Transcriptional feedback loops are central to the architecture of eukaryotic circadian clocks. Models of the Arabidopsis thaliana circadian clock have emphasized transcriptional repressors, but recently, Myb-like REVEILLE (RVE) transcription factors have been established as transcriptional activators of central clock components, including PSEUDO-RESPONSE REGULATOR5 (PRR5) and TIMING OF CAB EXPRESSION1 (TOC1). We show here that NIGHT LIGHT–INDUCIBLE AND CLOCK-REGULATED1 (LNK1) and LNK2, members of a small family of four LNK proteins, dynamically interact with morning-expressed oscillator components, including RVE4 and RVE8. Mutational disruption of LNK1 and LNK2 function prevents transcriptional activation of PRR5 by RVE8. The LNKs lack known DNA binding domains, yet LNK1 acts as a transcriptional activator in yeast and in planta. Chromatin immunoprecipitation shows that LNK1 is recruited to the PRR5 and TOC1 promoters in planta. We conclude that LNK1 is a transcriptional coactivator necessary for expression of the clock genes PRR5 and TOC1 through recruitment to their promoters via interaction with bona fide DNA binding proteins such as RVE4 and RVE8.

S-RNase is the female determinant of self-incompatibility (SI) in pear (Pyrus bretschneideri). After translocation to the pollen tube, S-RNase degrades rRNA and induces pollen tube death in an S-haplotype-specific manner. In this study, we found that the actin cytoskeleton is a target of P. bretschneideri S-RNase (PbrS-RNase) and uncovered a mechanism that involves phosphatidic acid (PA) and protects the pollen tube from PbrS-RNase cytotoxicity. PbrS-RNase interacts directly with PbrActin1 in an S-haplotype-independent manner, causing the actin cytoskeleton to depolymerize and promoting programmed cell death in the self-incompatible pollen tube. Proline 156 of PbrS-RNase is essential for the PbrS-RNase-PbrActin1 interaction, and the actin cytoskeleton-depolymerizing function of PbrS-RNase does not require its RNase activity. PbrS-RNase cytotoxicity enhances the expression of phospholipase D (PbrPLDδ1), resulting in increased PA levels in the incompatible pollen tube. PbrPLDδ1-derived PA initially prevents depolymerization of the actin cytoskeleton elicited by PbrS-RNase and delays the SI signaling that leads to pollen tube death. This work provides insights into the orchestration of the S-RNase-based SI response, in which increased PA levels initially play a protective role in incompatible pollen, until sustained PbrS-RNase activity reaches the point of no return and pollen tube growth ceases.

Calcium ions (Ca²⁺) play a crucial role in many key physiological processes; thus, the maintenance of Ca²⁺ homeostasis is of primary importance. Na⁺/Ca²⁺ exchangers (NCXs) play an important role in Ca²⁺ homeostasis in animal excitable cells. Bioinformatic analysis of the Arabidopsis genome suggested the existence of a putative NCX gene, Arabidopsis NCX-like (AtNCL), encoding a protein with an NCX-like structure and different from Ca²⁺/H⁺ exchangers and Na⁺/H⁺ exchangers previously identified in plant. AtNCL was identified to localize in the Arabidopsis cell membrane fraction, have the ability of binding Ca²⁺, and possess NCX-like activity in a heterologous expression system of cultured mammalian CHO-K1 cells. AtNCL is broadly expressed in Arabidopsis, and abiotic stresses stimulated its transcript expression. Loss-of-function atncl mutants were less sensitive to salt stress than wild-type or AtNCL transgenic overexpression lines. In addition, the total calcium content in whole atncl mutant seedlings was higher than that in wild type by atomic absorption spectroscopy. The level of free Ca²⁺ in the cytosol and Ca²⁺ flux at the root tips of atncl mutant plants, as detected using transgenic aequorin and a scanning ion-selective electrode, required a longer recovery time following NaCl stress compared with that in wild type. All of these data suggest that AtNCL encodes a Na⁺/Ca²⁺ exchanger-like protein that participates in the maintenance of Ca²⁺ homeostasis in Arabidopsis. AtNCL may represent a new type of Ca²⁺ transporter in higher plants.

Nitrogen (N) is an important element for plant growth, and a suitable N supply is crucial to ensure optimal yields from fruit trees. Frequently, application of N fertilizers to fruit trees is often excessive, which not only leads to environmental pollution, but also reduces the output from fruit trees through N toxicity. To evaluate the effects of different concentrations of nitrate on plant growth, root-morphological traits, and other nutritional element’s responses in pear, pear seedlings (Pyrus betulifolia Bunge) were treated with five levels of N. Both N-deficiency and an excess of N inhibited the growth and development of pear rootstocks. However, different visible symptoms were observed among treated leaves and roots. Leaf yellowing, the stimulation of root elongation, a decrease in nitrate reductase activity and chlorophyll content were observed under N-deficiency conditions. On the other hand, dark green leaves accompanied by coking, the suppression of root elongation, and a decrease in nitrate reductase activity and chlorophyll content were displayed under regimes of excess N. In addition, not only the N content, but also the content of other mineral nutrients was influenced by nitrate treatments. Taken together, these results suggested that a careful choice of N fertilizer supply is crucial to ensure normal growth and development in pear trees.