Pan-transcriptomic analysis reveals alternative splicing control of cold tolerance in rice.

Plants have evolved complex mechanisms to adapt to harsh environmental conditions. Rice (Oryza sativa) is a staple food crop that is sensitive to low temperatures. However, its cold stress responses remain poorly understood, thus limiting possibilities for crop engineering to achieve greater cold tolerance. In this study, we constructed a rice pan-transcriptome and characterized its transcriptional regulatory landscape in response to cold stress. We performed Iso-Seq and RNA-Seq of 11 rice cultivars subjected to a time-course cold treatment. Our analyses revealed that alternative splicing-regulated gene expression plays a significant role in the cold stress response. Moreover, we identified CATALASE C (OsCATC) and Os03g0701200 as candidate genes for engineering enhanced cold tolerance. Importantly, we uncovered central roles for the 2 serine-arginine-rich proteins OsRS33 and OsRS2Z38 in cold tolerance. Our analysis of cold tolerance and resequencing data from a diverse collection of 165 rice cultivars suggested that OsRS2Z38 may be a key selection gene in japonica domestication for cold adaptation, associated with the adaptive evolution of rice. This study systematically investigated the distribution, dynamic changes, and regulatory mechanisms of alternative splicing in rice under cold stress. Overall, our work generates a rich resource with broad implications for understanding the genetic basis of cold response mechanisms in plants.

MeSWEET15a/b genes play a role in the resistance of cassava (Manihot esculenta Crantz) to water and salt stress by modulating sugar distribution.

The sugar transporter SWEET plays a role in plant growth, carbon allocation, and abiotic stress resistance. We examined the function of SWEET in cassava (Manihot esculenta Crantz) under water and salt stress. Bioinformatics, subcellular localization, yeast deficient complementation, and virus-induced gene silencing (VIGS) were used to examine the function of SWEET in cassava. Twenty-eight MeSWEETs genes were found based on the conserved domain MtN3/saliva of SWEET transporters, two MeSWEET15a/b of them were identified by phylogenetic analysis, which were located on the cell membrane. They transfer sucrose, fructose, glucose, and mannitol from culture media to yeast cells, predominately transferring sucrose via bleeding fluid saps in plant. Leaf sucrose content was increased in MeSWEET15a/b-silenced cassava plants, resulting in changes in carbon distribution, with an increase in starch accumulation in the leaves and a decrease in starch accumulation in the roots. The silencing of MeSWEET15a/b genes led to tolerance to water and salt stress, consistent with a high accumulation of osmolytes, and low lipid membrane peroxidation. Changes in sugar distribution increased the expression of MeTOR and MeE2Fa in pTRV2-MeSWEET15a and pTRV2-MeSWEET15b cassava leaves. MeSWEET15a/b acts as pivotal modulators of sugar distribution and tolerance to water and high salt stress in cassava.

An Integration of MicroRNA and Transcriptome Sequencing Analysis Reveal Regulatory Roles of miRNAs in Response to Chilling Stress in Wild Rice.

A chromosome single segment substitution line (CSSL) DC90, which was generated by introgressing CTS-12, a locus derived from common wild rice (Oryza rufipogon Griff.), into the 9311 (Oryza sativa L. ssp. indica) background, exhibits a chilling tolerance phenotype under chilling stress. Here, an integration of microRNA (miRNA) deep sequencing and transcriptomic sequencing analysis was performed to explore the expression profiles of miRNAs and their target genes mediated by CTS-12 under chilling stress, and to reveal the possible regulatory mechanisms of miRNAs that are involved in chilling tolerance. Integration analysis revealed that a number of differentially expressed miRNAs (DEMs) and putative target genes with different expression patterns and levels were identified in 9311 and DC90 under chilling stress. KEGG enrichment analysis revealed that the target genes that are regulated by chilling-induced miRNAs are involved in the regulation of various biological processes/pathways, including protein biosynthesis, redox process, photosynthetic process, and chloroplast development in two genotypes. CRISPR/Cas9 editing of the target genes of the key DEMs in a chilling tolerant rice variety Zhonghua 11 (ZH11) found that LOC_Os11g48020 (OsGL1-11), one of the putative target genes of osa-miR1846a/b-5p and encoding a wax synthesis protein, is correlated with a chilling stress tolerance phenotype, implying osa-miR1846a/b-5p/OsGL1-11 plays an important role in CTS-12-mediated chilling stress tolerance regulatory pathway(s). Therefore, we speculate that the CTS-12 may regulate the key miRNA target genes in response to chilling stress by differential regulation of miRNAs in wild rice, thereby resulting in the variation of chilling tolerance phenotype between 9311 and DC90.

Enhancing Anti-Static Performance of Fibers by Construction of the Hybrid Conductive Network Structure on the Fiber Surface.

The hybrid antistatic agent SCNTs/OAA composed of sulfonated carbon nanotubes (SCNTs) and organic antistatic agent (OAA) was treated on the fiber surface to construct the hybrid conductive layer. Among them, SCNTs were synthesized through a simple method, and their chemical structure and morphology were characterized. SCNTs had good dispersibility due to the presence of sulfonic acid groups, which made SCNTs uniformly dispersed on the fiber surface. The SCNTs/OAA-treated fiber was hardly affected by relative humidity, because SCNTs form a continuous and uniform physical conductive network on the fiber surface. When the addition amount of SCNTs/OAA was 0.5~2 wt%, the fiber had excellent antistatic ability. Under the synergistic effect of SCNTs and OAA, the resistivity of SCNTs/OAA-treated fiber was almost not affected by fiber stretching.

Influence of Graphite/Graphene on the Tribological Behaviors of Self-Lubricating Fabric Composite.

Graphite/graphene particles were employed as functional fillers to modify hybrid polytetrafluoroethylene/polyisophthaloyl metaphenylene diamine (PTFE/Nomex) fabric-reinforced phenolic composites. The tribology behavior was investigated using a ball-on-disk wear tester, together with a 3D digital microscope. The graphite/graphene exhibits the synergetic effect from the results, which not only reduces the friction efficient but also improves the wear resistance of the composites. Moreover, the wear mechanisms were studied by the wear surface microstructure analysis. It is proposed that the synergetic effect includes mainly the positive rolling effect from the graphene and an improved load-carrying capacity brought by graphite. In addition, in order to obtain the optimized formulation to satisfy the bearing application, the influence of graphite and graphene content on the tribological property of the composites was studied in detail. On the basis of that, the application research was carried out on the bearing oscillating wear test, which will evaluate the engineering service life of the composite.

Protective functions of myricetin in LPS-induced cardiomyocytes H9c2 cells injury by regulation of MALAT1.

BACKGROUND: Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is a crucial mediator in response to inflammation. Myricetin protects cardiomyocytes against inflammatory injury. However, it's still unexplored whether myricetin exerted anti-inflammatory properties via MALAT1. The purpose of our study was to validate the cardio-protective function of myricetin against myocarditis and its underlying mechanism in vitro. METHODS: H9c2 cells were pre-incubated with myricetin before stimulation with lipopolysaccharide (LPS). Enforced silence of MALAT1 was achieved by transducing short hairpin (sh)-MALAT1 into H9c2 cells. Next, cell viability and apoptotic cells were detected with cell counting kit-8 (CCK-8) and Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) apoptosis detection kit, respectively. Western blot assay was conducted to examine apoptosis-relative proteins, pro-inflammatory factors, and signaling regulators. Quantitative real-time PCR (qRT-PCR) was performed to quantify pro-inflammatory factors and MALAT1 at mRNA levels. Enzyme-linked immune sorbent assay (ELISA) was employed to determine protein concentration of pro-inflammatory factors. RESULTS: Myricetin ameliorated LPS-elicited reduction of cell viability, augment of apoptosis, and overexpression of monocyte chemo-attractant protein-1 (MCP-1) and interleukin-6 (IL-6) in H9c2 cells. Meanwhile, phosphorylation of p65 and inhibitor of nuclear factor kappa B alpha (IκBα) were suppressed. Besides, myricetin enhanced the expression of MALAT1 which was originally down-regulated by LPS. However, the protective effects of myricetin against LPS-caused inflammatory lesions were abrogated in MALAT1-deficiency cells, with the restored phosphorylation of p65 and IκBα. CONCLUSION: Myricetin possessed an anti-inflammatory function against LPS-induced lesions in cardiomyocytes. Mechanically, myricetin up-regulated MALAT1, blocked LPS-evoked activation of nuclear factor-κB (NF-κB) inflammatory pathway, and, finally, exerted cardio-protective effects.

Validation of spectrophotometric determination of chlorogenic acid in fermentation broth and fruits.

A fast and accurate ultraviolet-visible (UV-vis) spectrophotometric method was established to determine the presence of chlorogenic acid (CGA) according to potassium ferricyanide-Fe (III) detection system. The reaction temperature and pH level greatly influenced the CGA absorption spectrum. Maximum UV-vis absorption wavelength occurred at 790 nm under the optimum conditions (at 25 °C and pH 7.0). The results of UV-vis were further assessed by comparing the results with those of high-performance liquid chromatography (HPLC). The proposed method showed a wide linear sensing range of 10.0-800.0 µg mL-1 (R2 = 0.9996) and a high degree of precision (%R.S.D. < 1.50) and recovery (%R.S.D. > 3.39). Results of CGA and HPLC methods correlated well with each other. The proposed accurate, rapid, sensitive, low-cost, and high-throughput method was successfully used to quantify CGA in fermentation broth and fruits (Mango). Therefore, it may be applied for measuring CGA in biological samples.

Improvement of oxytetracycline production mediated via cooperation of resistance genes in Streptomyces rimosus.

Increasing the self-resistance levels of Streptomyces is an effective strategy to improve the production of antibiotics. To increase the oxytetracycline (OTC) production in Streptomyces rimosus, we investigated the cooperative effect of three co-overexpressing OTC resistance genes: one gene encodes a ribosomal protection protein (otrA) and the other two express efflux proteins (otrB and otrC). Results indicated that combinational overexpression of otrA, otrB, and otrC (MKABC) exerted a synergetic effect. OTC production increased by 179% in the recombinant strain compared with that of the wild-type strain M4018. The resistance level to OTC was increased by approximately two-fold relative to the parental strain, thereby indicating that applying the cooperative effect of self-resistance genes is useful to improve OTC production. Furthermore, the previously identified cluster-situated activator OtcR was overexpressed in MKABC in constructing the recombinant strain MKRABC; such strain can produce OTC of approximately 7.49 g L-1, which represents an increase of 19% in comparison with that of the OtcR-overexpressing strain alone. Our work showed that the cooperative overexpression of self-resistance genes is a promising strategy to enhance the antibiotics production in Streptomyces.

Hydrothermal fabrication of hydroxyapatite/chitosan/carbon porous scaffolds for bone tissue engineering.

Porous carbon fiber felts (PCFFs) have great applications in orthopedic surgery because of the strong mechanical strength, low density, high stability, and porous structure, but they are biologically inert. To improve their biological properties, we developed, for the first time, the hydroxyapatite (HA)/chitosan/carbon porous scaffolds (HCCPs). HA/chitosan nanohybrid coatings have been fabricated on PCFFs according to the following stages: (i) deposition of chitosan/calcium phosphate precursors on PCFFs; and (ii) hydrothermal transformation of the calcium phosphate precursors in chitosan matrix into HA nanocrystals. The scanning electron microscopy images indicate that PCFFs are uniformly covered with elongated HA nanoplates and chitosan, and the macropores in PCFFs still remain. Interestingly, the calcium-deficient HA crystals exist as plate-like shapes with thickness of 10-18 nm, width of 30-40 nm, and length of 80-120 nm, which are similar to the biological apatite. The HA in HCCPs is similar to the mineral of natural bone in chemical composition, crystallinity, and morphology. As compared with PCFFs, HCCPs exhibit higher in vitro bioactivity and biocompatibility because of the presence of the HA/chitosan nanohybrid coatings. HCCPs not only promote the formation of bone-like apatite in simulated body fluid, but also improve the adhesion, spreading, and proliferation of human bone marrow stromal cells. Hence, HCCPs have great potentials as scaffold materials for bone tissue engineering and implantation.