Pollen viability Dotaz Zobrazit nápovědu
Research investigating the dynamics of male gametophyte (MG) development has proven to be challenging for the plant science community. Here we describe our protocol for separating Arabidopsis MG developmental stages, which is based on the centrifugation of pollen through a discontinuous Percoll concentration gradient. This Percoll gradient can be formed using a pipette, and it does not require a gradient maker. The purity of the isolated developing spores is as high as 70%, and in most separations it is well above 80%. Using this protocol, we can separate four different stages of pollen development-uninucleate microspore (UNM), bicellular pollen (BCP), tricellular immature pollen (TCP) and mature pollen grain (MPG). The duration of the separation procedure, excluding the cutting of flower inflorescences, is 6 h. This is reduced to 4 h when using a vacuum cleaning method to remove the MPGs before the Percoll density separation.
Sex determination in Rumex acetosa, a dioecious plant with a complex XY1 Y2 sex chromosome system (females are XX and males are XY1 Y2 ), is not controlled by an active Y chromosome but depends on the ratio between the number of X chromosomes and autosomes. To gain insight into the molecular mechanisms of sex determination, we generated a subtracted cDNA library enriched in genes specifically or predominantly expressed in female floral buds in early stages of development, when sex determination mechanisms come into play. In the present paper, we report the molecular and functional characterization of FEM32, a gene encoding a protein that shares a common architecture with proteins in different plants, animals, bacteria and fungi of the aerolysin superfamily; many of these function as β pore-forming toxins. The expression analysis, assessed by northern blot, RT-PCR and in situ hybridization, demonstrates that this gene is specifically expressed in flowers in both early and late stages of development, although its transcripts accumulate much more in female flowers than in male flowers. The ectopic expression of FEM32 under both the constitutive promoter 35S and the flower-specific promoter AP3 in transgenic tobacco showed no obvious alteration in vegetative development but was able to alter floral organ growth and pollen fertility. The 35S::FEM32 and AP3::FEM32 transgenic lines showed a reduction in stamen development and pollen viability, as well as a diminution in fruit set, fruit development and seed production. Compared with other floral organs, pistil development was, however, enhanced in plants overexpressing FEM32. According to these effects, it is likely that FEM32 functions in Rumex by arresting stamen and pollen development during female flower development. The aerolysin-like pore-forming proteins of eukaryotes are mainly involved in defence mechanisms against bacteria, fungi and insects and are also involved in apoptosis and programmed cell death (PCD), a mechanism that could explain the role of FEM32 in Rumex sex determination.
- MeSH
- bakteriální toxiny klasifikace genetika MeSH
- cytotoxické proteiny tvořící póry klasifikace genetika MeSH
- fylogeneze MeSH
- geneticky modifikované rostliny MeSH
- květy genetika růst a vývoj MeSH
- neplodnost rostlin genetika MeSH
- ovoce genetika růst a vývoj MeSH
- pyl genetika růst a vývoj MeSH
- regulace genové exprese u rostlin MeSH
- rostlinné proteiny klasifikace genetika MeSH
- Rumex genetika růst a vývoj MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- semena rostlinná genetika růst a vývoj MeSH
- stanovení celkové genové exprese metody MeSH
- tabák genetika růst a vývoj MeSH
- vývojová regulace genové exprese MeSH
- Publikační typ
- časopisecké články MeSH
Hydrogels belong to the group of materials with growing interest on the market of polymers. In this article, hydrogels based on Beetosan were obtained using ultraviolet (UV) radiation. Main component of hydrogel matrix-Beetosan-is chitosan obtained from naturally died honeybees. Such hydrogels were modified with active substances, that is, caffeine, bee pollen, Salvia officinalis (sage), and Aloe vera juice. Next, the analysis of cytotoxicity of hydrogels in relation to murine fibroblasts by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide and neutral red uptake assays were conducted. Furthermore, surface morphology, tensile strength, geometry, and roughness of hydrogels were characterized. Hydrogels did not show cytotoxicity to recommended L929 murine fibroblasts. These polymers did not affect adversely the growth and viability of these cells. Moreover, Beetosan hydrogels were characterized by flexibility as well as by diversified surface morphology that could indicate their high absorbency. Therefore these materials may be considered as useful for biomedical purposes with special emphasis on application as modern wound dressings that not only absorb wound exudate but also contain natural substances with therapeutic properties that is beneficial from the point of view of wound healing process.
- MeSH
- Aloe MeSH
- buněčné linie MeSH
- chitosan * chemie farmakologie MeSH
- fibroblasty cytologie metabolismus MeSH
- hojení ran účinky léků MeSH
- hydrogely * chemie farmakologie MeSH
- myši MeSH
- šalvěj lékařská MeSH
- testování materiálů * MeSH
- včely MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cationic amphipathic drugs, such as amiodarone, interact preferentially with lipid membranes to exert their biological effect. In the yeast Saccharomyces cerevisiae, toxic levels of amiodarone trigger a rapid influx of Ca(2+) that can overwhelm cellular homeostasis and lead to cell death. To better understand the mechanistic basis of antifungal activity, we assessed the effect of the drug on membrane potential. We show that low concentrations of amiodarone (0.1-2 microm) elicit an immediate, dose-dependent hyperpolarization of the membrane. At higher doses (>3 microm), hyperpolarization is transient and is followed by depolarization, coincident with influx of Ca(2+) and H(+) and loss in cell viability. Proton and alkali metal cation transporters play reciprocal roles in membrane polarization, depending on the availability of glucose. Diminishment of membrane potential by glucose removal or addition of salts or in pma1, tok1Delta, ena1-4Delta, or nha1Delta mutants protected against drug toxicity, suggesting that initial hyperpolarization was important in the mechanism of antifungal activity. Furthermore, we show that the link between membrane hyperpolarization and drug toxicity is pH-dependent. We propose the existence of pH- and hyperpolarization-activated Ca(2+) channels in yeast, similar to those described in plant root hair and pollen tubes that are critical for cell elongation and growth. Our findings illustrate how membrane-active compounds can be effective microbicidals and may pave the way to developing membrane-selective agents.
