ObjectiveTo explore the genetic relationship and reveal the genetic variations of 45 germplasm accessions of Artemisia argyi. MethodGenotyping-by-sequencing （GBS） was employed to mine single nucleotide-polymorphisms （SNPs） from the 45 germplasm accessions. Principal component analysis， phylogenetic analysis， population genetic structure analysis， and genetic variation analysis were conducted based on the SNPs. ResultA total of 111.91 Gb of data were obtained， with the Q20， Q30， and average GC content of 96.39%， 90.33%， and 39.37%， respectively. The comparison rate between clean reads and the reference genome was 70.24%-98.97%. A total of 22 399 Indels and 170 539 SNPs were obtained， and the 10^th pair of chromosomes had the most variation sites. The results of principal component analysis， cluster analysis， and genetic diversity analysis classified the 45 germplasm accessions into three groups. Group Ⅰ contained three germplasm accessions from Qichun County. The germplasm accessions in group Ⅱ were all wild. Group Ⅲ contained 31 germplasm accessions， with the most complex sources. Moreover， the 45 germplasm accessions can be classified into 3 subtypes， containing the genetic information from three ancestors. The results indicated rich genetic diversity of A. argyi from different sources， especially the germplasm accessions from Qichun County， Hubei province. ConclusionThis study provides theoretical support for breeding new varieties， developing specific SNP markers， and revealing the genetic relationship of A. argyi.

ObjectiveTo study the changes of dry matter accumulation， mineral element absorption and medicinal component accumulation in Belamcanda chinensis and to provide a theoretical basis for fertilization in the standardized cultivation of B. chinensis. MethodDuring the two-year growth period of B. chinensis， the samples were collected at 30-day intervals from day 90 after sowing for the measurement of dry matter accumulation and mineral element content in different organs as well as the flavonoid content in the rhizome. ResultAccording to the accumulation of dry matter in different organs， the two-year growth period of B. chinensis was divided into seven stages： seedling stage （before July in the first year）， first-year vegetative growth stage （August to October in the first year）， first-year late growth stage （November to January of the second year）， re-growth stage （February to March in the second year）， second-year vegetative growth stage （April to May in the second year）， reproductive growth stage （June to October in the second year）， second-year late growth stage （November of the second year to January of the third year）. During the whole growth period， the late growth stage of each year witnessed the expansion of the rhizome， and the accumulation of dry matter of the whole plant peaked after the reproductive growth stage. The content of flavonoids in the rhizome varied greatly among different stages. Specifically， the content of mangiferin， tectorigenin and irigenin was higher in the second-year vegetative growth stage， and that of tectoridin and iridin was higher in the first-year late growth stage and the second-year late growth stage. The content of irisflorentin was higher in the second-year vegetative growth stage and the second-year late growth stage than that in other stages. The total content of six flavonoids peaked in the second-year vegetative growth stage （April in the second year）. The correlation analysis showed that all the nine mineral elements had different degrees of positive correlations with tectorigenin， irigenin and irisflorentin. ConclusionIn the first year， B. chinensis presents slow dry matter accumulation and low demand for fertilizer， so the application of base fertilizer can be reduced. In the second year， especially after the reproductive growth， B. chinensis demands high fertilizer， so nitrogen， phosphorus and potassium fertilizers and other micronutrients should be applied as early as possible. In addition， the rhizome expands in October to November of the first year and November to December of the second year， when the supply of nitrogen and potassium to the rhizome should be ensured or beneficial elements should be supplemented to improve the quality of B. chinensis.

ObjectiveTo investigate the effects of different nitrogen application rates on the growth， yield and quality of Belamcanda chinensis and to provide a theoretical basis for the quality and production efficiency improvement of B. chinensis. MethodA field experiment was carried out on the basis of 120 kg·hm^-2 phosphate and potassium fertilizers， with five nitrogen fertilizer treatments： 0 （N₀）， 60 （N₆₀）， 120 （N₁₂₀）， 180 （N₁₈₀） and 240 （N₂₄₀） kg·hm^-2. The effects of different nitrogen application rates on the agronomic traits， yield， mineral element accumulation and flavonoid content of B. chinensis were studied. ResultApplication of nitrogen fertilizer was beneficial to dry matter accumulation and significantly increased the yield of rhizome. After the application of nitrogen fertilizer， the yield of rhizome increased by 48.75%-86.62%. The nitrogen application at 60-180 kg·hm^-2 increased the active components such as tectoridin， iridin and irisflorentin， while it was not conducive to the accumulation of tectorigenin and irigenin in the rhizome. The accumulation of Mn and Fe reached the highest when the nitrogen application rate was 120 and 60 kg·hm^-2， respectively. The principal component analysis of active component content showed that the comprehensive score was ranked as N₁₈₀>N₁₂₀>N₆₀>N₂₄₀>N₀. The correlation analysis indicated that the tectoridin content had a positive correlation with N accumulation and a negative correlation with Ca accumulation in B. chinensis. The content of tectorigenin was negatively correlated with N accumulation and positively correlated with Ca accumulation. The content of irigenin had positive correlations with Ca and Zn accumulation. ConclusionThe nitrogen application at 120-180 kg·hm^-2 can increase the rhizome yield and the content of tectoridin， iridin and irisflorentin in the rhizome， so as to improve the yield and quality of this herb.

ObjectiveTo investigate the effects of different potassium application rates on the growth， yield and quality of two-year-old Belamcanda chinensis and to provide a basis for potassium application to achieve high yield and quality of B. chinensis. MethodOn the basis of nitrogen and phosphorus fertilization at 120 kg·hm^-2， we designed five potassium treatments of 0 （K₀）， 60 （K₆₀）， 120 （K₁₂₀）， 180 （K₁₈₀） and 240 （K₂₄₀） kg·hm^-2 in a field experiment to study the effects of different potassium application rates on the agronomic traits， rhizome yield， flavonoid content and potassium fertilizer utilization efficiency of the two-year-old B. chinensis. ResultThe application of potassium fertilizer promoted the root development and tillering， which significantly improved the yield of B. chinensis. Compared with the K₀ treatment， the other potassium treatments increased the rhizome yield by 16.0%-44.0% and K₁₈₀ had the highest yield. The application of potassium fertilizer increased the accumulation of flavonoids in the rhizomes of B. chinensis. The total accumulation of six flavonoids （mangiferin， tectoridin， iridin， tectorigenin， irigenin and irisflorentin） increased by 33.2% in the K₁₂₀ treatment compared with that in the K₀ treatment. Furthermore， the application of potassium fertilizer promoted the accumulation of N， K， Mg， Fe and Zn. With the increase in potassium application rate， the potassium accumulation of B. chinensis first increased and then decreased， while the partial productivity and the absorptivity of roots and leaves gradually decreased. The agronomic efficiency of potassium fertilizer was higher at low to medium potassium application rates and decreased significantly at high potassium application rates. The results of the principal component analysis indicated that the ranking of the overall quality score of B. chinensis was K₁₂₀>K₁₈₀>K₂₄₀>K₆₀>K₀. ConclusionConsidering factors such as yield， quality and potassium utilization efficiency of two-year-old B. chinensis， the potassium application at 120-166 kg·hm^-2 is recommended for the production of B. chinensis in Tuanfeng County.

ObjectiveTo investigate the effect of the combined application of organic fertilizer and chemical fertilizer at different ratios on the growth， yield and quality of two-year-old Belamcanda chinensis and provide a scientific basis for fertilization to achieve high yield and quality of this medicinal plant. MethodIn a field plot experiment， we designed five fertilization patterns with organic fertilizer and chemical fertilizer at different ratios： OM₀ （pure chemical fertilizer）， OM₁ （30% organic fertilizer + chemical fertilizer， application of organic fertilizer at 2 567 kg·hm^-2）， OM₂ （50% organic fertilizer + chemical fertilizer， application of organic fertilizer at 4 283 kg·hm^-2）， OM₃ （70% organic fertilizer + chemical fertilizer， application of organic fertilizer at 6 000 kg·hm^-2）， and OM₄ （pure organic fertilizer， 8 567 kg·hm^-2）. The effect of different fertilization patterns on the agronomic properties， herb yield， flavonoid content， mineral element content， and fertilizer utilization rate of B. chinensis were determined. ResultThe combined application of organic fertilizer and chemical fertilizer at an appropriate ratio increased the leaf width and promoted tillering， thereby improving the yield of B. chinensis. The yield of OM₁ and OM₂ groups increased by 6.65% and 13.10%， respectively， compared with that of OM₀ group， while too high a proportion of organic fertilizer led to a decrease in yield. With organic fertilizer application， the content of mangiferin， tectoridin， iridin， tectorigenin， irigenin， and irisflorentin in the rhizomes of B. chinensis increased. Moreover， a certain proportion of organic fertilizer facilitated the accumulation of functional elements such as K， Fe and Cu. The results of the principal component analysis showed that the overall quality of B. chinensis was ranked in the order of OM₃ > OM₂ > OM₁ > OM₄ > OM₀. The OM₂ group had the maximum accumulation of N， P and K and the maximum partial factor productivity of fertilizer. ConclusionAccording to the agronomic traits， yield， quality indexes and fertilizer utilization rate of B. chinensis under different fertilization patterns， we recommended to apply 4 283 kg·hm^-2 organic fertilizer in combination with chemical fertilizer to the production of two-year-old B. chinensis， so as to improve the quality and yield of B. chinensis.

ObjectiveTo investigate the effects of different phosphorus application rates on the growth， yield and quality of two-year-old Belamcanda chinensis and provide a basis for phosphorus application in the high-quality and high-yielding cultivation of B. chinensis. MethodIn this study， we designed five phosphorus application rates of 0 （P₀）， 60 （P₆₀）， 120 （P₁₂₀）， 180 （P₁₈₀） and 240 （P₂₄₀） kg·hm^-2 on the basis of 120 kg·hm^-2 nitrogen and potassium fertilizers to investigate the effects of different phosphorus levels on the agronomic traits， yield， mineral content， flavonoid content and phosphorus utilization efficiency of B. chinensis through field experiments. ResultThe plant height and tillering number of B. chinensis gradually increased after phosphorus fertilization. The phosphorus application increased the rhizome yield by 37.59%-61.03% compared with P₀ treatment and the highest yield was achieved when phosphorus fertilizer was applied at 120 kg·hm^-2. The content of mangiferin and irisflorentin in the rhizomes gradually increased with the increase in phosphorus application and the moderate application of phosphorus fertilizer increased the content of tectoridin， iridin and irigenin. The highest Mn， Cu and Fe elements were achieved at 60 and 120 kg·hm^-2 of phosphorus fertilization， respectively. The overall quality scores of B. chinensis were ranked in the order of P₂₄₀>P₁₈₀>P₁₂₀>P₆₀>P₀. With the increase in phosphorus application， the phosphorus accumulation of B. chinensis first increased and then decreased， and the partial factor productivity， apparent utilization rate and agronomic efficiency of phosphorus fertilizer gradually decreased. ConclusionConsidering the yield， quality and phosphorus utilization efficiency of B. chinensis， it is recommended to apply phosphorus fertilizer at about 120 kg·hm^-2 in the production of B. chinensis in Tuanfeng county.

Coxsackievirus B type 3 (CVB3) is one of the major causative pathogens associated with viral meningitis and myocarditis, which are widespread in the human population and especially prevalent in neonates and children. These infections can result in dilated cardiomyopathy (DCM) and other severe clinical complications. There are no vaccines or drugs approved for the prevention or therapy of CVB3-induced diseases. During screening for anti-CVB3 candidates in our previous studies, we found that jiadifenoic acids C exhibited strong antiviral activities against CVB3 as well as other strains of Coxsackie B viruses (CVBs). The present studies were carried out to evaluate the antiviral activities of jiadifenoic acids C. Results showed that jiadifenoic acids C could reduce CVB3 RNA and proteins synthesis in a dose-dependent manner. Jiadifenoic acids C also had a similar antiviral effect on the pleconaril-resistant variant of CVB3. We further examined the impact of jiadifenoic acids C on the synthesis of viral structural and non-structural proteins, finding that jiadifenoic acids C could reduce VP1 and 3D protein production. A time-course study with Vero cells showed that jiadifenoic acids C displayed significant antiviral activities at 0-6 h after CVB3 inoculation, indicating that jiadifenoic acids C functioned at an early step of CVB3 replication. However, jiadifenoic acids C had no prophylactic effect against CVB3. Taken together, we show that jiadifenoic acids C exhibit strong antiviral activities against all strains of CVB, including the pleconaril-resistant variant. Our study could provide a significant lead for anti-CVB3 drug development.

Artemisia argyi (A. argyi), a plant with a longstanding history as a raw material for traditional medicine and functional diets in Asia, has been used traditionally to bathe and soak feet for its disinfectant and itch-relieving properties. Despite its widespread use, scientific evidence validating the antifungal efficacy of A. argyi water extract (AAWE) against dermatophytes, particularly Trichophyton rubrum, Trichophyton mentagrophytes, and Microsporum gypseum, remains limited. This study aimed to substantiate the scientific basis of the folkloric use of A. argyi by evaluating the antifungal effects and the underlying molecular mechanisms of its active subfraction against dermatophytes. The results indicated that AAWE exhibited excellent antifungal effects against the three aforementioned dermatophyte species. The subfraction AAWE6, isolated using D101 macroporous resin, emerged as the most potent subfraction. The minimum inhibitory concentrations (MICs) of AAWE6 against T. rubrum, M. gypseum, and T. mentagrophytes were 312.5, 312.5, and 625 μg·mL^-1, respectively. Transmission electron microscopy (TEM) results and assays of enzymes linked to cell wall integrity and cell membrane function indicated that AAWE6 could penetrate the external protective barrier of T. rubrum, creating breaches ("small holes"), and disrupt the internal mitochondrial structure ("granary"). Furthermore, transcriptome data, quantitative real-time PCR (RT-qPCR), and biochemical assays corroborated the severe disruption of mitochondrial function, evidenced by inhibited tricarboxylic acid (TCA) cycle and energy metabolism. Additionally, chemical characterization and molecular docking analyses identified flavonoids, primarily eupatilin (131.16 ± 4.52 mg·g^-1) and jaceosidin (4.17 ± 0.18 mg·g^-1), as the active components of AAWE6. In conclusion, the subfraction AAWE6 from A. argyi exerts antifungal effects against dermatophytes by disrupting mitochondrial morphology and function. This research validates the traditional use of A. argyi and provides scientific support for its anti-dermatophytic applications, as recognized in the Chinese patent (No. ZL202111161301.9).
Antifungal Agents/chemistry* ; Arthrodermataceae ; Artemisia/chemistry* ; Molecular Docking Simulation ; Mitochondria ; Microbial Sensitivity Tests