VFK1, a Vicia faba K(+) channel involved in phloem unloading.
In search of a K(+) channel involved in phloem transport we screened a Vicia faba cotyledon cDNA library taking advantage of a set of degenerated primers, flanking regions conserved among K(+) uptake channels. We cloned VFK1 (for Vicia faba K(+) channel 1) characterised by a structure known from the Shaker family of plant K(+) channels. When co-expressed with a KAT1 mutant in Xenopus oocytes, heteromers revealed the biophysical properties of a K(+) selective, proton-blocked channel. Northern blot analyses showed high levels of expression in cotyledons, flowers, stem and leaves. Using in situ PCR techniques we could localise the K(+) channel mRNA in the phloem. In the stem VFK1 expression levels were higher in the lower internodes. There channel transcripts increased in the light and thus under conditions of increased photosynthate allocation. VFK1 transcripts are elevated in sink leaves, and rise in source leaves during the experimental transition into sinks. Fructose- rather than sucrose- or glucose-feeding via the petiole induced VFK1 gene activity. We therefore monitored the fructose sensitivity of the sieve tube potential through cut aphid stylets. In response to an 1 h fructose treatment the sieve tube potential shift increased from 19 mV to 53 mV per 10-fold change in K(+) concentration. Under these conditions K(+) channels dominated the electrical properties of the plasma membrane. Based on the phloem localisation and expression patterns of VFK1 we conclude that this K(+) channel is involved in sugar unloading and K(+) retrieval.
Ache
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Julius-von-Sachs-Institut, Molekulare Pflanzenphysiologie und Biophysik, Lehrstuhl Botanik I, Universität Würzburg, D-97082 Würzburg, Germany.
Becker
D
D
Deeken
R
R
Dreyer
I
I
Weber
H
H
Fromm
J
J
Hedrich
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D003001:Cloning, Molecular; D004553:Electric Conductivity; D007887:Fabaceae / Q000166:cytology / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D005260:Female; D005632:Fructose / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D008027:Light; D008564:Membrane Potentials; D008954:Models, Biological; D008969:Molecular Sequence Data; D009865:Oocytes; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014158:Transcription, Genetic; D014981:Xenopus
D010940:Plant Proteins; D015221:Potassium Channels; C440748:VFK1 protein, Vicia faba; D005632:Fructose