SMS2, a Novel Allele of OsINV3, Regulates Grain Size in Rice.

Grain size has an important effect on rice yield. Although several key genes that regulate seed size have been reported in rice, their molecular mechanisms remain unclear. In this study, a rice small grain size 2 (sms2) mutant was identified, and MutMap resequencing analysis results showed that a 2 bp insertion in the second exon of the LOC_Os02g01590 gene resulted in a grain length and width lower than those of the wild-type Teqing (TQ). We found that SMS2 encoded vacuolar acid invertase, a novel allele of OsINV3, which regulates grain size. GO and KEGG enrichment analyses showed that SMS2 was involved in endoplasmic reticulum protein synthesis, cysteine and methionine metabolism, and propionic acid metabolism, thereby regulating grain size. An analysis of sugar content in young panicles showed that SMS2 reduced sucrose, fructose, and starch contents, thus regulating grain size. A haplotype analysis showed that Hap2 of SMS2 had a longer grain and was widely present in indica rice varieties. Our results provide a new theoretical basis for the molecular and physiological mechanisms by which SMS2 regulates grain size.

Toxic effects of cadmium on the physiological and biochemical attributes of plants, and phytoremediation strategies: A review.

Anthropogenic activities pose a more significant threat to the environment than natural phenomena by contaminating the environment with heavy metals. Cadmium (Cd), a highly poisonous heavy metal, has a protracted biological half-life and threatens food safety. Plant roots absorb Cd due to its high bioavailability through apoplastic and symplastic pathways and translocate it to shoots through the xylem with the help of transporters and then to the edible parts via the phloem. The uptake and accumulation of Cd in plants pose deleterious effects on plant physiological and biochemical processes, which alter the morphology of vegetative and reproductive parts. In vegetative parts, Cd stunts root and shoot growth, photosynthetic activities, stomatal conductance, and overall plant biomass. Plants' male reproductive parts are more prone to Cd toxicity than female reproductive parts, ultimately affecting their grain/fruit production and survival. To alleviate/avoid/tolerate Cd toxicity, plants activate several defense mechanisms, including enzymatic and non-enzymatic antioxidants, Cd-tolerant gene up-regulations, and phytohormonal secretion. Additionally, plants tolerate Cd through chelating and sequestering as part of the intracellular defensive mechanism with the help of phytochelatins and metallothionein proteins, which help mitigate the harmful effects of Cd. The knowledge on the impact of Cd on plant vegetative and reproductive parts and the plants' physiological and biochemical responses can help selection of the most effective Cd-mitigating/avoiding/tolerating strategy to manage Cd toxicity in plants.

3D carbonized wood-based integrated electrochemical immunosensor for ultrasensitive detection of procalcitonin antigen.

This work presents a novel 3D carbonized wood-based integrated electrochemical immunosensor for ultrasensitive detection of procalcitonin (PCT) antigen at picogram level, achieving a wide linear detection range for PCT concentrations range from 0.05 to 90 pg mL-1 with a low detection limit of 0.014 pg mL-1 (S/N = 3), outperforming the previous reports. 3D carbonized wood as a new immunosensor matrix is used for electrochemical PCT biosensing, improving the stability of electrode and overcoming the disadvantages of traditional glassy carbon electrode (GCE). It obtained an excellent detection result, due to it has abundant mutual crisscross microchannels that promote the reactants and electrons transfer, greatly amplify the current signal. This novel sandwich-type electrochemical immunosensor is composed of 3D carbonized wood, carboxylic multi-walled carbon nanotube (cMWCNT), Au@Co3O4 core-shell nanosphere and Au/single layer nitrogen-doped graphene (Au/SL-NG), when it is applied for PCT detection in real clinical samples, it exhibits high accuracy same as enzyme-linked immunosorbent assay (ELISA) method.

Preparation of Gas-Responsive Imprinting Hydrogel and Their Gas-Driven Switchable Affinity for Target Protein Recognition.

Novel gas-responsive imprinting hydrogels were fabricated by combining N,N'-dimethylaminoethyl methacrylate gas-sensitive monomers, N,N'-methylenebis(acrylamide) cross-linkers, and human serum albumin (HSA) template proteins via a free radical polymerization. The hydrogel exhibited a reversible gas-responsive property upon N2/CO2 exchange. This result was supported by the evidences from hydrogen nuclear magnetic resonance spectroscopy and scanning electron microscopy. By applying this property to sensing application, a CO2-responsive imprinted biosensor was originally designed on the surface of a glassy carbon electrode. The biosensor exhibited unique self-clean and self-recognition properties toward HSA proteins based on reversible conformational changes driven by N2/CO2 stimuli. Moreover, the proposed imprinted biosensor favored HSA proteins by showing satisfactory sensitivity and selectivity and a wider detection range with a low detection limit. As a rare example in imprint sensing, the biosensor was successfully applied to the HSA extraction from complex serum samples. With gas stimuli, the whole process was efficient, controllable, and harmless to the proteins. Thus, the developed biosensor may provide a new prospect in molecularly imprinted sensing applications.

H2O2-Based Method for Rapid Detection of Transgene-Free Rice Plants from Segregating CRISPR/Cas9 Genome-Edited Progenies.

Genome-editing techniques such as CRISPR/Cas9 have been widely used in crop functional genomics and improvement. To efficiently deliver the guide RNA and Cas9, most studies still rely on Agrobacterium-mediated transformation, which involves a selection marker gene. However, several limiting factors may impede the efficiency of screening transgene-free genome-edited plants, including the time needed to produce each life cycle, the response to selection reagents, and the labor costs of PCR-based genotyping. To overcome these disadvantages, we developed a simple and high-throughput method based on visual detection of antibiotics-derived H2O2 to verify transgene-free genome-edited plants. In transgenic rice containing hygromycin phosphotransferase (HPT), H2O2 content did not change in the presence of hygromycin B (HyB). In contrast, in transgenic-free rice plants with 10-h HyB treatment, levels of H2O2 and malondialdehyde, indicators of oxidative stress, were elevated. Detection of H2O2 by 3,3'-diaminobenzidine (DAB) staining suggested that H2O2 could be a marker to efficiently distinguish transgenic and non-transgenic plants. Analysis of 24 segregating progenies of an HPT-containing rice plant by RT-PCR and DAB staining verified that DAB staining is a feasible method for detecting transformants and non-transformants. Transgene-free genome-edited plants were faithfully validated by both PCR and the H2O2-based method. Moreover, HyB induced overproduction of H2O2 in leaves of Arabidopsis, maize, tobacco, and tomato, which suggests the potential application of the DAB method for detecting transgenic events containing HPT in a wide range of plant species. Thus, visual detection of DAB provides a simple, cheap, and reliable way to efficiently identify transgene-free genome-edited and HPT-containing transgenic rice.

Molecular characterization of the Oncidium orchid HDR gene encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase, the last step of the methylerythritol phosphate pathway.

Two pathways are used by higher plants for the biosynthesis of isoprenoid precursors: the mevalonate pathway in the cytosol and a 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway in the plastids, with 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (HDR) catalyzing the last step in the MEP pathway. In order to understand the contribution of MEP pathway in isoprenoid biosynthesis of Oncidium orchid, a full-length cDNA corresponding to HDR from the flower tissues of Oncidium Gower Ramsey was cloned. The deduced OncHDR amino acid sequence contains a plastid signal peptide at the N-terminus and four conserved cysteine residues. RT-PCR analysis of HDR in Oncidium flowering plants revealed ubiquitous expression in organs and tissues, with preferential expression in the floral organs. Phylogenetic analysis revealed evolutionary conservation of the encoding HDR protein sequence. The genomic sequence of the HDR in Oncidium is similar to that in Arabidopsis, grape, and rice in structure. Successful complementation by OncHDR of an E. coli hdr(-) mutant confirmed its function. Transgenic tobacco carrying the OncHDR promoter-GUS gene fusion showed expression in most tissues, as well as in reproductive organs, as revealed by histochemical staining. Light induced strong GUS expression driven by the OncHDR promoter in transgenic tobacco seedlings. Taken together, our data suggest a role for OncHDR as a light-activated gene.