Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.

OBJECTIVE: In Pakistan, traditional medicines are an important component of the medical system, with numerous varieties and great demands. However, due to the scattered resources and the lack of systematic collection and collation, adulteration of traditional Pakistani medicine (TPM) is common, which severely affects the safety of their medicinal use and the import and export trades. Therefore, it is urgent to systematically organize and unify the management of TPM and establish a set of standards and operable methods for the identification of TPM. METHODS: We collected and organized the information on 128 TPMs with regard to their medicinal parts, efficacy, usage, and genetic material, based on Pakistan Hamdard Pharmacopoeia of Eastern Medicine: Pharmaceutical Codex. The genetic information of TPM is summarized from national center for biotechnology information (NCBI) and global pharmacopoeia genome database (GPGD). Furthermore, we utilized bioinformatics technology to supplement the chloroplast genome (cp-genome) data of 12 TPMs. To build the web server, we used the Linux + Apache + MySQL + PHP (LAMP) system and constructed the webpage on a PHP: Hypertext Preprocessor (PHP) model view controller (MVC) framework. RESULTS: We constructed a new genomic database, the traditional Pakistani medicine genomic database (TPMGD). This database comprises five entries, namely homepage, medicinal species, species identification, basic local alignment search tool (BLAST), and download. Currently, TPMGD contains basic profiles of 128 TPMs and genetic information of 102 TPMs, including 140 cytochrome c oxidase subunit I (COI) sequences and 119 mitochondrial genome sequences from Bombyx mori, 1 396 internal transcribed spacer 2 (ITS2) sequences and 1 074 intergenic region (psbA-trnH) sequences specific to 92 and 83 plant species, respectively. Additionally, TPMGD includes 199 cp-genome sequences of 82 TPMs. CONCLUSION TPMGD is a multifunctional database that integrates species description, functional information inquiry, genetic information storage, molecular identification of TPM, etc. The database not only provides convenience for TPM information queries but also establishes the scientific basis for the medication safety, species identification, and resource protection of TPM.

The problems of non-standard use of pesticides, and excessive pesticide residues and excessive heavy metal, etc. are common in the productive process of safflower (Carthamus tinctorius) and they are the factors restricting the sustainable development of safflower industry. Pollution-free production is an effective measure to ensure high quality of safflower. This paper summarized the environment of production area, selection of improved varieties for the production in the local places and its characteristics, the standardized comprehensive agronomic management and pollution-free rational fertilization technology. Additionally, the pollution-free control of safflower pests and diseases should follow the principle of priority to prevention and comprehensive prevention. Agricultural, biological and physical control should be preferred to use, and be combined with safe and low toxicity of chemical control. The standardization and industrialization of safflower production were realized by the construction of a comprehensive control technology system of pests and diseases of pollution-free safflower, which promoted the healthy development of the safflower plantation industry and achieved pollution-free standards.

Rabdosia (Bl.) Hassk is a medicinal plant rich in enantiomers such as 15α-acetoxyl-6, 11α-epoxy-6α-hydroxy-20-oxo-6, 7-seco-ent-kaur-16-en-1, 7-olide and 15α-dihydroxy-6β-methoxy-6, 7-seco-6, 20-epoxy-1, 7-olide-ent-kaur-16-ene. It has high medicinal value and huge market potential, but the yield from wild Rabdosia (Bl.) Hassk species cannot meet the needs of marketization. Carrying out pollution-free cultivation of Rabdosia rubescens (Hemsl.) Hara and Rabdosia serra (Maxim.) Hara has become the most direct and effective measure to solve this problem in order to ensure the sustainable use of the resources of Rabdosia. This paper summarized a set of non- pollution cultivation system of R. rubescens and R. serra to achieve the"safe, effective, stable and controllable"target.

Chaenomelis Fructus is a traditional medicinal material derived from dry near-mature fruit of Chaenomeles speciosa (Sweet) Nakai. Chaenomelis Fructus has efficacy of leveling liver and relaxing tendons, harmonizing the stomach and resolving dampness. As the limit of the usage of available forest land, cultivated ginseng in the farmland would become the mainly Chaenomelis Fructus planting mode, meanwhile the non-polluted production technology would be the mainly development direction in the future. In this study, the non-polluted cultivation technology system of Chaenomelis Fructus was established based on the research results of field investigation in the cultivated regions. The system includes suitable planting regions selecting, planting method, field management, pest control and quality control. In order to promote the healthy and sustainable development of the cropland industry of Chaenomelis Fructus, the new varieties of good quality cultivated in farmland and the comprehensive prevention and control platform of disease and insect pest are put forward in this paper.

Anoectochilus, the fresh or dry whole grass of Anoectochilus roxburghii (Wall.) Lindl., has the effects of cooling blood, dispelling wind, dispelling dampness and resolving toxin, resulting in a wide range of clinical applications and large market demand. However, in the planting process, there are problems such as irregular planting management, excessive addition of pesticides, irregular processing links, even the phenomenon of selling adulteration, which seriously affects the quality of the medicine. In order to guarantee the quality and productivity of Chinese herbal medicines, this paper developed the standard operating procedures for the non-pollution Anoectochilus, including the non-pollution cultivation environment of Anoectochilus, planting methods, field management, pest control and quality control, etc., which provides guidance to promote the healthy development of the Anoectochilus industry.

Moutan Cortex and Paeonia Radix are also traditional Chinese herbal medicines. The market is in great demand. However, the market sales of Moutan Cortex and Paeonia Radix have problems such as pesticide residues, excessive heavy metals and degraded quality, which affect their safety and effectiveness. The establishment of a pollution-free and planting system can effectively reduce the pesticide residue and heavy metal content and improve the quality of the medicinal materials while ensuring the production of Moutan Cortex and Paeonia Radix. The article uses field data which based on P.suffruticosa and P.lactiflora cultivation, combined with growth habits and distribution of origin, this research has developed a pollution-free and planting technology system for P.suffruticosa and P.lactiflora. The system includes planting base regionalization, field management, fertilization, pest control and so on, which provides reference for P.suffruticosa and P.lactiflora.