Building a proton gradient across a biological membrane and between different tissues is a matter of great importance for plant development and nutrition. To gain a better understanding of proton distribution in the plant root apoplast as well as across the plasma membrane, we generated Arabidopsis plants expressing stable membrane-anchored ratiometric fluorescent sensors based on pHluorin. These sensors enabled noninvasive pH-specific measurements in mature root cells from the medium–epidermis interface up to the inner cell layers that lie beyond the Casparian strip. The membrane-associated apoplastic pH was much more alkaline than the overall apoplastic space pH. Proton concentration associated with the plasma membrane was very stable, even when the growth medium pH was altered. This is in apparent contradiction with the direct connection between root intercellular space and the external medium. The plasma membrane-associated pH in the stele was the most preserved and displayed the lowest apoplastic pH (6.0 to 6.1) and the highest transmembrane delta pH (1.5 to 2.2). Both pH values also correlated well with optimal activities of channels and transporters involved in ion uptake and redistribution from the root to the aerial part. In growth medium where ionic content is minimized, the root plasma membrane-associated pH was more affected by environmental proton changes, especially for the most external cell layers. Calcium concentration appears to play a major role in apoplastic pH under these restrictive conditions, supporting a role for the cell wall in pH homeostasis of the unstirred surface layer of plasma membrane in mature roots. Significance The pH on both sides of the plant plasma membrane was accurately measured in vivo using ratiometric fluorescent sensors. This enabled noninvasive access to membrane-associated pH and transmembrane delta pH values from the surface of the root up to the deepest cell layers beyond the Casparian strip barrier. We demonstrate that despite direct contact with the soil, the apoplastic pH close to the plasma membrane was maintained at values ranging from 6.0 to 6.4 in mature root cells, whereas the overall pH in the apoplastic space is far more acidic. Furthermore, we found that the cell wall plays a role in proton homeostasis in mature root.

A cDNA clone encoding human angiotensinogen was isolated from a cDNA library prepared from human liver mRNA and used to isolate the angiotensinogen gene. The complete exon sequence of this gene together with extensive intron and flanking sequences are reported. The human angiotensinogen gene contains five exons interrupted by four intervening sequences. We compared the intron-exon structure of this human gene with that of the rat gene or the genes coding for proteins such as alpha 1-antitrypsin and antithrombin III, whose primary amino acid sequences show similarities. The human angiotensinogen gene shows identical organization with the alpha 1-antitrypsin gene, but is different from the antithrombin III gene. The 5'-flanking sequence (-500 to -1 bp) of the human angiotensinogen gene was examined for hormone regulatory elements (HRE), which may be implicated in the interaction with the hormone receptor complexes.

Summary: A functional Shaker potassium channel requires assembly of four α‐subunits encoded by a single gene or various genes from the Shaker family. In Arabidopsis thaliana, AtKC1, a Shaker α‐subunit that is silent when expressed alone, has been shown to regulate the activity of AKT1 by forming heteromeric AtKC1–AKT1 channels. Here, we investigated whether AtKC1 is a general regulator of channel activity. Co‐expression in Xenopus oocytes of a dominant negative (pore‐mutated) AtKC1 subunit with the inward Shaker channel subunits KAT1, KAT2 or AKT2, or the outward subunits SKOR or GORK, revealed that the three inward subunits functionally interact with AtKC1 while the outward ones cannot. Localization experiments in plant protoplasts showed that KAT2 was able to re‐locate AtKC1 fused to GFP from endomembranes to the plasma membrane, indicating that heteromeric AtKC1–KAT2 channels are efficiently targeted to the plasma membrane. Functional properties of heteromeric channels involving AtKC1 and KAT1, KAT2 or AKT2 were analysed by voltage clamp after co‐expression of the respective subunits in Xenopus oocytes. AtKC1 behaved as a regulatory subunit within the heterotetrameric channel, reducing the macroscopic conductance and negatively shifting the channel activation potential. Expression studies showed that AtKC1 and its identified Shaker partners have overlapping expression patterns, supporting the hypothesis of a general regulation of inward channel activity by AtKC1 in planta. Lastly, AtKC1 disruption appeared to reduce plant biomass production, showing that AtKC1‐mediated channel activity regulation is required for normal plant growth.