Lonicera macranthoides Hand.-Mazz. is a valuable medicinal plant in China and is used worldwide. This study aimed to predict its suitable habitats in China using the MaxEnt model, and to assess the effects of environmental variables on indicator ingredients (chlorogenic acid, macranthoidin B, and dipsacoside B) via HPLC and chemometrics. Furthermore, to explore the molecular mechanisms underlying environment-quality relationships, preliminary indoor versus outdoor stress experiments were conducted, analyzing the expression of chlorogenic acid biosynthetic genes using qRT-PCR. The results showed that precipitation of the driest month was the most influential variable affecting distribution. Currently, suitable areas are mainly located between 21° N and 33° N. During the Last Glacial Maximum (LGM), habitats were more expansive, whereas they contracted during the Mid-Holocene (MH). Future projections indicated habitat loss under the SSP585 scenario, which was partially mitigated under the SSP126 scenario by 2090 S. Higher contents of chlorogenic acid and saponins were found in suitable habitats and were associated with soil, altitude, and precipitation. Notably, outdoor combined stress (low temperature and low sunshine) significantly regulated the expression of LmPAL, LmCHS, LmCHI, LmC4H, LmCCoAOMT, and LmANS. This study serves as a scientific basis for the conservation, sustainable cultivation, and stress-oriented breeding of L. macranthoides in China.

Glyphosate-resistant crops, developed using the EPSPS gene, are widely cultivated globally. Previous studies indicate that the TIPS-OsEPSPS gene provides glyphosate resistance in rice. In this research, the OsmEPSPS gene was utilized to develop glyphosate-resistant, maize, and soybean, and a rapid immunochromatographic strip (ICS) test was designed for specific detection of the OsmEPSPS protein. The ICS test employs a double-antibody sandwich format with two monoclonal antibodies: mAb #9, conjugated to colloidal gold for capture, and mAb #15, for detection at the test line. The ICS exhibited a limit of detection (LOD) of 0.031 μg/mL within 10 min. Additionally, the ICS could detect a concentration of 0.817 μg/mL within 1 min in both Eleusine indica and OsmEPSPS-overexpressing plants. The ICS is highly specific, sensitive, and stable, with an R2 value of 0.982. Notably, the ICS is capable of detecting EPSPS proteins with high homology to OsmEPSPS from Poaceae and other plants. In summary, this study developed a sensitive and specific test strip for OsmEPSPS detection, facilitating semiquantitative analysis on the basis of colorimetric response time and intensity. This ICS is not only suitable for the rapid identification of OsmEPSPS-overexpressing plants but also for phylogenetic analysis of EPSPS gene homology.

Glyphosate resistance is a critically important trait for genetically modified (GM) crops. Mutation of the rice EPSPS gene results in a high level of glyphosate resistance, presenting significant potential for the development of glyphosate-tolerant crops. The resistance of rice to glyphosate is correlated with the expression levels of resistance genes. Therefore, developing a convenient, stable, and sensitive method for quantifying the OsmEPSPS protein is crucial for the development of glyphosate-resistant crops. We developed a double-antibody sandwich quantitative ELISA (DAS-ELISA) using a specific monoclonal antibody (mAb) for OsmEPSPS capture and an HRP-conjugated anti-OsmEPSPS rabbit polyclonal antibody (pAb). The method could be used to detect OsmEPSPS within a linear range of 16-256 ng/mL with robust intra- and inter-batch duplicability (%CV values: 0.17%-7.24%). OsmEPSPS expression in the transgenic rice lines (54.44-445.80 μg/g) was quantified using the DAS-ELISA. Furthermore, the expression of the OsmEPSPS gene was validated through Western blotting. This study demonstrated the reliability and stability of the DAS-ELISA for OsmEPSPS detection in GM rice.

Correction for ‘Lonicera japonica Thunb. as a promising antibacterial agent for Bacillus cereus ATCC14579 based on network pharmacology, metabolomics, and in vitro experiments’ by Nan Xu et al., RSC Adv., 2023, 13, 15379–15390, https://doi.org/10.1039/D3RA00802A.

Correction for 'Lonicera japonica Thunb. as a promising antibacterial agent for Bacillus cereus ATCC14579 based on network pharmacology, metabolomics, and in vitro experiments' by Nan Xu et al., RSC Adv., 2023, 13, 15379-15390, https://doi.org/10.1039/D3RA00802A.

为促进金银花多酚组分释放,提高其抗氧化、抗过敏能力,该文利用黑曲霉对金银花进行固体发酵.结果显示,以总多酚、总黄酮含量为参考指标,选择6 d作为最优发酵时间.发酵0～6 d,黑曲霉发酵金银花醇提物DPPH·清除率由61.25％提升至84.57％,·OH清除率由23.43％提升至30.10％,ABTS+·清除率由32.09％提升至55.06％,总还原力由0.730 9提升至0.826 9;透明质酸酶抑制率由13.51％提升至64.86％.总多酚含量由76.60 mg/g提升至97.56 mg/g,总黄酮含量由20.76 mg/g提升至24.12 mg/g,总皂苷含量由3.50 mg/g提升至4.26 mg/g.同时,测得黑曲霉所产纤维素酶、β-葡萄糖苷酶和木聚糖酶酶活的最高值分别为11.70、15.78、32.39 IU/g.该研究表明黑曲霉固态发酵金银花,能够促进金银花中多酚物质释放.

Lonicera japonica Thunb. has attracted much attention for its treatment of bacterial and viral infectious diseases, while its active ingredients and potential mechanisms of action have not been fully elucidated.

In order to investigate the protective effect of the ethanol extract of Lonicera macranthoides on oxidative stress induced by D-galactose in mouse embryonic fibroblasts（MEF） and its potential mechanism, the active substances in the ethanol extract of Lonicera macranthoides were detected by extensive target metabolome sequencing and high performance liquid chromatography. The MEF cells were treated with 20 mg/mL D-galactose to establish a cell aging model. According to experimental groups, the MEF cells of the control group（0 mg/m L D-galactose）, model group（20mg/mL D-galactose）, Lonicera macranthoides ethanol extract group（100, 500, 1000 μg/m L ethanol extract of Lonicera macranthoides added with 20 mg/mL D-galactose） and positive control group（50 μg/m L Panax notoginseng saponin added with 20 mg/mL D-galactose） had been grew for 48 hours, observed the number of cells with ordinary light microscope, detected the inhibition rate of cells with CCK-8 method, detected the activity of superoxide dismutase（SOD）and the content of malondialdehyde（MDA） with biochemical method, detected the level of mitochondrial membrane potential（MMP） and mitochondrial reactive oxygen species（mtROS） with flow cytometry, and detected the expression of p53, p16. Besides, the expression of mitochondrial complex gene, Bcl-2, Caspase-3, and the expression of mitochondrial complex protein, Bcl-2, Caspase-3 were detected by Western Blot. The results showed that the ethanol extract of Lonicera macranthoides contained phenolic acids, flavonoids, organic acids, terpenoids and other active substances. Compared with the control group, the inhibition rate of cells in the model group was significantly increased andcompared with the model group, the ethanol extract group of Lonicera macranthoides and the positive control group effectively reduced the cell inhibition rate（P＜0.05）, significantly increased the number of cells（P＜0.05, P＜0.01） and SOD activity, decreased the content of MDA（P＜0.05, P＜0.01）, increased MMP, decreased the level of mtROS（P＜0.01）, effectively reduced the protein levels of p53, p16, Caspase-3, and increased the protein levels of Bcl-2 and NDUFV1（P＜0.05）. It showed that in MEF cells, the ethanol extract of Lonicera macranthoides can resist the oxidative stress induced by D-galactose by increasing the concentration of mitochondrial complex I protein and slowing down cell apoptosis.

Light has important effects on plant metabolism. However, the relationship between the chlorogenic acid (CGA) content and light in plants remains unclear. Here, we investigated the effects of shading treatment on gene expression and CGA content in Lonicera macranthoides Hand.-Mazz. (LM), a widely used medicinal plant. A total of 1891 differentially expressed genes (DEGs) were obtained in flower buds and 819 in leaves in response to light in shading treatment compared to the control sample by RNA-Seq. After shading treatment, the content of CGA in LM leaves decreased significantly by 1.78-fold, the carotenoid content increased, and the soluble sugar and starch contents significantly decreased. WGCNA and the expression of related genes verified by qRT‒PCR revealed that CGA synthesis pathway enzyme genes form a co-expression network with genes for carbohydrate synthesis, photosynthesis, light signalling elements, and transcription factor genes (TFs) that affect the accumulation of CGA. Through a virus-induced gene silencing (VIGS) system and CGA assay in Nicotiana benthamiana (NB), we determined that downregulation of NbHY5 expression decreased the CGA content in NB leaves. In this study, we found that light provides energy and material for the accumulation of CGA in LM, and light affects the expression of CGA accumulation-related genes. Our results show that different light intensities have multiple effects on leaves and flower buds in LM and are able to coregulate LmHY5 expression and CGA synthesis.

Cold acclimation is a complex biological process leading to the development of freezing tolerance in plants. In this study, we demonstrated that cold-induced expression of protease inhibitor FmASP in a citrus relative species kumquat ( Fortunella margarita (Lour.) Swingle) contributes to its freezing tolerance by regulating protein degradation. First, we found that only cold-acclimated kumquat plants, although with extensive leaf cellular damage during freezing, are able to resume their normal growth upon stress relief. To dissect the impact of cold acclimation on this extraordinary freezing tolerance, we performed protein abundance assay and quantitative proteomics analysis of kumquat leaves subjected to cold acclimation (4 °C), freezing treatment (−10 °C) and post-stress recovery (25 °C). FmASP and a few non-specific proteases were identified as differentially expressed proteins induced by cold acclimation and associated with stable protein abundance throughout the course of freezing treatment. FmASP was further characterized as a robust inhibitor that inhibits the degradation capacity of multiple proteases. In addition, heterogeneous expression of FmASP in Arabidopsis confirmed its positive function in freezing tolerance. Finally, we proposed a working model of FmASP and illustrated how this extracellular-localized protease inhibitor protects proteins from degradation and consequently maintains essential cellular function for freezing stress recovery. These findings revealed the important role of protease inhibition on freezing response and provide insights on how this role may help develop new strategies to enhance plant freezing tolerance. One-sentence summary A protease inhibitor of Fortunella margarita enhances protein stability and freezing tolerance by regulating non-specific protease degradation

本文旨在基于网络药理学方法分析金银花抗菌有效成分，预测其分子作用机制。从中药系统药理学数据库与分析平台TCSMP数据库以及文献中收集金银花有效成分及其作用的靶点，通过Uniprot蛋白数据转化为相对应的靶点基因。将有效成分和靶点基因导入Cytoscape3.7.2软件，构建金银花有效成分-靶点网络调控图。通过GeneCards数据库检索并筛选抗菌靶点，与金银花有效成分-靶点取交集得到金银花抗菌核心靶点。将金银花核心靶点基因上传至STRING数据库，构建抗菌靶点蛋白相互作用网络图。最后，通过DAVID数据库将金银花关键靶点进行GO富集分析和KEGG通路富集分析。结果显示，在金银花中共获得23种与抗菌活性有关的潜在化合物，获得了137个抗菌靶点。GO富集分析获得条目17个，其中生物过程10个、分子功能1个、细胞组成6个；它们参与了22条相关信号通路。预测槲皮素、山奈酚、木犀草素、β-谷甾醇和β-胡萝卜素等化合物是金银花抗菌的主要成分。这些化合物作用于MYC原癌基因蛋白（MYC proto-oncogene protein,MYC）、管内皮生长因子A（vascular endothelial growth factor A,VEGFA）、白细胞介素-6（interleukin-6,IL-6）、肿瘤坏死因子（tumor necrosis factor,TNF）等关键靶点，并参与调控TNF信号通路、MAPK信号通路、ErbB信号通路、HIF-1信号通路和Toll样受体信号通路等发挥其抗菌作用。结果表明，金银花具有多成分、多靶点、多通路的抗菌分子机制，为深入阐释金银花及其抗菌分子机制提供理论参考。

糖基转移酶Ⅰ(GT Ⅰ)家族是糖基转移酶中的一类,其主要功能涉及黄酮类化合物糖基的转移、淀粉和蔗糖的合成等.为揭示GTⅠ基因家族在粳稻生长发育以及抗逆中的作用,为粳稻GTⅠ基因家族成员的功能研究提供基础,为水稻的品种改良和提高水稻耐高温提供思路,采用生物信息学方法鉴定出30 个GTⅠ基因家族成员,并分析了粳稻GTⅠ基因家族成员的基因结构、蛋白质理化性质、染色体定位及基因进化、系统发育关系、保守结构域和蛋白质三维结构建模等.结果显示,粳稻的30 个GT Ⅰ基因家族成员在水稻12 条染色体上均有分布,其基因结构、保守基序和遗传进化相对保守,家族成员启动子均预测到脱落酸响应元件,家族蛋白质的三维结构保守含有 6 个β-折叠和7 个α-螺旋.通过转录组数据与实时荧光定量分析,GTⅠ基因家族大部分成员对高温胁迫的响应出现在高温处理后第6 小时,其中OS-GT29 表达量呈持续高表达.GTⅠ基因家族在拟南芥和粳稻功能存在相似.

Rhus chinensis Mill. (RCM) is the host plant of Galla chinensis, which is valued in traditional medicine. Environmental temperature directly determines the probability of gallnut formation and RCM growth. At present, there is no experiment to systematically analyse the stability of internal reference gene (RG) expression in RCM. In this experiment, leaves that did not form gallnuts were used as the control group, while leaves that formed gallnuts were used as the experimental group. First, we conducted transcriptome experiments on RCM leaves to obtain 45 103 differential genes and functional enrichment annotations between the two groups. On this basis, this experiment established a transcriptional gene change model of leaves in the process of gallnut formation after being bitten by aphids, and RCM reference candidate genes were screened from RNA sequencing (RNA‐seq) data. This study is based on RCM transcriptome data and evaluates the stability of 11 potential reference genes under cold stress (4 °C) and heat stress (34 °C), using three statistical algorithms (geNorm, NormFinder, and BestKeeper). The results show that GAPDH1 + PP2A2/UBQ are stable reference genes under heat stress, while GAPDH1 + ACT are the most stable under cold stress. This study is the first to screen candidate reference genes in RCM and could help guide future molecular studies in this genus.

Superoxide dismutase (SOD) activity is an important measure of plant stress tolerance used in cultivar improvement. At present, we are unaware of any widely available immunological reagents for the detection of SOD in Oryza sativa (common Asian rice) or other plants. In this study, we used insilico B-cell epitope prediction tools to generate peptides which were immunized into rabbits to yield polyclonal antibodies against Cu/Zn SODs. Immunoblotting demonstrated that the antibody specifically recognized both native and denatured Cu/Zn SODs in rice. In addition, this antibody can confirm the expression tendency of endogenous OsCu/Zn SODs under heat stress by immunoblotting, and has a positive reaction in tomato leaf extracts, as well as human Hela cells. Chloroplast content of Cu/Zn SODs in rice can be identified by ELISA indirect competition method using this antibody. These results suggest that this Cu/Zn SOD rabbit polyclonal antibody may be a useful tool for elucidating the biological functions of Cu/Zn SODs in plants.

Cytokinin plays a very important role in plants growth and development, and CRE1 has been identified as a cytokinin receptor. However, the biological function of CRE1 in cotton remains unclear. In this paper, GhCRE1 gene was cloned and its sequence was analyzed by bioinformatics. The results revealed that GhCRE1 had 11 exons and 10 introns, and its molecular weight, theoretical isoelectric point (pI) and number of amino acids differed from those of the homologous gene in Theobroma cacao and Arabidopsis thaliana, with three different protein domains. 15 unidentified proteins were found in potential interaction with CRE1 protein and 2 phosphorylation sites on CRE1 protein sequence were predicted by mutiple bioinformatics websites. Additionally, 8 cis-acting regulatory elements were detected on CRE1 promoter sequence, and found related to light signal and hormone. These results were conducive to unfolding the function of GhCRE1 in upland cotton.

芥菜(Brassica juncea L.)在中国种植广泛、历史悠久,是中国重要的特色蔬菜作物,近年来在乡村振兴中发挥着重要作用.中国芥菜种质资源丰富,选育的品种类型多样,包括根用芥菜、茎用芥菜、叶用芥菜以及薹用芥菜等.本综述阐述了中国芥菜的育种方法研究现状,介绍了审定、登记及保护的芥菜新品种的特征特性及来源,梳理了AFLP、SSR、RAPD等分子标记在芥菜种质资源评价、遗传多样性分析、杂交种种子纯度鉴定等方面的应用进展,以期为芥菜相关分子标记应用方面的研究提供信息.

Fatty acid desaturase-2 (FAD2) is a key enzyme in the production of polyunsaturated fatty acids in plants. RNAi technology can reduce the expression of FAD2 genes in Brassica napus seeds and acquire transgenic B. napus plants with a high oleic acid content, but the effect of seed-specific inhibition of FAD2 expression on B. napus seed metabolites is not clear. Here we use widely targeted metabolomics to investigate the metabolites of normal-oleic-acid rapeseed (OA) and high-oleic-acid rapeseed (HOA) seeds, resulting in a total of 726 metabolites being detected. Among them, 24 differential metabolites were significantly downregulated and 88 differential metabolites were significantly upregulated in HOA rapeseed. In further lipid profile experiments, more lipids in B. napus seeds were accurately quantified. The contents of glycolipids and phospholipids that contain C18:1 increased significantly and C18:2 decreased because FAD2 expression was inhibited. The changes in the expression of key genes in related pathways were also consistent with the changes in metabolites. The insertion site of the ihpRNA plant expression vector was reconfirmed through genomewide resequencing, and the transgenic event did not change the sequence of FAD2 genes. There was no significant difference in the germination rate and germination potential between OA and HOA rapeseed seeds because the seedspecific ihpRNA plant expression vector did not affect other stages of plant growth. This work provides a theoretical and practical guidance for subsequent molecular breeding of high OA B. napus.

本文以具有抗二化螟性状的Csu-miR260转基因水稻为试验材料,采用TaqMan探针实时荧光定量PCR(TaqMan RT-qPCR)和茎环实时荧光定量PCR(stem-loop RT-qPCR)分别对Csu-miR260转基因抗虫水稻中Csu-miR260插入序列和剪切形成的amiR260s的表达情况进行了定量检测.发现Csu-miR260插入序列和剪切形成的20 nt和21 nt两种长度amiR260s在转基因抗虫水稻苗期的根、茎、叶中表达,且无组织表达特异性.该试验解决了人工miRNA作物中amiRNA及前体检测引物的特异性问题,为人工miRNA植物干扰序列的表达分析提供技术参考,并为miRNA转基因作物遗传表达相关安全评价提供了数据参考.

MicroRNAs (miRNAs) are small, non-coding, regulatory RNAs that play important roles in abiotic stress responses in plants, but their regulatory roles in the adaptive response to heat stress at the booting stage in two rice varieties, 9311 and Nagina 22, remain largely unknown. In this study, 464 known miRNAs and 123 potential novel miRNAs were identified. Of these miRNAs, a total of 90 differentially expressed miRNAs were obtained with 9311 libraries as the control group, of which 54 were upregulated and 36 were downregulated. To gain insight into functional significance, 2773 potential target genes of these 90 differentially expressed miRNAs were predicted. GO enrichment analysis showed that the predicted target genes of differentially expressed miRNAs included NACs, LACs, CSD, and Hsp40. KEGG pathway analysis showed that the target genes of these differentially expressed miRNAs were significantly enriched in the plant hormone signal transduction pathway. The expression levels of 10 differentially expressed miRNAs and their target genes obtained by qRT-PCR were largely consistent with the sequencing results. This study lays a foundation for the elucidation of the miRNA-mediated regulatory mechanisms in rice at elevated temperatures.

利用SSR分子标记建立芥菜品种鉴定体系,旨在为我国芥菜育成品种鉴定提供高通量、快速鉴定方法.选取芥菜16个变种代表性品种用于SSR引物初筛,通过聚丙烯酰胺凝胶电泳从432对引物中筛选出84对条带清晰、稳定性好、多态性高的芥菜SSR引物,并对筛选出的引物5'端进行荧光标记.另外选取96份品种对上述84对引物进行复筛,利用基因分析仪对所有扩增片段大小进行分析,依据各引物的扩增稳定性、峰型易读取程度、多态性及染色体位置等,最终筛选出25对SSR引物作为核心引物,并为其设置参照品种以消除不同试验批次或平台等因素引起的误差.根据荧光颜色和扩增片段大小将25对核心引物分成6组,建立基于SSR标记的芥菜品种鉴定体系.利用该套核心引物构建了 189份芥菜品种的DNA指纹数据库,共检测到175个等位变异,平均每对引物检测到7个等位变异;基因多样性指数变化为0.239～0.870,平均为0.555;多态性信息含量在0.23～0.86之间,平均为0.493.本研究建立的基于毛细管电泳平台的芥菜SSR分子标记品种鉴定体系,可用于芥菜品种真实性快速鉴定、遗传多样性分析和DUS测试近似品种辅助筛选等.