[Mechanism and application prospects of microbial fertilizers in regulating quality formation of medicinal plants].

With the promotion of chemical fertilizer and pesticide reduction and green production of traditional Chinese medicines, microbial fertilizers have become a hot way to achieve the zero-growth of chemical fertilizers and pesticides, improve the yield and qua-lity of medicinal plants, maintain soil health, and promote the sustainable development of the planting industry of Chinese herbal medicines. Soil conditions and microenvironments are crucial to the growth, development, and quality formation of medicinal plants. Microbial fertilizers, as environmentally friendly fertilizers acting on the soil, can improve soil quality by replenishing organic matter and promoting the metabolism of beneficial microorganisms to improve the yield and quality of medicinal plants. In this regard, understanding the mechanism of microbial fertilizer in regulating the quality formation of medicinal plants is crucial for the development of herbal eco-agriculture. This study introduces the processes of microbial fertilizers in improving soil properties, participating in soil nutrient cycling, enhancing the resistance of medicinal plants, and promoting the accumulation of medicinal components to summarize the mechanisms and roles of bacterial fertilizers in regulating the quality formation of medicinal plants. Furthermore, this paper introduces the application of bacterial fertilizers in medicinal plants and makes an outlook on their development, with a view to providing a scientific basis for using microbial fertilizers to improve the quality of Chinese herbal medicines, improve the soil environment, promote the sustainable development of eco-agriculture of traditional Chinese medicine, and popularize the application of microbial fertilizers.

A Prenyltransferase Participates in the Biosynthesis of Anthraquinones in Rubia cordifolia.

Anthraquinones constitute the largest group of natural quinones, which are used as safe natural dyes and have many pharmaceutical applications. In plants, anthraquinones are biosynthesized through two main routes: the polyketide pathway and the shikimate pathway. The latter primarily forms alizarin-type anthraquinones, and the prenylation of 1,4-dihydroxy-2-naphthoic acid is the first pathway-specific step. However, the prenyltransferase responsible for this key step remains uncharacterized. In this study, the cell suspension culture of Madder (Rubia cordifolia), a plant rich in alizarin-type anthraquinones, was discovered to be capable of prenylating 1,4-dihydroxy-2-naphthoic acid to form 2-carboxyl-3-prenyl-1,4-naphthoquinone and 3-prenyl-1,4-naphthoquinone. Then, a candidate gene belonging to the UbiA superfamily, R. cordifolia  dimethylallyltransferase 1 (RcDT1), was shown to account for the prenylation activity. Substrate specificity studies revealed that the recombinant RcDT1 recognized naphthoic acids primarily, followed by 4-hydroxyl benzoic acids. The prenylation activity was strongly inhibited by 1,2- and 1,4-dihydroxynaphthalene. RcDT1 RNA interference significantly reduced the anthraquinones content in R. cordifolia callus cultures, demonstrating that RcDT1 is required for alizarin-type anthraquinones biosynthesis. The plastid localization and root-specific expression further confirmed the participation of RcDT1 in anthraquinone biosynthesis. The phylogenetic analyses of RcDT1 and functional validation of its rubiaceous homologs indicated that DHNA-prenylation activity evolved convergently in Rubiaceae via recruitment from the ubiquinone biosynthetic pathway. Our results demonstrate that RcDT1 catalyzes the first pathway-specific step of alizarin-type anthraquinones biosynthesis in R. cordifolia. These findings will have profound implications for understanding the biosynthetic process of the anthraquinone ring derived from the shikimate pathway.

Multi-omics revealed molecular mechanism of biphenyl phytoalexin formation in response to yeast extract-induced oxidative stress in Sorbus aucuparia suspension cells.

KEY MESSAGE: Yeast extract-induced oxidative stress in Sorbus aucuparia suspension cells leads to the biosynthesis of various hormones, which activates specific signaling pathways that augments biphenyl phytoalexin production. Pathogen incursions pose a significant threat to crop yield and can have a pronounced effect on agricultural productivity and food security. Biphenyl phytoalexins are a specialized group of secondary metabolites that are mainly biosynthesized by Pyrinae plants as a defense mechanism against various pathogens. Despite previous research demonstrating that biphenyl phytoalexin production increased dramatically in Sorbus aucuparia suspension cells (SASCs) treated with yeast extract (YE), the underlying mechanisms remain poorly understood. To address this gap, we conducted an in-depth, multi-omics analysis of transcriptome, proteome, and metabolite (including biphenyl phytoalexins and phytohormones) dynamics in SASCs exposed to YE. Our results indicated that exposure to YE-induced oxidative stress in SASCs, leading to the biosynthesis of a range of hormones, including jasmonic acid (JA), jasmonic acid isoleucine (JA-ILE), gibberellin A4 (GA4), indole-3-carboxylic acid (ICA), and indole-3-acetic acid (IAA). These hormones activated specific signaling pathways that promoted phenylpropanoid biosynthesis and augmented biphenyl phytoalexin production. Moreover, reactive oxygen species (ROS) generated during this process also acted as signaling molecules, amplifying the phenylpropanoid biosynthesis cascade through activation of the mitogen-activated protein kinase (MAPK) pathway. Key genes involved in these signaling pathways included SaBIS1, SaBIS2, SaBIS3, SaPAL, SaB4H, SaOMT, SaUGT1, SaLOX2, SaPR1, SaCHIB1, SaCHIB2 and SaCHIB3. Collectively, this study provided intensive insights into biphenyl phytoalexin accumulation in YE-treated SASCs, which would inform the development of more efficient disease-resistance strategies in economically significant cultivars.

Genome-Wide Identification and Characterization of the bHLH Gene Family and Its Response to Abiotic Stresses in Carthamus tinctorius.

The basic helix-loop-helix (bHLH) transcription factors possess DNA-binding and dimerization domains and are involved in various biological and physiological processes, such as growth and development, the regulation of secondary metabolites, and stress response. However, the bHLH gene family in C. tinctorius has not been investigated. In this study, we performed a genome-wide identification and analysis of bHLH transcription factors in C. tinctorius. A total of 120 CtbHLH genes were identified, distributed across all 12 chromosomes, and classified into 24 subfamilies based on their phylogenetic relationships. Moreover, the 120 CtbHLH genes were subjected to comprehensive analyses, including protein sequence alignment, evolutionary assessment, motif prediction, and the analysis of promoter cis-acting elements. The promoter region analysis revealed that CtbHLH genes encompass cis-acting elements and were associated with various aspects of plant growth and development, responses to phytohormones, as well as responses to both abiotic and biotic stresses. Expression profiles, sourced from transcriptome databases, indicated distinct expression patterns among these CtbHLH genes, which appeared to be either tissue-specific or specific to certain cultivars. To further explore their functionality, we determined the expression levels of fifteen CtbHLH genes known to harbor motifs related to abiotic and hormone responses. This investigation encompassed treatments with ABA, salt, drought, and MeJA. The results demonstrated substantial variations in the expression patterns of CtbHLH genes in response to these abiotic and hormonal treatments. In summary, our study establishes a solid foundation for future inquiries into the roles and regulatory mechanisms of the CtbHLH gene family.

Authenticating the geographical origin of the Chinese yam (Tiegun) with stable isotopes and multiple elements.

The Chinese yam, an important orphan crop with both high nutrient and health promoting value, is mainly produced in the Yellow-Huai-Hai plain near the river basins in China. The protected designation of origin (PDO)-labeled Chinese yam differs greatly from others in market acceptance and price, which has led to fakes and the need for reliable authentication methods. Hence, stable isotope ratios of δ 13C, δ 15N, δ 2D, and δ 18O and 44 multielemental contents were used to explore the authentication of geographical origins and the effect of environmental factors. Twenty-two elements and δ 15N were selected as the key variables to authenticate Chinese yams from three river basins as well as to authenticate them among traditional PDOs and others in the Yellow River basin. Moreover, six environmental factors, including the moisture index, maximum temperature, photosynthetically active radiation, soil organic carbon, total nitrogen and pH, were found to be highly related to these variances.

The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma.

Shikonin derivatives are natural naphthoquinone compounds and the main bioactive components produced by several boraginaceous plants, such as Lithospermum erythrorhizon and Arnebia euchroma. Phytochemical studies utilizing both L. erythrorhizon and A. euchroma cultured cells indicate the existence of a competing route branching out from the shikonin biosynthetic pathway to shikonofuran. A previous study has shown that the branch point is the transformation from (Z)-3''-hydroxy-geranylhydroquinone to an aldehyde intermediate (E)-3''-oxo-geranylhydroquinone. However, the gene encoding the oxidoreductase that catalyzes the branch reaction remains unidentified. In this study, we discovered a candidate gene belonging to the cinnamyl alcohol dehydrogenase family, AeHGO, through coexpression analysis of transcriptome data sets of shikonin-proficient and shikonin-deficient cell lines of A. euchroma. In biochemical assays, purified AeHGO protein reversibly oxidized (Z)-3''-hydroxy-geranylhydroquinone to produce (E)-3''-oxo-geranylhydroquinone followed by reversibly reducing (E)-3''-oxo-geranylhydroquinone to (E)-3''-hydroxy-geranylhydroquinone, resulting in an equilibrium mixture of the three compounds. Time course analysis and kinetic parameters showed that the reduction of (E)-3''-oxo-geranylhydroquinone was stereoselective and efficient in presence of NADPH, which determined that the overall reaction proceeded from (Z)-3''-hydroxy-geranylhydroquinone to (E)-3''-hydroxy-geranylhydroquinone. Considering that there is a competition between the accumulation of shikonin and shikonofuran derivatives in cultured plant cells, AeHGO is supposed to play an important role in the metabolic regulation of the shikonin biosynthetic pathway. Characterization of AeHGO should help expedite the development of metabolic engineering and synthetic biology toward production of shikonin derivatives.

[Overview of Chinese medicinal materials industry in 2021 and development suggestions].

This paper introduced the overview of the "eight trends" of Chinese medicinal materials(CMM) industry in 2021, analyzed the problems of CMM production, and put forward development suggestions. Specifically, "eight trends" could be summarized as follows.(1) The growing area of CMM tended to be stable, and some provinces began to release the local catalog of Dao-di herbs.(2) The protection process of new varieties accelerated, and a number of excellent varieties were bred.(3) The theory of ecological cultivation was further enriched, and the demonstration effect of ecological cultivation technology was prominent.(4) Some CMM realized complete mechanization and formed typical model cases.(5) The number of cultivation bases using the traceability platform increased, and provincial internet trading platforms were set up.(6) The construction of CMM industrial clusters accelerated, and the number of provincial-level regional brands increased rapidly.(7) Many new agricultural business entities were founded nationwide, and a variety of methods were used to drive the intensified development of CMM.(8) A number of local TCM laws were promulgated, and the management regulation of food and medicine homology substances catalogs was issued. On this basis, four suggestions for CMM production were proposed.(1) It is suggested to speed up the formulation of the national catalog of Dao-di herbs and carry out the certification of Dao-di herbs production bases.(2) Ecological planting of forest and grassland medicine should be further strengthened in terms of technical research and promotion based on the principle of ecological priority.(3) The basic work of disaster prevention should be paid more attention and technical measures for disaster mitigation should be developed.(4) The planted area of commonly used CMM should be incorporated into the national regular statistical system.

Study on the effect of compound cultivation on the growth feature and active ingredients content of Salvia miltiorrhiza.

We investigated the effects of the complex cultivation of Salvia miltiorrhiza on microbial communities, secretions, yield, and active ingredients, and the mechanism of action between microbial communities, secretions, and S. miltiorrhiza growth and development. Neither maize nor soybean was suitable to grow with S. miltiorrhiza, but sesame significantly increased salvinone content, the active ingredient of S. miltiorrhiza, and Tanshinone IIA, Tanshinone I, and Cryptotanshinone increased by 27.06%, 22.76%, and 26.41%, respectively, which increased the abundance and number of microbial communities in S. miltiorrhiza roots. 16S rRNA results showed that the most abundant bacterial phyla were Proteobacteria and Acidobacteriota, and their number increased with compound planting of sesame and S. miltiorrhiza. Salvia inter-root secretions affected the microbial community and Salvia growth and development, and lipids and lipid-like molecules significantly reduced Salvia yield and active ingredients. Overall, different plant secretions can lead to differences in the natural environment and Salvia root growth and development, and the composite planting of sesame with Salvia can improve inter-root microbial communities, enhance Salvia quality, and make fuller use of land resources.

[Strategic analysis on value accounting and realization of ecological products of Chinese materia medica].

Green agriculture has become the "wind vane" of agricultural reform in the new era. The development concept of Chinese materia medica(CMM) ecological agriculture and the emergence of new technologies provide momentum for the supply of high-quality green ecological products. As special ecological products, Dao-di herbs have medicinal, economic, ecological, and cultural values. The ecological planting of CMM and the value realization of ecological products are of great significance for the high-quality development of CMM agriculture and for prospering agriculture by improving quality. Therefore, this paper reviews the development status and typical cases, and puts forward the advantages of ecological product value realization of Dao-di herbs. On this basis, a value accounting system for ecological products of CMM was constructed from three aspects: supply services, ecological services, and cultural services. Finally, the suggestions for realizing the value of CMM ecological products are put forward.(1)It is suggested to strengthen the practice of ecological planting and production of CMM, and innovate the value realization model of ecological products.(2)The ecological planting bases of Dao-di herbs should be constructed to enhance the supply capacity of high-quality ecological products.(3)The quality and safety of ecological CMM should be improved to create the ecological planting brand of Dao-di herbs.(4)The ecological product value system and mechanism should be improved for realizing the value of CMM ecological products.(5)The education and publicity should be strengthened to raise the public awareness of the value of CMM ecological products.

[Chinese medicinal materials industry during the 14th Five-Year Plan period: trends and development suggestions].

This study steps through the future perspectives and gives the development suggestions of Chinese medicinal materials(CMM) industry by presenting the characteristics and open problems during the 13 th Five-Year Plan period. The development of CMM industry presents the following trends:(1)the development of Dao-di herbs speeds up with the increasing demand for high-quality CMM;(2)the mismatch between supply and demand is aggravating, which presses for supply-side structural reform;(3)ecological planting will become the core mode of production and bolster rural revitalization;(4)the demand for CMM with both medical and edible values keeps growing, and the antibiotic-free feed policy brings significant opportunities;(5)the "Internet Plus CMM" wave emerges, which promotes the construction of traceability system. Finally, we put forward the following suggestions for the sustainable development of CMM industry:(1)optimizing the layout for the production of Dao-di herbs according to local conditions;(2)strengthening the commercialization of the seeds and the breeding, multiplication, and extension of CMM to accelerate the realization of specialized seed production, mechanized seed processing, localized variety layout, and county-based unified seed supply;(3)ensuring the safety of pesticide use and accelerating the registration of special pesticides;(4)promoting both theoretical and practical research on ecological production of CMM;(5)publicizing the demonstration and popularization of CMM traceability system. Overall, significant progress has been achieved in the CMM industry during the 13 th Five-Year Plan period, and this industry is in a critical stage of high-quality development, facing both challenges and opportunities.

TMT-Based Quantitative Proteomics Analysis Reveals the Panoramic Pharmacological Molecular Mechanism of ß-Elemonic Acid Inhibition of Colorectal Cancer.

Colorectal cancer (CRC) is one of the most common cancers worldwide but has limited available therapeutic methods; therefore, there is a need to develop highly efficient prevention and treatment strategies. Here, we investigated the anti-cancer activity of ß-elemonic acid (EA) in CRC in vitro and in vivo. Our results showed that EA inhibited cell proliferation and migration in the CRC cell lines SW480 and HCT116. Moreover, EA significantly suppressed the growth of transplanted colorectal tumors in nude mice. Interestingly, high-throughput tandem mass tag (TMT)-based quantitative proteomics indicated that EA mainly targets tumor mitochondria and attenuates the translation of 54 mitochondrial ribosome proteins, many of which are discovered significantly upregulated in clinical CRC patients. More interestingly, EA at a low concentration (lower than 15 µg/ml) repressed the cell cycle by downregulating CDK1, CDK6, and CDC20, whereas at a high concentration (higher than 15 µg/ml), caused a non-apoptotic cell death-ferroptosis via downregulating ferritin (FTL) and upregulating transferrin (TF), ferroxidase (CP), and acyl-CoA synthetase long-chain family member 4 (ACSL4). This is the first report on the panoramic molecular mechanism of EA against CRC, which would make great contributions to developing a novel drug for colorectal cancer therapy.

[Contributions and strategies of eco-agriculture of Chinese medicine services for carbon dioxide peaking and carbon neutrality].

Carbon dioxide peaking and carbon neutrality have become hot issues of political and economic activities in China and abroad. The structure and development of various industries in China will be profoundly affected in the process of accomplishing "Dual Carbon" goals. Eco-agriculture of Chinese medicine(EACM) highlights the balance and sustainable development of the ecosystem while producing high-quality medicinal materials. With chemically synthesized fertilizers, pesticides, and growth regulators prohibited, EACM emphasizes the recycling of agricultural and sideline products and the reduction of waste output, which results in the minimal negative impact on the ecological environment. Therefore, it is typical agriculture with low-carbon sources and high-carbon sinks. This study reviewed the mechanism and potential of EACM in carbon dioxide peaking and carbon neutrality, analyzed the specific ways of EACM in reducing carbon sources and increasing carbon sinks based on the typical ecological planting pattern, and proposed the point of view to strengthen EACM as well as the "Dual Carbon" theory and research methods, so as to direct low-carbon and efficient deve-lopment. Furthermore, this study advocated to comprehensively promote the transformation of Chinese medicine production from chemical agriculture to eco-agriculture to improve the comprehensive benefits of contribution rate of carbon neutrality, explore and establish carbon sink compensation mechanism to ensure the sustainable and healthy development of EACM, and strengthen the training of EACM and "Dual Carbon" theory and technologies to continuously improve the capacity of EACM in sustainable development. This study is expected to provide a reference for the development of ecological functions in EACM and the development of economic functions through ecological functions.

[Problems and strategy of further development of Chinese medicinal materials with edible values].

"Medicine and food homology" is a precious health care concept rooted in the culture of Chinese medicine and plays an important role in the development of the national health industry. It is consistent with the current global trend that food and medicine are mutually penetrating. Accordingly, the Chinese medicinal materials with edible values have an increasing production scale. Especially, in the context that the development of Chinese medicinal materials is switching from pursuing quantity to quality, the food field has become the main market for the new production capacity of Chinese medicinal materials, which has presented a broad prospect. However, the quality standards of raw materials, production methods, and administration ways vary between the materials for edible and medicinal purposes. Specifically, the food for medicinal use on the market cannot meet the quality standards of medicinal mate-rials, while the medicinal materials fail to meet the taste requirements as food. As a result, these problems cause difficulties in market circulation and supervision. In this paper, we analyzed the formation of Chinese medicinal materials with edible values, compared the food with medicinal value, common food, and functional food, and analyzed the different quality requirements of Chinese medicinal materials used in different scenarios. Further, we advised the differential development of Chinese medicinal materials in different directions(edible or medicinal use) from production to supervision. Including:(1) In the variety registration of Chinese medicinal materials with edible values, the variety breeding direction should be announced according to the requirements that medicinal materials care more about the content of active ingredients and food use materials preferentially need to meet the requirements of edible palatability.(2) Differentiation can be reflected in the selection of cultivation mode and planting and processing technology of medicinal materials, The differential production technical specification of medicinal materials with edible values should be developed. Such as the "simulated cultivation" mode is encouraged in the plant of medicinal materials to ensure its quality and the strict management of inputs and sufficient cultivation years should be guaranteed. While for edible medicinal materials, more kinds of cultivation techniques can be selected according to their processing methods.(3) The market supervision of medicinal materials with edible values should be guided by the purpose of their sales and use, which depends on the accurately recognize of the relationship between the properties of medicinal materials with edible values and the situation of pharmacopeia collection.(4) During publicity, when used as ordinary food and health food medi-cinal materials, it should be noticed that the publicity of the product's efficacy must meet the requirements of corresponding regulations.

[Construction status and development strategy of GAP bases for Chinese herbal medicine].

Good agricultural practices(GAP) for Chinese herbal medicine(CHM) is the source of quality control in the production of CHM. To ensure the safety, efficacy, and quality of Chinese herbal and natural medicine, the international community has been exploring the implementation of GAP for CHM. The implementation of GAP for CHM has a far-reaching impact, and the scale planting of CHM has been expanding. However, the medicinal materials produced by the GAP bases for CHM still cannot fully meet the market demand, and the output and quality of CHM remain to be improved. This paper summarizes construction status of GAP bases for CHM.The 129 companies that passed the certification during the implementation of GAP for CHM included 196 GAP bases, forming the scale CHM production zone with Sichuan province, Yunnan province, Jilin province, Henan province, and Shandong province as the core.The total area of GAP bases for CHM in China is about 250 000-500 000 Mu(1 Mu≈667 m2), which is still less than 1% of the total production area of CHM. The international agricultural production quality management strategies are all market-oriented behaviors.Drawing on the international good agricultural and collection practice(GACP) and hazard analysis critical control point(HACCP) as well as the relevant policies of Chinese quality management of CHM, we put forward feasible suggestions for the further implementation of GAP for CHM:(1)The market rules and international experience should be followed and employed to promote GAP management of CHM and third-party implementation of GAP certification;(2)The sound development of GAP for CHM should be boosted with the HACCP management system and the revision of GAP for CHM;(3)The implementation of policies and standards should be stepped up to facilitate the building of a whole-course traceability system for the production of CHM with high quality and reasonable prices.

[Current status and prospects of implementing GAP in Chinese medicine injection materials].

Chinese medicine injections(CMIs) have higher requirements for quality consistency and controllability than other traditional Chinese medicine products. The implementation of Good Agricultural Practice of Medicinal Plants and Animals(GAP) is an important factor that guarantees the quality stability of raw Chinese medicinal materials and affects the quality of CMIs. Through literature review, data research, expert consultation, and statistical analysis, this paper analyzes the current status of GAP management of key CMIs and the impact of GAP management of raw medicinal materials on the quality consistency and controllability of CMIs. Furthermore, it demonstrates the rationality, necessity, and feasibility of the full implementation of GAP on the basis of CMIs safety re-evaluation.

Diverse Intercropping Patterns Enhance the Productivity and Volatile Oil Yield of Atractylodes lancea (Thunb.) DC.

Commercial cultivation of the medicinal plant Atractylodes lancea is significantly restricted by low survival rates and reduced yields. Intercropping can reasonably coordinate interspecific interactions, effectively utilize environmental resources, and increase survival and yield. We conducted a field experiment from 2014 to 2016 to analyze the advantages and effects of intercropping on A. lancea survival, growth traits, individual volatile oil content, and total volatile oil content. In addition to A. lancea monoculture (AL), five intercropping combinations were planted: Zea mays L. (ZM) + A. lancea, Tagetes erecta L. (TE) + A. lancea, Calendula officinalis L. (CO) + A. lancea, Glycine max (Linn.) Merr. (GM) + A. lancea, and Polygonum hydropiper L. (PH) + A. lancea. The survival and average rhizome weight of A. lancea was higher in the ZM, CO, and TE treatments than in the monoculture treatment, and the average plant height was higher in all intercropping treatments than in the monoculture. The volatile oil content of A. lancea from the ZM and CO treatments was significantly improved relative to that of monoculture plants. The volatile oil harvest was higher in the ZM, CO, and TE treatments than in the monoculture. We conclude that intercropping is an effective way to increase the survival and yield of A. lancea. Furthermore, intercropping with ZM, CO, and TE increases the harvest of four volatile oils from A. lancea.

[Cloning and functional verification of carboxyl CoA ligases(AeCCLs) in Arnebia euchroma].

Carboxyl CoA ligases(CCLs) is an important branch of adenylate synthetase gene family, which mainly has two-step catalytic reactions. Firstly, in the presence of adenosine triphosphate, it can catalyze the pyrophosphorylation of carboxylateswith diffe-rent structures to form corresponding acyl adenosine monophosphate intermediates. Secondly, adenosine monophosphate was replaced by free electrons in the mercaptan group of enzyme A or other acyl receptors by nucleophilic attack to form thioesters. In this study, on the basis of the transcriptome database of Arnebia euchroma, two genes were selected, named AeCCL5(XP_019237476.1) and AeCCL7(XP_019237476.1). Bioinformatics analysis showed that their relative molecular weights were 60.569 kDa and 60.928 kDa, theoretical PI were 8.59 and 8.92, respectively. They both have transmembrane domains but without signal peptide. By multiple sequence alignment and phylogenetic tree analysis, we found that the similarity between AeCCLs and other plant homologous proteins was not high, and the substrate binding sites of AeCCLs were not highly conserved. The reasons might be that the sequence and structure need to adapt to the changes of new substrates in the process of evolution. In this study, the full-length of AeCCL5 and AecCCL7 were cloned into the expression vector pCDFDuet-1. The proteins of AeCCL5 and AeCCL7 with His-tag were expressed in Escherichia coli. The proteins of AeCCL5 and AeCCL7 were purified by nickel column. In vitro enzymatic reactions proved that both AeCCL5 and AeCCL7 can participate in the upstream phenylpropane pathway of shikonin biosynthesisby catalyzing 4-coumaric acid to produce 4-coumarin-CoA, and then to synthesis p-hydroxybenzoic acid, which is an important precursor of shikonin biosynthesis in A. euchroma.

[Analysis of economic benefits of Chinese medicine eco-agriculture based on multiple stakeholders].

As the most advanced environment-friendly production model in the international society, ecological agriculture of Chinese materia medica(CMM) is the only way for the development of modern agriculture. With the proposal of the declaration on ecolo-gical agriculture of CMM, "Don't grab land from farmland, don't be enemies of grass and insects, don't be afraid of barren slopes and forests, and live up to the green and green mountains", the ecological planting of CMM has blossomed all over the country, and formed a scientific theory, technology and model. Based on the theory and method of economics, this paper expounds the comprehensive benefits and development advantages of ecological agriculture of CMM from the perspectives of farmers(producers), patients(consumers) and the country. From the perspective of medicinal farmers, the input and output income of conventional agriculture and ecological agriculture of CMM such as Panax ginseng, Astragalus propinquus, Atractylodes lancea, and Bupleurum chinense were compared, and it was found that ecological agriculture of CMM had obvious advantages in net income, average annual income and input-output ratio, which could better promote farmers' income. From the perspective of patients, according to the same dose, the content of active ingredients in ecologically planted CMMs is significantly higher than that in conventionally-planted herbs, and the amount of effective substances taken by patients is also higher, so as to achieve better therapeutic effect. At the national level, ecological planting of CMM is the key to ensuring the high-quality development of CMM industry, increasing farmers' income, ensuring the safety of people's drug use and promoting the sustainable development of agriculture. It is also an important part of realizing the harmonious development of economy, society and environment and promoting ecological civilization. In general, the declaration on ecological agriculture of CMM embo-dies the core characteristics and goals of ecological agriculture, and also points of the path and vision of ecological agriculture of CMM in the future. The declaration will guide production practice, promote the benefit of farmers, and lay the foundation for the sustainable development of CMM industry.

[Current situation of nitrogen application and its effects on yield and quality of Chinese materia medica].

Nitrogen fertilizer has been the long-lasting crucial component in cultivation of Chinese materia medica(CMM) and crops for its profound effects on enhancing the productivity. In consideration of its role in better production, intensive and excessive application of N fertilizer is often found in CMM cultivation. Therefore, firstly, this review summarized various concentrations of N application with regards to different CMM and districts from the literatures published in the last two decades. The recommended concentration of nitrogen application of forty seven CMM species were covered in this review. We found that the optimum rates of nitrogen fertili-zer for different medicinal plants species were varied in the range between 0-1 035.55 kg·hm~(-2). Most of the optimum rates of nitrogen fertilizer for CMM in published researches fell between 100-199 kg·hm~(-2). The optimum rate of nitrogen fertilizer is not only related to amount of nitrogen required for different medicinal plants but also to soil fertilities of different fields. In addition, we outlined the diffe-rent effects of proper and excessive nitrogen deposition on yield of CMM. Proper nitrogen deposition benefits the yield of CMM, howe-ver, excessive nitrogen use accounts for a decrease in CMM yield. We elucidated that nutritional content, water use efficiencies, and photosynthesis capacity were major influencing factors. Researches showed that proper nitrogen fertilizer could promote the water use efficiencies of plants and boost photosynthesis. Consequently, the yield of CMM can be enhanced after nitrogen deposition. However, negative effects of nitrogen fertilizer were also found on plant including producing toxic substances to the soil and causing severe pest damages. Lastly we analyzed the impact of N fertilizer application on secondary metabolites which accounts for a large part of active pharmaceutical ingredients of CMM. It usually caused an increase in nitrogen-containing secondary metabolites content and a decrease in non-nitrogen-containing secondary metabolites content respectively. The potential underlying mechanisms are the different synthetic pathways of these metabolites and the plant nutritional status. Synthesis of non-nitrogen-containing secondary metabolites like phenols can be inhibited after nitrogen application because of the competition of the same precursor substances between metabolites synthesis and plant growth. To sum up, impacts and mechanisms of nitrogen fertilizer on yield and quality enhancement of CMM were discussed in this review. Negative effects of excessive nitrogen application on CMM should be paid special attention in CMM cultivation and prescription fertilization based on the field soil quality is strongly recommended. Overall, this review aims to provides insights on improving the proper application of N fertilizer in the cultivation of CMM.

[Effects and mechanisms of nitrogen fertilizers on soil and tritrophic interactions in Chinese medicinal plants ecosystem].

Nitrogen is one of the most frequently used fertilizers in growth of Chinese medicinal plants(CMP). As in many other ecosystems, CMP ecosystem is also composed of plant-herbivore-natural enemy(tritrophic) interactions. Nitrogen fertilizer influences the growth and reproduction of CMP, and it is also able to heavily shape the ecosystem functions of CMP ecosystem through bottom-up forces. Understanding the specific effects of nitrogen fertilizer towards each trophic level will be beneficial to improve the resistance of CMP to herbivore and enhance the control efficiency of nature enemies to herbivore, and eventually, maximize the yield and quality of CMP. Most papers published on nitrogen use in plants focused mainly on the impact of nitrogen fertilization on CMP yield and quality. Influences of nitrogen application on CMP ecosystem get little attention at present. Therefore, this review summed up the potential effects of nitrogen fertilization on CMP ecosystem from perspectives of soil and tritrophic interactions. First of all, nitrogen fertilizer might decrease soil microbial biomass and altered the community structures of soil bacteria, fungi and protozoa. Negative effects of nitrogen fertilizer were found on biodiversity of soil bacteria and protozoa. Different fungi species respond differently to nitrogen fertili-zers. Nitrogen deposition can also decrease the soil pH. Decreases in soil microbial diversity and soil acidification can cause negative effects on CMP growth. In addition, nitrogen fertilizer could regulate the pest resistance of CMP including constitutive and inducible resistance. Both positive and negative effects of nitrogen application were found on pest resistance of CMP. Moreover, the development and predation of natural enemies were influenced by nitrogen deposition. Nitrogen influences natural enemies in many ways including plant volatiles, plant nutrient and structure and the supplementary food quality. Nectar and honeydew of plants and preys serve as important food source for natural enemies especially in early season when preys are still not available. Finally, the interactions between herbivores and their natural enemies were also shaped by nitrogen fertilizer in many aspects like increasing the nutritional content of prey and changing control efficiency of natural enemies. Some herbivores have evolved a strategy to sequester secondary metabolites which they absorbed from plant during their feeding. Studies showed that sequestration efficiency of secondary metabolites in prey could also be regulated by nitrogen. Parasitic, emergence, reproduction rate and longevity of parasites were found positively correlated with nitrogen deposition. Hopefully this study will shed light on practicable and economical application of nitrogen in cultivation of CMP.