Разработка программного обеспечения системы управления требованиями

Целью магистерской диссертации является – разработать программное обеспечение, которое позволяло бы пользователям сформировать простой и понятных документ, обеспечивающий единое виденье разрабатываемой ИС у всех заинтересованных лиц разного уровня (от руководителя до разработчика). В результате данной магистерской диссертации был предложен инструментарий, позволяющий студентам обучающихся по дисциплины «Инженерия требований к системам», изучить процесс формирования технических требований и их согласования с заинтересованными лицами (анг. stаckholdеr) и изучить жизненный цикл ИС.Thе purposе of thе mаstеr's thеsіs іs to dеvеlop softwаrе for rеquіrеmеnt mаnаgеmеnt sуstеm whіch could crеаtе pdf fіlе wіth аll tеchnіcаl documеntаtіon of futurе іnformаtіon sуstеm. The result of this magisterskoj thesis was proposed tool that allows students in the discipline "Engineering requirements", to study the process of formation technical requirements and coordination with stakeholders (eng. stаckholdеr) and study the life cycle of IP

Involvement of Arabidopsis thaliana endoplasmic reticulum KDEL-tailed cysteine endopeptidase 1 (AtCEP1) in powdery mildew-induced and AtCPR5-controlled cell death

Programmed cell death (PCD) is a prerequisite for successful development and it limits the spread of biotrophic pathogens in a rapid hypersensitive response at the site of infection. KDEL-tailed cysteine endopeptidases (KDEL CysEP) are a subgroup of papain-type cysteine endopeptidases expressed in tissues undergoing PCD. In Arabidopsis, three KDEL CysEPs (AtCEP1, AtCEP2, and AtCEP3) are expressed. We have previously shown that AtCEP1 is a factor of basal resistance to powdery mildew caused by the biotrophic ascomycete Erysiphe cruciferarum, and is expressed in spatiotemporal association with the late fungal development on Arabidopsis leaves. The endoplasmic reticulum-localized proenzyme of AtCEP1 was further visualized at the haustorial complex encased with callose. The AtCPR5 gene (CONSTITUTIVE EXPRESSION OF PR GENES 5) is a regulator of expression of pathogenesis related genes. Loss of AtCPR5 leads to spontaneous expression of chlorotic lesions which was associated with enhanced expression of AtCEP1. We used the atcpr5-2 mutant plants and the atcep1 atcpr5-2 double mutants harboring a non-functional reporter (PCEP1:: pre-pro-3xHA-EGFP-KDEL) for visualization of AtCEP1 promoter activity. We found the specific up-regulation of AtCEP1 in direct neighborhood of spreading leaf lesions thus likely representing cells undergoing PCD. Furthermore, we found a strong resistance of atcpr5 mutant plants against infection with E. cruciferarum. Loss of AtCEP1 had no obvious influence on the strong resistance of atcpr5-2 mutant plants against infection with E. cruciferarum. However, the area of necrotic leaf lesions associated with E. cruciferarum colonies was significantly larger in atcpr5-2 as compared to atcep1 atcpr5-2 double mutant plants. The presence of AtCEP1 thus contributes to AtCPR5-controlled PCD at the sites of powdery mildew infection

The formation of a camalexin-biosynthetic metabolon

Arabidopsis thaliana efficiently synthesizes the antifungal phytoalexin camalexin without apparent release of bioactive intermediates, such as indole-3-acetaldoxime, suggesting channeling of the biosynthetic pathway by formation of an enzyme complex. To identify such protein interactions, two independent untargeted co49 immunoprecipitation (co-IP) approaches with the biosynthetic enzymes CYP71B1 and CYP71A13 as baits were performed and the camalexin biosynthetic P450 enzymes were shown to co-purify. These interactions were confirmed by targeted co-IP and Förster resonance energy transfer measurements based on fluorescence lifetime microscopy (FRET-FLIM). Furthermore, interaction of CYP71A13 and Arabidopsis P450 Reductase 1 (ATR1) was observed. An increased substrate affinity of CYP79B2 in presence of CYP71A13 was shown, indicating allosteric interaction. Camalexin biosynthesis involves glutathionylation of an intermediary indole-3-cyanohydrin, synthesized by CYP71A12 and especially CYP71A13. It was demonstrated by FRET-FLIM and co-IP, that the glutathione transferase GSTU4, which is co-expressed with tryptophan- and camalexin-specific enzymes, was physically recruited to the complex. Surprisingly, camalexin concentrations were elevated in knock-out and reduced in GSTU4 overexpressing plants. This shows that GSTU4 is not directly involved in camalexin biosynthesis but rather has a role in a competing mechanism

Convergent evolution of plant pattern recognition receptors sensing cysteine-rich patterns from three microbial kingdoms

The Arabidopsis thaliana Receptor-Like Protein RLP30 contributes to immunity against the fungal pathogen Sclerotinia sclerotiorum. Here we identify the RLP30-ligand as a small cysteine-rich protein (SCP) that occurs in many fungi and oomycetes and is also recognized by the Nicotiana benthamiana RLP RE02. However, RLP30 and RE02 share little sequence similarity and respond to different parts of the native/folded protein. Moreover, some Brassicaceae other than Arabidopsis also respond to a linear SCP peptide instead of the folded protein, suggesting that SCP is an eminent immune target that led to the convergent evolution of distinct immune receptors in plants. Surprisingly, RLP30 shows a second ligand specificity for a SCP-nonhomologous protein secreted by bacterial Pseudomonads. RLP30 expression in N. tabacum results in quantitatively lower susceptibility to bacterial, fungal and oomycete pathogens, thus demonstrating that detection of immunogenic patterns by Arabidopsis RLP30 is involved in defense against pathogens from three microbial kingdoms

Biotic and Abiotic Stress Responses in Crop Plants

Agricultural productivity depends on increasingly extreme weather phenomena, and the use of germplasm that has to be continuously improved by plant breeders to become tolerant to various biotic and abiotic stresses. Molecular plant biologists try to understand the mechanisms associated with stress responses and provide knowledge that could be used in breeding programs. To provide a partial overview about our current understanding about molecular and physiological stress responses, and how this knowledge can be used in agriculture, we have edited a special issue on “Biotic and Abiotic Stress Responses in Crop Plants„. Contributions are from different fields including heat stress responses, stress responses during drought and salinity, as well as during flooding, and resistance and susceptibility to pathogenetic stresses and about the role of plant functional metabolites in biotic stress responses. Future research demand in particular areas of crop stress physiology is discussed, as well as the importance of translational research and investigations directly in elite crop plants and in the genetic resources available for breeding

Good Riddance? Breaking Disease Susceptibility in the Era of New Breeding Technologies

Despite a high abundance and diversity of natural plant pathogens, plant disease susceptibility is rare. In agriculture however, disease epidemics often occur when virulent pathogens successfully overcome immunity of a single genotype grown in monoculture. Disease epidemics are partially controlled by chemical and genetic plant protection, but pathogen populations show a high potential to adapt to new cultivars or chemical control agents. Therefore, new strategies in breeding and biotechnology are required to obtain durable disease resistance. Generating and exploiting a genetic loss of susceptibility is one of the recent strategies. Better understanding of host susceptibility genes (S) and new breeding technologies now enable the targeted mutation of S genes for genetic plant protection. Here we summarize biological functions of susceptibility factors and both conventional and DNA nuclease-based technologies for the exploitation of S genes. We further discuss the potential trade-offs and whether the genetic loss of susceptibility can provide durable disease resistance

Biotic and Abiotic Stress Responses in Crop Plants

While the demand for crop products continues to increase strongly, agricultural productivity is threatened by various stress factors, often associated with global warming. To sustain and improve yield, it is necessary to understand how plants respond to various stresses, and to use the generated knowledge in modern breeding programs. Most knowledge regarding the molecular mechanisms associated with stress responses has been obtained from investigations using the model plant Arabidopsis thaliana. Stress hormones, such as abscisic acid, jasmonic acid, and salicylic acid, have been shown to play key roles in defense responses against abiotic and biotic stresses. More recently, evidence that growth-regulating plant hormones are also involved in stress responses has been accumulating. Epigenetic regulation at the DNA and histone level, and gene regulation by small non-coding RNAs appear to be important as well. Many approaches have used mutant screens and next generation sequencing approaches to identify key players and mechanisms how plants respond to their environment. However, it is often unclear to which extent the elucidated mechanisms also operate in crops. This Special Issue Book, therefore, aims to close this gap and contains a number of contributions from labs that work both, on Arabidopsis and crops. The book includes contributions reporting how crop plant species respond to various abiotic stresses, such as drought, heat, cold, flooding, and salinity, as well as biotic stimuli during microbial infections. It contains reviews, opinions, perspectives, and original articles, and its focus is on our molecular understanding of biotic and abiotic stress responses in crops, highlighting, among other aspects, the role of stress hormones, secondary metabolites, signaling mechanisms, and changes in gene expression patterns and their regulation. Approaches and ideas to achieve stress tolerance and to maintain yield stability of agricultural crops during stress periods can be found in most chapters. These include also perspectives on how knowledge from model plants can be utilized to facilitate crop-plant breeding and biotechnology

Infection of barley with the parasitic fungus Blumeria graminis f.sp. hordei results in the induction of HvADH1 and HvADH2

Besides the established functions of alcohol dehydrogenase (ADH) in the flooding response and in seed and pollen metabolism there is increasing evidence for a role of the fermentative pathway in biotic interactions. We have recently shown that barley ADH may be involved in susceptibility to the parasitic fungus Blumeria graminis f.sp. hordei (Bgh). Here, the transcriptional regulation of the barley ADH genes HvADH1 and HvADH2 after challenge of susceptible barley leaves with Bgh is addressed. Bgh infection results in an induction of HvADH1 and HvADH2, whereas HvADH3 expression was not detectable in leaves. With the use of native polyacrylamide gels the iso-enzyme composition with and without challenge by Bgh was analyzed, showing an activation of HvADH1 and HvADH2 in Bgh treated leaves