Nitrogen (N) is a critical nutrient for plants but is often distributed unevenly in the soil. Plants therefore have evolved a systemic mechanism by which N starvation on one side of the root system leads to a compensatory and increased nitrate uptake on the other side. Here, we study the molecular systems that support perception of N and the long-distance signaling needed to alter root development. Rootlets starved of N secrete small peptides that are translocated to the shoot and received by two leucine-rich repeat receptor kinases (LRR-RKs). Arabidopsis plants deficient in this pathway show growth retardation accompanied with N-deficiency symptoms. Thus, signaling from the root to the shoot helps the plant adapt to fluctuations in local N availability.

The secreted peptide gene CLAVATA3 (CLV3) regulates stem cell fate in the shoot apical meristem in Arabidopsis thaliana plants, but the molecular structure of the active mature CLV3 peptide is controversial. Here, using nano-LC-MS/MS analysis of apoplastic peptides of A. thaliana plants overexpressing CLV3, we show that CLV3 is a 13-amino-acid arabinosylated glycopeptide. Post-translational arabinosylation of CLV3 is critical for its biological activity and high-affinity binding to its receptor CLV1.

SummaryPeptidomics is a challenging field in which to create a link between genomic information and biological function through biochemical analysis of expressed peptides, including precise identification of posttranslational modifications and proteolytic processing. We found that secreted peptides in Arabidopsis plants diffuse into the medium of whole-plant submerged cultures, and can be effectively identified by o-chlorophenol extraction followed by LC-MS analysis. Using this system, we first confirmed that a 12-amino-acid mature CLE44 peptide accumulated at a considerable level in the culture medium of transgenic plants overexpressing CLE44. Next, using an in silico approach, we identified a novel gene family encoding small secreted peptides that exhibit significant sequence similarity within the C-terminal short conserved domain. We determined that the mature peptide encoded by At1g47485, a member of this gene family, is a 15-amino-acid peptide containing two hydroxyproline residues derived from the conserved domain. This peptide, which we have named CEP1, is mainly expressed in the lateral root primordia and, when overexpressed or externally applied, significantly arrests root growth. CEP1 is a candidate for a novel peptide plant hormone.

SUMMARYLeucine-rich repeat receptor kinases (LRR-RKs) are the largest sub-family of transmembrane receptor kinases in plants. In several LRR-RKs, a loop-out region called an 'island domain', which intercepts the extracellular tandem LRRs at a position near the transmembrane domain, constitutes the ligand-binding pocket, but the absence of the island domain in numerous LRR-RKs raises questions about which domain recognizes the ligand in non-island domain LRR-RKs. Here, we used photoaffinity labeling followed by chemical and enzymatic digestion to show that BAM1, a CLV1/BAM-family LRR-RK whose extracellular domain comprises 22 consecutive LRRs, directly interacts with the small peptide ligand CLE9 at the LRR6-LRR8 region that is relatively distal from the transmembrane domain. Multiple sequence alignment and homology modeling revealed that the inner concave side of LRR6-LRR8 of CLV1/BAM-family LRR-RKs deviates slightly from the LRR consensus. In support of our findings, the clv1-4 mutant carries a missense mutation at the inner concave side of LRR6 of CLV1, and introduction of the corresponding mutation in BAM1 resulted in complete loss of ligand binding activity. Our results indicate that the ligand recognition mechanisms of plant LRR-RKs are more complex and diverse than anticipated.

A Sea-Bird Electronics (SBE 35) deep ocean reference thermometer is used with the SBE 9plus CTD system to calibrate the SBE 3 ocean thermometers of the CTD. The SBE 35 is standardized in water-triple-point and gallium-melting-point cells. The SBE 3 is calibrated with the SBE 35 under the assumption that discrepancies between SBE 3 and SBE 35 data are due to pressure sensitivity, the viscous heating effect, and time drift of the SBE 3. Based on the results of an in situ calibration, the pressure sensitivity and the viscous heating effect were evaluated for 11 SBE 3 thermometers. Three SBE 3s showed little pressure sensitivity, and eight had pressure sensitivities of 1–2 mK at 6000 dbar. The average viscous heating effect on the standard SBE 3 measurements was 0.5 mK. Both the accuracy and precision of the in situ calibrated SBE 3 data at depths greater than 2000 dbar were 0.4 mK relative to the SBE 35 reference.