Physiological and molecular mechanisms of ZnO quantum dots mitigating cadmium stress in Salvia miltiorrhiza.

This study delved into the physiological and molecular mechanisms underlying the mitigation of cadmium (Cd) stress in the model medicinal plant Salvia miltiorrhiza through the application of ZnO quantum dots (ZnO QDs, 3.84 nm). A pot experiment was conducted, wherein S. miltiorrhiza was subjected to Cd stress for six weeks with foliar application of 100 mg/L ZnO QDs. Physiological analyses demonstrated that compared to Cd stress alone, ZnO QDs improved biomass, reduced Cd accumulation, increased the content of photosynthetic pigments (chlorophyll and carotenoids), and enhanced the levels of essential nutrient elements (Ca, Mn, and Cu) under Cd stress. Furthermore, ZnO QDs significantly lowered Cd-induced reactive oxygen species (ROS) content, including H2O2, O2-, and MDA, while enhancing the activity of antioxidant enzymes (SOD, POD, APX, and GSH-PX). Additionally, ZnO QDs promoted the biosynthesis of primary and secondary metabolites, such as total protein, soluble sugars, terpenoids, and phenols, thereby mitigating Cd stress in S. miltiorrhiza. At the molecular level, ZnO QDs were found to activate the expression of stress signal transduction-related genes, subsequently regulating the expression of downstream target genes associated with metal transport, cell wall synthesis, and secondary metabolite synthesis via transcription factors. This activation mechanism contributed to enhancing Cd tolerance in S. miltiorrhiza. In summary, these findings shed light on the mechanisms underlying the mitigation of Cd stress by ZnO QDs, offering a potential nanomaterial-based strategy for enhancing Cd tolerance in medicinal plants.

Effects of natural extract from medicinal herbs on broilers experimentally infected with Eimeria tenella.

This study aimed to evaluate the effects of natural extracts from nine medicinal herbs (SMA) on the growth performance, immunity, and intestinal integrity of broilers experimentally infected with Eimeria tenella. A total of 252 one-day-old broiler chicks were divided into 7 groups with 3 replicates per group and 12 broilers per cage. The groups were uninfected-untreated blank control group (BC), infected-untreated negative control group (NC), SMA treatment groups, Chinese medicine positive control group (CM), and chemical drug positive control group (CD). The SMA groups were infected and fed a basal diet supplemented with 0.6 (SMA-L), 0.8 (SMA-M), and 1.0 (SMA-H) g/kg SMA. The CM and CD groups were infected and fed a basal diet supplemented with 15 g/kg Jiqiuchong San and 0.2 g/kg Diclazuril, respectively. Results showed that feeding SMA could significantly reduce the number of oocysts in infected chickens, especially 1.0 g/kg SMA, which exhibited moderate anticoccidial efficacy. When infected with E. tenella, the supplementation of 1.0 g/kg SMA increased the renal index; restored the hepatic, splenic, and bursal indexes to BC levels; increased the levels of immunoglobulin A (IgA), IgM, and IgY; and reduced the contents of tumor necrosis factor (TNF-α), interferon-γ (IFN-γ), interleukin-6 (IL-6), and IL-10 of the infected chickens. Moreover, treatment with 1.0 g/kg SMA alleviated the pathological changes in cecal tissue and increased the contents of zonula occludens-1 (ZO-1), occludin, claudin-1, and mucoprotein 2 (mucin-2) in cecal tissues of E. tenella-infected chickens. We found that 1.0 g/kg SMA reduced the number of oocysts, improved immunity, and alleviated intestinal barrier damage, which could improve the growth performance of infected chickens. Thus, SMA proved to be an effective natural extract against E. tenella and has the potential to be used as an efficient anticoccidial drug or additive.

Identification and characterization of a novel tyrosine aminotransferase gene (SmTAT3-2) promotes the biosynthesis of phenolic acids in Salvia miltiorrhiza Bunge.

Rosmarinic acid (RA) and salvianolic acid B (SAB) are main phenolic acids in Salvia miltiorrhiza Bunge have been widely used in the treatment of cardiovascular and cerebrovascular diseases due to their excellent pharmacological activity. RA is a precursor of SAB, and tyrosine transaminase (TAT, EC 2.6.1.5) is a crucial rate-limiting enzyme in their metabolism pathway. This study identified a novel TAT gene, SmTAT3-2, and found that it is a new transcript derived from unconventional splicing of SmTAT3. We used different substrates for enzymatic reaction with SmTAT1, SmTAT3 and SmTAT3-2. Subcellular localization of SmTAT1 and SmTAT3-2 was completed based on submicroscopic techniques. In addition, they were overexpressed and CRISPR/Cas9 gene edited in hairy roots of S. miltiorrhiza. Revealed SmTAT3-2 and SmTAT1 showed a stronger affinity for L-tyrosine than SmTAT3, localized in the cytoplasm, and promoted the synthesis of phenolic acid. In overexpressed SmTAT3-2 hairy roots, the content of RA and SAB was significantly increased by 2.53 and 3.38 fold, respectively, which was significantly higher than that of overexpressed SmTAT1 strain compared with EV strain. These findings provide a valuable key enzyme gene for the phenolic acids metabolism pathway and offer a theoretical basis for the clinical application.

Integrated Multiomics and Synergistic Functional Network Revealed the Mechanism in the Tolerance of Different Ecotypes of Salvia miltiorrhiza Bge. to Doxycycline Pollution.

Tetracycline pollution in soil irreversibly damages the biosafety of plants by inhibiting the mitochondrial function. Some traditional Chinese medicine (TCM) plants, such as Salvia miltiorrhiza Bunge, have a strong tolerance to mitochondrial damage. We comprehensively compared the doxycycline (DOX) tolerances of two ecotypes of S. miltiorrhiza in the Sichuan and Shandong provinces and found that the Sichuan ecotype had a lower yield reduction, more stable accumulation of medicinal ingredients, higher mitochondrial integrity, and a more robust antioxidant system. The synergetic response networks under DOX pollution of both ecotypes were constructed using RNA sequencing and ultrahigh-performance liquid chromatography-tandem mass spectrometry. The differentiation of the downstream pathways of aromatic amino acids (AAAs) produced variations in the DOX tolerance of S. miltiorrhiza in different regions. The Sichuan ecotype maintained redox homeostasis and xylem development by activating salvianolic acid and indole biosynthesis, while the Shandong ecotype balanced chemical and mechanical defenses by regulating the flavonoid biosynthesis. Rosmarinic acid, a downstream AAA molecule, maintains the mitochondrial homeostasis of plant seedlings under DOX pollution by targeting the ABCG28 transporter. We also highlight the significance of downstream AAA small molecules in guiding the development of bio-based environmental pollution remediation agents.

Identification of Salvia miltiorrhiza Bunge with high and low cadmium accumulation and insight into the mechanisms of cadmium accumulation.

Salvia miltiorrhiza Bunge is used as a Chinese herbal medicine (CHM) particularly its roots. No relevant reports at home and abroad have been made on the mechanism of cadmium (Cd) accumulation in S. miltiorrhiza. The Cd accumulation characteristics of S. miltiorrhiza from main cultivation areas in China were evaluated for the first time to obtain high and low Cd accumulation in S. miltiorrhiza roots. Results showed obvious differences in the Cd enrichment capacity of S. miltiorrhiza from different cultivation areas. We took the lead in identifying the germplasm resources of S. miltiorrhiza with high and low Cd accumulation, that is, S. miltiorrhiza roots from Pingyi Shangdong (SDPY) belongs to the resource with high Cd accumulation (SDPY-H) and that from Zhongjiang Sichuan (SCZJ) is the resources with low Cd accumulation (SCZJ-L) based on relevant physiological and biochemical indexes. Although the Cd content of SDPY-H was apparently higher than that from SCZJ-L, its translocation factor from root to aboveground part is significantly lower than that in SCZJ-L. Therefore, planting SCZJ-L is not only an economic and effective way to use Cd in slightly and moderately polluted soil, but also its aboveground part can be used for phytoremediation. Changes in Cd content before and after the use of transpiration inhibitor indicate that SDPY-H enriched Cd through the symplastic pathway, whereas SCZJ-L mainly enriched Cd through the apoplastic pathways. In addition, the role of the symplastic pathway in SCZJ-L is weaker than that in SDPY-H, which were preliminarily revealed by fluorescent quantitative polymerase chain reaction. The significant reduction of the SmNramps transcription expression amount is one of the reasons for the low Cd accumulation of SCZJ-L.

The complete chloroplast genome sequence of Curcuma flaviflora (Curcuma).

The complete chloroplast (cp) genome of Curcuma flaviflora, a medicinal plant in Southeast Asia, was sequenced. The genome size was 160 478 bp in length, with 36.3% GC content. A pair of inverted repeats (IRs) of 26 946 bp were separated by a large single copy (LSC) of 88 008 bp and a small single copy (SSC) of 18 578 bp, respectively. The cp genome contained 132 annotated genes, including 79 protein coding genes, 30 tRNA genes, and four rRNA genes. And 19 of these genes were duplicated in inverted repeat regions.

Optimization of extraction and antioxidant activity of polysaccharides from Salvia miltiorrhiza Bunge residue.

In this study, the process of extracting polysaccharides from Salvia miltiorrhiza Bunge residue was optimized by using a Box-Behnken design. Statistical analysis of the results showed that the linear and quadratic terms of the three variables of the extraction process had significant effects. The optimal conditions are as follows: extracting time of 2.6 h, extraction temperature of 89 °C, and ratio of water to raw material of 32 mL/g. Moreover, a new polysaccharide with antioxidant activity [i.e., SMWP-1 (∼5.27×10(5) Da)] was isolated from S. miltiorrhiza residue. The carbohydrate, uronic acid, and protein contents of SMWP-1 were 90.11%, 0.13%, and 0.53%, respectively. The SMWP-1 is composed of glucose, xylose, mannose, and galactose. The preliminary structural characterization of SMWP-1 was determined via Fourier transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM) analyses. This polysaccharide exhibited strong reducing power and free-radical scavenging activities in vitro against 2,2-diphenyl-1-picrylhydrazyl, superoxide anion, and hydroxyl. Therefore, SMWP-1 can be investigated further as a novel natural antioxidant.

Characterization, antioxidant and antitumor activities of polysaccharides from Salvia miltiorrhiza Bunge.

Polysaccharides were extracted from Salvia miltiorrhiza Bunge using response surface methodology (RSM) with ultrasonication. A Box-Behnken design was used to optimize the extraction parameters to maximize the polysaccharide extraction yield. The polysaccharide SMP-U1 was isolated and characterized; then the antioxidant and antiproliferation activities were evaluated in vitro. The modified optimal conditions were an ultrasonic power of 180 W, an extraction temperature of 54°C, and an extraction time of 32 min, achieving an extraction yield of 40.54±0.25%. The results indicate that SMP-U1 has significant antioxidant activity, scavenging the free radical 2,2-diphenyl-1-picrylhydrazyl. It has also exhibited effect on the proliferation of human breast carcinoma cells Bcap-37 and human esophageal carcinoma cells Eca-109, especially at a concentration of 0.30 mg/mL. In conclusion, SMP-U1 has remarkable in vitro antioxidant and antiproliferation activity, and has potential for application as a natural antioxidant or antitumor agent.

The physiological response of Artemisia annua L. to salt stress and salicylic acid treatment.

Salinity has a great influence on plant growth and distribution. A few existing reports on Artemisia annua L. response to salinity are concentrated on plant growth and artemisinin content; the physiological response and salt damage mitigation are yet to be understood. In this study, the physiological response of varying salt stresses (50, 100, 200, 300, or 400 mM NaCl) on A. annua L. and the effect of exogenous salicylic acid (0.05 or 0.1 mM) at 300-mM salt stress were investigated. Plant growth, antioxidant enzyme activity, proline, and mineral element level were determined. In general, increasing salt concentration significantly reduced plant growth. Superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were stimulated by salt treatment to a higher enzyme activity in treated plants than those in untreated plants. Content of proline had a visible range of increment in the salt-treated plants. Distribution of mineral elements was in inconformity: Na(+) and Ca(2+) were mainly accumulated in the roots; K(+) and Mg(2+) were concentrated in leaves and stems, respectively. Alleviation of growth arrest was observed with exogenous applications of salicylic acid (SA) under salt stress conditions. The activity of SOD and POD was notably enhanced by SA, but the CAT action was suppressed. While exogenous SA had no discernible effect on proline content, it effectively inhibited excessive Na(+) absorption and promoted Mg(2+) absorption. Ca(2+) and K(+) contents showed a slight reduction when supplemented with SA. Overall, the positive effect of SA towards resistance to the salinity of A. annua will provide some practical basis for A. annua cultivation.

A simple and reliable method for distinguishing danshen in salvia: simultaneous quantification of six active compositions by HPLC.

A simple and reliable method for distinguishing Danshen is important to evaluate the quality and clinical efficiency of these species. An HPLC method was developed for the determination of protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone, tanshinone I and tanshinone IIA in 23 samples of Salvia. The analytes were separated on an Agilent XDB C18 reversed-phase column coupled with a Phenomenex C18 guard column using a gradient elution of acetonitile-0.1% aqueous phosphoric acid as the mobile phase at a flow rate 0.8 mL/min and UV detection at 280 nm. The method allowing the simultaneous quantification of six major active compositions was optimized and validated for linearity, precision, accuracy and limits of detection (LOD) and quantification. The LOD ranged from 0.019 to 0.850 µg/mL (R(2) ≥ 0.9998). Accuracy, precision and reproducibility were all within the required limits. The average recovery between 96.49 and 102.16% and the relative standard deviations were <3.01%. Based on the six compositions content and clustering result, this research results suggest that these six major active compositions could be distinguishing markers for Danshen and non-Danshen.

[Neuroprotective effect of longistyline A against corticosterone-induced neurotoxicity in PC12 cells].

This study is to investigate the protective effect of longistyline A against corticosterone-induced neurotoxicity in PC12 cells. While PC12 cells were exposed to 100 micromol x L(-1) corticosterone for 48 h, cell survival rate was reduced and lactate dehydrogenase (LDH) release increased. In parallel, corticosterone caused significant elevations of DNA fragmentation, [Ca2+]i and caspase-3 activity. However, when the PC12 cells were incubated with longistyline A (4.0, 8.0 and 16.0 micromol x L(-1)) in the presence of 100 micromol x L(-1) corticosterone for 48 h, the effects were evidently alleviated, but dose-dependent manner was not obvious. In summary, longistyline A could generate a neuroprotective effect against corticosterone-induced neurotoxicity in PC12 cells possibly by decreasing [Ca2+]i and caspase-3 activity.

[Pollen viability and stigma receptivity of Salvia miltiorrhiza and its relative].

OBJECTIVE: To provide the basal data for artificial cross breeding of Chinese herb Salvia miltiorrhiza from 7 provinces in China and its 4 relatives. METHOD: The pollen viability was evaluated by TTC (2, 3, 5-triphenylte trazolium chloride) test and the stigma receptivity was evaluated by benzidine-H2O2 method. RESULT: The pollen viability of S. miltiorrhiza from 6 provinces in China and its 4 relatives deceased during time of pollen shedding. Their highest pollen viability was in 2 or 3 days after blooming. But the pollen viability of S. miltiorrhiza (wild and culture) from Hean province in China declined with time after blooming. The most obvious variation of the pollen viability was in S. miltiorrhiza from Shanxi province (RSD 71.3% ) and the least was in wild S. miltiorrhiza from Henan province (RSD 12.4%). The highest average pollen viability was wild S. miltiorrhiza (72.3%) from Henan province while the lowest was S. yunnanensis (38.8%). The stigmas of all the accessions had receptivity when blooming. The stigma receptivity of S. brevilabra was strong in 2 to 4 days after blooming, while the others had less change after blooming. The life span of pollen grains and stigmas could be maintained from 3 to 5 days. CONCLUSION: The optimum artificial pollination time of S. miltiorrhiza and its relatives was 2 to 3 days after blooming.

[Study on isozymes in six species of Curcuma].

OBJECTIVE: To explore genetic relationships of the 39 materials in six species of Curcuma. METHOD: The peroxidase isozyme (POD) and esterase isozyme (EST) were studied using vertical slab polyacrylamide gel electrophoresis (PAGE) technique, and the zymograms were analyzed using the software of NTSYSpc2. 1. RESULT: The interspecific zymogramatic differences were obvious. Each species possessed its own specific zymogram distinguishing form the others. In the analysis of EST isozyme, C. phaeocaulis, C. wenyujin, C. kwangsiensis and C. chuanhuangjiang had their own specific zymogram. In the analysis of POD isozyme, just C. phaeocaulis and C. kwangsiensis had their specific zymogram. CONCLUSION: The genetic relationships are not associated with the geographical distributions and the genetic relationship between C. longa and C. sichuanensis are very close.

[Principal component and cluster analysis of trace elements in Chinese herb Salvia miltiorrhiza and its relative species].

The contents of trace elements, including copper, zinc, iron, magnesium, calcium, chromium, lead, molybdenum, manganese, and cadmium in Chinese traditional herb S. miltiorrhiza and its relative species such as S. miltiorrhiza f. alba, S. evansiana, S. yunnanensis, S. przewarlskii, S. cavaleriei, S. cavaleriei var. simplicifolia, S. roborowskii, S. omeiana, S. tricuspis, S. brevilabra and S. cynica were determined by atomic absorption spectroscopy. The principal components analysis of SPSS was applied to the study of characteristic elements in S. miltiorrhiza and its relative species. Three principal components which accounted for 79.3% of total variance were extracted from the original data. The first factor accounted for 49.6% of the total variance, which means that iron, manganese, copper, zinc, cadmium and lead are the characteristic elements in S. miltiorrhiza and its relative species. The results of cluster analysis show that the samples could cluster reasonably into two groups. The samples of S. miltiorrhiza from different regions were classified into one group except S. miltiorrhiza from Zhejiang and S. miltiorrhiza f. alba. The other Salvia species were classified into another group except S. cavaleriei. The S. miltiorrhiza and other Salvia species can be distinguished by this method, whose accuracy of classification is 90%. The cluster analysis based on the contents of trace element in S. miltiorrhiza and its relative species provided a quick, accurate and simple method for authentication of herb Salvia miltiorrhiza.

[Studies on chromosome numbers of Salvia miltiorrhiza, S. flava and S. evansiana].

OBJECTIVE: To study the numbers of chromosome in Chinese herb Salvia miltiorrhiza from 7 provinces in China, and S. flava as well as S. evansiana from Yunnan province in China. METHOD: The young root was treated with the mixture of ice and water for 24 h, fixed with Carony's fixative for 6-12 h. After differentiating for 10-12 min with 1 mol x L(-1) hydrochloric acid at 60 'C and staining with carbol fuchsin,the section was observed under microscope. RESULT: Chromosome numbers of S. miltiorrhiza and S.flava were 2n = 2x = 16. The numbers of S. evansiana were 2n = 4x = 32. The basic numbers of the chromosomes were x = 8. And tetraploids were observed in S. miltiorrhiza from Sichuan provices and Shandong provices. CONCLUSION: The basic number of the chromosomes are x = 8. The chromosome numbers of S. miltiorrhiza, S.flava and S. evansiana are 16,16 and 32 respectively. As the chromosomes are the small or micro-small ones, it is difficult to use them for karyotype.

[Analysis on yield and quality of different Houttuynia cordata].

OBJECTIVE: To study the yield of different Houttuynia cordata and the changing regularity of their main officinal components in different periods, and provide a basis for harvesting in good time and breeding new varieties. METHOD: The main officinal composition and wet weight of overground parts per acreage of different accessions were determined periodically. The plot yields were tested when harvested and analyzed by analysis of variance. RESULT: The difference of the yields of different accessions was significant. The varying tendency of the content of main officinal component methyl nonyl ketone and its total content per acreage of different accessions were different. CONCLUSION: The integrate characters of Qionglai and Wangyu wild accessions are relatively better, worthy to be widely applied in production.

[Study on dry matter accumulations of Houttuynia cordata on heavy loam].

In this paper the characteristics of dry matter accumulation of Houttuynia cordata cultivated on three kinds of heavy loams were studied. The results showed that the accumulated curves of whole plant dry matter were somewhat different. During testing period, the accumulated curves of dry matter of Houttuynia cordata grown on clay silt loam and silt sand loam presented logistic ones, but those grown on sandy silt loam were still in line growth period. It was concluded that the harvest period of Houttuynia cordata on the sandy silt loam could be properly put off while only considered the yielding. The experiment on heavy loam also showed that the sandy field should be the better selection than others, but organic fertilizer should be enhanced as base fertilizer, with proper nitrogen as top application. This paper also discussed the dry matter accumulations of different organs of Houttuynia cordata.

[Isozymes variations among the germplasm resources of Houttuynia in Sichuan].

The peroxidase and esterase isozymes of the germplasm resources of Houttuynia in Sichuan were analyzed. The results showed that it existed plentiful isozymes variations among the germplasm resources of Houttuynia in Sichuan. The cultivated populations had the same isozymes band types as the wild ones. There were 6 different types of peroxidase isozymes bands and 10 different esterase isozymes band totally. Each peroxidase band type was composed of 4 to 6 bands, and esterase isozymes had 5 to 8 bands. There was no significant difference between Houttuynia cordata and Houttuynia emeiensis populations in their isozymes bands types. The results also showed that the number of peroxidase isozymes bands in northern areas was somewhat higher than that in southern areas. However, esterase isozymes band types had no geographic differences.