Effect the accumulation of bioactive constituents of a medicinal plant (Salvia Miltiorrhiza Bge.) by arbuscular mycorrhizal fungi community.

BACKGROUND: Arbuscular mycorrhizal fungi (AMF) form symbiotic relationships with various terrestrial plants and have attracted considerable interest as biofertilizers for improving the quality and yield of medicinal plants. Despite the widespread distribution of AMFs in Salvia miltiorrhiza Bunge's roots, research on the impact of multiple AMFs on biomass and active ingredient accumulations has not been conducted. In this study, the effects of five native AMFs (Glomus formosanum, Septoglomus constrictum, Rhizophagus manihotis, Acaulospora laevis, and Ambispora gerdemannii) and twenty-six communities on the root biomass and active ingredient concentrations of S. miltiorrhiza were assessed using the total factor design method. RESULTS: Thirty-one treatment groups formed symbiotic relationships with S. miltiorrhiza based on the pot culture results, and the colonization rate ranged from 54.83% to 89.97%. AMF communities had higher colonization rates and total phenolic acid concentration than single AMF, and communities also appeared to have higher root fresh weight, dry weight, and total phenolic acid concentration than single inoculations. As AMF richness increased, there was a rising trend in root biomass and total tanshinone accumulations (ATTS), while total phenolic acid accumulations (ATP) showed a decreasing trend. This suggests that plant productivity was influenced by the AMF richness, with higher inoculation benefits observed when the communities contained three or four AMFs. Additionally, the affinities of AMF members were also connected to plant productivity. The inoculation effect of closely related AMFs within the same family, such as G. formosanum, S. constrictum, and R. manihotis, consistently yielded lower than that of mono-inoculation when any combinations were applied. The co-inoculation of S. miltiorrhiza with nearby or distant AMFs from two families, such as G. formosanum, R. manihotis, and Ac. laevis or Am. gerdemannii resulted in an increase of ATP and ATTS by more than 50%. AMF communities appear to be more beneficial to the yield of bioactive constituents than the single AMF, but overall community inoculation effects are related to the composition of AMFs and the relationship between members. CONCLUSION: This study reveals that the AMF community has great potential to improve the productivity and the accumulation of bioactive constituents in S. miltiorrhiza, indicating that it is an effective way to achieve sustainable agricultural development through using the AMF community.

Genotype-Controlled Vertical Transmission Exerts Selective Pressure on Community Assembly of Salvia miltiorrhiza.

The plant's endophytic fungi play an important role in promoting host development and metabolism. Studies have shown that the factors affecting the assembly of the endophyte community mainly include host genotype, vertical transmission, and soil origin. However, we do not know the role of vertically transmitted endohytic fungi influences on the host-plant's endophytic community assembly. Salvia miltiorrhiza from three production areas were used as research objects; we constructed three production area genotypes of S. miltiorrhiza regenerated seedlings simultaneously. Based on high-throughput sequencing, we analyzed the effects of genotype, soil origin, and vertical transmission on endophytic fungal communities. The results show that the community of soil origins significantly affected the endophytic fungal community in the regenerated seedlings of S. miltiorrhiza. The influence of genotype on community composition occurs through a specific mechanism. Genotype may selectively screen certain communities into the seed, thereby exerting selection pressure on the community composition process of offspring. As the number of offspring increases gradually, the microbiota, controlled by genotype and transmitted vertically, stabilizes, ultimately resulting in a significant effect of genotype on community composition.Furthermore, we observed that the taxa influencing the active ingredients are also selected as the vertically transmitted community. Moreover, the absence of an initial vertically transmitted community in S. miltiorrhiza makes it more vulnerable to infection by pathogenic fungi. Therefore, it is crucial to investigate and comprehend the selection model of the vertically transmitted community under varying genotypes and soil conditions. This research holds significant implications for enhancing the quality and yield of medicinal plants and economic crops.

A Novel Based-Network Strategy to Identify Phytochemicals from Radix Salviae Miltiorrhizae (Danshen) for Treating Alzheimer's Disease.

Alzheimer's disease (AD) is a common age-related neurodegenerative disease that strikes millions worldwide. Herein, we demonstrate a new approach based on network target to identify anti-AD compounds from Danshen. Network pharmacology and molecular docking were employed to establish the DS-AD network, which mainly involved apoptosis of neuron cells. Then network scoring was confirmed via Connectivity Map analysis. M308 (Danshenxinkun D) was an anti-AD candidate with a high score (p < 0.01). Furthermore, we conducted ex vivo experiments with H2O2-treated PC12 cells to verify the neuroprotective effect of Salvia miltiorrhiza-containing plasma (SMP), and UPLC-Q-TOF/MS and RT-qPCR were performed to demonstrate the anti-AD activity of M308 from SMP. Results revealed that SMP could enhance cell viability and level of acetylcholine. AO/EB staining and Mitochondrial membrane potential (MMP) analysis showed that SMP significantly suppressed apoptosis, which may be due to anti-oxidative stress activity. Moreover, the effects of M308 and SMP on expressions of PSEN1, DRD2, and APP mRNA were consistent, and M308 can significantly reverse the expression of PSEN1 and DRD2 mRNA in H2O2-treated PC12 cells. The strategy based on the network could be employed to identify anti-AD compounds from Chinese herbs. Notably, M308 stands out as a promising anti-AD candidate for development.

Full-length transcriptome analysis of Coptis deltoidea and identification of putative genes involved in benzylisoquinoline alkaloids biosynthesis based on combined sequencing platforms.

KEY MESSAGE: The study carry out comprehensive transcriptome analysis of C. deltoidea and exploration of BIAs biosynthesis and accumulation based on UHPLC-MS/MS and combined sequencing platforms. Coptis deltoidea is an important medicinal plant with a long history of medicinal use, which is rich in benzylisoquinoline alkaloids (BIAs). In this study, Ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) and combined sequencing platforms were performed for exploration of BIAs biosynthesis, accumulation and comprehensive transcriptome analysis of C. deltoidea. By metabolism profiling, the accumulation of ten BIAs was analyzed using UHPLC-MS/MS and different contents were observed in different organs. From transcriptome sequencing result, we applied single-molecule real-time (SMRT) sequencing to C. deltoidea and generated a total of 75,438 full-length transcripts. We proposed the candidate biosynthetic pathway of tyrosine, precursor of BIAs, and identified 64 full length-transcripts encoding enzymes putatively involved in BIAs biosynthesis. RNA-Seq data indicated that the majority of genes exhibited relatively high expression level in roots. Transport of BIAs was also important for their accumulation. Here, 9 ABC transporters and 2 MATE transporters highly homologous to known alkaloid transporters related with BIAs transport in roots and rhizomes were identified. These findings based on the combined sequencing platforms provide valuable genetic information for C. deltoidea and the results of transcriptome combined with metabolome analysis can help us better understand BIAs biosynthesis and transport in this medicinal plant. The information will be critical for further characterization of C. deltoidea transcriptome and molecular-assisted breeding for this medicinal plant with scarce resources.

[Microecology and geoherbalism of traditional Chinese medicine].

Microecology was directly or indirectly involved in the growth and development, metabolism process, and component accumulation of traditional Chinese medicine(TCM) in various ways, which affected the formation and changes of the geoherbalism of TCM. It was one of the main tasks of traditional Chinese medical microecology(TCMM) to reveal the relationship among microecological structure and its change rule and the quality effect of TCM. The heterogeneity of soil environment caused by geographical and climatic factors, as well as the discreteness limitation caused by isolation factors such as distance and host selection, were the main causes of the differentiation of microecological geography of TCM. The microecology of TCM had important influences and contributions on the distinctive origin and quality of Dao-di herbs, which was mainly reflected in the formation of excellent germplasm(including disease and insect resistance, drought resistance, salt resistance, cold resistance, etc.), the increase of yield, the formation of medicinal parts, the metabolism and accumulation of effective components, the time limit of harvesting, and the toxicity, increasing efficiency or reducing toxicity of TCM in the processing, the changes of product efficiency after introduction, and the authenticity of fungus medicine. With the vigorous development of metabonomics and modern information technology, the following aspects would become the future research trends, including the microecologically mediated biogenic pathway of chemical components, the metabolic synthesis reactor of TCM based on the microecological quantitative effect relationship, the cultivation of genuine Chinese medicine based on reconstruction of microecological structure, the origin identification barcode traceability technology, and the toxicity reduction and efficiency enhancement technology of TCM based on the microecological.

[Research and prospect of Traditional Chinese Medical Microecology].

In the long-term evolution, microbes and hosts coexist widely, forming a symbiotic microecosystem and resulting the complex interactions of the metabolism. With the application of microecological theory in Chinese materia medica science, two main points have been accepted gradually. On the one hand, the prevention and treatment of human diseases by traditional Chinese medicines can be achieved through the correction and adjustment of the imbalance of the human microecosystem. On the other hand, the microecosystem can regulate the quality of traditional Chinese medicines in real time, and further affect their curative effect. Thus, a new discipline, Traditional Chinese Medical Microecology, has been gradually established. In this review, the background, theoretical structure, research directions, key problems and the relationship with human microecology of Traditional Chinese Medical Microecology were systematically summarized and prospected for promoting its development. Moreover, this review provides a reference protocol for further discoursing the microecological mechanism involving the efficacy of traditional Chinese medicines.

[DNA marker-assisted selection of medicinal plants (â ) .Breeding research of disease-resistant cultivars of Panax notoginseng].

The study is aimed to estimate the diversity of arbuscular mycorrhizal fungi (AMF) in the main producing areas of Salvia miltiorrhiza.Diversity of AMF was surveyed directly on spores isolated from the field soil, collected from 20 sites of 8 provinces. Identification of the AMF was made by observation of spore morphology. At least 27 recognized AMF species were identified in samples from field soil, belonging to seven genera of AMF-Acaulospora, Glomus, Funneliformis, Ambispora, Rhizophagus, Pacispora, and Claroideoglomus. Acaulospora and Glomus were the dominant genera, respectively including nine and eight species. A. laevis (90%), R.manihotis (80%), A. brieticulata (75%), A. tuberculata (70%) were the dominant species.Colonization rate was determined,colonization was easily found, but the colonization intensities were low, the colonization rate remained at 10.92%-25.93%. The similarity between provinces is generally low, and the similarity coefficients were from 0.20 to 0.57.

[Effects of Different AM Fungi on Quantity and Enzyme Activity of Rhizosphere Soil Microorganism of Paris polyphylla var. yunnanensis].

Objective: To investigate the rhizosphere soil microorganism and enzyme activity after Paris polyphylla var. yunnanensis inoculated Arbuscular Mycorrhizal( AM) fungi,which provide the technological condition for artificial cultivation of Paris polyphylla var. yunnanensis. Methods: By the method of combining the greenhouse pot inoculation trials with laboratory analysis, The effects of inoculation with 28 AM fungi on rhizosphere soil microorganism and soil enzyme activities of Paris polyphylla var. yunnanensis under sterilized soil conditions were observed. Results: There were certain mutualistic selection between Paris polyphylla var. yunnanensis and AM fungi. After induction treatment with AM fungi, different AM fungi had various effects on the quantity, microbial biomass carbon and soil enzyme activities of rhizosphere soil microorganism of Paris polyphylla var. yunnanensis. With application of AM fungi,the amount of culturable fungi from rhizosphere soil of Paris polyphylla var. yunnanensis were decreased,while the culturable bacteria and actinomycetes showed a rising trend,and increased the diversity index,the content of biomass carbon and protease,urease,phosphatase,catalase and invertase activities of rhizosphere soil microorganism. Conclusion: Applying AM fungi when artificial cultivate Paris polyphylla var. yunnanensis,by which can impel rhizosphere microorganism of Paris polyphylla var. yunnanensis to transform from low-fertile"fungus types"to high-fertile"bacterial forms",and raise fungi diversity and enzymatic activity of rhizosphere soil microorganism,it is helpful to maintain the stability and harmoniousness of microecosystem on rhizosphere of Paris polyphylla var. yunnanensis,which certify the efficiency and the possibility of spreading AM inoculum during artificial cultivation in the field.

[Isolation and Identification of Antagonistic Endophytic Actinomycetes Against Root Rot Disease in Ligusticum chuanxiong].

Objective: To get an understanding of the microflora of endophytic actinomycetes in Ligusticum chuanxiong,and to obtain the resource of antagonistic strains against Ligusticum chuanxiong root rot disease. Methods: Actinomycetes in stem nodes and rhizomes of Ligusticum chuanxiong were isolated and purified by a series of means,namely tissue block method, homogenate technique and plat streaking method. Further, dual culture and inhibition zone method were employed to test the antagonistic activity of these strains. To identify strains with potential precisely,both microscopic observation and DNA sequencing were conducted. Results: 83 strains of Ligusticum chuanxiong endophytic actinomycetes were identified, which consisted of 13 species groups. Among all of them,species identified as Streptomyces scopuliridis( KF600747. 1), Streptomyces griseorubiginosus( AB706352. 1) and Streptomyces agglomeratus( LC055413. 1) showed antagonistic activities against four kinds of pathogenic fungi of Ligusticum chuanxiong root rot disease. Conclusion: Actinomycetes which belong to Griseorubroviolaceus groups of streptomyce are potential biocontrol microbes to Ligusticum chuanxiong root rot,showing importance to the production of Chuanxiong Rhizoma.

[Correlation analysis between effectiveness of element in rhizonsphere soil and quality of Danshen (Salvia miltiorrhiza)].

In order to investigate the content and distribution of available element in the rhizonsphere soil of the growing areas of Salvia miltiorrhiza Bunge, the contents of available element (N,P,K,B,Cu,Zn,Fe,Mn) in 26 soil samples were tested and evaluated. The results showed that the contents of available P and Fe were very plentiful, available K, Cu and Zn were rich, available N and Mn were deficient, available B was extremely deficient in all growing areas of S. miltiorrhiza of eight provinces in China. The correlation analysis showed that the contents of eight kinds of available elements were varying degree correlation. The stepwise regression analysis between the contents of available elements of rhizonsphere soil and ten kinds of active ingredients of Danshen (Salviae Miltiorrhizae Radix et Rhizoma) were researched. The results showed that the rates of contribution of available N,B,Mn and Fe to quality of Danshen were relatively large and they were the significant factors, and the other factors did not show statistical significance. The recommended fertilizing strategies is that the usage of N,B and Mn fertilizers should be controlled according to different stages of growth of S. miltiorrhiza, and P fertilizer should be reduced in all growing areas of S. miltiorrhiza.

[Identification of Pathogens Causing Root Rot Disease on Ligusticum chuanxiong in Sichuan].

OBJECTIVE: To identify the pathogens causing root rot disease on Ligusticum chuanxiong from the main producing areas in Sichuan, and to provide gist of management of this disease in the future. METHODS: The diseased rhizomes of Ligusticurn chuanxiong with root rot symptom were sampled in two main producing areas in Sichuan. According to Koch's postulate, the pathogenicity on Ligusticum chuanxiong rhizomes of the isolates was confirmed. The representative isolates were identified based on their morphological characteristics and ribosomal DNA internal transcribed spacer(rDNA-ITS) sequences. RESULTS: The isolates(f2-16, 3-2, f4-19 and f5-7)which caused root rot disease held high homology reached 99%, 99%, 100% and 100% respectively, when compared with GenBank sequences GQ229075, JQ796755, JN232136 and FJ481024. CONCLUSION: The causing agents of root rot disease on Ligusticum chuanxiong belong to Fusarium solani, Fusarium oxysporum, Plectosphaerella cucumerina and Phoma glomerata. Pathogens from different regions are varied.

[A novel method for testing sterility of injections based on biothermodynamics].

This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.

Beneficial endophytic fungi improve the yield and quality of Salvia miltiorrhiza by performing different ecological functions.

Background: Endophytic fungi can enhance the growth and synthesis of secondary metabolites in medicinal plants. Salvia miltiorrhiza Bunge is frequently employed for treating cardiovascular and cerebrovascular ailments, with the primary bioactive components being salvianolic acid and tanshinone. However, their levels in cultivated S. miltiorrhiza are inferior to that of the wild herbs, so the production of high-quality medicinal herbs is sharply declining. Consequently, the utilization of beneficial endophytic fungi to improve the yield and quality of S. miltiorrhiza holds great significance for the cultivation of medicinal plants. Methods: In this study, nine non-pathogenic, endophytic fungal strains were introduced into sterile S. miltiorrhiza seedlings and cultivated both in vitro and in situ (the greenhouse). The effects of these strains on the growth indices, C and N metabolism, antioxidant activity, photosynthesis, and content of bioactive ingredients in S. miltiorrhiza were then evaluated. Results: The results showed that the different genera, species, or strains of endophytic fungi regulated the growth and metabolism of S. miltiorrhiza in unique ways. These endophytic fungi primarily exerted their growth-promoting effects by increasing the net photosynthetic rate, intercellular CO2 concentration, and the activities of sucrose synthase, sucrose phosphate synthase, nitrate reductase, and glutamine synthetase. They also enhanced the adaptability and resistance to environmental stresses by improving the synthesis of osmoregulatory compounds and the activity of antioxidant enzymes. However, their regulatory effects on the growth and development of S. miltiorrhiza were affected by environmental changes. Moreover, the strains that significantly promoted the synthesis and accumulation of phenolic acids inhibited the accumulation of tanshinones components, and vice versa. The endophytic fungal strains Penicillium meloforme DS8, Berkeleyomyces basicola DS10, and Acremonium sclerotigenum DS12 enhanced the bioaccumulation of tanshinones. Fusarium solani DS16 elevated the rosmarinic acid content and yields in S. miltiorrhiza. The strain Penicillium javanicum DS5 improved the contents of dihydrotanshinone, salvianolic acid B, and rosmarinic acid. The strains P. meloforme DS8 and B. basicola DS10 improved resistance. Conclusion: Various endophytic fungi affected the quality and yield of S. miltiorrhiza by regulating different physiological and metabolic pathways. This study also provides a novel and effective method to maximize the effects of beneficial endophytic fungi by selecting specific strains to design microbial communities based on the different ecological functions of endophytic fungi under varying environments and for specific production goals.

Modified acid polysaccharide derived from Salvia przewalskii with excellent wound healing and enhanced bioactivity.

Acid polysaccharide was extracted from Salvia przewalskii root powders (PSP), purified by diethylaminoethyl cellulose column (DEAE-52) and molecular sieve (PSP2). PSPm1 was obtained by modifying PSP2 with nitrite and phosphoric acid. The chemical structure of PSP2 and PSPm1 exhibited notable distinctions, primarily due to the absence of arabinose and promotion of glucuronic acid (GlcA). The structure of PSPm1 was deduced through the utilization of 1H, 13C, and 2-D NMR. The main chain was linked by α-D-Galp(1 â†’ 3)-α-Glcp-(1 â†’ fragments and →6)-ß-D-Galp fragments, with the presence of →4)-α-D-GlcpA-(1 â†’ 6)-ß-D-Galp-(1 â†’ ï¼Œ â†’ 4)-α-D-GalAp-(1 â†’ 2,4)-α-D-Rhap-(1 â†’ fragments and →6)-α-Glcp-(1 â†’ 2,4)-ß-D-Manp-(1 â†’ fragments. PSPm1 exhibited different immunoregulatory bioactivity in vitro, including haemostatic effects indicated by activated clotting time of 55.5 % reduction by the activated clotting time (ACT) test and wound healing function in vivo. PSPm1 also displayed better anti-tumor biological effects than unmodified. The structure-activity dissimilarity between PSP2 and PSPm1 primarily stems from variations in molecular weight (Mw), monosaccharide composition, and branching patterns. The modification of polysaccharides from the extract residues of Chinese medicinal materials may be a new form of drug supplements.

[Analysis of endophytic fungi community of Ligusticum chuanxiong using PCR-DGGE].

OBJECTIVE: To research the diversity of endophytic fungal communities among Ligusticum chuanxiong growing at 5 areas in Sichuan province, and illuminate the developing mechanism of geoherbs from the microecological perspective. METHOD: The PCR-DGGE and DNA sequencing techniques were used to analyze the endophytic fungi community of L. chuanxiong. RESULT: The community of endophytic fungi present difference among different growing areas. Though minor difference were found among individuals at the same area, similarity among individuals from the same growing areas were higher significantly than those from different growing areas. Compared with the other 4 growing areas, L. chuanxiong from Shiyang town, Dujiangyan city had more abundant endophytic fungi and low similarity to others, and which probably had special types of fungi. CONCLUSION: The abundant and stable endophytic fungal community is an important factor for the development of geoherb L. chuanxiong at Shiyang town, Dujiangyan city.

Effect of interfering TOR signaling pathway on the biosynthesis of terpenoids in Salvia miltiorrhiza Bge.

The TOR (Target of Rapamycin) signaling pathway, which takes TOR kinase as the core, regulates the absorption, distribution, and recycling of nutrients by integrating metabolic network and other signaling pathways, thus participating in the plant growth-defense trade-off. While terpenoids play an important role in plant growth, development, stress response, and signal transduction. The effect of the TOR signaling pathway on terpenoid biosynthesis in plants has yet to be studied in detail. In this study, the tissue culture seedlings of Salvia miltiorrhiza were treated with the TOR inhibitor AZD8055. The results show that the roots of the control group had begun to grow on the 8th day, while the seedlings treated with AZD8055 had no rooting signs. Combined with the expression changes of genes related to the TOR signaling pathway in the first 8 days, samples on the 3rd, 6th, and 8th days were selected for RNA-Seq analysis. Through RNA-Seq analysis, a total of 50,689 unigenes were obtained from the samples of these three periods, of which 4088 unigenes showed differential expression. The function enrichment and time-series analysis of differentially expressed genes (DEGs) showed that the main influence of the TOR signal pathway on plant growth-related processes was gradually transmitted with treatment time after TOR was inhibited. Pathway enrichment analysis of DEGs showed that the genes in the biosynthesis of terpenoids, such as diterpenoid and carotenoid biosynthetic pathways, could be regulated. Compared with other stages, DEGs related to terpenoid biosynthesis were mainly regulated in the S2 stage. In addition, the genes involved in terpenoid skeleton biosynthesis was also considerably enriched in the S2 stage, according to the results of gene set enrichment analysis (GSEA) of unigenes. Inhibition of the TOR signaling pathway may affect the biosynthesis of terpenoid signaling molecules, inhibit gibberellin's biosynthesis, and promote abscisic acid's biosynthesis. This study has discussed the effect of interfering with the TOR pathway on terpenoid biosynthesis in S. miltiorrhiza from the perspective of omics and provides new insight into the interaction between the terpenoid biosynthesis pathway and the growth-defense trade-off of medicinal plants.

Recombination of endophytic bacteria in asexual plant Ligusticum chuanxiong Hort. caused by transplanting.

Background: Long-term asexual reproduction can easily lead to the degradation of plant germplasm, serious diseases and insect pests, reduction of production and even catastrophic crop failure. "Mountain Breeding and Dam Cultivation" is the main cultivation mode of Ligusticum chuanxiong Hort., which successfully avoided the germplasm degradation caused by long-term asexual reproduction. The recombination of endophytic fungi of L. chuanxiong caused by off-site transplantation was considered to be an important reason for its germplasm rejuvenation. However, whether bacteria have the same regularity is not yet known. Methods: In this study, we carried out the experiment of cultivating propagation materials of L. chuanxiong in different regions and transplanting them to the same region. High-throughput sequencing was performed to analyze the bacterial communities in L. chuanxiong and its soil. Results: The results showed that after transplanting, the plant height, tiller number, fresh weight, etc. of L. chuanxiong in mountainous areas were significantly higher than those in dam areas. At the same time, significant changes had taken place in the endophytic bacteria in reproductive material stem nodes (Lingzi, abbreviated as LZ). The diversity and abundance of bacteria in dam area LZ (YL) are significantly higher than those in mountainous area LZ (ML). The relative abundance of bacteria such as Xanthobacteraceae, Micromonosporaceae, Beijerinkiaceae, Rhodanobacteria, in ML is significantly higher than YL, mainly classified in Proteobateria and Actinobacteriota. In addition, the abundance advantage of Actinobacteriota still exists in MY (underground mature rhizomes obtained by ML). Meanwhile, the bacterial community was different in different area of transplanting. The diversity of bacterial communities in dam soil (YLS) is significantly higher than that in mountain soil (MLS). MLS had more Acidobacteriota than YLS. Comparative analysis showed that 74.38% of bacteria in ML are found in MLS, and 87.91% of bacteria in YL are found in YLS. Conclusions: We can conclude that the community structure of endophytic bacteria recombined after the transplantation of L. chuanxiong, which was related to the bacterial community in soils. Moreover, after transplanting in mountainous areas, LZ accumulated more potentially beneficial Actinobacteriota, which may be an important reason for promoting the rejuvenation of germplasm in L. chuanxiong. However, this hypothesis requires more specific experiments to verify. This study provided a new idea that off-site transplanting may be a new strategy to restore vegetative plant germplasm resources.

Integrated analysis of lncRNAs, mRNAs, and TFs to identify network modules underlying diterpenoid biosynthesis in Salvia miltiorrhiza.

Long non-coding RNAs (lncRNAs) are transcripts of more than 200 nucleotides (nt) in length, with minimal or no protein-coding capacity. Increasing evidence indicates that lncRNAs play important roles in the regulation of gene expression including in the biosynthesis of secondary metabolites. Salvia miltiorrhiza Bunge is an important medicinal plant in China. Diterpenoid tanshinones are one of the main active components of S. miltiorrhiza. To better understand the role of lncRNAs in regulating diterpenoid biosynthesis in S. miltiorrhiza, we integrated analysis of lncRNAs, mRNAs, and transcription factors (TFs) to identify network modules underlying diterpenoid biosynthesis based on transcriptomic data. In transcriptomic data, we obtained 6,651 candidate lncRNAs, 46 diterpenoid biosynthetic pathway genes, and 11 TFs involved in diterpenoid biosynthesis. Combining the co-expression and genomic location analysis, we obtained 23 candidate lncRNA-mRNA/TF pairs that were both co-expressed and co-located. To further observe the expression patterns of these 23 candidate gene pairs, we analyzed the time-series expression of S. miltiorrhiza induced by methyl jasmonate (MeJA). The results showed that 19 genes were differentially expressed at least a time-point, and four lncRNAs, two mRNAs, and two TFs formed three lncRNA-mRNA and/or TF network modules. This study revealed the relationship among lncRNAs, mRNAs, and TFs and provided new insight into the regulation of the biosynthetic pathway of S. miltiorrhiza diterpenoids.

Transcriptomics combined with physiological analysis reveals the mechanism of cadmium uptake and tolerance in Ligusticum chuanxiong Hort. under cadmium treatment.

Introduction: Ligusticum chuanxiong Hort. is a widely used medicinal plant, but its growth and quality can be negatively affected by contamination with the heavy metal cadmium (Cd). Despite the importance of understanding how L. chuanxiong responds to Cd stress, but little is currently known about the underlying mechanisms. Methods: To address this gap, we conducted physiological and transcriptomic analyses on L. chuanxiong plants treated with different concentrations of Cd2+ (0 mg·L-1, 5 mg·L-1, 10 mg·L-1, 20 mg·L-1, and 40 mg·L-1). Results: Our findings revealed that Cd stress inhibited biomass accumulation and root development while activating the antioxidant system in L. chuanxiong. Root tissues were the primary accumulation site for Cd in this plant species, with Cd being predominantly distributed in the soluble fraction and cell wall. Transcriptomic analysis demonstrated the downregulation of differential genes involved in photosynthetic pathways under Cd stress. Conversely, the plant hormone signaling pathway and the antioxidant system exhibited positive responses to Cd regulation. Additionally, the expression of differential genes related to cell wall modification was upregulated, indicating potential enhancements in the root cell wall's ability to sequester Cd. Several differential genes associated with metal transport proteins were also affected by Cd stress, with ATPases, MSR2, and HAM3 playing significant roles in Cd passage from the apoplast to the cell membrane. Furthermore, ABC transport proteins were found to be key players in the intravesicular compartmentalization and efflux of Cd. Discussion: In conclusion, our study provides preliminary insights into the mechanisms underlying Cd accumulation and tolerance in L. chuanxiong, leveraging both physiological and transcriptomic approaches. The decrease in photosynthetic capacity and the regulation of plant hormone levels appear to be major factors contributing to growth inhibition in response to Cd stress. Moreover, the upregulation of differential genes involved in cell wall modification suggests a potential mechanism for enhancing root cell wall capabilities in isolating and sequestering Cd. The involvement of specific metal transport proteins further highlights their importance in Cd movement within the plant.

The Complete Chloroplast Genomes of Gynostemma Reveal the Phylogenetic Relationships of Species within the Genus.

Gynostemma is an important medicinal and food plant of the Cucurbitaceae family. The phylogenetic position of the genus Gynostemma in the Cucurbitaceae family has been determined by morphology and phylogenetics, but the evolutionary relationships within the genus Gynostemma remain to be explored. The chloroplast genomes of seven species of the genus Gynostemma were sequenced and annotated, of which the genomes of Gynostemma simplicifolium, Gynostemma guangxiense and Gynostemma laxum were sequenced and annotated for the first time. The chloroplast genomes ranged from 157,419 bp (Gynostemma compressum) to 157,840 bp (G. simplicifolium) in length, including 133 identical genes: 87 protein-coding genes, 37 tRNA genes, eight rRNA genes and one pseudogene. Phylogenetic analysis showed that the genus Gynostemma is divided into three primary taxonomic clusters, which differs from the traditional morphological classification of the genus Gynostemma into the subgenus Gynostemma and Trirostellum. The highly variable regions of atpH-atpL, rpl32-trnL, and ccsA-ndhD, the repeat unilts of AAG/CTT and ATC/ATG in simple sequence repeats (SSRs) and the length of overlapping regions between rps19 and inverted repeats(IRb) and between ycf1 and small single-copy (SSC) were found to be consistent with the phylogeny. Observations of fruit morphology of the genus Gynostemma revealed that transitional state species have independent morphological characteristics, such as oblate fruit and inferior ovaries. In conclusion, both molecular and morphological results showed consistency with those of phylogenetic analysis.