The transcription factor HBF1 directly activates expression of multiple flowering time repressors to delay rice flowering.

Flowering time (or heading date) is an important agronomic trait that determines the environmental adaptability and yield of many crops, including rice (Oryza sativa L.). Hd3a BINDING REPRESSOR FACTOR 1 (HBF1), a basic leucine zipper transcription factor, delays flowering by decreasing the expression of Early heading date 1 (Ehd1), Heading date 3a (Hd3a), and RICE FLOWERING LOCUS T 1 (RFT1), but the underlying molecular mechanisms have not been fully elucidated. Here, we employed the hybrid transcriptional factor (HTF) strategy to enhance the transcriptional activity of HBF1 by fusing it to four copies of the activation domain from Herpes simplex virus VP16. We discovered that transgenic rice lines overexpressing HBF1-VP64 (HBF1V) show significant delays in time to flower, compared to lines overexpressing HBF1-MYC or wild-type plants, via the Ehd1-Hd3a/RFT1 pathway, under both long-day and short-day conditions. Transcriptome deep sequencing analysis indicated that 19 WRKY family genes are upregulated in the HBF1V overexpression line. We demonstrate that the previously unknown gene, OsWRKY64, is a direct downstream target of HBF1 and represses flowering in rice, whereas three known flowering repressor genes, Days to heading 7 (DTH7), CONSTANS 3 (OsCO3), and OsWRKY104, are also direct target genes of HBF1 in flowering regulation. Taking these results together, we propose detailed molecular mechanisms by which HBF1 regulates the time to flower in rice. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-023-00107-7.

OsBIC1 Directly Interacts with OsCRYs to Regulate Leaf Sheath Length through Mediating GA-Responsive Pathway.

Cryptochrome 1 and 2 (CRY1 and CRY2) are blue light receptors involved in the regulation of hypocotyl elongation, cotyledon expansion, and flowering time in Arabidopsisthaliana. Two cryptochrome-interacting proteins, Blue-light Inhibitor of Cryptochrome 1 and 2 (BIC1 and BIC2), have been found in Arabidopsis. BIC1 plays critical roles in suppressing the physiological activities of CRY2, which include the blue light-dependent dimerization, phosphorylation, photobody formation, and degradation process, but the functional characterization of BIC protein in other crops has not yet been performed. To investigate the function of BIC protein in rice (Oryza sativa), two homologous genes of Arabidopsis BIC1 and BIC2, namely OsBIC1 and OsBIC2 (OsBICs), were identified. The overexpression of OsBIC1 and OsBIC2 led to increased leaf sheath length, whereas mutations in OsBIC1 displayed shorter leaf sheath in a blue light intensity-dependent manner. OsBIC1 regulated blue light-induced leaf sheath elongation through direct interaction with OsCRY1a, OsCRY1b, and OsCRY2 (OsCRYs). Longitudinal sections of the second leaf sheath demonstrated that OsBIC1 and OsCRYs controlled leaf sheath length by influencing the ratio of epidermal cells with different lengths. RNA-sequencing (RNA-seq) and quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) analysis further proved that OsBIC1 and OsCRYs regulated similar transcriptome changes in regulating Gibberellic Acids (GA)-responsive pathway. Taken together, these results suggested that OsBIC1 and OsCRYs worked together to regulate epidermal cell elongation and control blue light-induced leaf sheath elongation through the GA-responsive pathway.

A sensitive and rapid bacterial antibiotic susceptibility test method by surface enhanced Raman spectroscopy.

This study aims to develop a rapid bacterial antibiotic susceptibility test (AST) method by Bacteria-aptamer@AgNPs-surface enhanced Raman spectroscopy (SERS) and further evaluate the influence of different antibiotics on the Raman intensity of bacteria. The Raman intensity of Escherichia coli O157:H7 (E. coli O157:H7) and Staphylococcus aureus (S. aureus) in the presence of different concentrations of antibiotics in 2 h was detected by Bacteria-aptamer@AgNPs-SERS in this study. Our results found that the bacteria Raman signal peak at 735 cm-1 and the minimum inhibitory concentration (MIC) value was determined in 1 h according to Raman signals. In 2 h, the bacteria Raman signal growth at sub-MIC concentrations of four different kinds of antibiotics and the bacteria colony-forming unit (CFU) have similar enhancements. SERS utilizes special functions of rough metal surfaces and offers a huge enhancement of Raman intensities with reduced fluorescence backgrounds, which makes it an ultrasensitive tool of detection. This rapid AST method and the enhancement effect should be of value in search of new antibiotic drugs.

Biodegradable and Multifunctional Microspheres for Treatment of Hepatoma through Transarterial Embolization.

To improve the effectiveness of cancer treatment, this study aimed to develop biodegradable microspheres and to combine chemotherapy and radiotherapy via transcatheter arterial embolization/chemoembolization (TAE/TACE) with local radiation therapy for the slow release of chemotherapeutic agents. Microparticles were prepared by double emulsification using a biodegradable and biocompatible polymer Poly(D,l-lactide-co-glycolide) (PLGA). Since the microspheres contain a water-soluble Poly(vinylsulfonic acid) (PVSA) solution, the functional groups of this polymer dissociate into -SO3- in water. The positively charged doxorubicin can be loaded into beads, ensuring slow release. After 188Retin colloids were added into the microspheres, TACE was performed in a rat hepatocellular carcinoma model. Single photon emission computed tomography/computed tomography imaging and biodistribution analyses showed that the microspheres were still in the liver after 72 h. During 4 weeks of observation, ultrasound images showed that the Re/DOX@MS treatment had the most significant inhibitory effect on tumor growth. Biodegradable PLGA microspheres have the advantage of enabling local embolization therapy with reduced adverse effects. In the future, microspheres could serve as a drug delivery system for cancer treatment by combining therapeutic radionuclides and chemotherapeutic drugs, thereby improving treatment effects for hepatocellular carcinoma.

The OsABF1 transcription factor improves drought tolerance by activating the transcription of COR413-TM1 in rice.

Water deprivation causes substantial losses in crop yields around the world. In this study, we show that when overexpressed in transgenic rice (Oryza sativa), the bZIP transcription factor OsABF1 confers distinctly different drought-tolerance phenotypes when tethered to the transcriptional activator VP16 versus the transcriptional repressor EAR. We performed chromatin immunoprecipitation sequencing (ChIP-seq) and RNA sequencing (RNA-seq) assays on transgenic rice lines and determined that OsABF1 binds to DNA sequences containing an ACGT core motif. Analysis of the overlap between the RNA-sequencing and chromatin immunoprecipitation-sequencing data identified 242 OsABF1 target genes involved in multiple aspects of the drought response. Overexpression of one of these genes, COR413-TM1, which encodes a putative thylakoid membrane protein, resulted in a drought-tolerance phenotype without obvious side effects. In addition, OsABF1 directly regulates the expression of the protein phosphatase 2C (OsPP48 and OsPP108) and bZIP (OsbZIP23, OsbZIP46, and OsbZIP72) genes, thus forming a complex feedback circuit in the drought/abscisic acid signaling pathway.

OsMPH1 regulates plant height and improves grain yield in rice.

Plant height is a major trait affecting yield potential in rice. Using a large-scale hybrid transcription factor approach, we identified the novel MYB-like transcription factor OsMPH1 (MYB-like gene of Plant Height 1), which is involved in the regulation of plant height in rice. Overexpression of OsMPH1 leads to increases of plant height and grain yield in rice, while knockdown of OsMPH1 leads to the opposite phenotypes. Microscopy of longitudinal stem sections indicated that a change in internode cell length resulted in the change in plant height. RNA sequencing (RNA-seq) analysis of transgenic rice lines showed that multiple genes related to cell elongation and cell wall synthesis, which are associated with plant height and yield phenotypes, exhibited an altered expression profile. These results imply that OsMPH1 might be involved in specific recognition and signal transduction processes related to plant height and yield formation, providing further insights into the mechanisms underlying the regulation of plant height and providing a candidate gene for the efficient improvement of rice yield.

A Drought-Inducible Transcription Factor Delays Reproductive Timing in Rice.

The molecular mechanisms underlying photoperiod or temperature control of flowering time have been recently elucidated, but how plants regulate flowering time in response to other external factors, such as water availability, remains poorly understood. Using a large-scale Hybrid Transcription Factor approach, we identified a bZIP transcriptional factor, O. sativa ABA responsive element binding factor 1 (OsABF1), which acts as a suppressor of floral transition in a photoperiod-independent manner. Simultaneous knockdown of both OsABF1 and its closest homologous gene, OsbZIP40, in rice (Oryza sativa) by RNA interference results in a significantly earlier flowering phenotype. Molecular and genetic analyses demonstrate that a drought regime enhances expression of the OsABF1 gene, which indirectly suppresses expression of the Early heading date 1 (Ehd1) gene that encodes a key activator of rice flowering. Furthermore, we identified a drought-inducible gene named OsWRKY104 that is under the direct regulation of OsABF1 Overexpression of OsWRKY104 can suppress Ehd1 expression and confers a later flowering phenotype in rice. Together, these findings reveal a novel pathway by which rice modulates heading date in response to the change of ambient water availability.