The rice NUCLEAR FACTOR-YA5 and MICRORNA169a module promotes nitrogen utilization during nitrogen deficiency.

Nitrogen (N) is essential for plant growth and development. Therefore, understanding its utilization is essential for improving crop productivity. However, much remains to be learned about plant N sensing and signaling. Here, rice (Oryza sativa) NUCLEAR FACTOR-YA5 (OsNF-YA5) expression was tightly regulated by N status and induced under N-deficient conditions. Overexpression (OE) of OsNF-YA5 in rice resulted in increased chlorophyll levels and delayed senescence compared to control plants under normal N conditions. Agronomic traits were significantly improved in OE plants and impaired in knockout mutants under N-deficient conditions. Using a dexamethasone-inducible system, we identified the putative targets of OsNF-YA5 that include amino acid, nitrate/peptide transporters, and NITRATE TRANSPORTER 1.1A (OsNRT1.1A), which functions as a key transporter in rice. OsNF-YA5 directly enhanced OsNRT1.1A expression and N uptake rate under N-deficient conditions. Besides, overexpression of OsNF-YA5 also enhanced the expression of GLUTAMINE SYNTHETASE 1/2 (GS1/2) and GLUTAMINE OXOGLUTARATE AMINOTRANSFERASE 1/2 (GOGAT1/2), increasing free amino acid contents under N-deficient conditions. Osa-miR169a expression showed an opposite pattern with OsNF-YA5 depending on N status. Further analysis revealed that osa-miR169a negatively regulates OsNF-YA5 expression and N utilization, demonstrating that an OsNF-YA5/osa-miR169a module tightly regulates rice N utilization for adaptation to N status.

Seed-assembly-mediated fabrication and application of highly branched gold nanoshells having hollow and porous morphologies.

Highly branched gold nanoshells (BAuNSs) having hollow and porous morphologies have been fabricated via a seed-assembly-mediated strategy. Gold seed assemblies can be prepared by removal of SiO2nanotemplates with help of polyvinylpyrrolidone (PVP) molecules, which weakly link gold nanoparticles together even after SiO2etching. L-3,4-dihydroxy phenylalanine (L-DOPA) and AgNO3are employed as shape-directing agents to induce the anisotropic growth of gold. BAuNSs exhibit 7.4 and 4.4 times stronger activities than SiO2@Au nanoparticles in catalysis and surface-enhanced Raman scattering (SERS) applications, respectively, due to their large surface areas and numerous hot spots. It is necessary to find the optimal amount of gold deposition in fabrication to effectively utilize the hollow and porous morpologies of BAuNSs for catalysis and SERS applications. Overgrown nanobranches can fill the nanopores and nanogaps of BAuNSs, resulting in decrease of activities in applications. Overall, the seed-assembly-mediated fabrciation can be employed to produce plasmonic nanostructures having unique morphologies and high application activities.

Rice microRNA171f/SCL6 module enhances drought tolerance by regulation of flavonoid biosynthesis genes.

Plants have evolved sophisticated defense systems to enhance drought tolerance. These include the microRNA (miRNA) group of small noncoding RNAs that act as post-transcriptional regulators; however, details of the mechanisms by which they confer drought tolerance are not well understood. Here, we show that osa-MIR171f, a member of osa-MIR171 gene family, is mainly expressed in response to drought stress and regulates the transcript levels of SCARECROW-LIKE6-I (SCL6-I) and SCL6-II in rice (Oryza sativa). The SCL6 genes are known to be involved in shoot branching and flag leaf morphology. Osa-MIR171f-overexpressing (osa-MIR171f-OE) transgenic plants showed reduced drought symptoms compared with non-transgenic (NT) control plants under both field drought and polyethylene glycol (PEG)-mediated dehydration stress conditions. Transcriptome analysis of osa-MIR171f-OE plants and osa-mir171f-knockout (K/O) lines generated by clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) revealed that osa-mature-miR171a-f (osa-miR171) regulates the expression of flavonoid biosynthesis genes, consequently leading to drought tolerance. This upregulation in the osa-MIR171f-OE plants, which did not occur in NT control plants, was observed under both normal and drought conditions. Our findings indicate that osa-miR171 plays a role in drought tolerance by regulating SCL6-I and SCL6-II transcript levels.

Application of Upstream Open Reading Frames (uORFs) Editing for the Development of Stress-Tolerant Crops.

Global population growth and climate change are posing increasing challenges to the production of a stable crop supply using current agricultural practices. The generation of genetically modified (GM) crops has contributed to improving crop stress tolerance and productivity; however, many regulations are still in place that limit their commercialization. Recently, alternative biotechnology-based strategies, such as gene-edited (GE) crops, have been in the spotlight. Gene-editing technology, based on the clustered regularly interspaced short palindromic repeats (CRISPR) platform, has emerged as a revolutionary tool for targeted gene mutation, and has received attention as a game changer in the global biotechnology market. Here, we briefly introduce the concept of upstream open reading frames (uORFs) editing, which allows for control of the translation of downstream ORFs, and outline the potential for enhancing target gene expression by mutating uORFs. We discuss the current status of developing stress-tolerant crops, and discuss uORF targets associated with salt stress-responsive genes in rice that have already been verified by transgenic research. Finally, we overview the strategy for developing GE crops using uORF editing via the CRISPR-Cas9 system. A case is therefore made that the mutation of uORFs represents an efficient method for developing GE crops and an expansion of the scope of application of genome editing technology.

Characterization of dwarf and narrow leaf (dnl-4) mutant in rice.

Height and leaf morphology are important agronomic traits of the major crop plant rice (Oryza sativa). In previous studies, the dwarf and narrow leaf genes (dnl1, dnl2 and dnl3) have identified in rice. Using the Ac/Ds knockout system, we found a new dwarf and narrow leaf (dnl) mutant and identified mutated gene. The dnl-4 mutant showed reduced plant height and leaf blade width compared to the wild type, and increased leaf inclination. The morphological defects of the mutant were caused by the suppressed expression of the DNL-4 gene, which encodes a pfkB carbohydrate kinase protein. These results suggest that DNL-4 expression is involved in modulating plant height and leaf growth. Furthermore, DNL-4 expression also affects productivity in rice: the dnl-4 mutant exhibited reduced panicle length and grain width compared with the wild type. To understand DNL-4 function in rice, we analyzed the expression levels of leaf growth-related genes, such as NAL1, NAL7, and CSLD4, in the dnl-4 mutant. Expression of NAL1 and NAL7 was downregulated in the dnl-4 mutant compared to the wild type. The observation that DNL-4 expression corresponded with that of NAL1 and NAL7 is consistent with the narrow leaf phenotype of the dnl-4 mutant. These results suggest that DNL-4 regulates plant height and leaf structure in rice.

Photothermal structural modification of porous gold nanoshells via pulsed-laser irradiation: effects of laser wavelengths and surface conditions.

Pulsed lasers are promising candidates for fabricating plasmonic nanoparticles having unique structures, and there is a strong need for studies on the detailed effects of various experimental conditions on laser-induced fabrications. In this work, we demonstrate the effects of laser wavelengths and nanoparticle surface conditions, as well as laser fluences, in the structural modification of porous gold nanoshells induced by picosecond pulse irradiation. Laser wavelengths play a critical role in the modification because irradiating laser pulses excite not only porous gold nanoshells but also gold nanospheres, which have been produced via the melting of irradiated porous gold nanoshells. Significantly different localized surface plasmon resonances of gold nanospheres and porous gold nanoshells make the effect of laser wavelengths noticeable. The polyvinylpyrrolidone (PVP) concentration of colloid containing porous gold nanoshells also modifies the deformation of the structures. Gold nanostructures become positively charged by the irradiation, strengthening gold-PVP attraction. The stronger binding affinity of PVP is considered to reduce the deformation of irradiated porous gold nanoshells.

Laser-induced fabrication of porous gold nanoshells.

Pulse-laser irradiation is a promising approach to fabricate gold nanostructures with unique morphologies. Hollow and porous gold nanoshells with high surface-enhanced Raman scattering efficiency have been produced via irradiating SiO2@Au@SiO2 nanoparticles with nanosecond laser pulses; the synthetic strategy mainly relies on the laser-induced surface melting of gold nanoparticles.