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Methods of detection of viruses from cereals and grasses using SYBR Green I RT-qPCR
Jarošová, Jana ; Dráb, Tomáš ; Beoni, Eva ; Kumar, Jiban
The family Poaceae is one of the most important groups of plants in the terms of agricultural production. The family Poaceae also hosts many pathogens including >100 known virus species. The distribution of the viruses varies according to hosts, geographical and other conditions. In this methodology we provide assays for detection of nine cereal and grass viruses (Brome mosaic virus-BMV, Barley yellow dwarf virus-BYDV, Lolium latent virus-LoLV, Oat necrotic mottle virus-ONMV, Ryegrass mosaic virus-Rg- MV, Soil-borne cereal mosaic virus-SBCMV, Spartina mottle virus-SpMV, Wheat dwarf virus-WDV a Wheat streak mosaic virus-WSMV) by SYBR Green I RT-qPCR.
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Cryopreservation of sensitive potato genotypes
Faltus, Miloš ; Bilavčík, Alois ; Zámečník, Jiří
This methodology describes the preparation of explants prior to cryopreservation, dehydration of shoot tips, procedure and principles of sample cryopreservation, regeneration of control plants, including evaluation of explant survival and regeneration for sensitive potato genotypes which were formerly very difficult to cryopreserve. Ministry of agriculture of the Czech Republic is the user of this methodology and it applies it in the framework of the "National program for conservation and utilization of plant genetic resources, animals and micro-organisms important for nutrition and agriculture".
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The possibilities to combine biogas production and biowaste hydrothermal carbonization
Usťak, Sergej ; Honzík, Roman ; Muňoz, jakub
The objective of the methodology is to provide to farmers, agricultural consultants, owners and operators of biogas plants, to biowaste producers and processors, to experts in the field of soil reclamation and melioration, to investors and other interested parties on the concerned issues, the basic information about the potential at the combination of biogas production and biowaste hydrothermal carbonization. These options are evaluated on the basis of processing at accumulated energy intensity for the above combined processes and its comparison with conservative biogas production using the example biogas plant with installed electrical capacity of 800 kW. Part of the methodology is guided to evaluate the potential use of thermo- pressure hydrolysis unit as a device for digestate hydrothermal carbonization and other wastes in order to improve their performance, particularly in terms of employment in agriculture as highquality substrates for soil fertilization and for the purpose of soil carbon sequestration.
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The possibilities to use hydrothermally treated biowaste for air and water cleaning
Usťak, Sergej ; Muňoz, Jakub ; Usťaková, Marie
The aim of the methodology is to provide to farmers, agricultural consultants, biogas plants owners and operators, originators and processors of biowaste, experts in the field of reclamation and recultivation, investors and other interested parties on issues concerned with the basic information on the use of hydrothermally treated biowaste, especially hydrothermally treated residues from anaerobic fermentation for water and air cleaning. In the introduction, the methodology provides a basic overview on literature regarding to coal similar materials, methods for their preparation, especially the way of hydrothermal carbonization, its basic characteristics and its possibilities and perspectives of application in practice. Furthermore, the methodology describes the actual results obtained during the research project usable in practice and provides their agrochemical, environmental and economic evaluation in the case of application at soils. At the end summarizes the main potential applications and uses of new products based on hydrothermally modified residues as essential components of the filter medium for water and air cleaning.
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Determination of causal agent of ring rot of potato, Clavibacter michiganensis subsp. sepedonicus bacteria in breeding and multiplicated materials
Pánková, Iveta ; Krejzar, Václav
The methodology includes a method of artificial infection of plants and potato tubers as a prerequisite for the study of migration and localization of the pathogen in tubers and within the plant vascular tissues, time schedule and recommended size of samples of individual parts of plants and potato tubers during the growing season, depending on external conditions and potato genotype, how to deal with samples of the plants and potato tubers prior to treatment, method of processing samples of individual plant parts and tuber depending on the detection method and the optimization of methods for detection of the pathogen Clavibacter michiganensis subsp. sepedonicus. In compliance with the proposed methodological procedure should be interrupted vertical transmission of the pathogen in the process of breeding, the possibility of detection of the pathogen in breeding and propagation material and reduction of suspicious samples should be increased and the late positive determination of ring rot in the official phytosanitary and company checks abroad should be reduced.
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The use of microsatellite analysis for the characterisation of onion
Mitrová, Katarína ; Ovesná, Jaroslava
The object of the methodology is the process characterizing the autenticity of varieties of onion put through microsatellite analysis (SSR- Single Sequence Repeats) for the characterization of varieties of garlic into the practice. This method allows to identify varieties of onion and characterize genetic resources using DNA markers and determine their genetic similarity based on the length variability of short repeating sequences (microsatellites) in the characterization of varieties of onion, Allium cepa L. The principle of the method is amplification of the genome containing the microsatelite locus by polymerase chain reaction (PCR) using specific primers and subsequent analysis of the lengtht of the products and the identification of the DNA profile to the declared variety.
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The use of microsatellite analysis for the characterisation of garlic
Mitrová, Katarína ; Ovesná, Jaroslava ; Svobodová, Leona
The object of the methodology is the process characterizing the autenticity of varieties of garlic put through microsatellite analysis (SSR- Single Sequence Repeats) for the characterization of varieties of garlic into the practice. This method allows to identify varieties of garlic and characterize genetic resources using DNA markers and determine their genetic similarity based on the length variability of short repeating sequences (microsatellites) in the characterization of varieties of garlic, Allium sativum L. The principle of the method is amplification of the genome containing the microsatelite locus by polymerase chain reaction (PCR) using specific primers and subsequent analysis of the lengtht of the products and the identification of the DNA profile to the declared variety.
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Methodology for evaluation of pest resistance to zoocides by biological methods and antiresistance staregies
Kocourek, František ; Stará, Jitka ; Zichová, Tereza ; Hubert, Jan ; Nesvorná, Marta
The methodology contains description of biological methods for evaluation of pest resistance to zoocides arranged according to world standards. The methods are verified on case studies on pests with proved resistance to insecticides or with high risk of selection of resistance in Czechia, as pollen beetle, Colorado potato beetle, cabbage seedpod weevil, cabbage stem flea beetle and codling moth. The methodology contains survey of basic principles of insect resistance management and proposals of antiresistant strategies for pest mentioned in the case studies. The methods for evaluation of resistance are utilized for resistance monitoring performed by state administration or other organisations. Recommendations given in antiresistant strategies are utilized for pest management in winter rape, potatoes and apples.
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