Plant organs and tissues consist of many various cell types, often in different phases of their development. Such complex structures do not allow direct studies on behavior of individual cells. In contrast, populations of in vitro-cultured plant cells represent valuable tool for studying processes on a single-cell level, including cell morphogenesis. Here we describe characteristics of well-established model tobacco and Arabidopsis cell lines and provide detailed protocol on their cultivation, characterization, and genetic transformation.
Polar auxin transport is a crucial process for control and coordination of plant development. Studies of auxin transport through plant tissues and organs showed that auxin is transported by a combination of phloem flow and the active, carrier-mediated cell-to-cell transport. Since plant organs and even tissues are too complex for determination of the kinetics of carrier-mediated auxin uptake and efflux on the cellular level, simplified models of cell suspension cultures are often used, and several tobacco cell lines have been established for auxin transport assays. However, there are very few data available on the specificity and kinetics of auxin transport across the plasma membrane for Arabidopsis thaliana suspension-cultured cells. In this report, the characteristics of carrier-mediated uptake (influx) and efflux for the native auxin indole-3-acetic acid and synthetic auxins, naphthalene-1-acetic and 2,4-dichlorophenoxyacetic acids (NAA and 2,4-D, respectively) in A. thaliana ecotype Landsberg erecta suspension-cultured cells (LE line) are provided. By auxin competition assays and inhibitor treatments, we show that, similarly to tobacco cells, uptake carriers have high affinity towards 2,4-D and that NAA is a good tool for studies of auxin efflux in LE cells. In contrast to tobacco cells, metabolic profiling showed that only a small proportion of NAA is metabolized in LE cells. These results show that the LE cell line is a useful experimental system for measurements of kinetics of auxin carriers on the cellular level that is complementary to tobacco cells.
- MeSH
- Arabidopsis cytologie růst a vývoj metabolismus MeSH
- biologický transport MeSH
- buněčné kultury MeSH
- fenotyp MeSH
- hypokotyl cytologie růst a vývoj metabolismus MeSH
- kotyledon cytologie růst a vývoj metabolismus MeSH
- kyselina 2,4-dichlorfenoxyoctová metabolismus MeSH
- kyseliny indoloctové metabolismus MeSH
- kyseliny naftalenoctové metabolismus MeSH
- listy rostlin cytologie růst a vývoj metabolismus MeSH
- metabolom MeSH
- regulátory růstu rostlin metabolismus MeSH
- semenáček cytologie růst a vývoj metabolismus MeSH
- tabák cytologie růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Intercellular flow of the phytohormone auxin underpins multiple developmental processes in plants. Plant-specific pin-formed (PIN) proteins and several phosphoglycoprotein (PGP) transporters are crucial factors in auxin transport-related development, yet the molecular function of PINs remains unknown. Here, we show that PINs mediate auxin efflux from mammalian and yeast cells without needing additional plant-specific factors. Conditional gain-of-function alleles and quantitative measurements of auxin accumulation in Arabidopsis and tobacco cultured cells revealed that the action of PINs in auxin efflux is distinct from PGP, rate-limiting, specific to auxins, and sensitive to auxin transport inhibitors. This suggests a direct involvement of PINs in catalyzing cellular auxin efflux.
- MeSH
- ABC transportéry genetika metabolismus MeSH
- Arabidopsis cytologie fyziologie metabolismus růst a vývoj MeSH
- biologický transport MeSH
- buněčná membrána metabolismus MeSH
- financování organizované MeSH
- ftalimidy farmakologie MeSH
- gravitropismus MeSH
- HeLa buňky MeSH
- kinetika MeSH
- kořeny rostlin fyziologie MeSH
- kultivované buňky MeSH
- kyseliny indoloctové metabolismus MeSH
- kyseliny naftalenoctové metabolismus MeSH
- lidé MeSH
- membránové transportní proteiny genetika metabolismus MeSH
- mutace MeSH
- proteiny huseníčku genetika metabolismus MeSH
- Saccharomyces cerevisiae genetika MeSH
- tabák MeSH
- transfekce MeSH
- transformace genetická MeSH
- Check Tag
- lidé MeSH
