The auxin-induced K(+) channel gene Zmk1 in maize functions in coleoptile growth and is required for embryo development.

The transcript level and in turn protein density of the K(+)-uptake channel ZMK1 in maize (Zea mays) coleoptiles is controlled by the phytohormone auxin. ZMK1 is involved in auxin-regulated coleoptile elongation as well as gravi- and phototropism. To provide unequivocal evidence for the role of ZMK1 in these elementary processes we screened for maize plants containing a Mutator-tagged Zmk1 gene. In a site-selected approach, we were able to identify three independent alleles of Mutator-transposon insertions in Zmk1. zmk1-m1::Mu1 plants were characterised by a Mu1 transposon inside intron 1 of ZMK1. When we analysed the Zmk1-transcript abundance in growing coleoptiles of these homozygous mutants, however, we found the K(+)-channel allele overexpressed. In consequence, elevated levels of K(+)-channel transcripts resulted in a growth phenotype as expected from more efficient K(+)-uptake, representing a central factor for turgor formation. Following Zmk1 expression during maize embryogenesis, we found this K(+)-channel gene constitutively expressed throughout embryo development and upregulated in late stages. In line with a vital role in embryogenesis, the mutations of exon 2 and intron 2 of Zmk1-zmk1-m2::Mu8 and zmk1-m3::MuA2-caused a lethal, defective-kernel phenotype. Thus, these results demonstrate the central role of the auxin-regulated K(+)-channel gene Zmk1 in coleoptile growth and embryo development.

Differential expression of K+ channels between guard cells and subsidiary cells within the maize stomatal complex.

Grass stomata are characterized by dumbbell-shaped guard cells forming a complex with a pair of specialized epidermal cells, the subsidiary cells. Stomatal movement is accomplished by a reversible exchange of potassium and chloride between these two cell types. To gain insight into the molecular machinery involved in K+ transport within the stomatal complex of Zea mays, we determined the spatial and temporal expression pattern of potassium channels in the maize leaf. KZM2 and ZORK were isolated and identified as new members of the plant Shaker K+ channel family. Northern blot analysis identified fully developed leaves as the predominant site of KZM2 expression. Following enzymatic digestion and separation of leaf tissue into epidermal, mesophyll, and vascular fractions, KZM2 and ZORK transcripts were localized in the epidermis. Using a collection of individually isolated guard cell or subsidiary cell protoplasts, ZORK transcripts were found in both cell types while KZM2 was exclusively expressed in the guard cell population. The previously identified K+ channel genes ZMK1 and KZM1 were expressed in subsidiary cells and guard cells, respectively, whereas ZMK2 transcripts were not detected. These data indicate that the interaction between subsidiary cells and guard cells is based on overlapping as well as differential expression of K+ channels in the two cell types of the maize stomatal complex.

The K+ channel KZM1 mediates potassium uptake into the phloem and guard cells of the C4 grass Zea mays.

In search of K(+) channel genes expressed in the leaf of the C(4) plant Zea mays, we isolated the cDNA of KZM1 (for K(+) channel Zea mays 1). KZM1 showed highest similarity to the Arabidopsis K(+) channels KAT1 and KAT2, which are localized in guard cells and phloem. When expressed in Xenopus oocytes, KZM1 exhibited the characteristic features of an inward-rectifying, potassium-selective channel. In contrast to KAT1- and KAT2-type K(+) channels, however, KZM1 currents were insensitive to external pH changes. Northern blot analyses identified the leaf, nodes, and silks as sites of KZM1 expression. Following the separation of maize leaves into epidermal, mesophyll, and vascular fractions, quantitative real-time reverse transcriptase-PCR allowed us to localize KZM1 transcripts predominantly in vascular strands and the epidermis. Cell tissue separation and KZM1 localization were followed with marker genes such as the bundle sheath-specific ribulose-1,5-bisphosphate carboxylase, the phloem K(+) channel ZMK2, and the putative sucrose transporter ZmSUT1. When expressed in Xenopus oocytes, ZmSUT1 mediated proton-coupled sucrose symport. Coexpression of ZmSUT1 with the phloem K(+) channels KZM1 and ZMK2 revealed that ZMK2 is able to stabilize the membrane potential during phloem loading/unloading processes and KZM1 to mediate K(+) uptake. During leaf development, sink-source transitions, and diurnal changes, KZM1 is constitutively expressed, pointing to a housekeeping function of this channel in K(+) homeostasis of the maize leaf. Therefore, the voltage-dependent K(+)-uptake channel KZM1 seems to mediate K(+) retrieval and K(+) loading into the phloem as well as K(+)-dependent stomatal opening.