摘要：
BACKGROUND: Many medical graduate students lack a thorough understanding of decision curve analysis (DCA), a valuable tool in clinical research for evaluating diagnostic models. METHODS: This article elucidates the concept and process of DCA through the lens of clinical research practices, exemplified by its application in diagnosing liver cancer using serum alpha-fetoprotein levels and radiomics indices. It covers the calculation of probability thresholds, computation of net benefits for each threshold, construction of decision curves, and comparison of decision curves from different models to identify the one offering the highest net benefit. RESULTS: The paper provides a detailed explanation of DCA, including the creation and comparison of decision curves, and discusses the relationship and differences between decision curves and receiver operating characteristic curves. It highlights the superiority of decision curves in supporting clinical decision-making processes. CONCLUSION: By clarifying the concept of DCA and highlighting its benefits in clinical decisionmaking, this article has improved researchers' comprehension of how DCA is applied and interpreted, thereby enhancing the quality of research in the medical field.
摘要：
Type 2 diabetes mellitus (T2DM) is a chronic and complex disease, and traditional drugs have many side effects. The active compound dihydromyricetin (DHM), derived from natural plants, has been shown in our previous study to possess the potential for reducing blood glucose levels; however, its precise molecular mechanism remains unclear. In the present study, network pharmacology and transcriptomics were performed to screen the molecular targets and signaling pathways of DHM disturbed associated with T2DM, and the results were partially verified by molecular docking, RT-PCR, and Western blotting at in vivo levels. Firstly, the effect of DHM on blood glucose, lipid profile, and liver oxidative stress in db/db mice was explored and the results showed that DHM could reduce blood glucose and improve oxidative stress in the liver. Secondly, GO analysis based on network pharmacology and transcriptomics results showed that DHM mainly played a significant role in anti-inflammatory, antioxidant, and fatty acid metabolism in biological processes, on lipoprotein and respiratory chain on cell components, and on redox-related enzyme activity, iron ion binding, and glutathione transferase on molecular functional processes. KEGG system analysis results showed that the PI3K-Akt signaling pathway, IL17 signaling pathway, HIF signaling pathway, MAPK signaling pathway, AGE-RAGE signaling pathway in diabetic complications, and TNF signaling pathway were typical signaling pathways disturbed by DHM in T2DM. Thirdly, molecular docking results showed that VEGFA, SRC, HIF1A, ESR1, KDR, MMP9, PPARG, and MAPK14 are key target genes, five genes of which were verified by RT-PCR in a dose-dependent manner. Finally, Western blotting results revealed that DHM effectively upregulated the expression of AKT protein and downregulated the expression of MEK protein in the liver of db/db mice. Therefore, our study found that DHM played a therapeutic effect partially by activation of the PI3K/AKT/MAPK signaling pathway. This study establishes the foundation for DHM as a novel therapeutic agent for T2DM. Additionally, it presents a fresh approach to utilizing natural plant extracts for chemoprevention and treatment of T2DM.
作者机构：
[Huarong Huang; Yi Xu] College of Agronomy and Biotechnology, China Agricultural University, Beijing, China;[Yini Wu; Tiansu Tan] College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, China;[Feng Chen] College of Horticulture, Anhui Agricultural University, Hefei, China
通讯机构：
[Yi Xu] C;College of Agronomy and Biotechnology, China Agricultural University, Beijing, China
摘要：
Cultivating perennial energy crops (PECs) not only enhances soil organic carbon (SOC) sequestration but also provides a reliable feedstock for bioenergy production. Nevertheless, the lack of exploration of the SOC sequestration rate and its spatiotemporal dynamics at a global scale hinders the industrial-scale implementation of PECs cultivation. In this study, we conducted a comprehensive global meta-analysis to quantify the SOC sequestration rate and elucidate its spatiotemporal dynamics. The results showed that PECs cultivation globally achieves a SOC sequestration rate of 0.21 Mg ha−1 yr−1. The crop age and soil depth were identified as the most important driving factors of the SOC sequestration rate. The SOC sequestration rate of PECs cultivation initially increased with crop age but eventually declined, reaching its peak at 9 years. Notably, the SOC sequestration rate was higher in deeper soil layers, particularly within 30–60 cm, compared to the topsoils (0–30 cm). Our results provide compelling evidence that PECs have promising potential for enhancing SOC sequestration, particularly in deep soils over the long term. These results contribute to the formulation of relevant policies aimed at effectively deploying PECs to facilitate the transition to renewable energy sources and achieve carbon neutrality.
作者机构：
[Liu, Gang; Liu, Naiyuan; Liu, Zhuangzhuang; Zhao, Jinfeng; Fang, Jun; Wang, Keyu] Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Hunan, Peoples R China.;[Liu, Gang; Liu, Naiyuan; Liu, Zhuangzhuang; Zhao, Jinfeng; Fang, Jun; Wang, Keyu] Hunan Engn Lab Pollut Control & Waste Utilizat Swi, Changsha 410128, Peoples R China.;[Yao, Hao] Changsha IMADEK Intelligent Technol Co Ltd, Changsha, Peoples R China.;[Pan, Junting] Chinese Acad Agr Sci, Inst Agr Resources & Reg Planning, State Key Lab Efficient Utilizat Arid & Semiarid A, Beijing 100081, Peoples R China.
通讯机构：
[Fang, J ] H;Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Hunan, Peoples R China.;Hunan Engn Lab Pollut Control & Waste Utilizat Swi, Changsha 410128, Peoples R China.
关键词：
Meta genomic analysis;Network analysis;Rice straw-mushroom compost;Sawdust-rice husk compost
摘要：
This study aimed to assess the impact of microbial agent and different compost material, on physicochemical parameters dynamic change, nitrogen-transfer gene/bacterial community interaction network during the pig manure composting. Incorporating a microbial agent into rice straw-mushroom compost reduced the NH(3) and total ammonia emissions by 25.52% and 14.41%, respectively. Notably, rice straw-mushroom with a microbial agent reduced the total ammonia emissions by 37.67%. NH(4)(+)-N and pH emerged as primary factors of phylum-level and genus-level microorganisms. Microbial agent increased the expression of narG, nirK, and nosZ genes. Rice straw-mushroom elevated the content of amoA, nirK, nirS, and nosZ genes. Alcanivorax, Luteimonas, Pusillimonas, Lactobacillus, Aequorivita, Clostridium, Moheibacter and Truepera were identified as eight core microbial genera during the nitrogen conversion process. This study provides a strategy for reducing ammonia emissions and analyzes the potential mechanisms underlying compost processes.
作者机构：
[Peng, Zhihao; Chen, Jinjun; Zou, Zhaoxia; Wei, Chao; Cai, Jisen] College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410000, China;[Zou, Zhaoxia] School of Public Health & Laboratory Medicine, Hunan University of Medicine, Huaihua, 418000, China. Electronic address: zhaoxia.zou@outlook.com;[Chen, Jinjun] Hunan Provincial Engineering Technology Research Center for Cell Mechanics and Function Analysis, Changsha, 418000, China. Electronic address: jinjunchen@hunau.edu.cn
摘要：
Spider venom boasts extensive peptide diversity, constituting a natural biochemical arsenal for defense and predation. The new family HvAMPs, including 9 homologous members, were identified from the unnormalized cDNA library of Heteropoda venatoria venom gland by Sanger sequencing. The putative mature peptide is composed of 22 aliphatic amino acid residues. The mature peptides of HvAMP1 and HvAMP5, with 3 different amino acids, were synthesized and both were shown to adopt an amphipathic α-helical structure and amphipathicity in SDS buffer by CD spectroscopy. In comparison to HvAMP1, HvAMP5 exhibits higher antibacterial activity, particularly against Gram-positive bacteria, coupled with reduced hemolytic activity and cytotoxicity. Results from SYTO 9/PI staining indicate that HvAMP5 acts by disrupting bacterial cell membranes. Analysis of the relationships between structures and functions suggests that HvAMP5 enhances antibacterial activity and reduces mammalian cell toxicity by increasing positive charge and proline substitution. The three residues variation can augment the electrostatic attraction of antibacterial peptides to the bacterial phospholipid bilayer. The present study suggests that the HvAMPs may exert lytic action against cells of different origins to increase cellular and tissue barrier permeability to facilitate spider's defense or predation. Moreover, HvAMP5 holds promise as a novel antibacterial agent for treating Gram-positive bacterial infections. Simultaneously, the numerous diverse amino acid residue substitutions within the HvAMP family offer a template for future study.
作者机构：
[Xiong, Xiaoran; Pan, Weisong; Kong, Zhicheng] College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, Hunan, China;[Xiong, Xiaoran; Pan, Weisong; Kong, Zhicheng] Hunan Engineering Research Center for Cell Mechanics and Functional Analysis, Changsha 410128, Hunan, China;[Wu, Chuan] Hunan Novomore Biotechnology Co., Ltd., Changsha 410000, Hunan, China
关键词：
molecular farming;plant bioreactor;protein purification;recombinant protein
摘要：
Plant bioreactor is a new production platform for expression of recombinant protein, which is one of the cores of molecular farming. In this study, the anti DYKDDDDK (FLAG) antibody was recombinantly expressed in tobacco (Nicotiana benthamiana) and purified. FLAG antibody with high affinity was obtained after immunizing mice for several times and its sequence was determined. Based on this, virus vectors expressing heavy chain (HC) and light chain (LC) inoculated into Nicotiana benthamiana leaves by using Agrobacterium-mediated delivery. Accumulation of the HC and LC was analyzed by SDS/PAGE followed by Western blotting probed with specific antibodies from 2 to 9 days postinfiltration (dpi). Accumulation of the FLAG antibody displayed at 3 dpi, and reached a maximum at 5 dpi. It was estimated that 66 mg of antibody per kilogram of fresh leaves could be obtained. After separation and purification, the antibody was concentrated to 1 mg/mL. The 1:10 000 diluted antibody can probe with 1 ng/mL FLAG fused antigen well, indicating the high affinity of the FLAG antibody produced in plants. In conclusion, the plant bioreactor is able to produce high affinity FLAG antibodies, with the characteristics of simplicity, low cost and highly added value, which contains enormous potential for the rapid and abundant biosynthesis of antibodies.
关键词：
RNA barcode segments;SARS-CoV-2 variants and related lineages;HCoVs;Genetic tests;Complete genome sequences
摘要：
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen responsible for coronavirus disease 2019 (COVID-19), continues to evolve, giving rise to more variants and global reinfections. Previous research has demonstrated that barcode segments can effectively and cost-efficiently identify specific species within closely related populations. In this study, we designed and tested RNA barcode segments based on genetic evolutionary relationships to facilitate the efficient and accurate identification of SARS-CoV-2 from extensive virus samples, including human coronaviruses (HCoVs) and SARSr-CoV-2 lineages. Nucleotide sequences sourced from NCBI and GISAID were meticulously selected and curated to construct training sets, encompassing 1733 complete genome sequences of HCoVs and SARSr-CoV-2 lineages. Through genetic-level species testing, we validated the accuracy and reliability of the barcode segments for identifying SARS-CoV-2. Subsequently, 75 main and subordinate species-specific barcode segments for SARS-CoV-2, located in ORF1ab, S, E, ORF7a, and N coding sequences, were intercepted and screened based on single-nucleotide polymorphism sites and weighted scores. Post-testing, these segments exhibited high recall rates (nearly 100%), specificity (almost 30% at the nucleotide level), and precision (100%) performance on identification. They were eventually visualized using one and two-dimensional combined barcodes and deposited in an online database (http://virusbarcodedatabase.top/). The successful integration of barcoding technology in SARS-CoV-2 identification provides valuable insights for future studies involving complete genome sequence polymorphism analysis. Moreover, this cost-effective and efficient identification approach also provides valuable reference for future research endeavors related to virus surveillance.
摘要：
Cadmium (Cd) is a highly toxic heavy metal that causes serious damage to plant and human health. Phytolacca acinosa Roxb. has a large amount of aboveground biomass and a rapid growth rate, and it has been identified as a novel type of Cd hyperaccumulator that can be harnessed for phytoremediation. However, the molecular mechanisms underlying the response of P. acinosa to Cd2+ stress remain largely unclear. In this study, the phenotype, biochemical, and physiological traits of P. acinosa seeds and seedlings were analyzed under different concentrations of Cd2+ treatments. The results showed higher Cd2+ tolerance of P. acinosa compared to common plants. Meanwhile, the Cd2+ content in shoots reached 449 mg/kg under 10 mg/L Cd2+ treatment, which was obviously higher than the threshold for Cd hyperaccumulators. To investigate the molecular mechanism underlying the adaptability of P. acinosa to Cd stress, RNA-Seq was used to examine transcriptional responses of P. acinosa to Cd stress. Transcriptome analysis found that 61 genes encoding TFs, 48 cell wall-related genes, 35 secondary metabolism-related genes, 133 membrane proteins and ion transporters, and 96 defense system-related genes were differentially expressed under Cd2+ stress, indicating that a series of genes were involved in Cd2+ stress, forming a complex signaling regulatory mechanism. These results provide new scientific evidence for elucidating the regulatory mechanisms of P. acinosa response to Cd2+ stress and new clues for the molecular breeding of heavy metal phytoremediation.
作者机构：
[Zhang, Ya; Zhou, Leyin; Zeng, Hao] College of Plant Protection, Hunan Agricultural University, Changsha 410128, China;[Wang, Chong] College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China;[Yang, Xiao] Key Laboratory of Land Surface Pattern and Simulation, Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences, Beijing 100101, China;[Liu, Shuangqing] College of Plant Protection, Hunan Agricultural University, Changsha 410128, China. Electronic address: liushuangqing@hunau.edu.cn
通讯机构：
[Shuangqing Liu] C;College of Plant Protection, Hunan Agricultural University, Changsha 410128, China
摘要：
Phenazine-1-carboxylic acid (PCA) is a new type of agrochemical used to prevent plant diseases, but its effects on aquatic organisms are unclear. To comprehensively assess the impacts of PCA for aquatic organisms and its associated environmental risks, this study investigated, taking zebrafish as the research object, the toxicological mechanism of PCA by means of optical microscopy, hematoxylin and eosin (HE) staining, ultrastructural observation, physiological and biochemical testing, transcriptome sequencing, metabolome analysis, fluorescence quantitative PCR and molecular simulation. The results indicated that PCA was detrimental to zebrafish embryos, larvae and adults, with LC(50) values at 96h of 3.9093mg/L, 8.5075mg/L, and 13.6388mg/L, respectively. PCA caused abnormal spontaneous movement, slowed the heart rate, delayed hatching, shortened the body length, slowed growth, and caused malformations. PCA mainly affected the brain, liver, heart, and ovaries. PCA distorted cell morphology, damaged mitochondrial membranes, disintegrated mitochondrial ridges, and dissociated nuclear membranes. PCA inhibited the enzyme activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX), decreased the malondialdehyde (MDA) content and disrupted antioxidant effects. The results of omics studies confirmed that PCA interfered with the transcriptional and metabolic network of zebrafish, downregulating most genes and metabolites. PCA mainly affected functions related to mitochondrial steroids, lipids, sterols, oxidoreductase activity and pathways involving cofactors, steroids, porphyrin, cytochromes, which specifically bound to targets such as panx3, agmat, and ace2. PCA was moderately toxic to zebrafish, and its usage should be strictly controlled to reduce toxic effects on aquatic organisms. The results of this study provide a new insights for ecotoxicology research.
期刊：
Current Microbiology,2024年81(1):1-8 ISSN：0343-8651
通讯作者：
Pan, H;Pan, Hu;Tian, Y
作者机构：
[Dai, Yan-na; Pan, H; Zhang, Yi-fan; Pan, Hu; Pu, Ji-feng] Tibet Acad Agr & Anim Husb Sci, Inst Agr Qual Stand & Testing, Lhasa 850032, Peoples R China.;[Tian, Yun; Liu, Hu-hu; Tian, Y; Pan, Hu; Pan, H] Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Peoples R China.;[Dai, Yan-na; Pan, H; Zhang, Yi-fan; Pan, Hu; Pu, Ji-feng] Agr & Livestock Prod Engn Technol Res Ctr Tibet Au, Lhasa 850032, Peoples R China.;[Yang, Xiao-feng] Sichuan Acad Agr Sci, Inst Agr Qual Stand & Testing Technol, Chengdu 610066, Peoples R China.
通讯机构：
[Pan, H ] T;[Tian, Y ; Pan, H] H;Tibet Acad Agr & Anim Husb Sci, Inst Agr Qual Stand & Testing, Lhasa 850032, Peoples R China.;Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Peoples R China.;Agr & Livestock Prod Engn Technol Res Ctr Tibet Au, Lhasa 850032, Peoples R China.
摘要：
A novel Gram-stain-negative, aerobic, rod-shaped bacterium named T808(T) was isolated from an alpine soil in Qamdo, Tibet, PR China. Strain T808(T) grew at 5-30degree celsius, pH 5.0-9.0 (optimum, 25degree celsius and pH 7.0-8.0) with 0-2% (w/v) NaCl (optimum, 0%). The 16S rRNA gene sequences of strain T808(T) showed the highest similarity with Pararhizobium herbae CCBAU83011(T) (98.8%), followed by Pararhizobium polonicum F5.1(T) (98.7%), Pararhizobium giardinii H152(T) (98.5%), Rhizobium gei ZFJT-2( T) (98.4%), and Pararhizobium antarcticum NAQVI59(T) (97.5%). The highest digital DNA-DNA hybridization (dDDH), core-proteome average amino acid identity (cpAAI) and average nucleotide identity (ANI) values between strain T808(T) and related strains were estimated as 28.0%, 92.1% and 84.4%, respectively. Phylogenetic analysis based on 16S rRNA, core-proteome and whole-genome indicated that strain T808(T) belonged to the genus Pararhizobium. The genome size was 6.24 Mbp with genomic DNA G + C content of 60.1%. The major cellular fatty acids were Summed feature 8 (C-18:1 omega 7c or C-18:1 omega 6c), C-16:0 and C-19:0 cyclo omega 8c. The polar lipids were diphosphatidyl glycerol, phosphatidyl glycerol, phosphatidyl ethanolamine, phosphatidyl choline and unidentified aminophospholipid. The isoprenoid quinone were ubiquinone-10 and ubiquinone-9. Based on phenotypic, phylogenetic, and genotypic data, strain T808(T) is considered to represent a novel species of the genus Pararhizobium, for which the name Pararhizobium qamdonense sp. nov. is proposed. The type strain is T808(T) (= JCM 36247( T) = CICC 25216( T)). According to phylogenetic coherence based on 16S rRNA, core-proteome and whole-genome, it is also proposed that the type strain Rhizobium gei Shi et al. 2016 should be reclassified as Pararhizobium gei comb. nov., the type strain is ZFJT-2( T) (= CCTCC AB 2013015( T) = KCTC 32301( T) = LMG 27603( T)).
摘要：
BACKGROUND AND PURPOSE: The P2X3 receptor, a trimeric ionotropic purinergic receptor, has emerged as a potential therapeutic target for refractory chronic cough (RCC). Nevertheless, gefapixant/AF-219, the only marketed P2X3 receptor antagonist, might lead taste disorders by modulating the human P2X2/3 (hP2X2/3) heterotrimer. Hence, in RCC drug development, compounds exhibiting strong affinity for the hP2X3 homotrimer and a weak affinity for the hP2X2/3 heterotrimer hold promise. An example of such a molecule is sivopixant/S-600918, a clinical Phase II RCC candidate with a reduced incidence of taste disturbance compared to gefapixant. Sivopixant and its analogue, (3-(4-([3-chloro-4-isopropoxyphenyl]amino)-3-(4-methylbenzyl)-2,6-dioxo-3,6-dihydro-1,3,5-triazin-1(2H)-yl)propanoic acid (DDTPA), exhibit both high affinity and high selectivity for hP2X3 homotrimers, compared with hP2X2/3 heterotrimers. The mechanism underlying the druggable site and its high selectivity remains unclear. EXPERIMENTAL APPROACH: To analyse mechanisms that distinguish this drug candidate from other inhibitors of the P2X3 receptors we used a combination of chimera construction, site covalent occupation, metadynamics, mutagenesis and whole-cell recording. KEY RESULTS: The high affinity and selectivity of sivopixant/DDTPA for hP2X3 receptors was determined by the tri-symmetric site located close to the upper vestibule. Substitution of only four amino acids inside the upper body domain of hP2X2 with those of hP2X3, enabled the hP2X2/3 heterotrimer to exhibit a similar level of apparent affinity for sivopixant/DDTPA as the hP2X3 homotrimer. CONCLUSION AND IMPLICATIONS: From the receptor-ligand recognition perspective, we have elucidated the molecular basis of novel RCC clinical candidates' cough-suppressing properties and reduced side effects, offering a promising approach to the discovery of novel drugs that specifically target P2X3 receptors.
摘要：
Recycling livestock manure in agroecosystems can maintain crop production, improve soil fertility, and reduce environmental losses. However, there has been no comprehensive assessment of synergies and trade-offs in the food-energy-soil-environment nexus under manure application. Here, we evaluate the sustainability of maize production under four fertilization regimes (mineral, mineral and manure mixed, manure, and no fertilization) from the aspect of food security, energy output, soil quality, and environmental impact based on a five-year field experiment. Manure and mineral mixed fertilization maintained grain and straw quantity and quality compared with mineral fertilization. Manure and mineral mixed fertilization increased stem/leaf ratio and field residue index by 9.1-28.9% and 4.5-17.9%, respectively. Manure also maintained the theoretical ethanol yield but reduced the straw biomass quality index by increasing ash. Further, manure application increased the soil quality index by 40.5% and reduced N(2)O emissions by 55.0% compared with mineral fertilization. Manure application showed the highest sustainability performance index of 19, followed by mineral (15), mixed (13), and without fertilization (8). In conclusion, manure application maintains food production and energy output, enhances soil quality, and reduces environmental impact, thereby improving the sustainability of maize production.
作者：
Yan Zhao;Xianyu Tang;Shitou Xia*;Jieying Nong;Kan Cui;...
期刊：
Journal of Fungi,2024年10(4):271- ISSN：2309-608X
通讯作者：
Shitou Xia
作者机构：
These authors contributed equally to this work.;[Yan Zhao; Xianyu Tang; Jieying Nong] Hunan Provincial Key Laboratory of Phytohormones and Growth Development, Hunan Agricultural University, Changsha 410128, China;Author to whom correspondence should be addressed.;[Kan Cui] Institute of Plant Protection, Hunan Academy of Agricultural Sciences, Changsha 410125, China;[Shitou Xia] Hunan Provincial Key Laboratory of Phytohormones and Growth Development, Hunan Agricultural University, Changsha 410128, China<&wdkj&>Author to whom correspondence should be addressed.
通讯机构：
[Shitou Xia] H;Hunan Provincial Key Laboratory of Phytohormones and Growth Development, Hunan Agricultural University, Changsha 410128, China<&wdkj&>Author to whom correspondence should be addressed.
关键词：
S. sclerotiorum;pathogenicity;histone methylation;mycotoxins;stress
摘要：
Histone post-translational modification is one of the main mechanisms of epigenetic regulation, which plays a crucial role in the control of gene expression and various biological processes. However, whether or not it affects fungal virulence in Sclerotinia sclerotiorum is not clear. In this study, we identified and cloned the histone methyltransferase Defective in methylation 5 (Dim5) in S. sclerotiorum, which encodes a protein containing a typical SET domain. SsDim5 was found to be dynamically expressed during infection. Knockout experiment demonstrated that deletion of SsDim5 reduced the virulence in Ssdim5-1/Ssdim5-2 mutant strains, accompanied by a significant decrease in H3K9 trimethylation levels. Transcriptomic analysis further revealed the downregulation of genes associated with mycotoxins biosynthesis in SsDim5 deletion mutants. Additionally, the absence of SsDim5 affected the fungus’s response to oxidative and osmotic, as well as cellular integrity. Together, our results indicate that the H3K9 methyltransferase SsDim5 is essential for H3K9 trimethylation, regulating fungal virulence throug mycotoxins biosynthesis, and the response to environmental stresses in S. sclerotiorum.
摘要：
In traditional Chinese medicine, Angelica dahurica is a valuable herb with numerous therapeutic applications for a range of ailments. There have not yet been any articles on the methodical assessment and choice of the best reference genes for A. dahurica gene expression studies. Real-time quantitative PCR (RT-qPCR) is widely employed as the predominant method for investigating gene expression. In order to ensure the precise determination of target gene expression outcomes in RT-qPCR analysis, it is imperative to employ stable reference genes. In this study, a total of 11 candidate reference genes including SAND family protein (SAND), polypyrimidine tract-binding protein (PTBP), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), TIP41-like protein (TIP41), cyclophilin 2 (CYP2), elongation factor 1 α (EF1α), ubiquitin-protein ligase 9 (UBC9), tubulin β-6 (TUB6), thioredoxin-like protein YLS8 (YLS8), and tubulin-α (TUBA) were selected from the transcriptome of A. dahurica. Subsequently, three statistical algorithms (geNorm, NormFinder, and BestKeeper) were employed to assess the stability of their expression patterns across seven distinct stimulus treatments. The outcomes obtained from these analyses were subsequently amalgamated into a comprehensive ranking using RefFinder. Additionally, one target gene, phenylalanine ammonia-lyase (PAL), was used to confirm the effectiveness of the selected reference genes. According to the findings of this study, the two most stable reference genes for normalizing the expression of genes in A. dahurica are TIP41 and UBC9. Overall, our research has determined the appropriate reference genes for RT-qPCR in A. dahurica and provides a crucial foundation for gene screening and identifying genes associated with the biosynthesis of active ingredients in A. dahurica.
摘要：
Cadmium (Cd) contamination is a widespread environmental issue. There is a lack of knowledge about the impacts of applying arbuscular mycorrhizal fungi (AMF) and biochar, either alone or in their combination, on alleviating Cd phytotoxicity in Ligustrum lucidum. Therefore, a pot experiment was conducted in a greenhouse, where L. lucidum seedlings were randomly subjected to four regimes of AMF treatments (inoculation with sterilized AMF, with Rhizophagus irregularis, Diversispora versiformis, alone or a mixture of these two fungi), and two regimes of biochar treatments (with or without rice-husk biochar), as well as three regimes of Cd treatments (0, 15, and 150mgkg(-1)), to examine the responses of growth, photosynthetic capabilities, soil enzymatic activities, nutritional concentrations, and Cd absorption of L. lucidum plants to the interactive effects of AMF, biochar, and Cd. The results demonstrated that under Cd contaminations, AMF alone significantly increased plant total dry weight, soil pH, and plant nitrogen (N) concentration by 84%, 3.2%, and 13.2%, respectively, and inhibited soil Cd transferring to plant shoot by 42.2%; biochar alone significantly enhanced net photosynthetic rate, soil pH, and soil catalase of non-mycorrhizal plants by 16.4%, 9%, and 11.9%, respectively, and reduced the soil Cd transferring to plant shoot by 44.7%; the additive effect between AMF and biochar greatly enhanced plant total dry weight by 101.9%, and reduced the soil Cd transferring to plant shoot by 51.6%. Furthermore, dual inoculation with D. versiformis and R. irregularis conferred more benefits on plants than the single fungal species did. Accordingly, amending Cd-contaminated soil with the combination of mixed-fungi inoculation and biochar application performed the best than either AMF or biochar alone. These responses may have been attributed to higher mycorrhizal colonization, soil pH, biomass accumulation, and biomass allocation to the roots, as well as photosynthetic capabilities. In conclusion, the combined use of mixed-fungi involving D. versiformis and R. irregularis and biochar addition had significant synergistic effects on enhancing plant performance and reducing Cd uptake of L. lucidum plants in Cd-contaminated soil.
