Ultraviolet metalens for photoacoustic microscopy with an elongated depth of focus.

Ultraviolet photoacoustic microscopy (UV-PAM) can achieve in vivo imaging without exogenous markers and play an important role in pathological diagnosis. However, traditional UV-PAM is unable to detect enough photoacoustic signals due to the very limited depth of focus (DOF) of excited light and the sharp decrease in energy with increasing sample depth. Here, we design a millimeter-scale UV metalens based on the extended Nijboer-Zernike wavefront-shaping theory which can effectively extend the DOF of a UV-PAM system to about 220 µm while maintaining a good lateral resolution of 1.063 µm. To experimentally verify the performance of the UV metalens, a UV-PAM system is built to achieve the volume imaging of a series of tungsten filaments at different depths. This work demonstrates the great potential of the proposed metalens-based UV-PAM in the detection of accurate diagnostic information for clinicopathologic imaging.

SPL9 mediates freezing tolerance by directly regulating the expression of CBF2 in Arabidopsis thaliana.

BACKGROUND: Freezing stress inhibits plant development and causes significant damage to plants. Plants therefore have evolved a large amount of sophisticated mechanisms to counteract freezing stress by adjusting their growth and development correspondingly. Plant ontogenetic defense against drought, high salt, and heat stresses, has been extensively studied. However, whether the freezing tolerance is associated with ontogenetic development and how the freezing signals are delivered remain unclear. RESULTS: In this study, we found that the freezing tolerance was increased with plant age at the vegetative stage. The expressions of microRNA156 (miR156) and SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 9 (SPL9), playing roles in regulation of ontogenetic development, were induced by cold stress. Overexpression of SPL9 (rSPL9) promoted the expression of C-REPEAT BINDING FACTOR 2 (CBF2) and hereafter enhanced the freezing tolerance. Genetic analysis indicated that the effect of rSPL9 on freezing tolerance is partially restored by cbf2 mutant. Further analysis confirmed that SPL9 directly binds to the promoter of CBF2 to activate the expression of CBF2, and thereafter increased the freezing tolerance. CONCLUSIONS: Therefore, our study uncovers a new role of SPL9 in fine-tuning CBF2 expression and thus mediating freezing tolerance in plants, and implies a role of miR156-SPL pathway in balancing the vegetative development and freezing response in Arabidopsis.

The miR159-MYB33-ABI5 module regulates seed germination in Arabidopsis.

Drought stress restricts crop productivity and exacerbates food shortages. The plant hormone, abscisic acid (ABA), has been shown to be a pivotal player in the regulation of drought tolerance and seed germination in plants. ABA accumulates under abiotic stresses to promote miR159 expression. miR159 is an ancient and conserved plant miRNA that plays diverse roles in plant development, seed germination, and drought response in Arabidopsis. Our previous studies demonstrated that miR159 regulates the vegetative phase change by repressing the ABI5 activation and thereafter preventing hyperactivation of miR156. However, whether the miR159-MYB33-ABI5 module plays a role in seed germination and drought response, and if so, how they interact genetically, remain largely unexplored. Here, we show that loss-of-function of miR159 (mir159ab) confers enhanced drought tolerance and hypersensitivity of seed germination to ABA. Genetic analyses demonstrated that loss-of-function mutation in the ABI5 gene suppresses the hypersensitivity of mir159ab to ABA, and the insensitivity of myb33 seeds to ABA treatment is ABI5 dependent. ABI5 functions downstream of MYB33 and miR159 in response to ABA. Therefore, our results uncover a new role for the miR159-MYB33-ABI5 module in the regulation of drought response and seed germination in plants.

Function of Nuclear Pore Complexes in Regulation of Plant Defense Signaling.

In eukaryotes, the nucleus is the regulatory center of cytogenetics and metabolism, and it is critical for fundamental biological processes, including DNA replication and transcription, protein synthesis, and biological macromolecule transportation. The eukaryotic nucleus is surrounded by a lipid bilayer called the nuclear envelope (NE), which creates a microenvironment for sophisticated cellular processes. The NE is perforated by the nuclear pore complex (NPC), which is the channel for biological macromolecule bi-directional transport between the nucleus and cytoplasm. It is well known that NPC is the spatial designer of the genome and the manager of genomic function. Moreover, the NPC is considered to be a platform for the continual adaptation and evolution of eukaryotes. So far, a number of nucleoporins required for plant-defense processes have been identified. Here, we first provide an overview of NPC organization in plants, and then discuss recent findings in the plant NPC to elaborate on and dissect the distinct defensive functions of different NPC subcomponents in plant immune defense, growth and development, hormone signaling, and temperature response. Nucleoporins located in different components of NPC have their unique functions, and the link between the NPC and nucleocytoplasmic trafficking promotes crosstalk of different defense signals in plants. It is necessary to explore appropriate components of the NPC as potential targets for the breeding of high-quality and broad spectrum resistance crop varieties.

Cold Stress Response Mechanisms in Anther Development.

Unlike animals that can escape threats, plants must endure and adapt to biotic and abiotic stresses in their surroundings. One such condition, cold stress, impairs the normal growth and development of plants, in which most phases of reproductive development are particularly susceptible to external low temperature. Exposed to uncomfortably low temperature at the reproductive stage, meiosis, tapetal programmed cell death (PCD), pollen viability, and fertilization are disrupted, resulting in plant sterility. Of them, cold-induced tapetal dysfunction is the main cause of pollen sterility by blocking nutrition supplements for microspore development and altering their timely PCD. Further evidence has indicated that the homeostatic imbalances of hormones, including abscisic acid (ABA) and gibberellic acid (GA), and sugars have occurred in the cold-treated anthers. Among them, cold stress gives rise to the accumulation of ABA and the decrease of active GA in anthers to affect tapetal development and represses the transport of sugar to microspores. Therefore, plants have evolved lots of mechanisms to alleviate the damage of external cold stress to reproductive development by mainly regulating phytohormone levels and sugar metabolism. Herein, we discuss the physiological and metabolic effects of low temperature on male reproductive development and the underlying mechanisms from the perspective of molecular biology. A deep understanding of cold stress response mechanisms in anther development will provide noteworthy references for cold-tolerant crop breeding and crop production under cold stress.

ABI5 acts downstream of miR159 to delay vegetative phase change in Arabidopsis.

Vegetative development constitutes a critical phase in plant development, and it is regulated by an evolutionarily conserved miR156-SPL pathway. Previous studies have shown that miR159 acts to prevent the hyperactivation of miR156 to regulate the timing of vegetative phase change in Arabidopsis. However, whether miR159 integrates into the abscisic acid (ABA) signaling pathway to control vegetative phase change remains unexplored, since miR159 also plays an important regulatory role in ABA response. Here, we show that the expression of ABI5 (ABA INSENSITIVE5), a crucial regulator in the ABA signaling pathway, is significantly elevated in the loss-of-function mutant of miR159 (mir159ab). Loss of function in ABI5 (abi5) promotes juvenile-to-adult transition, whereas overexpression of ABI5 delays this transition under short-day conditions. Genetic analyses indicated that the effect of mir159ab on vegetative phase change is ABI5 dependent. Further analysis confirmed that MYB33, a major target of miR159, promotes the transcription of ABI5 by directly binding to its promoter. ABI5 functions upstream of miR156 to promote juvenile development by affecting the expression of genes in the miR156-SPL pathway. Therefore, our study uncovers a new role of ABI5 in vegetative development in plants, and implies a role of ABA signaling in vegetative development in Arabidopsis.

Repression of miR156 by miR159 Regulates the Timing of the Juvenile-to-Adult Transition in Arabidopsis.

Temporally regulated microRNAs have been identified as master regulators of developmental timing in both animals and plants. In plants, vegetative development is regulated by a temporal decrease in miR156 level, but how this decreased expression is initiated and then maintained during shoot development remains elusive. Here, we show that miR159 is required for the correct timing of vegetative development in Arabidopsis thaliana Loss of miR159 increases miR156 level throughout shoot development and delays vegetative development, whereas overexpression of miR159 slightly accelerated vegetative development. The repression of miR156 by miR159 is predominantly mediated by MYB33, an R2R3 MYB domain transcription factor targeted by miR159. Loss of MYB33 led to subtle precocious vegetative phase change phenotypes in spite of the significant downregulation of miR156. MYB33 simultaneously promotes the transcription of MIR156A and MIR156C, as well as their target, SPL9, by directly binding to the promoters of these three genes. Rather than acting as major players in vegetative phase change in Arabidopsis, our results suggest that miR159 and MYB33 function as modifiers of vegetative phase change; i.e., miR159 facilitates vegetative phase change by repressing MYB33 expression, thus preventing MYB33 from hyperactivating miR156 expression throughout shoot development to ensure correct timing of the juvenile-to-adult transition in Arabidopsis.

The Important Function of Mediator Complex in Controlling the Developmental Transitions in Plants.

Developmental transitions in plants are tightly associated with changes in the transcriptional regulation of gene expression. One of the most important regulations is conferred by cofactors of RNA polymerase II including the mediator complex, a large complex with a modular organization. The mediator complex recruits transcription factors to bind to the specific sites of genes including protein-coding genes and non-coding RNA genes to promote or repress the transcription initiation and elongation using a protein-protein interaction module. Mediator complex subunits have been isolated and identified in plants and the function of most mediator subunits in whole life cycle plants have been revealed. Studies have shown that the Mediator complex is indispensable for the regulation of plant developmental transitions by recruiting age-, flowering-, or hormone-related transcription factors. Here, we first overviewed the Mediator subunits in plants, and then we summarized the specific Mediator subunits involved in developmental transitions, including vegetative phase change and floral transition. Finally, we proposed the future directions to further explore their roles in plants. The link between Mediator subunits and developmental transitions implies the necessity to explore targets of this complex as a potential application in developing high quality crop varieties.

Crop Pollen Development under Drought: From the Phenotype to the Mechanism.

Drought stress induced pollen sterility is a harmful factor that reduces crop yield worldwide. During the reproductive process, the meiotic stage and the mitotic stage in anthers are both highly vulnerable to water deficiency. Drought at these stages causes pollen sterility by affecting the nature and structure of the anthers, including the degeneration of some meiocytes, disorientated microspores, an expanded middle layer and abnormal vacuolizated tapeta. The homeostasis of the internal environment is imbalanced in drought-treated anthers, involving the decreases of gibberellic acid (GA) and auxin, and the increases of abscisic acid (ABA), jasmonic acid (JA) and reactive oxygen species (ROS). Changes in carbohydrate availability, metabolism and distribution may be involved in the effects of drought stress at the reproductive stages. Here, we summarize the molecular regulatory mechanism of crop pollen development under drought stresses. The meiosis-related genes, sugar transporter genes, GA and ABA pathway genes and ROS-related genes may be altered in their expression in anthers to repair the drought-induced injures. It could also be that some drought-responsive genes, mainly expressed in the anther, regulate the expression of anther-related genes to improve both drought tolerance and anther development. A deepened understanding of the molecular regulatory mechanism of pollen development under stress will be beneficial for breeding drought-tolerant crops with high and stable yield under drought conditions.

Silencing of miR156 confers enhanced resistance to brown planthopper in rice.

MAIN CONCLUSION: Silencing of miR156 in rice confers enhanced resistance to brown planthopper through reducing JA and JA-Ile biosynthesis. Rice brown planthopper (BPH, Nilaparvata lugens Stål) threatens the sustainability of rice production and global food security. Due to the rapid adaptation of BPH to current germplasms in rice, development of novel types of resistant germplasms becomes increasingly important. Plant ontogenetic defense against pathogen and herbivores offers a broad spectrum and durable resistance, and has been experimentally tested in many plants; however, the underlying molecular mechanism remains unclear. miR156 is the master regulator of ontogeny in plants; modulation of miR156 is, therefore, expected to cause corresponding changes in BPH resistance. To test this hypothesis, we silenced miR156 using a target mimicry method in rice, and analyzed the resistance of miR156-silenced plants (MIM156) to BPH. MIM156 plants exhibited enhanced resistance to BPH based on analyses of honeydew excretion, nymph survival, fecundity of BPH, and the survival ratio of rice plants after BPH infestation. Molecular analysis indicated that the expression of MPK3, MPK6, and WRKY70, three genes involved in BPH resistance and jasmonic acid (JA) signaling, was altered in MIM156 plants. The JA and bioactive jasmonoyl-isoleucine levels and the expression of genes involved in JA biosynthesis were significantly reduced in MIM156 plants. Restoration of JA level by exogenous application increased the number of BPH feeding on MIM156 plants and reduced its resistance to BPH. Our findings suggest that miR156 negatively regulates BPH resistance by increasing JA level in rice; therefore, modulation of miR156-SPLs' pathway may offer a promising way to breed rice varieties with enhanced resistance against BPH and elite agronomically important traits.