Sequencing and analysis of the complete mitochondrial genomes of Toona sinensis and Toona ciliata reveal evolutionary features of Toona.

BACKGROUND: Toona is a critical genus in the Meliaceae, and the plants of this group are an asset for both restorative and restorative purposes, the most flexible of which are Toona sinensis and Toona ciliata. To concentrate on the advancement of mitochondrial(Mt) genome variety in T.sinensis and T.ciliata, the Mt genomes of the two species were sequenced in high throughput independently, after de novo assembly and annotation to construct a Mt genome map for comparison in genome structure. Find their repetitive sequences and analyze them in comparison with the chloroplast genome, along with Maximum-likelihood(ML) phylogenetic analysis with 16 other relatives. RESULTS: (1) T. sinensis and T.ciliata are both circular structures with lengths of 683482 bp and 68300 bp, respectively. They share a high degree of similarity in encoding genes and have AT preferences. All of them have the largest Phe concentration and are the most frequently used codons. (2) Both of their Mt genome are highly preserved in terms of structural and functional genes, while the main variability is reflected in the length of tRNA, the number of genes, and the value of RSCU. (3) T. siniensis and T. ciliata were detected to have 94 and 87 SSRs, respectively, of which mononucleotides accounted for the absolute proportion. Besides, the vast majority of their SSRs were found to be poly-A or poly-T. (4)10 and 11 migrating fragments were identified in the comparison with the chloroplast genome, respectively. (5) In the ML evolutionary tree, T.sinensis and T.ciliata clustered individually into a small branch with 100% support, reflecting two species of Toona are very similarly related to each other. CONCLUSIONS: This research provides a basis for the exploitation of T.sinensis and T.ciliata in terms of medicinal, edible, and timber resources to avoid confusion; at the same time, it can explore the evolutionary relationship between the Toona and related species, which does not only have an important practical value, but also provides a theoretical basis for future hybrid breeding of forest trees, molecular markers, and evolutionary aspects of plants, which has great scientific significance.

Characterization of the 1-Deoxy-D-xylulose 5-Phosphate synthase Genes in Toona ciliata Suggests Their Role in Insect Defense.

The first enzyme, 1-Deoxy-D-xylulose-5-phosphate synthase (DXS), in the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway for isoprenoid precursor biosynthesis has been reported to function differently according to species. However, the current state of knowledge about this gene family in Toona ciliata is limited. The TcDXS gene family was identified from the whole genome of T. ciliata by firstly using bioinformatics analysis. Then, the phylogenetic tree was built and the promoter cis-elements were predicted. Six DXS genes were identified and divided into three groups, which had similar domains and gene structure. They are located on five different chromosomes and encode products that do not vary much in size. An analysis of the cis-acting elements revealed that TcDXS genes possessed light, abiotic stress, and hormone responsive elements. Ultimately, TcDXS1/2/5 was cloned for an in-depth analysis of their subcellular localization and expression patterns. The subcellular localization results of TcDXS1/2/5 showed that they were located in the chloroplast envelope membranes. Based on tissue-specific analyses, TcDXS1/2/5 had the highest expression in mature leaves. Under Hypsipyla robusta stress, their different expressions indicated that these genes may have insect-resistance functions. This research provides a theoretical basis for further functional verification of TcDXSs in the future, and a new concept for breeding pest-resistant T. ciliata.

Genome-Wide Identification and Expression Analysis of the Aux/IAA Gene Family of the Drumstick Tree (Moringa oleifera Lam.) Reveals Regulatory Effects on Shoot Regeneration.

Auxin plays a critical role in organogenesis in plants. The classical auxin signaling pathway holds that auxin initiates downstream signal transduction by degrading Aux/IAA transcription repressors that interact with ARF transcription factors. In this study, 23 MoIAA genes were identified in the drumstick tree genome. All MoIAA genes were located within five subfamilies based on phylogenetic evolution analysis; the gene characteristics and promoter cis-elements were also analyzed. The protein interaction network between the MoIAAs with MoARFs was complex. The MoIAA gene family responded positively to NAA treatment, exhibiting different patterns and degrees, notably for MoIAA1, MoIAA7 and MoIAA13. The three genes expressed and functioned in the nucleus; only the intact encoding protein of MoIAA13 exhibited transcriptional activation activity. The shoot regeneration capacity in the 35S::MoIAA13-OE transgenic line was considerably lower than in the wild type. These results establish a foundation for further research on MoIAA gene function and provide useful information for improved tissue culture efficiency and molecular breeding of M. oleifera.

Selection and validation of reference genes for measuring gene expression in Toona ciliata under different experimental conditions by quantitative real-time PCR analysis.

BACKGROUND: Before studying gene expression of different organisms, it is important to determine the best reference gene. At present, the most accurate method of detecting gene expression is quantitative real-time PCR (RT-qPCR). With this method, reference genes that are stable in different biological systems and under different conditions can be obtained. Toona ciliata Roem (T. ciliata). is a valuable and fast-growing timber specie. In this study, 20 reference genes were identified using RT-qPCR, as a primary prerequisite for future gene expression analysis. Four different methods, geNorm, NormFinder, BestKeeper, and RankAggreg were used to evaluate the expression stability of the 20 candidate reference genes in various tissues under different conditions. RESULTS: The experimental results showed that TUB-α was the most stably expressed reference gene across all samples and UBC17 was the most stable in leaves and young stems under Hypsipyla robusta (H. robusta) and methyl jasmonate (MeJA) treatments. In addition, PP2C59 and UBC5B were the best-performing genes in leaves under H. robusta treatment, while HIS1 and ACT7 were the best reference genes in young stems. The two best reference genes were 60S-18 and TUB-α after treatment at 4 °C. The expression of HIS6 and MUB1 was the most stable under PEG6000 treatment. The accuracy of the selected reference genes was verified using the transcription factor MYB3 (TcMYB3) gene. CONCLUSIONS: This is the first report to verify the best reference genes for normalizing gene expression in T. ciliata under different conditions, which will facilitate future elucidation of gene regulations in this species.

Composition and Leaching Toxicity of Hydrochloric Acid Pickling Sludge Generated from the Hot-Dip Galvanized Steel Industry.

Steel hydrochloric acid pickling sludge (SHPS), containing the heavy metals Fe, Zn, and Ni and a high chloride salt content, is considered a type of hazardous solid waste because of its potential harm to human health and the environment. In addition, the SHPS yield is large, but the main treatment currently used is only safe for landfills. Although studying the composition and leaching toxicity of SHPS is of great importance, only a small amount of related literature is available. This paper can help compensate for this deficiency. SHPS is analyzed from the aspects of its formation mechanism, pH, moisture content, elemental concentration, phase composition, microstructure, and leaching toxicity. The results show that its pH ranges from 2.25 to 11.11, and the moisture content ranges from 45.47% to 83.34%. Additionally, the concentration of Fe is the highest, with values from 29.80% to 50.65%, while other alkali metal elements, namely, Ca, K, and Na, have values of 0.36% to 23.07%, 0.02% to 19.82%, and 0.38% to 3.31%, respectively. Heavy metal elements, namely, Zn, Ni, Mn, Cr, and Pb, have values of 0.02% to 14.88%, 0.001% to 0.05%, 0.03% to 0.38%, 0.01% to 0.09%, and 0.02% to 0.19%, respectively. Anions, namely, SO4 2-, Cl-, F-, and NO3 -, have contents of 0.09% to 0.34%, 0.54% to 5.73%, 0.001% to 0.04%, and 0.01% to 0.15%, respectively. X-ray diffraction (XRD) analysis shows that Fe and Zn are mainly present in oxides, Ca is present as CaO and CaCO3, and chlorine is present in NaCl. Moreover, scanning electron microscopy (SEM) analysis shows that the microscopic structure consists mainly of bright and fluffy irregular spheres; stripes; flakes; and dark, very small irregular particles. The leaching toxicity test based on HJ/T 299-2007 (China) was performed, where SHPS samples were treated with a mixed solution of sulfuric acid, nitric acid, and pure water (pH = 3.20 ± 0.05) at a liquid-to-solid ratio of 10:1 for a period of 18 h. The leachate was filtered and analyzed for Cr, Ni, Mn, Zn, etc. The leaching results indicate that Zn and Ni are the main elements that cause SHPS to be hazardous to the environment. These research results can provide a reference for later researchers studying the effective treatment of SHPS, such as more effective treatments for reducing toxicity and resource utilization.

Transcriptome profiling of Toona ciliata young stems in response to Hypsipyla robusta Moore.

Toona ciliata is a traditional woody plant that can be used as a medicinal material in China. The extracts of its roots, stems, leaves, and flowers all have a wide range of bioactive compounds. However, T. ciliata has been facing an unresolved pest problem caused by Hypsipyla robusta Moore (HRM), which seriously affects its growth and development. In this study, the expression level of TcMYB3 gene reached the maximum (28-fold) at 12 h and transcriptome sequencing of young stems eaten by HRM for 0, 3, 12, and 21 h were performed. A large number of differentially expressed genes (DEGs) were identified including jointly up-regulated genes (263) and down-regulated genes (378). JA synthesis and signaling transduction, terpene biosynthesis, and MAPKs signaling pathway were analyzed in depth and found that TcOPR3, TcJAR1, TcJAZs, and TcTPS9 genes possessed anti-insect potential. Moreover, MYB and ERF transcription factor (TF) families were significantly strengthened to the point that they may participate in induced defense mechanisms in T. ciliata. These data not only provide insights into the molecular mechanisms in resistance of T. ciliata to HRM but also helps to explore the new biocontrol strategies against insects in eco-friendly woody plants.

Evolution and Expression Patterns of the Fructose 1,6-Bisphosptase Gene Family in a Miracle Tree (Neolamarckia cadamba).

Neolamarckia cadamba (N. cadamba) is a fast-growing tree species with tremendous economic and ecological value; the study of the key genes regulating photosynthesis and sugar accumulation is very important for the breeding of N. cadamba. Fructose 1,6-bisphosptase (FBP) gene has been found to play a key role in plant photosynthesis, sugar accumulation and other growth processes. However, no systemic analysis of FBPs has been reported in N. cadamba. A total of six FBP genes were identifed and cloned based on the N. cadamba genome, and these FBP genes were sorted into four groups. The characteristics of the NcFBP gene family were analyzed such as phylogenetic relationships, gene structures, conserved motifs, and expression patterns. A cis-acting element related to circadian control was first found in the promoter region of FBP gene. Phylogenetic and quantitative real-time PCR analyses showed that NcFBP5 and NcFBP6 may be chloroplast type 1 FBP and cytoplasmic FBP, respectively. FBP proteins from N. cadamba and 22 other plant species were used for phylogenetic analyses, indicating that FBP family may have expanded during the evolution of algae to mosses and differentiated cpFBPase1 proteins in mosses. This work analyzes the internal relationship between the evolution and expression of the six NcFBPs, providing a scientific basis for the evolutionary pattern of plant FBPs, and promoting the functional studies of FBP genes.

Reduction-Magnetic Separation of Pickling Sludge by Biomass Pyrolysis Reducing Gas.

The neutralization process of carbon steel pickling wastewater produces a large amount of steel hydrochloric acid pickling sludge (SHPS), and improper treatment of this sludge poses a serious threat to the environment. Considering that SHPS contains a large amount of iron oxide and given the huge demand for iron concentrate in China's ironmaking industry, refining iron oxide in SHPS into iron concentrate will have great environmental and economic benefits. This paper proposes a new method that uses biomass (corncob) to replace conventional coal-based reductants for the recovery of iron components in SHPS to simultaneously utilize two kinds of solid waste resources. Factors that affect the iron recovery rate and iron grade of SHPS, such as the reaction temperature, corncob dosage, residence time, and magnetic field strength, were studied using a fixed bed and a magnetic separator. These studies were combined with thermodynamic analysis, thermogravimetric analysis, X-ray diffraction, inductively coupled plasma-mass spectrometry, gas chromatography, etc. The results showed that when the reaction temperature was 680 °C, the corncob dosage was 5%, the residence time was 20 min, and the magnetic field strength was 200 mT, the recovery rate of iron reached 91.83%, and the iron grade of the recovered products was 67.72%, meeting the level I requirements in GB/T 32545-2016. Based on this result, a process involving SHPS reduction roasting with corncob pyrolysis reducing gas-magnetic separation was established to recover iron from SHPS. This process not only effectively utilizes the iron oxide in SHPS by converting it into iron concentrate powder for the ironmaking industry but also proves that the pyrolysis gas of corncob has good reduction ability.

[The study on oral carcinoma-associated fibroblast promoting the proliferation of lingual carcinoma cell line].

OBJECTIVE: The aim of this study was in vitro to investigate the effects of oral carcinoma-associated fibroblasts (CAFs) on the proliferation of lingual carcinoma cells. METHODS: The interaction model between primary oral CAFs and a lingual carcinoma cell line Tca8113 was established for this study project. The effects of CAFs on the viability and cell cycle of Tca8113 were investigated through morphological observation, MTT assay and flow cytometry. RESULTS: After oral CAFs interacted with Tca8113 directly, Tca8113 showed the cellular shape changes in morphology; compared with normal fibroblasts (NFs), CAFs enhanced the viability of Tca8113 (P<0.05), and increased the percentage of carcinoma cells in S phase and G2 phase (48.1% vs 40.0%). CONCLUSION: Oral CAFs can promote the proliferation of the lingual carcinoma cell line Tca8113 in vitro, and may play a key role in the progression of oral squamous cell carcinoma (OSCC).

[Topical thalidomide having good effect on refractory chronic discoid lupus erythematosus: a case report].

Systemic thalidomide has good effect on refractory chronic discoid lupus erythematosus (CDLE) whereas topical regimen of it has not been reported previously. In this paper, a case of refractory CDLE cured with topical thalidomide in clinic was reported and the feasibility of topical therapy was reviewed.

[Investigation of the degradation and the mechanisms of artificial basement membrane co-cultured with oral carcinoma-associated fibroblasts].

OBJECTIVE: To investigate the degradation of artificial basement membrane (matrigel) co-cultured with oral carcinoma-associated fibroblasts (CAFs) and its possible mechanism. METHODS: CAFs and normal fibroblasts (NFs) were incubated on matrigel for 24, 48, 72 h. Equivalent amounts of conditioned medium were collected and assayed for total protein, hydroxyproline and matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) activity by gelatin zymography. RESULTS: Oral CAFs were superior to oral NFs in total protein and hydroxyproline density, CAFs present more pro-MMP-2 and activated MMP-2. CONCLUSION: CAFs were superior to NFs in degradation of matrigel. CAFs might play a key role in the reconstitution of extracellular matrix and the progression of tumor.