discussing the data on protein and RNA components of the translation machinery that directly interact with microtubules or microtubule motor proteins. © 2018, Pleiades Publishing, Ltd

First report of Septotinia populiperda on potato tubers in Russia

[No abstract available

Interactions between the Translation Machinery and Microtubules

Microtubules are components of eukaryotic cytoskeleton that are involved in the transport of various components from the nucleus to the cell periphery and back. They also act as a platform for assembly of complex molecular ensembles. Ribonucleoprotein (RNP) complexes, such as ribosomes and mRNPs, are transported over significant distances (e.g. to neuronal processes) along microtubules. The association of RNPs with microtubules and their transport along these structures are essential for compartmentalization of protein biosynthesis in cells. Microtubules greatly facilitate assembly of stress RNP granules formed by accumulation of translation machinery components during cell stress response. Microtubules are necessary for the cytoplasm-to-nucleus transport of proteins, including ribosomal proteins. At the same time, ribosomal proteins and RNA-binding proteins can influence cell mobility and cytoplasm organization by regulating microtubule dynamics. The molecular mechanisms underlying the association between the translation machinery components and microtubules have not been studied systematically; the results of such studies are mostly fragmentary. In this review, we attempt to fill this gap by summarizing and discussing the data on protein and RNA components of the translation machinery that directly interact with microtubules or microtubule motor proteins. © 2018, Pleiades Publishing, Ltd

On the interaction of ribosomal protein RPL22e with microtubules

Microtubule (MT) protein preparations often contain components of the translation machinery, including ribosome proteins. To understand the biological meaning of it we studied the interaction of ribosomal protein RPL22e with the MT. We found that bacteria expressed purified RPL22e-GFP-6His did co-sediment with brain tubulin MTs with 1.3 µM dissociation coefficient. Such a KD is comparable to some specific MT-associated proteins. Distinct in vitro interaction of RPL22e-GFP with MTs was also observed by TIRF microscopy. In real-time assay, RPL22e-GFP molecules stayed bound to MTs for several seconds, and 15% of them demonstrated random-walk along MTs with diffusion coefficient 0.03 µ2/s. Deletion of basic areas of RPL22e did not have an impact on KD, and deletion of acidic tail slightly increased association with MTs. Interestingly, the deletion of acidic tail increased diffusion coefficient as well. The interaction of RPL22e with MTs is hardly noticeable in vivo in cultured cells, probably since a significant part of the protein is incorporated into the ribosomes. The mobility of ribosomal protein on the MTs probably prevents its interfering with MT-dependent transport and could ameliorate its transport to the nucleus. © 2019 International Federation for Cell Biolog

NEW FUNGAL POTATO PATHOGENS: RESISTANCE TO FUNGICIDES

New pathogenic to potato fungal strains were found on potato tubers grown in the Kostroma and Moscow regions in 2018. These strains were isolated into axenic cultures and analyzed by cultural, morphological and molecular methods. They were identified as Ilyonectria crassa and Septotinia populiperda . The pathogenicity of these species has been proven by the Koch triad. The slices of potato tubers were inoculated with macroconidia of I. crassa and mycelium of S. populiperda . Mycelium from the slices were isolated into axenic culture. These fungal strains were completely identical to the parent strains. Previously, these species of fungi were not reported as potato pathogens. Resistance of I. crassa and S. populiperda to fungicides fludioxonil (preparation Maxim), azoxystrobin (Quadris), colloidal silver (Zeroxxe) and difenoconazole (Score) were tested. I. crassa was sensitive to Maxim and Quadris; S.populiperda to Maxim and Zeroxxe

Fungal pathogens of tomato in South-Western Russia (Krasnodar Territory)

During a study of fungal diseases of tomato in the South of Russia (Krasnodar Territory) 56 fungal isolates associated with tomato fruits were obtained. Most of them belonged to the species Alternaria alternata. Alternaria solani, Fusarium equiseti, Phomopsis phaseoli, Chaetomium cochliodes, Clonostachys sp., Irpex lacteus, Colletotrichum coccodes were also identified. Laboratory experiments revealed that Clonostachys sp., C. сochliodes, P. phaseoli, I. lacteus, and F. equiseti developed well on the fruit’s slices. Fusarium equiseti was the only species that can penetrate the tomato through epidermis and infect entire fruit. The most effective fungicide against F. equiseti was difenoconazole (EC50 = 0.08 mg/L); pencycuron was also effective (EC50 = 32.5 mg/L). Thiabendazole completely inhibited the growth of F. equiseti at the concentration 100 mg/L (EC50 = 47 mg/L).При изучении грибных болезней томата в Краснодарском крае из пораженных плодов были выделены в чистую культуру 56 штаммов грибов. При анализе видовой принадлежности коллекционных изолятов культурально-морфологическими и молекулярными методами оказалось, что большая их часть принадлежала виду Alternaria alternata. Также были идентифицированы Alternaria solani, Fusarium equiseti, Phomopsis phaseoli, Chaetomium cochliodes, Clonostachys sp., Irpex lacteus, Colletotrichum coccodes. Лабораторные эксперименты по заражению ломтиков плодов томата показали, что Clonostachys sp., C. сochliodes, P. phaseoli, I. lacteus, F. equiseti способны их успешно заражать. F. equiseti оказался единственным видом из исследованных, способным заражать неповрежденные плоды, проникая через эпидермис. Оценка восприимчивости F. equiseti к фунгицидам выявила, что наибольшей эффективностью отличался дифеноконазол (EC50 = 0.08 mg/l). Пенцикурон также показал хорошую эффективность (EC50 32.5 mg/l). Тиабендазол полностью ингибировал рост колонии F. equiseti при концентрации 100 mg/l (EC50 = 47 mg/l)

DISTRIBUTION OF THIABENDAZOLE RESISTANT HELMINTHOSPORIUM SOLANI STRAIN

Helminthosporium solani strains were isolated from potato tubers that were grown in Russia, Germany and the Netherlands. Thiabendazole was effective against the sensitive strains of H. solani (EC50< 7.3 mg/l); however, 8 studied strains from Russia and the Netherlands were found to be extremely resistant to it (EC50 > 1000 mg/l). The sequence of their β-tubulin gene contained a SNP mutation in 198 codon or 200 codon, translating to Gln (CAG) instead of Glu (GAG) or Tyr (TAC) instead of Phe (TTC), respectively. Resistant to thiabendazole strains were found in distant regions of Russia (Moscow, Kostroma, Kaluga district) and in the Netherlands. Resistance to thiabendazole of the Russian, European, and American strains had the same genetic background and was conferred by the same mutations

The occurrence of thiabendazole-resistant isolates of Helminthosporium solani on potato seed tubers in Russia

Silver scurf is a potato tuber disease caused by Helminthosporium solani, which has become an increasing problem in many regions of Russia. The fungicide thiabendazole is widely used to control silver scab. Its target of action is the beta-tubulin gene. Single nucleotide mutations in this gene lead to a multiple increase in the resistance of the strain to the fungicide. The resistance of 37 Helminthosporium solani strains to the fungicide thiabendazole was tested in vitro. Tested strains were isolated from potato tubers grown in seven regions of Russia. In this study, the presence of thiabendazole-resistant strains was shown in populations of the European part of Russia, but not for the Far Eastern part. In the Far Eastern part, potatoes are grown from seeds of local production and there is practically no exchange with the European part of Russia. In the present study, we obtain sequences of the beta-tubulin gene and tested the resistance of strains affecting potatoes in the Far Eastern part of Russia to thiabendazole to show the absence of resistant strains. © 2020, Deutsche Phytomedizinische Gesellschaft

Resistance of Helminthosporium solani strains to selected fungicides applied for tuber treatment

Helminthosporium solani strains were isolated from potato tubers collected in Russia or taken from imported German and Dutch seed tubers. Sequences of the nuclear ribosomal genes and internal transcribed spacers (ITS) for all 24 tested strains were identical and had 100% similarity to the sequences from GenBank identified as Helminthosporium solani. The obtained molecular data confirmed the morphological identification based on the width and length of conidia, the shape of conidiophores and the colony morphology. Screening for resistance to the fungicides Score 250 SC (active ingredient difenoconazole 250 g/l), Quadris (azoxystrobin 250 g/l), Tecto 500 SC (thiabendazole 500g/l), Zeroxxe [colloidal silver particles (3 g/l) stabilized with amphoteric surfactant] was done. Agar blocks with pure cultures of the fungal strains were placed in the centre of Petri dishes containing malt agar amended with fungicide concentrations of 0.1, 1, 10, 100 and 1000 mg/l (accounted for the concentration of the active ingredient). Malt agar free of fungicide was used as the control. Growth inhibition of 50% (EC50) compared to the control was detected based on the dose-response curves. Difenoconazole (EC50 < 0.12mg/l) and colloidal silver (EC50 < 76 mg/l) were the most effective fungicides. No strains resistant to the aforementioned fungicides were found. In most cases, azoxystrobin was effective against H. solani (EC50 < 7 mg/l), but there were several strains with high resistance to this fungicide (EC50 > 100 mg/l). Thiabendazole appears to be effective against the sensitive strains of H. solani (EC50 < 7.3 mg/l); however, six studied strains from Russia and the Netherlands were found to be extremely resistant to it (EC50 > 1000 mg/l). The sequence of their β-tubulin gene contained a SNP mutation in the 198 codon or 200 codon, translating to Gln (CAG) instead of Glu (GAG) or Tyr (TAC) instead of Phe (TTC), respectively. Thus, the resistance to thiabendazole of the Russian, European and American strains had the same genetic background and was conferred by the same mutations. © 2017, Edizioni ETS. All rights reserved

ДЕТЕКЦИЯ COLLETOTRICHUM COCCODES С ПОМОЩЬЮ ПЦР В РЕАЛЬНОМ ВРЕМЕНИ

Colletotrichum coccodes causes dangerous diseases of potato and tomato, known as Anthracnose and Black Spot. Morphologically, they are often difficult to distinguish from diseases caused by other microorganisms. On the green tomato fruits disease may be asymptomatic. It appears only on ripe red fruits. For quick and accurate diagnosis and identification of the pathogen a test system for real-time PCR is proposed. To develop a test system the nucleotide sequence of the glycerol-3-phosphate dehydrogenase gene was determined in 4 different strains of C. coccodes. Based on the results obtained and the analysis of similar sequences of other species available in the GenBank Database, the C. coccodes species-specific primers and probe were designed. To test the specificity of the created test system, PCR was performed with DNA isolated from pure cultures of 15 different types of parasitic and saprotrophic fungi associated with tomato and potato plants (Fusarium oxysporum, F. verticillium, Phomopsis phaseoli, Alternaria alternatа, Helminthosporium solani, Colletotrichum coccodes, Phellinus ferrugineovelutinus, Stemphylium vesicarium, Helminthosporium solani, Phomopsis phaseoli, Neonectria radicicola, Rhizoctonia solani, Penicillium sp., Cladosporium fulvum, C. cladosporioides). The presence of Colletotrichum coccodes DNA was determined at a 20–27 threshold cycle. The remaining types of fungi were determined after 40 cycles or were not detected at all. The test system allows confidently detect in the analyzed PCR-mixture concentrations of C. coccodes exceeding 0.01 ng/mm3. Using the test system created was investigated the presence of C. coccodes in tomato leaves with symptoms of fungal diseases and potato tubers without external symptoms of the disease. Leaves with fungal lesion symptoms were collected from 2 different fields in the Krasnodar region, tubers – from the fields in the Kostroma, Moscow, Kaluga, Nizhny Novgorod regions. A single leaf of tomato containing C. coccodes DNA was found while the presence of C. coccodes DNA tubers grown in the Kostroma, Moscow and Kaluga regions was detected in 5 samples.Фитопатогенный гриб Colletotrichum coccodes вызывает опасные заболевания картофеля и томата, известные как антракноз и черная пятнистость клубней. По морфологическим признакам их часто трудно отличить от заболеваний, вызываемых другими микроорганизмами; на зеленых плодах томата болезнь может протекать бессимптомно, проявляясь только на созревших красных плодах. Для быстрой и точной диагностики возбудителя предлагается тест-система для ПЦР в реальном времени. Для разработки тест-системы была определена последовательность нуклеотидов гена глицеролтрифосфатдегидрогеназы 45 штаммов C. coccodes, выделенных из клубней картофеля в разных регионах России. На основании полученных результатов и анализа аналогичных последовательностей других видов, имеющихся в базе данных GenBank, были сконструированы видоспецифичные для C. coccodes праймеры и зонд. Для проверки специфичности созданной тест-системы проводили ПЦР с ДНК, выделенной из чистых культур 15 различных видов паразитических и сапротрофных грибов, ассоциированных с растениями томата и картофеля (Fusarium oxysporum, F. verticillium, Phomopsis phaseoli, Alternaria alternatа, Helminthosporium solani, Colletotrichum coccodes, Phellinus ferrugineovelutinus, Stemphylium vesicarium, Helminthosporium solani, Phomopsis phaseoli, Neonectria radicicola, Rhizoctonia solani, Penicillium sp., Cladosporium fulvum, C. cladosporioides). Присутствие ДНК Colletotrichum coccodes определялось при пороговом цикле 20–27, тогда как остальные виды определялись после 40 циклов или не детектировались. Тест-система позволяет уверенно детектировать в анализируемой ПЦР-смеси концентрации ДНК C. coccodes, превышающие 0.01 нг/мм3. С помощью разработанной тест-системы было исследовано присутствие C. coccodes в листьях томата с симптомами поражения грибными болезнями и в клубнях картофеля без внешних симптомов заболевания. Листья с симптомами грибного поражения были собраны с двух разных полей в Краснодарском крае, клубни – с полей в Костромской, Московской, Калужской, Нижегородской областях. В Краснодарском крае был обнаружен один лист томата, содержащий ДНК C. coccodes; достоверное присутствие ДНК этого патогена было выявлено в 5 образцах клубней, выращенных в Костромской, Московской, Калужской областях