A high-throughput procedure for the identification of genes contributing to plant defence mechanisms

Attempted host cell invasion by pathogenic fungi is known to trigger cell polarisation. Barley MLO is a member of a family of heptahelical membrane proteins unique to plants and is thought to regulate polarised SNARE protein-dependent secretory processes. Recessively inherited mutations in Mlo mediate resistance against all isolates (broad-spectrum resistance) of the biotrophic barley powdery mildew fungus, Blumeria graminis f. sp. hordei (Bgh). It is assumed that the fungus utilizes MLO to suppress defence-associated secretion for successful host cell entry. In this study, a genetic screen was established based on gene silencing by double-stranded RNA interference (dsRNAi). This was accomplished by transient expression of inverted repeat DNAs in single epidermal barley cells. To identify novel components of MLO-modulated defence processes to Bgh, approximately 700 inverted repeat DNA constructs, derived from barley epidermal cDNAs, were generated and ballistically delivered into epidermal cells of both Mlo and mlo genotypes. Two dsRNAi constructs were identified permitting enhanced Bgh entry in a resistant mlo genotype. Both target the same isoform of an actin depolymerising factor, designated HvADF3. ADFs exist throughout all eukaryotic kingdoms and function in actin filament turnover (treadmilling), thereby contributing to actin cytoskeleton dynamics. Transient ectopic expression of HvADF3 in mlo epidermal cells also permitted enhanced Bgh invasion. Ectopic expression of HvADF3 or dsRNAi-mediated gene silencing resulted in the disappearance of phalloidin-stainable actin filaments, consistent with actin serving as substrate of ADFs. Closer inspection of the topology of the HvADF3 dsRNAi constructs revealed that the presumed silencing effect might be triggered by undesired overexpression. However, due to published pharmacological evidence implicating actin filament reorganisation in plant defence responses, the role of HvADF3 was further investigated. HvADF3 overexpression greatly reduced callose accumulation at pathogen entry sites, a common local stress response at the plant cell wall known to be sensitive to actin-depolymerising drugs. Replacements of an N-terminal serine residue, previously shown to be a phosphorylation target of ADF activity, to Ala or Asp enhanced and reduced Bgh entry rates, respectively, upon transient expression. Ectopic expression of six out of nine tested Arabidopsis ADF genes in barley phenocopied the effect of HvADF3 overexpression, demonstrating that multiple heterologous ADF isoforms can enhance pathogen entry. Although ectopic ADF expression impaired broad-spectrum resistance to Bgh, race-specific resistance (R) triggered by R genes Mlg, Mla1, or Mla6 was not affected. This is consistent with previous genetic data indicating separate pathways for broad-spectrum and R gene-mediated immune responses. Resistance to two tested inappropriate powdery mildews, Bg f sp tritici and Erysiphe pisi, that colonise in nature monocotyledonous wheat and dicotyledonous pea hosts respectively, was impaired upon HvADF3 overexpression. Interestingly, this impairment required the presence of Mlo, suggesting that both inappropriate powdery mildews potentially modulate barley defence reactions via MLO. To identify presumed common components of pathogen-triggered and developmentally controlled cell polarity, a range of Arabidopsis mutants with defects in leaf hair (trichome) development were tested for altered infection phenotypes to diverse powdery mildew fungi. Preliminary data indicate that the absence of an actin-related protein (ARP) 2/3 complex component, CROOKED, enhances pathogen entry of inappropriate powdery mildew species

Virtual commissioning of robotized workplace for cold spraying

Diplomová práca sa zaoberá virtuálnym sprevádzkovaním robotizovaného pracoviska pre studený nástrek. Teoretická časť práce obsahuje prehľad súčasného stavu poznania v oblasti virtuálneho uvedenia do prevádzky, teórie tvorby užívateľských prostredí HMI, zaoberá sa princípmi programovacieho jazyka Rapid ako nástrojom pre ovládanie priemyselných robotov a analýzou komponentov robotizovaného pracoviska. V praktickej časti je definovaná štruktúra navrhnutého užívateľského programu a užívateľského prostredia, ktoré sú vytvorené pomocou softvérových nástrojov Robotstudio a Screenmaker. Posledná časť podrobne rozoberá kroky počas implementovania vytvoreného programu spolu s HMI do prevádzky spojeného s testovaním funkcionalít na reálnom zariadení pre studený nástrek. Súčasťou diplomovej práce je tiež užívateľský manuál vytvorený pre obsluhu priemyselného robota.The diploma thesis deals with the virtual commissioning of robotized workplace for cold spraying. The theoretical part of the thesis contains an overview of the current state of knowledge in the field of virtual commissioning, the theory of crating HMI user environments, deals principles of the Rapid programming language as a tool for controlling industrial robots and analysis of robotic workplace components. The practical part defines the structure of the proposed user program and user environment, which are created using software tools Robotstudio and Screenmaker. The last part discusses in detail the steps during the implementation of the created program together with the HMI into commission, which is associated with the testing of functionalities on a real device for cold spraying. The diploma thesis also includes a user manual created for operating an industrial robot.

Valeology

ВАЛЕОЛОГИ

Hygiene

ГИГИЕНАУЧЕБНЫЕ ПОСОБИЯГИГИЕНА ТРУДАПРОФЕССИОНАЛЬНАЯ ГИГИЕНАОКРУЖАЮЩЕЙ СРЕДЫ ГИГИЕНАГИГИЕНА ПИТАНИЯГИГИЕНА ДЕТЕЙ И ПОДРОСТКОВГИГИЕНА ПОМЕЩЕНИЙВ учебном пособии рассматриваются вопросы гигиены лечебно-профилактических организаций, труда, детей и подростков, окружающей среды, питания, населенных мест и жилища

Hygiene

ГИГИЕНАУЧЕБНЫЕ ПОСОБИЯГИГИЕНА ТРУДАПРОФЕССИОНАЛЬНАЯ ГИГИЕНАОКРУЖАЮЩЕЙ СРЕДЫ ГИГИЕНАГИГИЕНА ПИТАНИЯГИГИЕНА ДЕТЕЙ И ПОДРОСТКОВГИГИЕНА ПОМЕЩЕНИЙВ учебном пособии рассматриваются вопросы по методологии гигиены, гигиене лечебно-профилактических учреждений, труда, детей и подростков, окружающей среды, населенных мест и жилища, питания, а также гигиена жаркого климата

Laboratory classes on hygiene

ГИГИЕНАЛАБОРАТОРНЫЕ ЗАНЯТИЯГИГИЕНА ПИТАНИЯГИГИЕНА ПОМЕЩЕНИЙПОСОБИЯИНОСТРАННЫЕ СТУДЕНТЫЛабораторные занятия предназначены для иностранных студентов и включают 35 тем по гигиене

Environmental medicine

ПОСОБИЯЭКОЛОГИЯЭКОЛОГИЧЕСКАЯ МЕДИЦИНАМЕДИЦИНАОКРУЖАЮЩАЯ СРЕДА И ЗДРАВООХРАНЕНИЕОКРУЖАЮЩЕЙ СРЕДЫ ЗАГРЯЗНЕНИЕОКРУЖАЮЩЕЙ СРЕДЫ ВОЗДЕЙСТВИЕ, БОЛЕЗНИПИЩЕВЫХ ПРОДУКТОВ ЗАГРЯЗНЕНИЕВ пособии освещены вопросы влияния загрязнения окружающей среды на здоровье человека

Radiation medicine

ПОСОБИЯРАДИАЦИОННАЯ ГИГИЕНАTUTORIALRADIATION MEDICINEThe content of the tutorial "Radiation medicine" for students of high medical educational establishments corresponds with the basic educational plan and program, proved by Ministry of Health Care of the Republic of Belarus. The tutorial is prepared for students of medical, pharmaceutical, stomatological, medical-preventive, medical-diagnostic faculties of institutes of higher education. Соответствует основному учебному плану и программе, утвержденным Министерством здравоохранения Республики Беларусь. Для студентов медицинских, фармацевтических, стоматологических, лечебно-профилактических, лечебно-диагностических факультетов высших учебных заведений