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Rice is the primary staple food for more than 50% of the world’s population. Narrowing the gap between the maximum potential food crop yield and actual yield is critical for improving the current yield, resource use efficiency, and global food security. Here, we examined the fertilizer use efficiency (FUE), radiation use efficiency (RUE), and temperature production efficiency (TPE) of four management treatments (super high yield [SH], high yield and high efficiency [HH], farmer level [FP], and basic production level [CK]). SH and HH treatments significantly reduced the yield gap by 22.4 and 9.5%, respectively. The large yield gap between HH and FP was mainly attributed to high specific leaf weight at the jointing stage (7.5–7.7 mg·cm−2), and the high leaf area maintained during tillering–jointing stages (35.4–37.6 m2·m−2·per day). Compared with FP, HH increased the specific leaf weight in the heading stage (8.2–8.4 mg·cm−2), relative crop growth rate, net assimilation rate (NAR), and mean leaf area index (> 2.6). Moreover, compared with FP, HH significantly increased partial factor productivity (PFP) of nitrogen, FUE, TPE, and RUE owing to greater yield and NAR after the full heading stage. Although the HH yield was 93.32% that of SH, HH increased PFP of fertilizer (12.5%), fertilizer nitrogen (9.07%), and fertilizer K2O (36.34%), and required 26% less fertilizer than SH. The findings of this study could facilitate high-efficiency rice production and bridging of yield gaps.

To explore suitable synthesis parameters of efficient graft copolymerization on wool, the disulfide bonds in wool chains were reduced to sulfhydryl groups by the treatment of thioglycolic acid to wool, and then wool-g-poly(glycidyl methacrylate) was prepared by graft copolymerization of wool with glycidyl methacrylate by building a redox system between sulfhydryl groups and ammonium persulfate. The wool-g-poly(glycidyl methacrylate) was characterized by the analyses of Fourier transform infrared spectroscopy, scanning electron microscopy, thermogravimetric and x-ray diffraction. Influences of molar concentrations of monomer and initiator as well as reaction temperature on grafting ratios were investigated. The experimental results showed that the sulfhydryl groups/ammonium persulfate redox system built could effectively initiate graft polymerization of lipophilic glycidyl methacrylate on the wool surface in an aqueous medium. The suitable process parameters of graft copolymerization were: 0.6 mmol/L molar concentration of glycidyl methacrylate, 3.0 mmol/L molar concentration of ammonium persulfate, and reaction temperature of 50 ℃. At the suitable parameters, the grafting ratio of wool-g-poly(glycidyl methacrylate) showed a maximum value of 195.8 %, indicating an efficient graft copolymerization was achieved. The copolymerization could reduce the degree of crystallinity of wool. The wool-g-poly(glycidyl methacrylate) had higher thermal stability than raw wool at a temperature of below 406 ℃ by performing the thermogravimetric analysis. Based on the results, we successfully built new sulfhydryl groups on the wool/ammonium persulfate redox system, prepared the wool-g-poly(glycidyl methacrylate), and this work will lay an important foundation for the application of wool-g-poly(glycidyl methacrylate) in the textile field.

Objectives. The study is aimed at exploring the effect of the controlled release of the glial-derived neurotrophic factor (GDNF) on nerve regeneration. Methods. The PLGA/chitosan composite nerve conduit was used to bridge the dissected trunk of the rat facial nerve. GDNF microcapsules were loaded into the nerve conduit. Nine weeks after surgery, the facial nerve zygomatic and buccal branches were labeled with fluorescent indicators. The incorrectly grown facial neurons were reversed and counted. The facial nerve functional recovery was assessed by measuring the maximum evoked potential. Results. The nerve conduit was filled with different regenerating factors, such as the GDNF, GDNF microcapsules, or saline (control). The number of incorrectly regenerated neurons was lower with the nerve conduits filled with GDNF microcapsules than with those supplied with just the GDNF. However, neither the GDNF nor GDNF microcapsules affected the number of regenerated neurons. The functional recovery of the facial nerve was the best, with the nerve conduit filled with GDNF microcapsules closest to the healthy uncut facial nerve. Conclusion. The stable slow-release GNDF microcapsule inside the biodegradable nerve conduit can reduce the extent of incorrect growth of the facial nerve neuron when bridging the dissected rat facial nerve trunk. The technique has a good effect on functional nerve recovery.

qCT7, a novel QTL for increasing seedling cold tolerance in rice, was fine-mapped to a 70.9-kb region on chromosome 7, and key OsWRKY115 was identified in transgenic plants. Cold stress caused by underground cold-water irrigation seriously limits rice productivity. We systemically measured the cold-responsive traits of 2,570 F2 individuals derived from two widely cultivated rice cultivars, Kong-Yu-131 and Dong-Nong-422, to identify the major genomic regions associated with cold tolerance. A novel major QTL, qCT7, was mapped on chromosome 7 associated with the cold tolerance and survival, using whole-genome re-sequencing with bulked segregant analysis. Local QTL linkage analysis with F

The Green Revolution gene sd1 has been used extensively in modern rice breeding, especially in indica cultivars. However, elite sd1 alleles and related germplasm resources used for japonica rice breeding have not been identified, and extensive efforts are needed for japonica rice breeding to obtain new dwarfing sources. Data from MBKbase-Rice revealed seven sd1 haplotypes in indica and four in japonica rice. Two new sd1 alleles were identified in indica rice. In 295 japonica accessions from northeast Asia, except for the weak functional allele SD1-EQ, sd1-r was the major allele, reducing plant height in comparison with SD1-EQ. Japonica germplasm resources carrying reported sd1 alleles were identified by genotype searching and further verified by literature search, genealogical analysis, and dCaps markers. Pedigrees and geographic distribution showed that sd1-r is an excellent allele widely used in northern China and Tohoku in Japan, and sd1-j is commonly used in east China and Kyushu in Japan. Dongnong- and Xiushui-series cultivars carrying sd1-r and sd1-j, respectively, are essential branches of the backbone parents of Chinese japonica rice, Akihikari and Ce21, with the largest number of descendants and derived generations. In semi-dwarf japonica rice breeding, sd1-d was introgressed into Daohuaxiang 2 (DHX2). Dwarf and semi-dwarf lines carrying sd1-d were selected and designated as 1279 and 1280, respectively, after withstanding typhoon-induced strong winds and heavy rains in 2020, and are anticipated to become useful intermediate materials for future genetic research and breeding. This work will facilitate the introduction, parental selection, and marker-assisted breeding, and provide a material basis for the next step in identifying favorable genes that selected together with the sd1 alleles in japonica backbone parents.

Background Low-temperature chilling is a major abiotic stress leading to reduced rice yield and is a significant environmental threat to food security. Low-temperature chilling studies have focused on physiological changes or coding genes. However, ceRNA mechanism in rice at low temperatures has not been reported. Therefore, to establish the ceRNA network and elucidate the transcriptional regulatory network and physiological mechanism of rice leaves in response to low-temperature stress, experiments were conducted using two rice (Oryza sativa. L) varieties with significantly different cold tolerances. Results The cold-tolerant variety has prolonged and relatively stable regulation of antioxidant enzymes (superoxide dismutase [SOD] and peroxidase [POD] catalase [CAT], and total antioxidant capacity) and osmotic substances (Proline [PRO], Soluble sugars [SS], and Soluble protein [SP]), which can remove harmful substances to maintain plant cell stability. Further screening based on VIP (variable importance in the projection) identified SOD and POD as two of the most essential antioxidant phenotypes. Furthermore, by combining antioxidant physiological indices with whole transcriptome data through weighted gene co-expression network analysis (WGCNA), the highly correlated modules were black and green, significantly associated with critical antioxidant indices (SOD and POD). Enrichment analysis revealed that black module genes were significantly enriched in the redox pathway. This module hub gene included UDP-glucosyltransferase family protein (Os05g0527000), sesquiterpene synthase (Os08g0167800), indole-3-glycerophosphatase gene (Os03g0797500), encoding oxidoreductase gene (Os04g0339400), and unknown genes (Os05g0212900). The green module genes were significantly enriched in the linoleic acid metabolic pathway. This module hub gene included WRKY transcription factor (Os11g0685700),abscisic acid (ABA) signal transduction pathway-related gene plasma membrane hydrogen-ATPase (Os02g0825600), Beta-Ig-containing H3 structural domain protein leucine-rich repeat-like receptor kinase (Os02g0615800), and two unknown genes (Os03g0103950 and Os08g0288050). Therefore, we selected module hub and significantly enriched pathway genes to construct key competing endogenous ceRNA networks. Both modules’ networks were integrated to obtain the ceRNA network of six lncRNAs regulating three co-expressed mRNAs through four miRNAs and six lncRNAs regulating two co-expressed mRNAs through four miRNAs. Finally, we performed preliminary gene sequence difference analysis, subcellular localization, and phenotypic validation of mutants for the putative glutamate protein OsGLR1.2 (Os02g0787600), which was previously thought to be associated with Ca2+ transport. Conclusions The results of this study reveal the characteristics of the response of rice to low temperature and provide insight into the mechanisms of regulation of rice to low temperature.

As one of the most important part of the ideal plant type of japonica rice, leaf shape affects the photosynthesis and carbohydrate accumulation directly. Mining and using new leaf shape related genes/QTLs can further enrich the theory of molecular breeding and accelerate the breeding process of japonica rice. In the present study, 2 RILs and a natural population with 295 japonica rice varieties were used to map QTLs for flag leaf length (FL), flag leaf width (FW) and flag leaf area (FLA) by linkage analysis and genome-wide association study (GWAS) through 2 years. A total of 64 QTLs were detected by 2 ways, and pleiotropic QTLs qFL2 (Chr2_33,332,579) and qFL10 (Chr10_10,107,835; Chr10_10,230,100) consisted of overlapping QTLs mapped by linkage analysis and GWAS through 2 years were identified. The candidate genes LOC_Os02g54254, LOC_Os02g54550, LOC_Os10g20160, LOC_Os10g20240, LOC_Os10g20260 were obtained, filtered by linkage disequilibrium (LD), and haplotype analysis. LOC_Os10g20160 (SD-RLK-45) showed outstanding characteristics in quantitative real-time PCR (qRT-PCR) analysis in leaf development period, belongs to S-domain receptor-like protein kinases gene and probably to be a main gene regulating flag leaf width of japonica rice. The results of this study provide valuable resources for mining the main genes/QTLs of japonica rice leaf development and molecular breeding of japonica rice ideal leaf shape.

The C (Cys) 2H (His) 2-type transcription factor is one of the most important transcription factors in plants and plays a regulatory role in the physiological responses of rice to abiotic stresses. A novel rice C2H2-type zinc finger protein, abscisic acid (ABA)-drought-reactive oxygen species (ROS) 3 (OsADR3), was found to confer drought stress tolerance by enhancing antioxidant defense and regulating OsGPX1. Overexpression of OsADR3 in rice increased tolerance to drought stress by increasing ROS scavenging ability and ABA sensitivity. In contrast, CRISPR/Cas9-mediated knockout of osadr3 increased the sensitivity of rice to drought and oxidative stress. An exogenous ROS-scavenging reagent restored the drought-stress tolerance of osadr3-CRISPR plants. Global transcriptome analysis suggested that OsADR3 increased the expression of OsGPX1 under drought stress. Electrophoretic mobility shift, yeast one-hybrid, and dual-luciferase reporter assays revealed that OsADR3 modified the expression of OsGPX1 by directly binding to its promoter. Knockdown of OsGPX1 repressed ROS scavenging ability under drought stress in OsADR3-overexpression plants. These findings suggest that OsADR3 plays a positive regulatory role in drought-stress tolerance by inducing antioxidant defense and associated with the ABA signaling pathway in rice.

β-ketoacyl-CoA synthase is a key enzyme in the biosynthesis of over-long-chain fattty acids; thus, it plays a crucial role in plant resistance to stress. Herein, 33 OsKCSs were classified into five subfamilies based on phylogenetic analysis and domain structure in rice. Nonsynonymous to synonymous substitution ratio (Ka/Ks) analysis showed that duplicated genes were possibly under strong purifying selection pressure. Cis-element analysis revealed that OsKCSs may be regulated by different transcription factors. SNP distribution of OsKCSs in 295 germplasm accessions seemed associated with functional disparity under environmental stress. Additionally, 10 of the 33 OsKCSs were not expressed in different rice tissues. RNA-Seq and qRT-PCR analysis demonstrated that seven OsKCSs (OsKCS5, OsKCS6, OsKCS14, OsKCS15, OsKCS4, OsKCS30, and OsKCS21) had significantly different expression levels under 0.5% NaCl, 0.5% Na2CO3, and a 10-day incubation at 17 °C. Our results provide a solid foundation for understanding the roles of OsKCS genes and the mechanism by which they regulate stress resistance.

Transcranial alternating current stimulation (tACS) applies a sinusoidal oscillating current to modulate intrinsic oscillatory activity. Relevant studies of tACS have indicated that tACS can increase spontaneous brain activity in the occipital area. However, few studies have compared the effects of tACS with different current intensities on spontaneous brain activity in the occipital region. In this study, 10-Hz tACS was delivered to the occipital region at different current intensities (i.e., 1 and 2 mA). We investigated the effect of the tACS on both eyes-open and eyes-closed resting-state electroencephalography (EEG). A total of 20 subjects and fifteen subjects were recruited to participate in the 1-mA tACS experiment and the 2-mA tACS experiment, respectively. Ten subjects participated in both experiments. The experimental results demonstrated that both 1-mA tACS and 2-mA tACS could increase occipital resting-state EEG activities. For the eyes-open condition, alpha activity elicited by 2-mA tACS increased significantly greater than that elicited by 1-mA tACS, while 1-mA tACS could produce greater alpha activity compared to 2 mA for the eyes-closed condition. These results suggested that the optimal current intensity might be different for the eyes-open and eyes-closed resting-state conditions, laying a foundation for the subsequent study of occipital tACS on task-state EEG activities.

Caffeic acid O-methyltransferase (COMT) is one of the core enzymes involved in lignin synthesis. However, there is no systematic study on the rice COMT gene family. We identified 33 COMT genes containing the methyltransferase-2 domain in the rice genome using bioinformatic methods and divided them into Group I (a and b) and Group II. Motifs, conserved domains, gene structure and SNPs density are related to the classification of

Nitrogen-based nutrients are the main factors affecting rice growth and development. As the nitrogen (N) application rate increased, the nitrogen use efficiency (NUE) of rice decreased. Therefore, it is important to understand the molecular mechanism of rice plant morphological, physiological, and yield formation under low N conditions to improve NUE. In this study, changes in the rice morphological, physiological, and yield-related traits under low N (13.33 ppm) and control N (40.00 ppm) conditions were performed. These results show that, compared with control N conditions, photosynthesis and growth were inhibited and the carbon (C)/N and photosynthetic nitrogen use efficiency (PNUE) were enhanced under low N conditions. To understand the post-translational modification mechanism underlying the rice response to low N conditions, comparative phosphoproteomic analysis was performed, and differentially modified proteins (DMPs) were further characterized. Compared with control N conditions, a total of 258 DMPs were identified under low N conditions. The modification of proteins involved in chloroplast development, chlorophyll synthesis, photosynthesis, carbon metabolism, phytohormones, and morphology-related proteins were differentially altered, which was an important reason for changes in rice morphological, physiological, and yield-related traits. Additionally, inconsistent changes in level of transcription and protein modification, indicates that the study of phosphoproteomics under low N conditions is also important for us to better understand the adaptation mechanism of rice to low N stress. These results provide insights into global changes in the response of rice to low N stress and may facilitate the development of rice cultivars with high NUE by regulating the phosphorylation level of carbon metabolism and rice morphology-related proteins.

Nitrogen is an important nutrient for plant growth and essential metabolic processes. Roots integrally obtain nutrients from soil and are closely related to the growth and development of plants. In this study, the morphological analysis of rice root tissues collected at different time points under low-nitrogen and normal nitrogen conditions demonstrated that, compared with normal nitrogen treatment, the root growth and nitrogen use efficiency (NUE) of rice under low-nitrogen treatment were significantly improved. To better understand the molecular mechanisms of the rice root system’s response to low-nitrogen conditions, a comprehensive transcriptome analysis of rice seedling roots under low-nitrogen and control conditions was conducted in this study. As a result, 3171 differentially expressed genes (DEGs) were identified. Rice seedling roots enhance NUE and promote root development by regulating the genes related to nitrogen absorption and utilization, carbon metabolism, root growth and development, and phytohormones, thereby adapting to low-nitrogen conditions. A total of 25,377 genes were divided into 14 modules using weighted gene co-expression network analysis (WGCNA). Two modules were significantly associated with nitrogen absorption and utilization. A total of 8 core genes and 43 co-expression candidates related to nitrogen absorption and utilization were obtained in these two modules. Further studies on these genes will contribute to the understanding of low-nitrogen adaptation and nitrogen utilization mechanisms in rice.

To provide the basis for thermal conductivity regulation of vermicular graphite cast iron (VGI), a new theoretical method consisting of shape interpolation, unit cell model and numerical calculation was proposed. Considering the influence of the graphite anisotropy and interfacial contact thermal conductivity (ICTC), the effective thermal conductivity of a series of unit cell models was calculated by numerical calculation based on finite difference. The effects of microstructure on effective thermal conductivity of VGI were studied by shape interpolation. The experimental results were in good agreement with the calculated ones. The effective thermal conductivity of VGI increases in power function with the decrease in graphite shape parameter, and increases linearly with the increase in graphite volume fraction and thermal conductivity of matrix. When the graphite volume fraction increases by 1%, the thermal conductivity of nodular cast iron increases by about 0.18 W/(m·K), while that of gray cast iron increases by about 3 W/(m·K). The thermal conductivity of cast iron has the same sensitivity to the thermal conductivity of matrix regardless of the graphite shape parameter. The thermal conductivity of matrix increased by 15 W/(m·K) and the thermal conductivity of cast iron increased by about 12 W/(m·K). Moreover, the more the graphite shape deviates from the sphere, the greater the enhancement effect of graphite anisotropy on thermal conductivity than the hindrance effect of interface between graphite and matrix. This work can provide guidance for the development of high thermal conductivity VGI and the study of thermal conductivity of composites containing anisotropic dispersed phase particles with complex shapes.

The relationships of cooling rate with microstructure and thermal conductivity of vermicular graphite cast iron (VGI) cylinder block were studied, which are important for design and optimization of the casting process of VGI cylinder blocks. Cooling rates at different positions in the cylinder block were calculated based on the cooling curves recorded with a solidification simulation software. The metallographic structure and thermal conductivity were observed and measured using optical microscopy (OM), scanning electrical microscopy (SEM) and laser flash diffusivity apparatus, respectively. The effects of the cooling rate on the vermicularity, total and average areas of all graphite particles, and the pearlite fraction in the VGI cylinder block were investigated. It is found that the vermicularity changes in parabola trend with the increase of cooling rate. The total area of graphite particles and the cooling rate at eutectoid stage can be used to predict pearlite fraction well. Moreover, it is found that the thermal conductivity at room temperature is determined by the average area of graphite particles and pearlite fraction when the range of vermicularity is from 80% to 93%. Finally, the quantitative models are established to calculate the vermicularity, pearlite fraction, and thermal conductivity of the VGI cylinder block.

With the expansion of the epidemic, online multimedia teaching has become a common trend. The reasoning model of multimedia teaching evaluation is a useful tool to infer the result of teaching effects and predict the tendency. However, the ambiguity in the linguistic-valued evaluation leads to reasoning problems always in the context with uncertainty. To make the reasoning model better deal with multiple and multidimensional reasoning problems in uncertainty environment, while considering both positive evidence and negative evidence at the same time, this paper mainly focuses on a linguistic truth-valued intuitionistic fuzzy layered aggregation (LTV-IFLA) reasoning method. First, based on the layered linguistic truth-valued intuitionistic fuzzy lattice (LTV-IFL), we realize aggregating the linguistic truth-valued information through the layered average aggregation (LAA) operator presented by this paper. Furthermore, a layered weighted average aggregation (LWAA) operator is proposed to consider setting different weights to achieve personalization of the reasoning results. Finally, a multiple multidimensional reasoning model which simulates the reasoning of human language is presented to illustrate the method’s rationality and validity.

Nitrogen is not only a macronutrient essential for crop growth and development, but also one of the most critical nutrients in farmland ecosystem. Insufficient nitrogen supply will lead to crop yield reduction, while excessive application of nitrogen fertilizer will cause agricultural and eco-environment damage. Therefore, mining low-nitrogen tolerant rice genes and improving nitrogen use efficiency are of great significance to the sustainable development of agriculture. This study was conducted by Genome-wide association study on a basis of two root morphological traits (root length and root diameter) and 788,396 SNPs of a natural population of 295 rice varieties. The transcriptome of low-nitrogen tolerant variety (Longjing 31) and low-nitrogen sensitive variety (Songjing 10) were sequenced between low and high nitrogen treatments. A total of 35 QTLs containing 493 genes were mapped. 3085 differential expressed genes were identified. Among these 493 genes, 174 genes showed different haplotype patterns. There were significant phenotype differences among different haplotypes of 58 genes with haplotype differences. These 58 genes were hypothesized as candidate genes for low nitrogen tolerance related to root morphology. Finally, six genes (Os07g0471300, Os11g0230400, Os11g0229300, Os11g0229400, Os11g0618300 and Os11g0229333) which expressed differentially in Longjing 31 were defined as more valuable candidate genes for low-nitrogen tolerance. The results revealed the response characteristics of rice to low-nitrogen, and provided insights into regulatory mechanisms of rice to nitrogen deficiency.

Drought stress at jointing and booting phases of plant development directly affects plant growth and productivity in rice. Limited by natural factors, the jointing and booting stages in rice varieties are known to overlap in high-latitude areas that are more sensitive to water deficit. However, the regulation of photosynthetic carbon fixation and sucrose metabolism in rice leaves under different degrees of drought stress remains unclear. In this study, rice plants were subjected to three degrees of drought stress (−10, −25, −and 40 kPa) for 15 days during the jointing-booting stage, we investigated photosynthetic carbon sequestration and sucrose metabolism pathways in rice leaves and analyzed key genes and regulatory networks using transcriptome sequencing in 2016. And we investigated the effects of drought stress on the growth periods of rice with overlapping growth periods in 2016 and 2017. The results showed that short-term drought stress promoted photosynthetic carbon fixation. However, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activity significantly decreased, resulting in a significant decrease in photosynthetic rate. Drought stress increased the maximum activity of fructose-1,6-bisphosphate aldolase (FBA). FBA maintains the necessary photosynthetic rate during drought stress and provides a material base after the resumption of irrigation in the form of controlling the content of its reaction product triose phosphate. Drought stress significantly affected the activities of sucrose synthase (SuSase) and sucrose phosphate synthase (SPS). Vacuoles invertase (VIN) activity increased significantly, and the more severe the drought, the higher the VIN activity. Severe drought stress at the jointing-booting stage severely restricted the growth process of rice with overlapping growth stages and significantly delayed heading and anthesis stages. Transcriptome analysis showed that the number of differentially expressed genes was highest at 6–9 days after drought stress. Two invertase and four β-amylase genes with time-specific expression were involved in sucrose-starch metabolism in rice under drought stress. Combined with weighted gene co-expression network analysis, VIN and β-amylase genes up-regulated throughout drought stress were regulated by OsbZIP04 and OsWRKY62 transcription factors under drought stress. This study showed that any water deficit at the jointing-booting stage would have a serious effect on sucrose metabolism in leaves of rice with overlapping growth stages.

Bortezomib, a proteasome inhibitor, proved potent in the treatment of recurrent multiple myeloma or mantle cell lymphoma. However, slow progress was made when it was applied to treat solid tumors. We discovered that different head and neck squamous cell carcinoma (HNSCC) cell lines had significantly different sensitivities to bortezomib, and also demonstrated that individual relatively sensitive HNSCC cell lines had fewer ΔNp63α expressions. Based on these findings, we speculated that ΔNp63α may be a key factor in the resistance of HNSCC cells to bortezomib. ΔNp63α knockdown made HNSCC more sensitive to bortezomib, while ΔNp63α overexpression made it more resistant. RNA sequencing (RNA-seq) analysis of ΔNp63α-knockdown cells revealed clear alterations in the subset of genes that were associated with oxidative stress and antioxidant defense. The gene CYGB was downregulated significantly. CHIP-seq detection showed that CYGB was the transcriptional regulatory site of ΔNp63α. CHIP-PCR showed evidence of ΔNp63α binding. The detection of the dual-luciferase reporter gene demonstrated that ΔNp63α significantly enhanced the CYGB promoter activity. Furthermore, we confirmed that CYGB plays a role in clearing excess ROS induced by bortezomib to inhibit HNSCC apoptosis. Consequently, ΔNp63α regulated the expression of CYGB in HNSCC. CYGB was the target of transcription regulation of ΔNp63α. It reduced apoptosis by clearing excess ROS produced by bortezomib, and thus exerted drug resistance.

Water shortages and nitrogen (N) fertilizer overuse limit japonica rice production in Northeastern China. The interactions between water-saving irrigation and nitrogen management on rice root and shoot growth is still our research focus. Here, japonica rice (DN425) was subjected to the irrigation methods W1 (flooding irrigation), W2 [mild alternate wetting and drying irrigation (AWD); −10 kPa], W3 (severe AWD; −30 kPa), and different N fertilizer ratios were applied in different growth stages, namely, N1 (6:3:1:0), N2 (5:3:1:1), and N3 (4:3:2:1). From jointing to full heading stages, the highest photosynthate production capacity and root activity were obtained under W1N2. AWD markedly affected the root system and resulted in root senescence at later growth stages. Grain yield and N utilization efficiency were closely and positively correlated with the relative water content, crop growth rate (CGR), leaf area duration (LAD), the increase rate of root length density, root surface area density, and root volume density (RVD) from the jointing to full heading stages. This positive correlation was also observed in the increased rate of root bleeding sap (RBS) under W1N2 and CGR under W2N3. From full heading to maturity stages, N2 could promote root growth, LAD, and CGR under AWD to a greater extent than those under the other treatments. Water use efficiency (WUE) and N uptake efficiency (NUpE) were both negatively associated with the decreased rate of RVD, root dry weight (RDW), and RBS. They were closely and positively correlated with the increased rate of RDW and CGR. Our results suggested that W2N2 treatment delayed root senescence, maintained leaf photosynthesis, optimized the crop growth rate from full heading to maturity stages, and improved grain yield. The optimal grain yield, WUE, and NUpE were achieved at the irrigation water amount and topdressing fertilizer ratio of 41.40–50.34 × 102 and 31.20–34.83 kg ha–1, respectively.

Objectives: The optimal surgical approach to treat recurrent laryngeal nerve (RLN) infiltration by differentiated thyroid cancer (DTC) remains a subject of debate. This study explored the feasibility and efficiency of immediate ansa cervicalis nerve (ACN)-to-RLN anastomosis for the management of RLN infiltration by DTC. Material and Methods: Fifty-three patients who underwent immediate ACN-to-RLN anastomosis during DTC extirpation were enrolled in the present study. Thirty-seven cases presented with unilateral vocal cord paralysis before the operation (Group A), and another 16 patients presented with normal vocal cord mobility preoperatively (Group B). Multidimensional assessments, videostroboscopy, voice assessment, and laryngeal electromyography (LEMG) were performed preoperatively and postoperatively. Results: All videostroboscopy, voice assessment and LEMG parameters in Group A deteriorated 1 month after the operation and improved 1 year after the operation compared with preoperative data. In Group B, all parameters 1 year after the operation improved significantly compared with the corresponding parameters 1 month after the operation. LEMG in Group A and B provided substantial evidence for the maturation of neural regeneration from ACN and demonstrated that the laryngeal muscles were reinnervated successfully by this procedure. Conclusions: If the RLN is infiltrated by DTC, immediate ACN-to-RLN anastomosis during complete excision of DTC could restore satisfactory phonatory function and does not compromise oncological radicality.

Background Plant height is a key factor in the determination of rice yield since excessive height can easily cause lodging and reduce yield. Therefore, the identification and analysis of plant height-related genes to elucidate their physiological, biochemical, and molecular mechanisms have significant implications for rice breeding and production. Results High-throughput quantitative trait locus (QTL) sequencing analysis of a 638-individual F2:3 mapping population resulted in the identification of a novel height-related QTL (qPH9), which was mapped to a 2.02-Mb region of Chromosome 9. Local QTL mapping, which was conducted using 13 single nucleotide polymorphism (SNP)-based Kompetitive allele-specific PCR (KASP) markers for the qPH9 region, and traditional linkage analysis, facilitated the localization of qPH9 to a 126-kb region that contained 15 genes. Subsequent haplotype and sequence analyses indicated that OsPH9 was the most probable candidate gene for plant height at this locus, and functional analysis of osph9 CRISPR/Cas9-generated OsPH9 knockout mutants supported this conclusion. Conclusion OsPH9 was identified as a novel regulatory gene associated with plant height in rice, along with a height-reducing allele in ‘Dongfu-114’ rice, thereby representing an important molecular target for rice improvement. The findings of the present study are expected to spur the investigation of genetic mechanisms underlying rice plant height and further the improvement of rice plant height through marker-assisted selection.

Rice (Oryza sativa L.) is an important crop in China. Although it is known that its yield is restricted by nitrogen (N) supply, the response of the root system to N supply specifically has not been systematically explored. This study aimed to investigate the effect of N uptake on grain yield to clarify the relationships between root morphophysiological traits and N uptake, and to understand relation between phytohormones and root morphophysiological traits. Two N-efficient absorption cultivars (NEAs) and two N-inefficient absorption cultivars (NIAs) were grown in the field, and three N conditions, deficient N (60 kg ha–1), intermediate N (180 kg ha–1), and sufficient N (240 kg ha–1), were applied during the growing season. The results showed higher dry matter and grain yield in NEAs than in NIAs, which was mainly attributed to increased N uptake in the mid- and late growth stages under all N conditions. And NEAs have different root regulation methods to obtain higher N accumulation and yield under different N supply conditions. Under lower N conditions, compared with NIAs, NEAs shown greater total root length, root oxidation activity, and root active absorbing surface area and smaller root diameter owing to higher indole-3-acetic acid and cytokinin content and lower 1-aminocyclopropane-1-carboxylic acid content in the early growth stages to respond to low N stress faster, laying a morphophysiological basis for its high N-uptake capacity in the mid- and late growth stages. Under higher N conditions, NEAs had higher root oxidation activity and root active absorbing surface area for N uptake and yield formation owing to higher abscisic acid and cytokinin content in the mid- and late growth stages, which improved the seed setting rate, thereby increasing the rice grain yield. These results suggest that NEAs can optimize the morphophysiological characteristics of roots through phytohormone regulation to adapt to different nutrient conditions, thereby promoting N accumulation and yield formation in rice.

Background This study investigated the role of lncRNA CASC15 in laryngeal squamous cell carcinoma (LSCC). Methods This study included 58 LSCC patients. Both tumor (LSCC) and adjacent (within 3 cm around tumors) non-tumor tissues from 3 different sites of each patient were collected. CCK-8 assay was used to determine cell proliferation. The expression levels of proteins and mRNAs were determined by Western blotting analysis and qRT-PCRs, respectively. Results CASC15 was upregulated in LSCC and high expression levels of CASC15 predicted poor survival. In LSCC tissues, CASC15 was negatively correlated with miR-365 but positively correlated with cyclin D1. In LSCC cells, overexpression of CASC15 resulted in downregulation of miR-365 and upregulation of cyclin D1. Overexpression of miR-365 did not affect the expression of CASC15 but downregulated cyclin D1. Overexpression of Cyclin D1 did not affect the expression of miR-365 and CASC15. Overexpression of CASC15 and cyclin D1 led to promoted, while overexpression of miR-365 led to inhibited LSCC cell proliferation. In addition, overexpression of miR-365 reduced the effects of overexpression of CASC15. Conclusion Therefore, CASC15 upregulates cyclin D1 by downregulating miR-365 in LSCC to promote cell proliferation.

Background Skeletal muscle atrophy and fibrosis are pathological conditions that contribute to morbidity in numerous conditions including aging, cachexia, and denervation. Muscle atrophy is characterized as reduction of muscle fiber size and loss of muscle mass while muscle fibrosis is due to fibroblasts activation and excessive production of extracellular matrix. Purinergic receptor P2Y2 has been implicated in fibrosis. This study aims to elucidate the roles of P2Y2 in sleketal muscle atrophy and fibrosis. Methods Primary muscle fibroblasts were isolated from wild type and P2Y2 knockout (KO) mice and their proliferating and migrating abilities were assessed by CCK-8 and Transwell migration assays respectively. Fibroblasts were activated with TGF-β1 and assessed by western blot of myofibroblast markers including α-SMA, CTGF, and collagen I. Muscle atrophy and fibrosis were induced by transection of distal sciatic nerve and assessed using Masson staining. Results P2Y2 KO fibroblasts proliferated and migrated significantly slower than WT fibroblasts with or without TGF-β1.The proliferation and ECM production were enhanced by P2Y2 agonist PSB-1114 and inhibited by antagonist AR-C118925. TGF-β1 induced fibrotic activation was abolished by P2Y2 ablation and inhibited by AKT, ERK, and PKC inhibitors. Ablation of P2Y2 reduced denervation induced muscle atrophy and fibrosis. Conclusions P2Y2 is a promoter of skeletal muscle atrophy and activation of fibroblasts after muscle injury, which signaling through AKT, ERK and PKC. P2Y2 could be a potential intervention target after muscle injury.

Salinity tolerance plays an important role in direct seeding cultivation since tolerance to salinity at the bud burst stage of rice directly affects seedling formation and yield. However, information on the quantitative trait loci (QTL) associated with salinity tolerance at the bud burst stage is limited. Here, a recombinant inbred line (RIL) population, derived from a cross between Kongyu131 (salinity-sensitive) and Xiaobaijingzi (salinity-tolerant), was used to identify QTLs for salinity tolerance at the bud burst stage using a high-density genetic map. Four QTLs were identified. The threshold LOD score for QTL identification was 3.0. Among the four QTLs detected, two QTLs namely qRSL3 (PVE, 22.59%) and qRRL3 (PVE, 21.21%) were considered to be one QTL, named qST3, which was the major QTL related to salinity tolerance on chromosome 3. This major QTL was located within an interval of 2.55 Mb and contained 253 candidate genes. Using RNA-seq data, 51 differentially expressed genes (DEGs) were detected among the candidate region. According to Rice Annotation Project and quantitative real-time PCR (qRT-PCR) analysis, Os03g0664800 was accepted as the most likely candidate gene for qST3. These results will be useful for improving salinity tolerance of rice and for developing rice cultivars suitable for direct seeding in salinized soil.

Background Rice (Oryza sativa L.) is one of the most widely grown food crops, and its yield and quality are particularly important for a warm-saturated diet. Cold stress restricts rice growth, development, and yield; however, the specific mechanism of cold tolerance in rice remains unknown. Results The analysis of leaf physiological and photosynthetic characteristics showed that the two rice varieties were significantly affected by cold stress, but the cold-tolerant variety KY131 had more stable physiological characteristics, maintaining relatively good photosynthetic capacity. To better explore the transcriptional regulation mechanism and biological basis of rice response to cold stress, a comprehensive analysis of the rice transcriptome and lipidome under low temperature and control temperature conditions was carried out. The transcriptomic analysis revealed that lipid metabolism, including membrane lipid and fatty acid metabolism, may be an important factor in rice cold tolerance, and 397 lipid metabolism related genes have been identified. Lipidomics data confirmed the importance of membrane lipid remodeling and fatty acid unsaturation for rice adaptation to cold stress. This indicates that the changes in the fluidity and integrity of the photosynthetic membrane under cold stress lead to the reduction of photosynthetic capacity, which could be relieved by increased levels of monogalactosyldiacylglycerol that mainly caused by markedly increased expression of levels of 1,2-diacylglycerol 3-beta-galactosyltransferase (MGD). The upregulation of phosphatidate phosphatase (PAP2) inhibited the excessive accumulation of phosphatidate (PA) to produce more phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylglycerol (PG), thereby preventing of membrane phase transition under cold stress. In addition, fatty acid β-oxidation is worth further study in rice cold tolerance. Finally, we constructed a metabolic model for the regulatory mechanism of cold tolerance in rice, in which the advanced lipid metabolism system plays a central role. Conclusions Lipidome analysis showed that membrane lipid composition and unsaturation were significantly affected, especially phospholipids and galactolipids. Our study provides new information to further understand the response of rice to cold stress.

Background Cold stress caused by low temperatures is an important factor restricting rice production. Identification of cold-tolerance genes that can stably express in cold environments is crucial for molecular rice breeding. Results In this study, we employed high-throughput quantitative trait locus sequencing (QTL-seq) analyses in a 460-individual F2:3 mapping population to identify major QTL genomic regions governing cold tolerance at the seedling stage in rice. A novel major QTL (qCTS6) controlling the survival rate (SR) under low-temperature conditions of 9°C/10 days was mapped on the 2.60-Mb interval on chromosome 6. Twenty-seven single-nucleotide polymorphism (SNP) markers were designed for the qCST6 region based on re-sequencing data, and local QTL mapping was conducted using traditional linkage analysis. Eventually, we mapped qCTS6 to a 96.6-kb region containing 13 annotated genes, of which seven predicted genes contained 13 non-synonymous SNP loci. Quantitative reverse transcription PCR analysis revealed that only Os06g0719500, an OsbZIP54 transcription factor, was strongly induced by cold stress. Haplotype analysis confirmed that +376 bp (T>A) in the OsbZIP54 coding region played a key role in regulating cold tolerance in rice. Conclusion We identified OsbZIP54 as a novel regulatory gene associated with rice cold-responsive traits, with its Dongfu-104 allele showing specific cold-induction expression serving as an important molecular variation for rice improvement. This result is expected to further exploration of the genetic mechanism of rice cold tolerance at the seedling stage and improve cold tolerance in rice varieties by marker-assisted selection.

Objectives/hypothesis This study explored the feasibility and efficiency of main branch of ansa cervicalis nerve (ACN)-to-recurrent laryngeal nerve (RLN) anastomosis for management of paroxysmal laryngospasm due to unilateral vocal cord paralysis (UVCP). Methods Thirteen patients who underwent main branch of ACN-to-RLN anastomosis for management of paroxysmal laryngospasm due to UVCP were enrolled in the present study. Multidimensional assessments, including videostroboscopy, voice assessment, and laryngeal electromyography (LEMG), were performed preoperatively and postoperatively. Results This series was limited to UVCP with iatrogenic causes, including thyroidectomy, cervical spine surgery, and thoracic surgery. After main branch of ACN-to-RLN anastomosis, all cases showed significant airway improvement, and laryngospasm was completely abolished in 92.3% (12 of 13) of cases. Videostroboscopy showed that the bulging and paradoxical adduction of the affected vocal cord during a sniff were abolished immediately after operation, and there was no significant difference in vocal fold position or glottal closure before versus after the operation. LEMG showed that the postoperative recruitment and amplitude of voluntary motor unit potential in the affected thyroarytenoid muscle during a sniff were significantly decreased compared to preoperative values, and postoperative recruitment showed significant improvement during phonation compared to that preoperatively. Voice assessment showed that there were no significant differences in overall grade, roughness, breathiness, jitter (local), shimmer (local), noise-to-harmonics ratio, or maximum phonation time after the operation compared to the preoperative values. Conclusions Main branch of ACN-to-RLN anastomosis could have long-lasting efficacy in the management of paroxysmal laryngospasm due to UVCP, with no apparent compromise of voice quality. Level of evidence 4 Laryngoscope, 130:2412-2419, 2020.

Gold nano rods (GNRs) have showed cytotoxicity to cancer cells. At the same time, it shows little effects on non-tumor cells. Between GNRs and sub-cellular organelles, the understanding of interaction plays a very important role to determine the intracellular mechanisms. The purpose of what we done is to explain the effects of the surface properties of GNRs on specific cancer cell death. Three GNR samples with different aspect ratios were finely prepared by the seed-mediated growth method. Then the intracellular transport and the in vitro/vivo mechanisms of cancer cell death were studied by transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), laser light scattering, and flow cytometry (FCM). It was found that GNRs700 exhibited the largest photothermal conversion efficiency. However, the GNR660 with or without light stimulation exhibited the highest cytotoxicity against cancer cells, which was contradict to the general knowledge. Detailed intracellular investigations showed that the lysosome was the key sub-organelle affecting the GNR function. Further experiments revealed that cytotoxicity was strongly affected by the GNR's surface potential. This potential was actually related to the density of surface cationic molecules, which further regulated lysosomal membrane penetration. The results obtained herein indicated that the physicochemical properties of the surface potential mediated the specific toxicity of GNRs against tumours.

Background: There has been increased interest in the research of proteasome inhibitors for more than two decades. Hotspots in this field are constantly changing. Objective: This study aimed to investigate trends in proteasome inhibitors research from 1992 to 2018 and compare the contributions of such research from different countries and authors. Methods: We used Excel 2013 and VoSviewer to analyze bibliometric data on the subject of proteasome inhibitors, including the number of publications, citations frequency, H-index, and country contributions and hotspots (keywords of popular scientific fields). Results: A total of 3646 articles were included. The USA contributed the largest percentage of articles (1742), with the most citations (90666) and the highest H-index (139). The journal Blood had the most articles. Dana Farber Cancer Institute and Millennium Pharmaceuticals Incorporation were the most contributive institutions. Keywords could be divided into three clusters: Basic experiment, clinical research, and others. Conclusion: The number of proteasome inhibitors articles has been increasing for the past 27 years. The USA made the largest contribution in this field. Recent studies on the topic of “carfilzomib” are relatively new and should be closely followed in proteasome inhibitors research.

The geometric structures, electronic and mechanical properties of the high vacancy concentration intermetallic FeAl (experimental value: 3.3 at.% at 1451 K) were investigated by first-principles ca...

Formation of Poly(Glycidyl Methacrylate) (PGMA) films on bacterial cellulose (BC) supports (aerogels) was achieved via activators regenerated by electron transfer for atom transfer radical polymerization (ARGET ATRP). Epoxy groups on PGMA were used for covalent coupling with catalase. Immobilized catalases on BC@PGMA were characterized by Fourier transform infrared spectroscopy (FTIR) and Typical scanning electron microscopy (SEM). The immobilized catalase amount reached a high value of 116 mg/g. Furthermore, the thermal, pH and storage stabilities of the immobilized catalase were improved significantly. After 10 use cycles, the BC@PGMA-catalase still retained approximately 63.5 % of the initial activity. This work demonstrates the potential of hierarchical nanomaterials for improving enzyme performance, leveraging the benefits of both nano- and macroscale supports. These results demonstrate that the BC@PGMA-catalase has high stability and good reusability, suggesting that the BC aerogel modified with PGMA may find applications in biotechnology and as biocatalyst.

OBJECTIVE To evaluate the extent of airway improvement and voice quality in patients with bilateral vocal fold paralysis (BVFP) who underwent selective laryngeal reinnervation surgery. METHODS Seven patients with BVFP caused by thyroid surgeries were enrolled. They underwent selective laryngeal reinnervation surgery. Videostroboscopy data, voice perceptual data (grade [G]), acoustic data, laryngeal electromyography data, and pulmonary function test data were obtained preoperatively and postoperatively. RESULTS Videostroboscopic videos showed that six patients could achieve moderate-to-maximal abduction in the bilateral vocal folds during inspiration, whereas all patients achieved adduction in the bilateral vocal folds during phonation at 4 to 7 months postoperatively. G score was decreased significantly versus preoperative values (P < 0.05), and vocal functional parameters were improved significantly at 12 months postoperatively (P < 0.05). The aerodynamic parameter of maximum phonation time was significantly longer than the preoperative value (P < 0.05). Most parameters in pulmonary function test recovered to normal reference levels as early as 3 months postoperatively, whereas maximal inspiratory pressure (PImax) values were still slightly lower than normal levels 12 months after surgery. All of these parameters improved significantly versus preoperative values. Electromyographic data at 12 months postoperatively showed full interference potentials in bilateral posterior cricoarytenoid muscles during inspiration and full interference potentials in bilateral thyroarytenoid muscles during phonation in all patients. Moderate electric potentials were seen in left interarytenoid muscle in one failed patient. CONCLUSION This new selective laryngeal reinnervation procedure can achieve physiological movements of the bilateral vocal folds in selected patients with BVFP. LEVEL OF EVIDENCE 4. Laryngoscope, 129:2669-2673, 2019.

Background Tongue squamous cell carcinoma (TSCC) is the second most common malignancy in oral carcinoma. lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) was regarded as an oncogenic factor in various carcinomas. However, its underlying molecular mechanisms in the development and progression of TSCC have not been well featured till now. Methods The expressions of MALAT1, miR-140-5p and p21 (RAC1)-activated kinase 1 (PAK1) mRNA were measured by RT-qPCR assay. The protein level of PAK1 was determined by western blot analysis. Cell viability was detected by Cell Counting Kit-8 assay. Transwell chamber was used to detect cell migratory and invasive capability. Luciferase reporter assay, RNA-binding protein immunoprecipitation (RIP) assay and biotin pull-down assay were applied to evaluate the relationship between MALAT1, miR-140-5p and PAK1. Xenograft experiments were performed to assess the effect and mechanism of MALAT1 in TSCC tumor growth. Results The expression of MALAT1 and p21 (RAC1)-activated kinase 1 (PAK1) was upregulated and microRNA-140-5p (miR-140-5p) expression was downregulated in TSCC tissues and cells. MALAT1 knockdown induced miR-140-5p expression by direct interaction. Moreover, MALAT1 knockdown inhibited proliferation, migration, and invasion by upregulating miR-140-5p expression in TSCC cells. Additionally, PAK1 was identified as a direct target of miR-140-5p. Also, MALAT1 knockdown inhibited PAK1 expression by upregulating miR-140-5p in TSCC cells. Furthermore, miR-140-5p overexpression curbed the proliferation, migration, and invasion of TSCC cells by targeting PAK1. Finally, MALAT1 knockdown inhibited tumor growth by upregulating miR-140-5p and downregulating PAK1 in mouse xenograft models of TSCC. Conclusion MALAT1 contributed to TSCC progression via miR-140-5p-PAK1 regulatory axis, highlighting a potential target for TSCC management.

Caffeic acid O-methyltransferase (COMT) is one of the core enzymes involved in lignin synthesis. However, there is no systematic study on the rice COMT gene family. We identified 33 COMT genes containing the methyltransferase-2 domain in the rice genome using bioinformatic methods and divided them into Group I (a and b) and Group II. Motifs, conserved domains, gene structure and SNPs density are related to the classification of OsCOMTs. The tandem phenomenon plays a key role in the expansion of OsCOMTs. The expression levels of fourteen and thirteen OsCOMTs increased or decreased under salt stress and drought stress, respectively. OsCOMTs showed higher expression levels in the stem. The lignin content of rice was measured in five stages; combined with the expression analysis of OsCOMTs and multiple sequence alignment, we found that OsCOMT8, OsCOMT9 and OsCOMT15 play a key role in the synthesis of lignin. Targeted miRNAs and gene ontology annotation revealed that OsCOMTs were involved in abiotic stress responses. Our study contributes to the analysis of the biological function of OsCOMTs, which may provide information for future rice breeding and editing of the rice genome.

ObjectivesThis study aimed to investigate the status of the current knowledge about laryngopharyngeal reflux disease (LPRD) among Chinese otolaryngologists.DesignMulti-centre cross-sectional survey.Setting220 medical centres in different regions of China.ParticipantsA total of 2254 otolaryngologists from 220 medical centres in China who were successfully on-site surveyed between November 2019 and December 2020.Main outcome measuresAwareness about LPRD included knowledge about risk factors, symptoms, laryngoscope signs, related diseases, current diagnostic methods and treatments.ResultsThe percentage of participants who had heard of LPRD was 96.4%, with academic conferences as the most common source of information (73.3%). The most commonly known risk factor, symptom, laryngoscope sign, related disease, diagnostic method and treatment were alcohol consumption (44.0%), pharyngeal foreign body sensation (66.9%), hyperaemia (52.4%), pharyngolaryngitis (54.8%), pH monitoring (47.6%) and medication (82.1%), respectively. Only 28.3% of all participants knew that 24 h pH or multichannel intraluminal impedance pH monitoring was the most accurate diagnostic test. As many as 73.1% of all participants knew that proton pump inhibitors were the first-line treatment drugs. An analysis of the overall status of awareness using a scoring system suggested that otolaryngologists were better aware owing to more access, working at 3A hospitals, and postgraduate or above educational background (all pConclusionAlthough the majority of Chinese otolaryngologists had heard of LPRD, their overall awareness about the disease was not encouraging. More efforts are needed to increase the knowledge about LPRD among this group of physicians.Trial registration numberChiCTR1900025581

Low-temperature tolerance during the bud-bursting stage is an important characteristic of direct-seeded rice. The identification of cold-tolerance quantitative trait loci (QTL) in species that can stably tolerate cold environments is crucial for the molecular breeding of rice with such traits. In our study, high-throughput QTL-sequencing analyses were performed in a 460-individual F2:3 mapping population to identify the major QTL genomic regions governing cold tolerance at the bud-bursting (CTBB) stage in rice. A novel major QTL, qCTBB9, which controls seed survival rate (SR) under low-temperature conditions of 5°C/9 days, was mapped on the 5.40-Mb interval on chromosome 9. Twenty-six non-synonymous single-nucleotide polymorphism (nSNP) markers were designed for the qCTBB9 region based on re-sequencing data and local QTL mapping conducted using traditional linkage analysis. We mapped qCTBB9 to a 483.87-kb region containing 58 annotated genes, among which six predicted genes contained nine nSNP loci. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis revealed that only Os09g0444200 was strongly induced by cold stress. Haplotype analysis further confirmed that the SNP 1,654,225 bp in the Os09g0444200 coding region plays a key role in regulating the cold tolerance of rice. These results suggest that Os09g0444200 is a potential candidate for qCTBB9. Our results are of great significance to explore the genetic mechanism of rice CTBB and to improve the cold tolerance of rice varieties by marker-assisted selection.

Over-application of nitrogen (N) fertilizer in fields has had a negative impact on both environment and human health. Domesticated rice varieties with high N use efficiency (NUE) reduce fertilizer requirements, enabling sustainable agriculture. Genome-wide association study (GWAS) analysis of N absorption and utilization traits under low and high N conditions was performed to obtain 12 quantitative trait loci (QTLs) based on genotypic data including 151,202 single-nucleotide polymorphisms (SNPs) developed by re-sequencing 267 japonica rice varieties. Eighteen candidate genes were obtained by integrating GWAS and transcriptome analyses; among them, the functions of OsNRT2.4, OsAMT1.2, and OsAlaAT genes in N transport and assimilation have been identified, and OsJAZ12 and OsJAZ13 also play important roles in rice adaptation to abiotic stresses. A NUE-related candidate gene, OsNAC68, was identified by quantitative real-time PCR (qRT-PCR) analyses. OsNAC68 encodes a NAC transcription factor and has been shown to be a positive regulator of the drought stress response in rice. Overexpression of OsNAC68 significantly increased rice NUE and grain yield under deficient N conditions, but the difference was not significant under sufficient N conditions. NUE and grain yield significantly decreased under both N supply conditions in the osbnac68 mutant. This study provides crucial insights into the genetic basis of N absorption and utilization in rice, and a NUE-related gene, OsNAC68, was cloned to provide important resources for rice breeding with high NUE and grain yield.

Low-temperature tolerance during the bud-bursting stage is an important characteristic of direct-seeded rice. The identification of cold-tolerance quantitative trait loci (QTL) in species that can stably tolerate cold environments is crucial for the molecular breeding of rice with such traits. In our study, high-throughput QTL-sequencing analyses were performed in a 460-individual F2:3 mapping population to identify the major QTL genomic regions governing cold tolerance at the bud-bursting (CTBB) stage in rice. A novel major QTL, qCTBB9, which controls seed survival rate (SR) under low-temperature conditions of 5°C/9 days, was mapped on the 5.40-Mb interval on chromosome 9. Twenty-six non-synonymous single-nucleotide polymorphism (nSNP) markers were designed for the qCTBB9 region based on re-sequencing data and local QTL mapping conducted using traditional linkage analysis. We mapped qCTBB9 to a 483.87-kb region containing 58 annotated genes, among which six predicted genes contained nine nSNP loci. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis revealed that only Os09g0444200 was strongly induced by cold stress. Haplotype analysis further confirmed that the SNP 1,654,225 bp in the Os09g0444200 coding region plays a key role in regulating the cold tolerance of rice. These results suggest that Os09g0444200 is a potential candidate for qCTBB9. Our results are of great significance to explore the genetic mechanism of rice CTBB and to improve the cold tolerance of rice varieties by marker-assisted selection.