Investigation of the Inhibitory Effects of Illicium verum Essential Oil Nanoemulsion on Fusarium proliferatum via Combined Transcriptomics and Metabolomics Analysis.

Fusarium proliferatum is the main pathogen that causes Panax notoginseng root rot. The shortcomings of strong volatility and poor water solubility of Illicium verum essential oil (EO) limit its utilization. In this study, we prepared traditional emulsion (BDT) and nanoemulsion (Bneo) of I. verum EO by ultrasonic method with Tween-80 and absolute ethanol as solvents. The chemical components of EO, BDT, and Bneo were identified by gas chromatography-mass spectrometry (GC-MS) and the antifungal activity and mechanism were compared. The results show that Bneo has good stability and its particle size is 34.86 nm. The contents of (-) -anethole and estragole in Bneo were significantly higher than those in BDT. The antifungal activity against F. proliferatum was 5.8-fold higher than BDT. In the presence of I. verum EO, the occurrence of P. notoginseng root rot was significantly reduced. By combining transcriptome and metabolomics analysis, I. verum EO was found to be involved in the mutual transformation of pentose and glucuronic acid, galactose metabolism, streptomycin biosynthesis, carbon metabolism, and other metabolic pathways of F. proliferatum, and it interfered with the normal growth of F. proliferatum to exert antifungal effects. This study provide a theoretical basis for expanding the practical application of Bneo.

The hypothalamic steroidogenic pathway mediates susceptibility to inflammation-evoked depression in female mice.

BACKGROUND: Depression is two-to-three times more frequent among women. The hypothalamus, a sexually dimorphic area, has been implicated in the pathophysiology of depression. Neuroinflammation-induced hypothalamic dysfunction underlies behaviors associated with depression. The lipopolysaccharide (LPS)-induced mouse model of depression has been well-validated in numerous laboratories, including our own, and is widely used to investigate the relationship between neuroinflammation and depression. However, the sex-specific differences in metabolic alterations underlying depression-associated hypothalamic neuroinflammation remain unknown. METHODS: Here, we employed the LPS-induced mouse model of depression to investigate hypothalamic metabolic changes in both male and female mice using a metabolomics approach. Through bioinformatics analysis, we confirmed the molecular pathways and biological processes associated with the identified metabolites. Furthermore, we employed quantitative real-time PCR, enzyme-linked immunosorbent assay, western blotting, and pharmacological interventions to further elucidate the underlying mechanisms. RESULTS: A total of 124 and 61 differential metabolites (DMs) were detected in male and female mice with depressive-like behavior, respectively, compared to their respective sex-matched control groups. Moreover, a comparison between female and male model mice identified 37 DMs. We capitalized on biochemical clustering and functional enrichment analyses to define the major metabolic changes in these DMs. More than 55% of the DMs clustered into lipids and lipid-like molecules, and an imbalance in lipids metabolism was presented in the hypothalamus. Furthermore, steroidogenic pathway was confirmed as a potential sex-specific pathway in the hypothalamus of female mice with depression. Pregnenolone, an upstream component of the steroid hormone biosynthesis pathway, was downregulated in female mice with depressive-like phenotypes but not in males and had considerable relevance to depressive-like behaviors in females. Moreover, exogenous pregnenolone infusion reversed depressive-like behaviors in female mice with depression. The 5α-reductase type I (SRD5A1), a steroidogenic hub enzyme involved in pregnenolone metabolism, was increased in the hypothalamus of female mice with depression. Its inhibition increased hypothalamic pregnenolone levels and ameliorated depressive-like behaviors in female mice with depression. CONCLUSIONS: Our study findings demonstrate a marked sexual dimorphism at the metabolic level in depression, particularly in hypothalamic steroidogenic metabolism, identifying a potential sex-specific pathway in female mice with depressive-like behaviors.

Control of pathogenic fungi on Panax notoginseng by volatile oils from the food ingredients Allium sativum and Foeniculum vulgare.

To screen natural drugs with strong inhibitory effects against pathogenic fungi related to P. notoginseng, the antifungal activities of garlic and fennel EOs were studied by targeting P. notoginseng disease-associated fungi, and the possible action mechanisms of garlic and fennel EOs as plant fungicides were preliminarily discussed. At present, the antifungal mechanism of EOs has not been fully established. Therefore, understanding the antifungal mechanism of plant EOs is helpful to address P. notoginseng diseases continuous cropping disease-related obstacles and other agricultural cultivation problems. First, the Oxford cup method and chessboard were used to confirm that the EOs and oxamyl had a significant inhibitory effect on the growth of Fusarium oxysporum. F. oxysporum is the main pathogen causing root rot of P. notoginseng and the preliminary study on the antifungal mechanisms of the EOs against F. oxysporum showed that the inhibition of EOs mainly affects cell membrane permeability and cell processes and affects the enzyme activities of micro-organism, to achieve antifungal effects. Finally, an in vivo model verified that both two EOs could significantly inhibit the occurrence of root rot caused by F. oxysporum.

Anticoagulant activity analysis and origin identification of Panax notoginseng using HPLC and ATR-FTIR spectroscopy.

INTRODUCTION: Panax notoginseng is one of the traditional precious and bulk-traded medicinal materials in China. Its anticoagulant activity is related to its saponin composition. However, the correlation between saponins and anticoagulant activities in P. notoginseng from different origins and identification of the origins have been rarely reported. OBJECTIVES: We aimed to analyze the correlation of components and activities of P. notoginseng from different origins and develop a rapid P. notoginseng origin identification method. MATERIALS AND METHODS: Pharmacological experiments, HPLC, and ATR-FTIR spectroscopy (variable selection) combined with chemometrics methods of P. notoginseng main roots from four different origins (359 individuals) in Yunnan Province were conducted. RESULTS: The pharmacological experiments and HPLC showed that the saponin content of P. notoginseng main roots was not significantly different. It was the highest in main roots from Wenshan Prefecture (9.86%). The coagulation time was prolonged to observe the strongest effect (4.99 s), and the anticoagulant activity was positively correlated with the contents of the three saponins. The content of ginsenoside Rg1 had the greatest influence on the anticoagulant effect. The results of spectroscopy combined with chemometrics show that the variable selection method could extract a small number of variables containing valid information and improve the performance of the model. The variable importance in projection has the best ability to identify the origins of P. notoginseng; the accuracy of the training set and the test set was 0.975 and 0.984, respectively. CONCLUSION: This method is a powerful analytical tool for the activity analysis and identification of Chinese medicinal materials from different origins.

Root-Associated Microbiomes of Panax notoginseng under the Combined Effect of Plant Development and Alpinia officinarum Hance Essential Oil.

Essential oils (EOs) have been proposed as an alternative to conventional pesticides to inhibit fungal pathogens. However, the application of EOs is considerably limited due to their highly volatile nature and unpredictable effects on other microbes. In our study, the composition of bacterial and fungal communities from the rhizosphere soil of P. notoginseng under four treatment levels of Alpinia officinarum Hance EO was characterized over several growth stages. Leaf weight varied dramatically among the four EO treatment levels after four months of growth, and the disease index at a low concentration (0.14 mg/g) of EO addition was the lowest among the P. notoginseng growth stages. The content of monomeric saponins was elevated when EO was added. Bacterial and fungal diversity in the absence of plants showed a decreasing trend with increasing levels of EO. Bacterial diversity recovery was more correlated with plant growth than was fungal diversity recovery. Compared with the control (no EO addition), a low concentration of EO significantly accumulated Actinomycota, including Acidothermus, Blastococcus, Catenulispora, Conexibacter, Rhodococcus, and Sinomonas, after one month of plant-microbial interaction. Overall, the results showed that both the plant growth stage and EOs drive changes in the microbial community composition in the rhizosphere of P. notoginseng. Plant development status had a stronger influence on bacterial diversity than on fungal diversity. EO had a more significant effect on fungal community composition, increasing the dominance of Ascomycota when EO concentration was increased. Under the interaction of P. notoginseng growth and EO, a large number of bacterial genera that have been described as plant growth-promoting rhizobacteria (PGPR) responded positively to low concentrations of EO application, suggesting that EO may recruit beneficial microbes in the root zone to cope with pathogens and reduce root rot disease. These results offer novel insights into the relationship between EO application, altered microbial communities in the plant roots, plant growth stage, and disease occurrence.

Chemical Constituents of the Essential oil from Cuminum cyminum L. and Its Antifungal Activity against Panax notoginseng Pathogens.

Cuminum cyminum L. (Cumin) is a flavoring agent that is commonly used worldwide, and is rich in essential oil. Essential oils (Eos) have been intensively investigated in regard to their potential for disease control in plants, which is provided a chance for the blossom of green pesticides. The chemical components of Cumin essential oil (CEO) were revealed by GC/MS, such as cuminaldehyde (44.53 %), p-cymene (12.14 %), (-)-ß-pinene (10.47 %) and γ-terpinene (8.40 %), and found they can inhibit the growth of P. notoginseng-associated pathogenic fungi in vitro and the inhibitory effect of cuminaldehyde was similar to that of hymexazol. SEM and TEM images demonstrated that cuminaldehyde and CEO increased cell permeability and disrupted membrane integrity. The expression of disease-related genes of Fusarium oxysporum showed that CEO induced the expression of most genes, which disrupted biosynthesis, metabolism and signaling pathways. These studies verified the potential of CEO as a plant fungicide that is environmentally friendly and provided ideas for developing new products for controlling root diseases that affect P. notoginseng.

Biological Activities of Two Essential Oils from Pogostemon cablin and Eupatorium fortunei and Their Major Components against Fungi Isolated from Panax notoginseng.

Panax notoginseng (Burkill) F.H.Chen (Araliaceae), of which the dry root and rhizome are precious traditional Chinese medicine, suffers severely from diseases during planting. Essential oils (EOs) with antimicrobial activity are a possibility for the development of green pesticides. We extracted EOs from Pogostemon cablin (Blanco) Benth. and Eupatorium fortunei Turcz., respectively and tested their inhibitory rates on fungi isolated from diseased P. notoginseng by the Oxford cup method. The compounds of the EO were identified by GC/MS and the minimum inhibitory concentrations (MICs) of the EOs and their main components were evaluated by the 96-well plate method. We also mixed P. cablin EO, E. fortunei EO and hymexazol in pairs to explore whether their combinations produce stronger antifungal effects than individual components. Finally, we evaluated the effects of the EOs against Fusarium oxysporum in vivo. P. cablin EO and E. fortunei EO exhibited different antifungal activities against fungi, with the inhibitory rates of 21.02 %-100 % and 54.84 %-100 % and MICs of 0.07-0.88 mg/mL and 0.20-1.17 mg/mL, respectively. Pogostone (24.96 %) and thymol (15.64 %) were the major compounds of P. cablin EO and E. fortunei EO, respectively, and they exhibited stronger antifungal activities than EOs, with MICs of 0.008-0.078 mg/mL and 0.12-0.31 mg/mL, respectively. Moreover, hymexazol was mixed with E. fortunei EO, and the inhibitory effect against Cylindrocarpon destructans was enhanced with a synergistic effect. The disease incidence and disease index of EO treatments decreased significantly in vivo. Based on our study, P. cablin EO and E. fortunei EO have great potential to be developed into green fungicides for use in agriculture to control diseases of P. notoginseng.

Fungicidal Activity of Essential Oils from Cinnamomum cassia against the Pathogenic Fungi of Panax notoginseng Diseases.

The frequent disease of Panax notoginseng caused by the pathogenic fungi in field cultivation has become the major threaten to the sustainable development of it. The present study was conducted to find natural agent with potential inhibition against pathogen. Therefore, the inhibitory effects of Cinnamomum cassia (L.) J.Presl essential oils (EOs) against P. notoginseng associated pathogenic fungi were conducted both in vitro and in vivo experiments. The results of the Oxford cup test revealed that C. cassia dry bark EO (50 mg/mL) had significant inhibitory activity on the growth of all tested fungi, and the growth of various pathogens was completely inhibited, except for that of Fusarium solani. Therefore, the constituents of C. cassia EOs were analyzed by GC/MS, and the research demonstrated that the main constituents of C. cassia dry bark EO were trans-cinnamaldehyde (75.65 %), (E)-2-methoxycinnamaldehyde (6.08 %), cinnamaldehyde (3.47 %) and cinnamyl acetate (1.02 %). The MIC results showed that C. cassia dry bark EO and the main compounds had good antifungal effect on the tested strains, and the inhibitory effect was similar to that of hymexazol (chemical pesticide). By analyzing the value of the fraction inhibitory concentration index (FICI), additive effects, irrelevant effects and synergistic effects were observed after the mixture of hymexazol against various pathogens. Moreover, in vivo model showed that C. cassia dry bark EO could reduce the occurrence of anthrax in P. notoginseng. To widen the resources of C. cassia available, the compositions of both C. cassia fresh bark and leaf EOs were also tested and many common compositions existed among them. Taken together, it was concluded that C. cassia EO had the potential use in the field to reduce the pathogenic disease.

Antifungal Effect of Essential Oils from Five Kinds of Rutaceae Plants - Avoiding Pesticide Residue and Resistance.

Panax notoginseng root is a traditional Chinese herb, of which the yield and quality have been seriously affected by microorganisms, and is commonly used to treat various kinds of bleeding. In this experiment, the effects of the antifungal properties of essential oils (EOs) from five kinds of Rutaceae plants on the growth of three kinds of pathogens were studied to develop natural, environmentally friendly antifungal agents. Citrus medica EO was found to have stronger inhibitory effects on the growth of pathogenic fungi in vitro than other EOs with the Oxford cup method, of which the chemical composition was further investigated by GC/MS. The major components were d-limonene (22.79 %) and γ-terpinene (9.71 %). The antifungal activities were evaluated by MIC and FIC assays. In these assays, C. medica EO, d-limonene and γ-terpinene were effective against three pathogens of P. notoginseng with MIC values ranging from 0.12 to 12.05 mg/mL. The association between hymexazol and C. medica EO showed a high synergistic effect with lower FIC index values (FICi=0.31-2.00). Furthermore, C. medica EO was further assessed in P. notoginseng planted in a continuous cropping soil (CCS) and was found to reduce the disease incidence and disease severity compared with P. notoginseng planted in CCS only without EO addition. This finding suggested that C. medica EO has potential as a natural environmentally antifungal agent against pathogens of P. notoginseng, ensuring its safety.

Monitoring Antifungal Agents of Artemisia annua against Fusarium oxysporum and Fusarium solani, Associated with Panax notoginseng Root-Rot Disease.

Root rot of Panax notoginseng has received great attention due to its threat on the plantation and sustainable utilization of P. notoginseng. To suppress the root-rot disease, natural ingredients are of great importance because of their environment friendly properties. In this study, we found that the methanol extract from Artemisia annua leaves has strong antifungal effects on the growth of Fusarium oxysporum and Fusarium solani resulting into root-rot disease. Essential oil (EO) thereof was found to be the most active. GC-MS analysis revealed 58 ingredients and camphor, camphene, ß-caryophyllene, and germacrene D were identified as the major ingredients. Further antifungal assays showed that the main compounds exhibit various degrees of inhibition against all the fungi tested. In addition, synergistic effects between A. annua EO and chemical fungicides were examined. Finally, in vivo experiments were conducted and disclosed that P. notoginseng root rot could be largely inhibited by the petroleum ether extract from A. annua, indicating that A. annua could be a good source for controlling P. notoginseng root-rot.

Two New Triterpenoids from the Roots of Codonopsis pilosula.

Pseudolarolides U and V, two new triterpenoids, and four biogenetically related compounds, pseudolarolides E, F, K, and P were isolated from the roots of Codonopsis pilosula (Campanulaceae). Their structures were determined by spectroscopic data. The regulation of Sirtuin 1 (SIRT1) activity by all the isolated compounds was evaluated.

Role of Phenolic Acids from the Rhizosphere Soils of Panax notoginseng as a Double-Edge Sword in the Occurrence of Root-Rot Disease.

Chemical agents in the rhizosphere soils of plants might have an influence on root-rot disease, which therefore might reveal the mechanism of root rot in Panax notoginseng (P. notoginseng). With this hypothesis the alterations of phenolic acids (PAs) in the rhizosphere soils of P. notoginseng after pathogen infection were determined. The effects of PAs on the growth of Fusarium oxysporum (F. oxysporum), a fungal pathogenic factor for P. notoginseng, as well as production of fusaric acid, a wilting agent for the plants, were also examined. The results indicate the presence of five PAs (ferulic acid, syringic acid, p-hydroxybenzoic acid, p-coumaric acid, and vanillic acid) in the rhizosphere soils of P. notoginseng, whose contents in the rhizosphere soils of healthy plants are higher than those of the diseased ones. Further we found that individual PA could inhibit the mycelium growth and spore production of F. oxysporum, but stimulate fusaric acid production as well, disclosing the double-edge sword role of PAs in the occurrence of root rot of P. notoginseng and paving the way for the intervention of P. notoginseng root rot via balancing PAs.

Effects of Essential Oils from Zingiberaceae Plants on Root-Rot Disease of Panax notoginseng.

Replanting obstacles of Panax notoginseng caused by complex factors, including pathogens, have received great attention. In this study, essential oils (EOs) from either Alpinia officinarum Hance or Amomum tsao-ko (Zingiberaceae) were found to inhibit the growth of P. notoginseng-associated pathogenic fungi in vitro. Subsequent GC-MS analysis revealed the chemical profiles of two plant derived EOs. Linalool and eucalyptol were found to be abundant in the EOs and tested for their antifungal activities. In addition, the synergistic effects of A. tsao-ko EOs and hymexazol were also examined. These findings suggested that Zingiberaceae EOs might be a good source for developing new green natural pesticides fighting against root-rot of P. notoginseng.

[Rapid Prediction Study of Total Flavonids Content in Panax notoginseng Using Infrared Spectroscopy Combined with Chemometrics].

The variation on origin and growth environment could make a holistic impact on the secondary metabolites and quality of traditional Chinese medicine. In recent years, the origin of Panax notoginseng is spread from the genuine producing area of Wenshan to surrounding cities. The content of three saponins, as an indicator, is to ensure the quality of Panax notoginseng in Chinese pharmacopoeia. However, a single indicator is limited to comprehensive quality evaluation of Panax notoginseng. In this study, the total flavonoids content of Panax notoginseng was determinated by ultraviolet-visible (UV-Vis) spectrophotometry, Fourier transform infrared (FTIR) spectroscopy combined with chemometrics, as a rapid prediction model of total flavonoids content, was establish to provide some basic information for rapid and holistic quality assessment of Panax notoginseng. A total of 96 UV-Vis and FTIR spectra of Panax notoginseng originated from 12 regions were collected. The UV-Vis spectra of samples were recorded at 268 nm, and the content of total flavonoids was calculated based on standard linear equation of rutin. Pre-processing data were calculated with first (1D) and second derivative (2D), Savitsky-Golay smoothing with seven, nine, and eleven points. 2/3 of the 96 individuals were selected to form the training set by using Kennard-stone algorithm, and the rest were used as prediction set. Training set data were used to establish the orthogonal signal correction-partial least squares regression (OSC-PLSR) model and the 1/7 cross-validation method was used for screening optimal numbers of principal component, the prediction set was utilized to verify the accuracy and reliability of the OSC-PLSR model. Results showed that: (1) The correlation coefficient r of standard rutin was 0.9997, and the linear concentration range was from 5.6 to 72.0 µg·mL(-1), namely, there were good correlation between the absorbance and concentration. (2) The Panax notoginseng contained higher content of total flavonoids (more than 7 mg·g(-1)) in three genuie producing areas of Wenshan, Luoping county and Shilin county. (3) After the same points of Savitsky-Golay smoothing, the model predictive ability of 2D is better than that of 1D, and the predictive ability of different processing model has an obvious difference. (4) In all prediction models, the 2D+SG 7+OSC-PLSR (R(2)(pre)=0.976 1, RMSEP=0.325 2) and 2D+SG 11+OSC-PLSR (R(2)(pre)=0.946 9, RMSEP=0.382 0) model showed an excellent predictive effect, the value of RMSEP was below 0.4, and the predicted values were close to the detection values. The result indicated that FTIR combined with OSC-PLSR could accurately predict the content of total flavonoids. It could provide a rapid, simple, and effective method for the holistic quality control of Panax notoginseng.

[Rapid prediction of the content of polyphyllin in various species of Paris by infrared spectrometry].

Polyphyllin is the main active constituent in Paris which was a traditional Chinese medicine. In order to evaluate the quality of Paris rapidly and ensure the efficacy in clinical therapy, we quantified the contents of polyphyllin Ⅰ, polyphyllin Ⅱ and polyphyllin Ⅶ using infrared spectroscopy with partial least squares regression(PLSR). The method for evaluating the quality of Paris was established. Infrared spectra of 78 samples from various species in different origins were collected. The contents of polyphyllin Ⅰ, Ⅱ and Ⅶ were determined by high performance liquid chromatography(HPLC). The HPLC data were combined with the spectral data to predict the contents of three polyphyllin rapidly. Multiplicative signal correction(MSC), standard normal variate(SNV), orthogonal signal correction(OSC), first derivative and second derivative were utilized for the spectral preprocessing. Then, the optimized spectral data were used to establish the quantitative prediction model based on PLSR. The results showed that the best spectral pretreatment of polyphyllin Ⅰ and Ⅱ were MSC+OSC+2nd Der and that of polyphyllin Ⅶ was MSC+SNV+OSC+2nd Der. In the quantitative calibration model, the determination coefficients (R²) of polyphyllin Ⅰ, polyphyllin Ⅱ and polyphyllin Ⅶ were 0.930 8, 0.934 8 and 0.912 3, respectively while the Root mean square error of estimation(RMSEE) were 1.855 0, 0.632 3 and 0.001 6 mg•g⁻¹, respectively. In the verification model, the R² of polyphyllin Ⅰ, polyphyllin Ⅱ and polyphyllin Ⅶ were 0.948 8, 0.703 6 and 0.801 7, respectively, and the root mean square error of prediction(RMSEP)were 1.704 6, 1.227 8 and 0.002 0 mg•g⁻¹, respectively. Because of the predictive value of quantitative model was closed to the real value, the effect of the model was good. The model of polyphyllin Ⅰ and Ⅱ were better than that of polyphyllin Ⅶ. The developed method was non-destructive, fast, and accurate. It was feasible to determine the content of polyphyllin in Paris.

[Study on the Genetic Relationship of Panax Notoginseng and Its Wild Relatives Based on Fourier Translation Infrared Spectroscopy].

Wild relatives play a very important role in enriching germplasm resources and improving the quality and yield of cultivated species. In this paper, the genetic relationship between Panax notoginseng and its wild relatives has been investigated by using Fourier transform infrared (FTIR) spectroscopy in order to provide theoretical bases in the variety improvement of P. notoginseng as well as the development and utilization of germplasm resources. The FTIR spectra of P. notoginseng and its wild relatives (P. japonicus var. major, P. stipuleanatus, P. vietnamensis, P. japonicus var. bipinnatifidus) as well as Panax notoginsenosides were collected. The original infrared spectra of P. notoginseng and its wild relatives were pretreated by automatic baseline correction, smoothing, ordinate normalization and second derivative. The genetic relationship between P. notoginseng and its wild relatives has been studied together with the aid of principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA). By comparing the infrared spectra of P. notoginseng with that of panax notoginsenosides, some common peaks such as 3 400, 2 930, 1 635, 1 385, 1 075 and 927 cm-1 has been found. It showed that the peak heights of P. notoginseng samples may relate with the content of panax notoginsenosides. The original infrared spectra of P. notoginseng are similar to its wild relatives and the absorption peaks of the functional groups of C­H, CO, O­H, C­N and C­O were presented. There were some differences in the fingerprint region (1 800～500 cm-1) of the second derivative spectra of these five species samples. The characteristic absorption peaks such as 1 385 and 784 cm-1 has an obviously differentiation. Then the fingerprint region of second derivative spectra is subjected to be analyzed by PCA and PLS-DA. By comparing the 3D score plots of these two methods, the classification result of PLS-DA is significantly better than PCA. In addition, the result of HCA which based on the six principal components of PLS-DA has shown that P. japonicus var. major and P. vienamensis have close relationship with P. notoginseng while P. stipuleanatus and P. japonicus var. bipinnatifidus are far from P. notoginseng. The use of Fourier transform infrared spectroscopy combined with chemometrics methods could effectively investigate the genetic relationship between P. notoginseng and its wild relatives. Furthermore, it could provide reference for the research of medicinal plants.

A molecular network-based pharmacological study on the protective effect of Panax notoginseng rhizomes against renal ischemia-reperfusion injury.

Objective: We aimed to explore the protective effect of Panax notoginseng rhizomes (PNR) on renal ischemia and reperfusion injury (RIRI) and the underlying molecular network mechanism based on network pharmacology and combined systemic experimental validation. Methods: A bilateral RIRI model was established, and Cr, SCr, and BUN levels were detected. Then, the PNR was pretreated 1 week before the RIRI model was prepared. To determine the effects of the PNR in RIRI, histopathological damage and the effect of PNRs to the kidney was assessed, using TTC, HE, and TUNEL staining. Furthermore, the underlying network pharmacology mechanism was detected by screening drug-disease intersection targets from PPI protein interactions and GO and KEGG analysis, and the hub genes were screened for molecular docking based on the Degree value. Finally, the expression of hub genes in kidney tissues was verified by qPCR, and the protein expression of related genes was further detected by Western blot (WB). Results: PNR pretreatment could effectively increase Cr level, decrease SCr and BUN levels, reduce renal infarct areas and renal tubular cell injury areas, and inhibit renal cell apoptosis. By using network pharmacology combined with bioinformatics, we screened co-targets both Panax notoginseng (Sanchi) and RIRI, acquired ten hub genes, and successfully performed molecular docking. Of these, pretreatment with the PNR reduced the mRNA levels of IL6 and MMP9 at postoperative day 1 and TP53 at postoperative day 7, and the protein expression of MMP9 at postoperative day 1 in IRI rats. These results showed that the PNR could decrease kidney pathological injury in IRI rats and inhibit apoptotic reaction and cell inflammation so as to improve renal injury effectively, and the core network mechanism is involved in the inhibition of MMP9, TP53, and IL-6. Conclusion: The PNR has a marked protective effect for RIRI, and the underlying mechanism is involved in inhibiting the expression of MMP9, TP53, and IL-6. This striking discovery not only provides fruitful evidence for the protective effect of the PNR in RIRI rats but also provides a novel mechanic explanation.

[On-farm conservation and utilization of paddy rice, wheat and maize landrace varieties in 15 unique ethnic groups in Yunnan, China].

On-farm conservation and utilization of crop landraces have been proposed as means of conserving plant germplasm, in contrast to ex situ germplasm conservation, but little is known about the effectiveness of this approach. This paper reports the findings from a survey conducted in 15 unique ethnic groups of the Yunnan Province on the conservation and utilization of paddy rice, wheat (including wheat, barley, oats, and rye) and maize landrace varieties through participatory rural appraisal (PRA) and questionnaires. The surveyed regions covered 306 villages (the village group) from 237 administrative villages in 124 towns (township) distributed in 36 counties of 11 prefectures (city) in Yunnan Province. The survey showed that 44.8%, 77.5%, and 37.3% of the visited villages had lost their paddy rice, wheat, and maize landraces, respectively. A total of 901 landraces were collected, including 371 rice, 119 wheat, and 411 maize varieties, respectively. There were 2.9 on-farm varieties on average per village, 3.3, 8.0 and 5.2 varieties on average per 100 households, 1 000 peasants and 100 Ha farmland areas, respectively. Among the 306 villages, two villages (Laomian and Qingkou) maintained the highest crop diversity with 18 varieties (including 10 rice and 8 maize varieties) and 14 paddy rice varieties. Also, on-farm varieties in different ethnic groups varied significantly from each other, ranging from 16 to 120 varieties per group. The diversity of paddy rice varieties was the highest, ranging from 1 to 72, and the diversity was mainly distributed in southern, southwest Yunnan of tropical, subtropical ethnic regions. The wheat varieties ranged from 0 to 47 and distributed in northern, northwest Yunnan of high altitude, temperature, and cold ethnic regions. The maize varieties ranged from 4 to 40. These patterns of variation in on-farm varieties are directly associated with traditional culture and custom for the ethnic minorities and reflect the lack of improved varieties for the local special environments. These findings confirm the role of on-farm conservation in plant genetic resources in China and provide baseline information for better on-farm conservation and utilization of plant germplasm in China.

[Analysis of genetic variation in rice paddy landraces across 30 years as revealed by microsatellite DNA markers].

To reveal the genetic variation of rice paddy landraces across 30 years, we compared the genetic variation of between 6 paddy rice landraces grown in Yuanyang Hani's terraced fields in Yuanyang County, Yunnan Province in the 1970s (past-grown landraces) and 6 paired ones that have been grown during the past decade (current-grown landraces) using 60 SSR markers. The results showed that one to four alleles were amplified in 60 loci and 159 alleles in all the landraces tested. The number of alleles from the current-grown landraces decreased by 7 alleles compared to the past-grown landraces. The average number of alleles (Na), effective number of alleles (Ne), locus polymorphism information content (PIC), and genotype diversity (H') of the past-grown landraces were higher than those of the current-grown landraces, with Na of 2.567>2.450, Ne of 2.052>1.968, PIC of 0.469>0.439, and H' of 0.768>0.722. The average genetic similarity coefficient (GS) of the past-grown landraces was 0.437 with a range from 0.200 to 0.700 based on the 60 SSR markers, and the average GS of the current-grown landraces was 0.473 with a range from 0.117 to 0.667. In conclusion, the genetic diversity in current-grown landraces was decreased compared to the past-grown landraces, and the degree of variation in some of the allele locus varied in different rice landraces as a result of 30 years' natural and artificial selection.

Integrated physiological, metabolomic, and proteome analysis of Alpinia officinarum Hance essential oil inhibits the growth of Fusarium oxysporum of Panax notoginseng.

Fusarium oxysporum is the main pathogen of Panax notoginseng root rot, and chemical fungicides remain the primary measures to control the disease. Plant essential oil (EO) is a volatile plant secondary metabolic product that does not produce any residue to replace chemical pesticide. To comprehensively understand the antifungal mechanism of Alpinia officinarum Hance EO, the physiological indicators, proteome and metabolome were analyzed using F. oxysporum spores and hyphae treated with different EO concentrations. The cell membrane was damaged after both low and high concentrations of EO treatment, along with leakage of the cell contents. To resist the destruction of membrane structure, fungi can increase the function of steroid biosynthesis and expression of these catalytic enzymes, including squalene monooxygenase (SQLE), sterol 14alpha-demethylase (CYP51, CYP61A), delta14-sterol reductase (TM7SF2, ERG4), methylsterol monooxygenase (MESO1), and sterol 24-C-methyltransferase (SMT1). Furthermore, the tricarboxylic acid cycle (TCA) was influenced by inhibiting the expression of glutamate synthase (GLT1), 4-aminobutyrate aminotransferase (ABAT), and succinate-semialdehyde dehydrogenase (gabD); increasing malate and gamma-aminobutyric acid (GABA); and decreasing citrate content. The spore germination rate and mycelia growth were decreased because the expression of cohesin complex subunit SA-1/2 (IRR1) and cohesion complex subunit (YCS4, BRN1, YCG1) were inhibited. Particularly, under high EO concentrations, cyclin-dependent kinase (CDC28) and DNA replication licensing factor (MCM) were further inhibited to disrupt the cell cycle and meiosis, thus affecting cell division. The results of this study will enrich the understanding of the antifungal mechanism of EOs and provide an important basis to develop new plant-derived fungicides.