A Comprehensive Analysis to Elucidate the Effects of Spraying Mineral Elements on the Accumulation of Flavonoids in Epimedium sagittatum during the Harvesting Period.

The harvesting period is a critical period for the accumulation of flavonoids in the leaves of the important medicinal plant Epimedium sagittatum. In this study, we conducted an experiment on E. sagittatum leaves sprayed with mineral elements with the aim of improving the quality of the herbal leafage during the harvesting period. We elucidated the changes in flavonoids (icariin, epimedin A, epimedin B, and epimedin C) in E. sagittatum leaves. The sum of main flavonoids content reached a maximum (11.74%) at 20 days after the high-concentration Fe2+ (2500 mg·L-1) treatment. We analyzed the FT-IR spectra characteristics of E. sagittatum leaf samples using the FT-IR technique, and constructed an OPLS-DA model and identified characteristic peaks to achieve differentiated identification of E. sagittatum. Further, widely untargeted metabolomic analysis identified different classes of metabolites. As the most important characteristic flavonoids, the relative contents of icariin, icaritin, icariside I, and icariside II were found to be up-regulated by high-Fe2+ treatment. Our experimental results demonstrate that high-concentration Fe2+ treatment is an effective measure to increase the flavonoids content in E. sagittatum leaves during the harvesting period, which can provide a scientific basis for the improvement of E. sagittatum leaf cultivation agronomic measures.

Study on the Regulation of Exogenous Hormones on the Absorption of Elements and the Accumulation of Secondary Metabolites in the Medicinal Plant Artemisia argyi Leaves.

As an important medicinal plant, we still do not know the effect of exogenous hormones on absorption of elements and accumulation of secondary metabolites in Artemisia argyi leaves. In this work, we analyzed the difference in 21 elements absorbed by A. argyi leaves under three exogenous hormone (MeJA, SA and ABA) treatments, and also clarified the correlation between 21 elements and eight bioactive components. Different hormone treatments changed the absorption and enrichment of elements, and the composition also changed significantly. The contents of eight bioactive components changed significantly under different hormone treatments. When A. argyi was stimulated by exogenous hormones, the content of secondary metabolites was adjusted in the leaves through changes in the absorption and enrichment of elements. The widely untargeted metabolomic analysis further confirmed that ABA changes the metabolic direction of secondary metabolites in A. argyi leaves and stimulates the biosynthesis of multiple secondary metabolites including phenylpropanoids, flavonoids, terpenoids, alkaloids and others. These results provide a new perspective for the changes in element absorption and the mechanism of secondary metabolic components in A. argyi leaves under exogenous hormone treatments, and also deepen people's understanding of the interaction mechanism between medicinal plants and hormones.

Drought Stress Stimulates the Terpenoid Backbone and Triterpenoid Biosynthesis Pathway to Promote the Synthesis of Saikosaponin in Bupleurum chinense DC. Roots.

Bupleurum chinense is an important medicinal plant in China; however, little is known regarding how this plant transcribes and synthesizes saikosaponins under drought stress. Herein, we investigated how drought stress stimulates the transcriptional changes of B. chinense to synthesize saikosaponins. Short-term drought stress induced the accumulation of saikosaponins, especially from the first re-watering stage (RD_1 stage) to the second re-watering stage (RD_2 stage). Saikosaponin-a and saikosaponin-d increased by 84.60% and 75.13%, respectively, from the RD_1 stage to the RD_2 stage. Drought stress also stimulated a rapid increase in the levels of the hormones abscisic acid, salicylic acid, and jasmonic acid. We screened 49 Unigenes regarding the terpenoid backbone and triterpenoid biosynthesis, of which 33 differential genes were significantly up-regulated during drought stress. Moreover, one P450 and two UGTs are possibly involved in the synthesis of saikosaponins, while some transcription factors may be involved in regulating the expression of key enzyme genes. Our study provides a reference for the cultivation of B. chinense and a practical means to ensure the quality (safety and effectiveness) of B. chinense for medicinal use, as well as insights into the modernization of the China Agriculture Research System.

Osthole Induces Apoptosis and Inhibits Proliferation, Invasion, and Migration of Human Cervical Carcinoma HeLa Cells.

PURPOSE: To study the effect of osthole extract on proliferation, migration, invasion, and apoptosis of human cervical carcinoma HeLa cells and investigate its underlying mechanism. METHODS: HeLa cells were exposed to osthole at various concentrations. Cell viability, migration, and invasion were detected by MTT assay, scratch wound-healing assay, and invasion assay, respectively. The proportion of cells undergoing apoptosis was analyzed by flow cytometry. Western blot and RT-qPCR were performed to determine changes in the expression of key factors in the Wnt/ß-catenin signaling pathway. RESULTS: The osthole extract effectively inhibited the proliferation, migration, and invasion potential of HeLa cells in a dose-dependent manner. The rate of apoptosis induction in HeLa cells treated with the osthole extract for 48 h was significantly higher than that of the untreated controls. Outcomes of the western blotting analysis and RT-qPCR showed that the expression of ß-catenin, c-Myc, cyclin D1, survivin, and MMP-9 was significantly inhibited. CONCLUSION: Osthole could significantly inhibit the malignant behavior of HeLa cells and induce cellular apoptosis. Inactivation of the Wnt/ß-catenin signaling pathway by osthole may be a mechanism to control cancer metastasis.

Isolation of endophytic bacteria from Rehmannia glutinosa Libosch and their potential to promote plant growth.

In order to study the growth promoting potential of endophytic bacteria from Rehmannia glutinosa Libosch, a total of 25 different bacteria belonging to 7 genera were identified by 16S rRNA gene sequencing, including Bacillus, Micrococcus, Lysinibacillus, Brevibacterium, Halomonas, Kocuria and Terribacillus. In this study, thirteen bacterial strains were found to solubilize inorganic phosphate, with the isolate Kocuria rosea (EH15) having the highest phosphorus dissolution activity (3.70 µg/mL). Twelve isolates were positive for nitrogen fixation abilities. Twenty-two strains produced indole-3-acetic acid (IAA) in the presence of L-tryptophan, and eleven of the twenty-two isolates synthesized IAA in the absence of L-tryptophan. The strain K. rosea (EH15) was capable of producing the highest IAA amount (15.36 and 7.98 mg/L) in Luria Bertani (LB) broth containing 0.2% L-tryptophan and lacking L-tryptophan, respectively. Ten isolates had siderophore production abilities with Bacillus amyloliquefacieus EH10 (0.26) and Brevibacterium frigoritolerans EH13 (0.32) showing high siderophore production characteristics. Five bacteria endogenous were selected to evaluate the growth parameters of Brassica napus L. and all isolates exhibited a significantly greater increase in seedling height, root length, fresh weight and dry weight, than the control plants. The greatest improvement appeared in the case of co-inoculation of EH10 and EH15, except in dry weight, and the biggest enhancement in dry weight occurred in the strain EH15. In general, these endophytic bacteria indicate a potential as microbial fertilizers to promote the growth of R. glutinosa Libosch.

[Cloning and Expression Analysis of Acetyl-CoA C-acetyltransferase Gene in Isodon rubescens].

Objective: To clone the acetyl-CoA C-acetyl transferase( AACT) gene from Isodon rubescens, and to analyze the bioinformatics and expression of the gene. Methods: According to the IrAACT gene sequence of Isodon rubescens transcriptome,a pair of primers was designed, and the ORF of cDNA sequence was obtained by reverse transcription PCR. Bioinformatic analysis of this gene and its corresponding protein were performed. Real-time quantitative PCR( q PCR) was used to detect the relative expression levels of IrAACT different tissues of Isodon rubescens. Results: The IrAACT cDNA sequence contained a 1 254 bp open reading frame and encoded a predicted protein of 417 amino acids. IrAACT had extensive homology with AACTs from other plant species, such as Salvia miltiorrhiza, et al. Bioinformatic analysis showed that IrAACT-encoding protein contained the thiolase Ⅱ catalytic domain. q PCR analysis showed that the expression of IrAACT was tissue-specific, and accumulation of transcripts was greater in flowers and leaves, followed by stems, roots and callus. Conclusion: It is the first time to report IrAACT gene and its relative expression level. The results will provide a groundwork for studying the function of IrAACT in terpenoid biosynthesis of Isodon rubescens.

Prodigiosin found in Serratia marcescens y2 initiates phototoxicity in the cytomembrane

Background: Light can be absorbed by bacterial pigment and affects its growth. Prodigiosin is a red pigment found in various bacterial species. The purpose of this study was to investigate the impacts of light on prodigiosin production, biomass formation, and membrane integrity of Serratia marcescens y2. Results: S. marcescens y2 grew better and produced more intracellular prodigiosin in darkness than in illumination. The pigment leakage ratio from cells was detected more in light than in darkness conditions. Ethidium bromide uptake assay could visually prove the prodigiosin-related loss of membrane integrity under illumination. A higher concentration of malondialdehyde (MDA) was detected in light-treated culture than in darkness. Tests of different light treatments (red, yellow, blue and green) showed that the maximum extracellular pigment and the minimum biomass formation and intracellular pigment were obtained in green light. Conclusions: Prodigiosin could absorb light, and then initiate phototoxicity damage of the cytomembrane.

Fine mapping TaFLW1, a major QTL controlling flag leaf width in bread wheat (Triticum aestivum L.).

INTRODUCTION: Flag leaf width (FLW) is directly related to photosynthetic capacity and yield potential in wheat. In a previous study, Qflw.nau-5A controlling FLW was detected on chromosome 5A in the interval possessing Fhb5 for type I Fusarium head blight (FHB) resistance using a recombinant inbred line population derived from Nanda2419 × Wangshuibai. MATERIALS AND METHODS: Qflw.nau-5A near-isogenic line (NIL) with the background of Mianyang 99-323 and PH691 was developed and evaluated. FLW inheritance was investigated using two F2 populations developed from crossing the Qflw.nau-5A NILs with their recurrent parents. One hundred ten and 28 recombinants, which included 10 and 5 types of recombinants, were identified from 2816 F2 plants with Mianyang 99-323 background and 1277 F2 plants with PH691 background, respectively, and phenotyped in field trials for FLW and type I FHB resistance. Deletion bin mapping was applied to physically map Qflw.nau-5A. RESULTS AND CONCLUSIONS: The introduction of Wangshuibai Qflw.nau-5A allele reduced the FLW up to 3 mm. In the F2 populations, Qflw.nau-5A was inherited like a semi-dominant gene, and was therefore designated as TaFLW1. The FLW of the recombinant lines displayed a distinct two-peak distribution. Recombinants with wider leaves commonly have Mianyang 99-323 or PH691 chromatin in the 0.2 cM Xwmc492-Xwmc752 interval that resided in the 5AL12-0.35-0.57 deletion bin, and recombinants with narrow leaves were Wangshuibai genotype in this interval. Phenotypic recombination between FLW and type I FHB resistance was identified, implying TaFLW1 was in close linkage with Fhb5. These results should aid wheat breeders to break the linkage drag through marker-assisted selection and assist in the map-based cloning of TaFLW1.

[Effect of N, P, K on yield and quality of Rabdosia rubescens].

OBJECTIVE: To research the three fertilizer of N, P, K impacting Rabdosia rubescens on the yield and quality. METHODS: The optimum design of three factors with D-saturation design plan of N, P, K was adopted in the pot experiment. RESULTS: The fertilizers of N, P, K made an obvious improvement in the growth, yield and quality of Rabdosia rubescens. CONCLUSIONS: The N fertilizer plays an important role in the growth of plants. The K fertilizer makes a great impact on the accumulating of oridonin. The N, P, K fertilizers applied reasonably can make the Rabdosia rubescens high yields and improve its quality.