plant methods
Dotaz
Zobrazit nápovědu
Vypracovala sa jednoduchá, citlivá a reprodukovateľná metóda dôkazu extracelulárnej β-galaktozidázy. Kalusovou kultúrou, resp. koreňom klíčnych rastlín sekrétovaná β-galaktozidáza hydrolyzuje substrát (6-bróm-2-naftyl-β-D-galaktopyranozid) na β-D-galaktózu a 6-bróm-2-naftol, ktorý po simultánnej azokopulácii s hexazotovaným p-rozanilínom, resp. bázickým fuksínom vytvára červenohnedú vo vode takmer nerozpustnú zlúčeninu (dÍEizóniovú soI). Zafarbenie pod kalusovou kultúrou a okolo nej, ako aj koreňa iďíčnych rastlín je prejavom činnosti extracelulárnej β-galaktozidázy Študovanými objektmi. Intenzita zafarbenia je mierou ich enzýmovej aktivity. Intracelulárne lokalizovaný enzým má majoritné, kým extracelulárna β-galaktozidáza minoritné zastúpenie
A simple, sensitive and reproducible method of detection of extracellular β-galactosidase was worked out. β-Galactosidase secreted by the callus culture, or the roots of sprouting plants, hydrolyzes the substrate (6-bromo-2-naphthyl β-D-galactopyranoside) to produce β-D-galactose and 6-bromo-2-naphthol, which after simultaneous azocoupling with hexazotized p-rosaniline, or basic fuchsine, produce a reddish brown compound, nearly insoluble in water (a diazonium salt). The colouring under the callus culture and around it, as well as the colouring of the roots of the sprouting plants, is a manifestation of the activity of extracellular β-galactosidase by the objects under study. The intensity of the colouring is a measure of their enzymatic activity. The share of intracellularly localized enzyme represents the majority and the share of extracellular β-galactosidase the minority.
Agrobacterium tumefaciens is widely used as a versatile tool for development of stably transformed model plants and crops. However, the development of Agrobacterium based transient plant transformation methods attracted substantial attention in recent years. Transient transformation methods offer several applications advancing stable transformations such as rapid and scalable recombinant protein production and in planta functional genomics studies. Herein, we highlight Agrobacterium and plant genetics factors affecting transfer of T-DNA from Agrobacterium into the plant cell nucleus and subsequent transient transgene expression. We also review recent methods concerning Agrobacterium mediated transient transformation of model plants and crops and outline key physical, physiological and genetic factors leading to their successful establishment. Of interest are especially Agrobacterium based reverse genetics studies in economically important crops relying on use of RNA interference (RNAi) or virus-induced gene silencing (VIGS) technology. The applications of Agrobacterium based transient plant transformation technology in biotech industry are presented in thorough detail. These involve production of recombinant proteins (plantibodies, vaccines and therapeutics) and effectoromics-assisted breeding of late blight resistance in potato. In addition, we also discuss biotechnological potential of recombinant GFP technology and present own examples of successful Agrobacterium mediated transient plant transformations.
3rd ed. 304 s.
Pomocou nerozpustného chromogénneho substrátu (čierny gelasfér) sa vypracovala jednoduchá, citlivá a reprodukovateľná metóda dôkazu extracelulárnych proteolytických enzýmov. Extracelu- lárne proteolytické enzýmy (produkované kalusovou alebo suspenznou kultúrou) hydrolyzujú čierny gelasfér. Zafarbenie okolo kalusovej kultúry a pod ňou je znakom činnosti kalusovou kultúrou produkovaných extracelulárnych proteolytických enzýmov. Intenzita zafarbenia kultivačného mé- dia je mierou aktivity extracelulárnych proteolytických enzýmov suspenznej kultúry.
By means of an insoluble chromogenic substrate (black gelasphere), a simple, sensitive and repro- ducible method of detection of extracellular proteolytic enzymes was worked out. Extracellular proteolytic enzymes (produced by a callus or suspension culture) hydrolyse black gelasphere. Colouring around the callus culture and under it is a sign of the activity of extracellular proteolytic enzymes produced by the callus culture. The intensity of colouring of the cultivating medium is a measure of the activity of extracellular proteolytic enzymes of the suspension culture.
There are various preparatory techniques for light microscopy permitting access to the inner structure of plant body and its development. Minute objects might be processed as whole-mount preparations, while voluminous ones should be separated into smaller pieces. Hereby we summarize some of the "classical" techniques to cut more voluminous objects into slices and access their inner structure either for simple anatomical analysis or for further processing (e.g., histochemistry, immunohistochemistry, in situ hybridization, enzyme histochemistry).
115 s. ; 26 cm
- Konspekt
- Lékařské vědy. Lékařství
- NLK Obory
- veřejné zdravotnictví
- farmacie a farmakologie
- management, organizace a řízení zdravotnictví
- alternativní lékařství
- NLK Publikační typ
- publikace WHO
- MeSH
- chemický průmysl MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- nemoci úst prevence a kontrola MeSH
- nemoci z povolání prevence a kontrola MeSH
- nemoci zubů prevence a kontrola MeSH
- průzkumy a dotazníky MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
Histochemical methods allow for identification and localization of various components within the tissue. Such information on the spatial heterogeneity is not available with biochemical methods. However, there is limitation of the specificity of such detection in context of complex tissue, which is important to consider, and interpretations of the results should regard suitable control treatments if possible. Hereby we present set of selected simple staining and histochemical methods with comments based on our laboratory experience.
