PhyB-dependent phosphorylation of mitogen-activated protein kinase cascade MKK2-MPK2 positively regulates red light-induced stomatal opening.

Red light induces stomatal opening by affecting photosynthesis, metabolism and triggering signal transductions in guard cells. Phytochrome B (phyB) plays a positive role in mediating red light-induced stomatal opening. However, phyB-mediated red light guard cell signalling is poorly understood. Here, we found that phyB-mediated sequential phosphorylation of mitogen-activated protein kinase kinase 2 (MAPKK2, MKK2) and MPK2 in guard cells is essential for red light-induced stomatal opening. Mutations in MKK2 and MPK2 led to reduced stomatal opening in response to white light, and these phenotypes could be observed under red light, not blue light. MKK2 interacted with MPK2 in vitro and in plants. MPK2 was directly phosphorylated by MKK2 in vitro. Red light triggered the phosphorylation of MKK2 in guard cells, and MKK2 phosphorylation was greatly reduced in phyB mutant. Simultaneously, red light-stimulated MPK2 phosphorylation in guard cells was inhibited in mkk2 mutant. Furthermore, mkk2 and mpk2 mutants exhibit significantly smaller stomatal apertures than that of wild type during the stomatal opening stage in the diurnal stomatal movements. Our results indicate that red light-promoted phyB-dependent phosphorylation of MKK2-MPK2 cascade in guard cells is essential for stomatal opening, which contributes to the fine-tuning of stomatal opening apertures under light.

An Improved Method for Monitoring Multiscale Plant Species Diversity of Alpine Grassland Using UAV: A Case Study in the Source Region of the Yellow River, China.

Plant species diversity (PSD) is essential in evaluating the function and developing the management and conservation strategies of grassland. However, over a large region, an efficient and high precision method to monitor multiscale PSD (α-, ß-, and γ-diversity) is lacking. In this study, we proposed and improved an unmanned aerial vehicle (UAV)-based PSD monitoring method (UAVB) and tested the feasibility, and meanwhile, explored the potential relationship between multiscale PSD and precipitation on the alpine grassland of the source region of the Yellow River (SRYR), China. Our findings showed that: (1) UAVB was more representative (larger monitoring areas and more species identified with higher α- and γ-diversity) than the traditional ground-based monitoring method, though a few specific species (small in size) were difficult to identify; (2) UAVB is suitable for monitoring the multiscale PSD over a large region (the SRYR in this study), and the improvement by weighing the dominance of species improved the precision of α-diversity (higher R 2 and lower P values of the linear regressions); and (3) the species diversity indices (α- and ß-diversity) increased first and then they tended to be stable with the increase of precipitation in SRYR. These findings conclude that UAVB is suitable for monitoring multiscale PSD of an alpine grassland community over a large region, which will be useful for revealing the relationship of diversity-function, and helpful for conservation and sustainable management of the alpine grassland.

RIC7 plays a negative role in ABA-induced stomatal closure by inhibiting H2O2 production.

When plants encounter environmental stresses, phytohormone abscisic acid (ABA) accumulates quickly and efficiently reduces water loss by inducing stomatal closure. Reactive oxygen species (ROS) is an important regulator in ABA-induced stomatal closure, and ROS generation is modulated by multiple components in guard-cell ABA signaling. ROP interactive CRIB-containing protein 7 (RIC7) has been found to negatively regulate ABA-induced stomatal closure. However, the molecular details of the RIC7 function in this process are unclear. Here, by using two RIC7 overexpressing mutants, we confirmed the negative role of RIC7 in ABA-induced stomatal closure and found that guard cells of RIC7 overexpressing mutants generated less H2O2 than the wild type with ABA treatment, which were consistent with the reduced expression levels of ROS generation related NADPH oxidase genes AtRBOHD and AtRBOHF, and cytosolic polyamine oxidase genes PAO1 and PAO5 in the RIC7 overexpressing mutants. Furthermore, external applied H2O2 failed to rescue the defects of stomatal closure in RIC7 overexpressing mutants. These results suggest that RIC7 affects H2O2 generation in guard cells, and the function of H2O2 is dependent on RIC7 in ABA-induced stomatal closure, indicative of interdependency between RIC7 and H2O2 in ABA guard-cell signaling.

SnRK2.6 interacts with phytochrome B and plays a negative role in red light-induced stomatal opening.

Light is an important environmental factor for plant growth and development. Phytochrome B (phyB), a classical red/far-red light receptor, plays vital role in controlling plant photomorphogenesis and light-induced stomatal opening. Phytohormone abscisic acid (ABA) accumulates rapidly and triggers a series of physiological and molecular events during the responses to multiple abiotic stresses. Recent studies showed that phyB mutant synthesizes more ABA and exhibits improved tolerance to salt and cold stress, suggesting that a crosstalk exists between light and ABA signaling pathway. However, whether ABA signaling components mediate responses to light remains unclear. Here, we showed that SnRK2.6 (Sucrose Nonfermenting 1-Related Protein Kinase 2.6), a key regulator in ABA signaling, interacts with phyB and participates in light-induced stomatal opening. First, we checked the interaction between phyB and SnRK2s, and found that SnRK2.2/2.3/2.6 kinases physically interacted with phyB in yeast and in vitro. We also performed co-IP assay to support that SnRK2.6 interacts with phyB in plant. To investigate the role of SnRK2.6 in red light-induced stomatal opening, we obtained the snrk2.6 mutant and overexpression lines, and found that snrk2.6 mutant exhibited a significantly larger stomatal aperture under red light treatment, while the two independent overexpression lines showed significantly smaller stomatal aperture, indicative of a negative role for SnRK2.6 in red light-induced stomatal opening. The interaction of SnRK2.6 with red light receptor and the negative role of SnRK2.6 in red light-induced stomatal opening provide new evidence for the crosstalk between ABA and red light in guard cell signaling.