Penicillium polonicum-mediated green synthesis of silver nanoparticles: Unveiling antimicrobial and seed germination advancements.

Silver nanoparticles (AgNPs), widely recognized for their nanoscale geometric size and unique properties, such as large specific surface area, high permeability, and high safety, were synthesized using the endophytic fungus Penicillium polonicum PG21 through a green approach. Four key synthesis factors-48 h, 45 °C, pH 9.0, and 80 mM AgNPs concentration-were optimized. Characterization via ultraviolet-visible spectroscopy, transmission electron microscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction revealed the AgNPs as approximately 3-25 nm spherical particles with numerous functional groups ensuring stability. AgNPs were tested against various fungal and bacterial plant pathogens, including Botrytis cinerea (EB-1), Alternaria alternata (EB-2, EB-3), Fusarium solani (RG-1), Williamsia serinedens (SL-1), Sphingopyxis macrogoltabida (SL-2), Bacillus velezensis (SL-3), and Pseudomonas mediterranea (SL-4), causing agricultural challenges. PG21-synthesized AgNPs exhibited inhibition rates against all tested fungi, with 60 µg/mL AgNPs demonstrating optimal inhibition rates. Notably, EB-1 experienced a significant growth inhibition, reaching an inhibition rate reached of 74.22 ± 1.54%. Conversely, RG-1 exhibited the smallest inhibitory effect at 48.13 ± 0.92%. The effect of AgNPs on safflower seed germination and growth revealed notable increases in shoot length, fresh weight, stem length, and number of lateral roots-1.4, 1.4, 1.33, and 10.67 times higher than the control, respectively, at an AgNPs concentration of 80 µg/mL. In conclusion, green-synthesized AgNPs demonstrate pathogen toxicity, showcasing potential applications in disease management for industrial crops and promoting plant growth.

First Report of Fusarium falciforme Causing Root and Rhizome Rot of Epimedium sagittatum in China.

Epimedium sagittatum (Sieb.et Zucc.) Maxim., a perennial herb, is an important medicinal plant, rich in flavonoids, and widely used in the treatment of sexual dysfunction, rheumatic disease, and cancers (Tan et al. 2016). In July 2022, a disease of root and rhizome was found on E. sagittatum aged 1-8 years in a planting area (266 ha) of Zhumadian City (32°58'12" N, 114°37'48" E), Henan Province, China. The disease incidence per field (660 m2) was around 10-15% in six randomly surveyed fields planted with about 10,000 E. sagittatum plants each. Symptoms included leaf yellowing, root and rhizome browning, rotting and necrosis, and eventually the whole plant wilted and died. Fifteen plants with symptoms were sampled to isolate the pathogen. Symptomatic tissues were cut into small pieces of 5×5 mm, surface sterilized with 75% ethanol for 30 s, followed by three rinses with sterile double-distilled water (ddH2O). The pieces were then surface disinfected with 0.1% HgCl2 for 30 s, rinsed three times with sterile ddH2O, placed onto potato dextrose agar (PDA) plates and incubated at 28°C in the dark for 5 days. Twelve deferent Fusarium spp. colonies were purified by excising hyphal tip onto PDA for cultivation. Pathogenicity test of all strains was performed. Only isolate GY2 could result in root and rhizome rot of host plant. Colonies of GY2 on PDA had abundant white aerial mycelia with yellow halo. Macroconidia were hyaline, falciform, with a slightly curved apical cell and blunt basal cell, 29.7~45.0 (average 38.3) × 4.5~6.6 (average 5.3) µm (n =50), with 2-3 septa. Microconidia were oval, or reniform, hyaline, 8.4~26.5 (average 16.5) × 2.7~6.0 (average 4.5) µm (n =50), with 0-2 septa. Morphological characteristics of isolate GY2 were consistent with those of the Fusarium solani species complex (FSSC) (Chehri et al. 2015). For molecular identification, a region of the translation elongation factor 1-α (TEF) and RNA polymerase second largest subunit (RPB2) of GY2 were PCR-amplified and sequenced using the primers EF1-728F/986R (Carbone et al. 1999) and RPB2-5f2/7cr (O'Donnell et al. 2010), respectively. The TEF and RPB2 sequences (GenBank accession nos. OR978135.1 and OR978136.1) of GY2 were concatenated for a phylogenetic analysis using the Bayesian method (Zhang et al. 2020). The phylogenetic tree revealed that isolate GY2 clustered with F. falciforme with a credibility value of 99%. Morphological and molecular results support identification of isolate GY2 as F. falciforme. A pathogenicity test was performed on 4-year-old healthy plants grown in pots. Twenty healthy plants were inoculated by pouring a 200 mL conidial suspension (1×106 conidia/mL) around the rhizome. Control plants received 200 mL of sterile ddH2O. All treatments were maintained in a greenhouse at 25±1°C and 80% relative humidity. The assay was conducted three times. After 20 days, similar symptoms as those in the field were observed on the inoculated plants, whereas controls remained asymptomatic. Fusarium falciforme was reisolated from the symptomatic plants and showed the same morphological and molecular characteristics as isolate GY2, fulfilling Koch's postulates. Fusarium falciforme was reported to cause root rot of tobacco (Qiu et al. 2023) and industrial hemp (Paugh et al. 2022). However, this is the first report of F. falciforme causing root and rhizome rot of E. sagittatum. Our study will contribute to the development of strategies for the effective management of this disease on E. sagittatum.

First Report of Alternaria alternata Causing Leaf blight on Epimedium sagittatum (Sieb. et Zucc.) Maxim. in China.

Epimedium sagittatum (Sieb.et Zucc.) Maxim. is an important material of traditional Chinese medicine because of the rich content of flavonoids that are used to treat osteoporosis, liver cancer, and sexual dysfunction (Liu et al. 2013). A leaf blight was observed on E. sagittatum in Zhumadian City, China (32°58'12" N, 114°37'48" E, continental monsoon climate) in June 2021. Survey indicated that about 18% of the plants were infected in a 266-ha commercial planting area. The initial symptoms were white patches with tan borders, irregular in outline, with small black particles visible on the center of the lesions. In a week or so, patches extended throughout the leaf, and then leaves withered. Thirty leaves with symptoms collected from five different sites were cut into 5×5 mm pieces, and then surface-sterilized with 75% ethanol for 15 s followed by rinsing with double distilled water (ddH2O) three times. The pieces were then disinfested with 0.1% HgCl2 solution for 30 s, and rinsed with ddH2O, then placed onto potato-dextrose agar medium (PDA) and incubated in the dark for 3 d at 28°C. Eight fungal isolates were purified; of these, only the isolate HY2-1 infected the host plant and was selected for further morphological characterization. The colonies of HY2-1 were olive green with loose aerial hyphae on PDA. Conidiophores were single or branched, producing brown conidia in short chains. Conidia were obclavate, obpyriform, or ellipsoidal, 15.9-47.3 µm × 7.6-16.6 µm (n=50) and pale brown or dark brown with a short cylindrical beak at the tip that contained 1-5 transverse septa and 0-4 longitudinal septa. Morphological characteristics of the isolate were identical with those of Alternaria species (Huang et al. 2022). For molecular identification, the internal transcribed spacers (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Weir et al. 2012), major allergen Alt a 1(Alt a 1) and translation elongation factor 1-α gene (TEF) (Lawrence et al. 2013) were amplified and sequenced using the primers ITS4/5, GDF/GDR, Alt-F/R, and EF1-728F/986R, respectively. The results of the sequencing were uploaded to GenBank as ITS (OR418487), GAPDH (OR419792), Alt a 1 (OR419794), and TEF (OR419796), respectively. Phylogenetic analyses were performed by concatenating all the sequenced loci using the Bayesian method in Phylosuite (Zhang et al.2020). The phylogenetic tree indicated that the isolate belongs to the A. alternata clade with a bootstrap value of 75%. The pathogen was identified as A. alternata based on the morphological and molecular results. To satisfy Koch's postulates, a conidial suspension (106 conidia/mL) of the HY2-1 was prepared with ddH2O to infect the healthy plants. Ninety healthy leaves on 30 plants in pots were punctured using a sterilized needle, and then inoculated by spraying the conidial suspension on the wounded leaves in a greenhouse at 25°C and 80% relative humidity. The control plants were sprayed with ddH2O. The plants showed similar symptoms to the original infected plant 15 d after inoculation. The controls showed no symptoms. A pure culture of A. alternata was isolated and identified again as previously described. Leaf blight caused by A. alternata has been reported on Taro (Liu et al. 2020), Toona ciliata (Wang et al. 2023), etc. To our knowledge, this is the first report of E. sagittatum leaf blight caused by A. alternata in China. The results will help to develop effective control strategies for leaf blight on E. sagittatum.

First Report of Colletotrichum fructicola Causing Anthracnose on Epimedium sagittatum (Sieb. et Zucc.) Maxim. in China.

Epimedium sagittatum (Sieb.et Zucc.) Maxim., belonging to the family Berberidaceae and genus Epimedium, is a perennial herb widely studied for its anti-osteoporosis, anti-cancer, and anti-sexual-dysfunction effects in Asian countries (Tan et al. 2016; Zhang et al. 2016). High levels of bioactive chemicals in Epimedium spp. has endowed it with important clinical and commercial values (Liu et al. 2013). In September 2021, a leaf disease was found in Zhumadian City, China (32°58'12" N, 114°37'48" E). Survey statistics indicated that disease prevalence in a 266-ha planting area was approximately 29.6%. The lesions appeared at the leaf tips, gradually enlarged, and were brown with a yellow halo. Further, the lesions were dry with distributed black spots. Thirty infected leaves collected from five sites within the planting base . The collected leaves were cut into 5×5 mm pieces , surface-sterilized in 75% alcohol for 15 s, triple washed with sterile ddH2O, disinfested with 0.1% HgCl2 solution for 30 s (Liu et al. 2021), triple washed again with sterilized ddH2O, and then placed onto PDA and incubated in the dark for 3 d at 28°C. Subsequently, five fungal strains were purified; among them, only the isolate HY3-2 infected the host plant and was selected for further morphological characterization. The colonies of HY3-2 initially appeared white, their mycelia became gray at the center after 4 d, and orange-red conidial clumps appeared in them after 7 d. Conidia (10.0-19.5 µm × 4.5-5.6 µm, n=50) were single celled, nearly spherical or stick-shaped and colorless. Morphological characteristics of the isolate were consistent with those of Colletotrichum species. Additionally, glycerol-3-phosphate dehydrogenase (gapdh), actin (act), calmodulin (cal), ß-tubulin 2 (tub2), and chitin synthase-1 (chs-1), (Weir et al. 2012) were amplified and sequenced using the primers GDF/GDR, ACT-512F/783R, CL1C/CL2C, T1/Bt2b, and CHS-79F/354R, respectively for molecular identification. The resulting sequences were deposited in GenBank: gapdh (ON351609), act (ON351608), tub2 (ON351610), chs-1 (ON532788), and cal (ON532787). Phylogenetic analyses were performed by concatenating all the sequenced loci using the Bayesian method (Zhang et al. 2020). The phylogenetic tree showed that the isolate belongs to C. fructicola clade with a credibility value of 85%.To satisfy Koch's postulates, a conidial suspension (106 conidia/mL) of the isolate HY3-2 were prepared with sterile ddH2O to infect the leaves. Ninety healthy leaves from 30 plants in pots were punctured using a sterilized needle (Huang et al. 2022), and inoculated by spraying the conidial suspension on the leaves in a greenhouse at 25°C and 80% relative humidity. In the control plants, the suspension was replaced with water. After 7 d, the inoculated plants showed symptoms similar to those of the original infected plant, whereas the control showed no symptoms. C. fructicola was isolated and identified again as previously described. A pathogenicity test was also conducted in the field using the same method as that used in the greenhouse in July 2022, the results of which were consistent with those of the greenhouse. In China, C. fructicola has been reported on Walnut (Wang et al. 2022), Punica granatum (Hu et al. 2023) and others. To our knowledge, this is the first report of C. fructicola causing anthracnose in E. sagittatum in China. This report provides an important basis for further disease control research.

A Comprehensive Analysis to Elucidate the Effects of Spraying Mineral Elements on the Accumulation of Flavonoids in Epimedium sagittatum during the Harvesting Period.

The harvesting period is a critical period for the accumulation of flavonoids in the leaves of the important medicinal plant Epimedium sagittatum. In this study, we conducted an experiment on E. sagittatum leaves sprayed with mineral elements with the aim of improving the quality of the herbal leafage during the harvesting period. We elucidated the changes in flavonoids (icariin, epimedin A, epimedin B, and epimedin C) in E. sagittatum leaves. The sum of main flavonoids content reached a maximum (11.74%) at 20 days after the high-concentration Fe2+ (2500 mg·L-1) treatment. We analyzed the FT-IR spectra characteristics of E. sagittatum leaf samples using the FT-IR technique, and constructed an OPLS-DA model and identified characteristic peaks to achieve differentiated identification of E. sagittatum. Further, widely untargeted metabolomic analysis identified different classes of metabolites. As the most important characteristic flavonoids, the relative contents of icariin, icaritin, icariside I, and icariside II were found to be up-regulated by high-Fe2+ treatment. Our experimental results demonstrate that high-concentration Fe2+ treatment is an effective measure to increase the flavonoids content in E. sagittatum leaves during the harvesting period, which can provide a scientific basis for the improvement of E. sagittatum leaf cultivation agronomic measures.

Study on the Regulation of Exogenous Hormones on the Absorption of Elements and the Accumulation of Secondary Metabolites in the Medicinal Plant Artemisia argyi Leaves.

As an important medicinal plant, we still do not know the effect of exogenous hormones on absorption of elements and accumulation of secondary metabolites in Artemisia argyi leaves. In this work, we analyzed the difference in 21 elements absorbed by A. argyi leaves under three exogenous hormone (MeJA, SA and ABA) treatments, and also clarified the correlation between 21 elements and eight bioactive components. Different hormone treatments changed the absorption and enrichment of elements, and the composition also changed significantly. The contents of eight bioactive components changed significantly under different hormone treatments. When A. argyi was stimulated by exogenous hormones, the content of secondary metabolites was adjusted in the leaves through changes in the absorption and enrichment of elements. The widely untargeted metabolomic analysis further confirmed that ABA changes the metabolic direction of secondary metabolites in A. argyi leaves and stimulates the biosynthesis of multiple secondary metabolites including phenylpropanoids, flavonoids, terpenoids, alkaloids and others. These results provide a new perspective for the changes in element absorption and the mechanism of secondary metabolic components in A. argyi leaves under exogenous hormone treatments, and also deepen people's understanding of the interaction mechanism between medicinal plants and hormones.

Drought Stress Stimulates the Terpenoid Backbone and Triterpenoid Biosynthesis Pathway to Promote the Synthesis of Saikosaponin in Bupleurum chinense DC. Roots.

Bupleurum chinense is an important medicinal plant in China; however, little is known regarding how this plant transcribes and synthesizes saikosaponins under drought stress. Herein, we investigated how drought stress stimulates the transcriptional changes of B. chinense to synthesize saikosaponins. Short-term drought stress induced the accumulation of saikosaponins, especially from the first re-watering stage (RD_1 stage) to the second re-watering stage (RD_2 stage). Saikosaponin-a and saikosaponin-d increased by 84.60% and 75.13%, respectively, from the RD_1 stage to the RD_2 stage. Drought stress also stimulated a rapid increase in the levels of the hormones abscisic acid, salicylic acid, and jasmonic acid. We screened 49 Unigenes regarding the terpenoid backbone and triterpenoid biosynthesis, of which 33 differential genes were significantly up-regulated during drought stress. Moreover, one P450 and two UGTs are possibly involved in the synthesis of saikosaponins, while some transcription factors may be involved in regulating the expression of key enzyme genes. Our study provides a reference for the cultivation of B. chinense and a practical means to ensure the quality (safety and effectiveness) of B. chinense for medicinal use, as well as insights into the modernization of the China Agriculture Research System.

Genome-wide identification and characterization of the HD-Zip gene family and expression analysis in response to stress in Rehmannia glutinosa Libosch.

The HD-Zip family of transcription factors is unique to the plant kingdom, and play roles in modulation of plant growth and response to environmental stresses. R. glutinosa is an important Chinese medicinal material. Its yield and quality are susceptible to various stresses. The HD-Zip transcription factors is unique to the plant, and roles in modulation of plant growth and response to environmental stresses. However, there is no relevant research on the HD-ZIP of R. glutinosa. In this study, 92 HD-Zip transcription factors were identified in R. glutinosa, and denominated as RgHDZ1-RgHDZ92. Members of RgHDZ were classified into four groups (HD-ZipI-IV) based on the phylogenetic relationship of Arabidopsis HD-Zip proteins, and each group contains 38, 18, 17, and 19 members, respectively. Expression analyses of RgHDZ genes based on transcriptome data showed that the expression of these genes could be induced by the endophytic fungus of R. glutinosa. Additionally, we showed that RgHDZ genes were differentially expressed in response to drought, waterlogging, temperature, and salinity treatments. This study provides important information for different expression patterns of stress-responsive HD-Zip and may contribute to the better understanding of the different responses of plants to biotic and abiotic stresses, and provide a molecular basis for the cultivation of resistant varieties of R. glutinosa.

[Identification of miR160 family and their target genes in Rehmannia glutinosa in response to endophytic fungus GG22 infection].

This study aims to reveal the possible role of miR160 family in Rehmannia glutinosa in response to the infection of endophytic fungus Fusarium oxysporum GG22. Specifically, miR160 precursors and mature miR160 were retrieved from the small RNA database yielded by high-throughput sequencing. RNAfold was used to analyze the precursor structure, and DNAMAN and MEGA to analyze conservation and evolution of miR160 precursors and mature miR160. The target genes of miR160 were predicted and annotated, and the interaction was analyzed. Based on degradome sequencing, the target genes were further identified. The results showed that miR160 precursors had intact stem-loop structures. The precursor and mature sequences were conserved, particularly the 3 rd-16 th bases of the 5'-terminal. According to the phylogenetic tree, R. glutinosa had close evolutionary relationship with Arabidopsis thaliana, Oryza sativa, Salvia miltiorrhiza, and Sesamum indicum. A total of 22 target genes of miR160 were predicted and most of them were auxin response factor(ARF) genes. The target genes were involved in the Gene Ontology(GO) terms of biological processes, cellular components, and molecular functions. According to the degradome sequencing results, four target genes of miR160 were ARF(ARF18, ARF22) genes. R. glutinosa regulated its growth in response to the infection of endophytic fungus by changing the expression of miR160 and the target genes. qRT-PCR result of the differentially expressed rgl-miR160a and rgl-miR160a-3p was consistent with the sequencing result. This study clarifies the molecular mechanism of R. glutinosa in response to GG22 stress, laying a theoretical basis for the improvement and future research of R. glutinosa.

De novo genome assembly of the potent medicinal plant Rehmannia glutinosa using nanopore technology.

Rehmannia glutinosa is a potent medicinal plant with a significant importance in traditional Chinese medicine. Its root is enriched with various bioactive molecules mainly iridoids, possessing important pharmaceutical properties. However, the molecular biology and evolution of R. glutinosa have been largely unexplored. Here, we report a reference genome of R. glutinosa using Nanopore technology, Illumina and Hi-C sequencing. The assembly genome is 2.49 Gb long with a scaffold N50 length of 70 Mb and high heterozygosity (2%). Since R. glutinosa is an autotetraploid (4n = 56), the difference between each set of chromosomes is very small, and it is difficult to distinguish the two sets of chromosomes using Hi-C. Hence, only one set of the genome size was mounted to the chromosome level. Scaffolds covering 52.61% of the assembled genome were anchored on 14 pseudochromosomes. Over 67% of the genome consists of repetitive sequences dominated by Copia long terminal repeats and 48,475 protein-coding genes were predicted. Phylogenetic analysis corroborates the placement of R. glutinosa in the Orobanchaceae family. Our results indicated an independent and very recent whole genome duplication event that occurred 3.64 million year ago in the R. glutinosa lineage. Comparative genomics analysis demonstrated expansion of the UDP-dependent glycosyltransferases and terpene synthase gene families, known to be involved in terpenoid biosynthesis and diversification. Furthermore, the molecular biosynthetic pathway of iridoids has been clarified in this work. Collectively, the generated reference genome of R. glutinosa will facilitate discovery and development of important pharmacological compounds.

Isolation of endophytic bacteria from Rehmannia glutinosa Libosch and their potential to promote plant growth.

In order to study the growth promoting potential of endophytic bacteria from Rehmannia glutinosa Libosch, a total of 25 different bacteria belonging to 7 genera were identified by 16S rRNA gene sequencing, including Bacillus, Micrococcus, Lysinibacillus, Brevibacterium, Halomonas, Kocuria and Terribacillus. In this study, thirteen bacterial strains were found to solubilize inorganic phosphate, with the isolate Kocuria rosea (EH15) having the highest phosphorus dissolution activity (3.70 µg/mL). Twelve isolates were positive for nitrogen fixation abilities. Twenty-two strains produced indole-3-acetic acid (IAA) in the presence of L-tryptophan, and eleven of the twenty-two isolates synthesized IAA in the absence of L-tryptophan. The strain K. rosea (EH15) was capable of producing the highest IAA amount (15.36 and 7.98 mg/L) in Luria Bertani (LB) broth containing 0.2% L-tryptophan and lacking L-tryptophan, respectively. Ten isolates had siderophore production abilities with Bacillus amyloliquefacieus EH10 (0.26) and Brevibacterium frigoritolerans EH13 (0.32) showing high siderophore production characteristics. Five bacteria endogenous were selected to evaluate the growth parameters of Brassica napus L. and all isolates exhibited a significantly greater increase in seedling height, root length, fresh weight and dry weight, than the control plants. The greatest improvement appeared in the case of co-inoculation of EH10 and EH15, except in dry weight, and the biggest enhancement in dry weight occurred in the strain EH15. In general, these endophytic bacteria indicate a potential as microbial fertilizers to promote the growth of R. glutinosa Libosch.

[Transcriptome analysis reveals genes involved in biosynthesis of secondary metabolism in Cornus officinalis].

In order to explore genetic basis for the biosynthesis of secondary metabolism,the transcriptome of Cornus officinalis was sequenced by the new generation of high-throughput sequencing technology,A total of 96 032 unigenes were assembled with an average length of 590.53 bp. Among them, 35 478 unigenes were annotated in the public databases NR,Swissprot,COG,GO,KOG,Pfam and KEGG. Based on the assignment of KEGG pathway, 84 involved in ridoid biosynthesis and 487 unigenes involved in others secondary metabolites biosynthesis were found. Additionally,53 unigenes and 72 unigenes were predicted to have potential functions of cytochome P450 and UDP- glycosyltransferases based on the annotation result, which may encode responsible for secondary metabolites modification. This study was the first comprehensive transcriptome analysis for C. officinalis, and the candidate genes involved in the biosynthesis of secondary metabolites were obtained. The transcriptome data constitutes a much more abundant genetic resource that can be utilized to benefit further molecular biology studies on C. officinalis.

[Cloning and Expression Analysis of Acetyl-CoA C-acetyltransferase Gene in Isodon rubescens].

Objective: To clone the acetyl-CoA C-acetyl transferase( AACT) gene from Isodon rubescens, and to analyze the bioinformatics and expression of the gene. Methods: According to the IrAACT gene sequence of Isodon rubescens transcriptome,a pair of primers was designed, and the ORF of cDNA sequence was obtained by reverse transcription PCR. Bioinformatic analysis of this gene and its corresponding protein were performed. Real-time quantitative PCR( q PCR) was used to detect the relative expression levels of IrAACT different tissues of Isodon rubescens. Results: The IrAACT cDNA sequence contained a 1 254 bp open reading frame and encoded a predicted protein of 417 amino acids. IrAACT had extensive homology with AACTs from other plant species, such as Salvia miltiorrhiza, et al. Bioinformatic analysis showed that IrAACT-encoding protein contained the thiolase Ⅱ catalytic domain. q PCR analysis showed that the expression of IrAACT was tissue-specific, and accumulation of transcripts was greater in flowers and leaves, followed by stems, roots and callus. Conclusion: It is the first time to report IrAACT gene and its relative expression level. The results will provide a groundwork for studying the function of IrAACT in terpenoid biosynthesis of Isodon rubescens.

Selection for high oridonin yield in the Chinese medicinal plant Isodon (Lamiaceae) using a combined phylogenetics and population genetics approach.

Oridonin is a diterpenoid with anti-cancer activity that occurs in the Chinese medicinal plant Isodon rubescens and some related species. While the bioactivity of oridonin has been well studied, the extent of natural variation in the production of this compound is poorly known. This study characterizes natural variation in oridonin production in order to guide selection of populations of Isodon with highest oridonin yield. Different populations of I. rubescens and related species were collected in China, and their offspring were grown in a greenhouse. Samples were examined for oridonin content, genotyped using 11 microsatellites, and representatives were sequenced for three phylogenetic markers (ITS, rps16, trnL-trnF). Oridonin production was mapped on a molecular phylogeny of the genus Isodon using samples from each population as well as previously published Genbank sequences. Oridonin has been reported in 12 out of 74 species of Isodon examined for diterpenoids, and the phylogeny indicates that oridonin production has arisen at least three times in the genus. Oridonin production was surprisingly consistent between wild-collected parents and greenhouse-grown offspring, despite evidence of gene flow between oridonin-producing and non-producing populations of Isodon. Additionally, microsatellite genetic distance between individuals was significantly correlated with chemical distance in both parents and offspring. Neither heritability nor correlation with genetic distance were significant when the comparison was restricted to only populations of I. rubescens, but this result should be corroborated using additional samples. Based on these results, future screening of Isodon populations for oridonin yield should initially prioritize a broad survey of all species known to produce oridonin, rather than focusing on multiple populations of one species, such as I. rubescens. Of the samples examined here, I. rubescens or I. japonicus from Henan province would provide the best source of oridonin.

Isolation, synthesis and structures of ginsenoside derivatives and their anti-tumor bioactivity.

Protopanaxatriol saponins obtained with AB-8 macroporous resin mainly consisted of ginsenosides Rg(1) and Re. A novel mono-ester of ginsenoside-Rh(1) (ginsenoside-ORh(1)) was synthesized through further enzymatic hydrolysis and octanoyl chloride modifications. A 53% yield was obtained by a facile synthetic method. The structures were identified on the basis of 1D-NMR and 2D-NMR, as well as ESI-TOF-MS mass spectroscopic analyses. The isolated and synthetic compounds were applied in an anti-tumor bioassay, in which ginsenoside ORh(1) showed moderate effects on Murine H22 Hepatoma Cells.

[Analysis of biochemical indices and efficient ingredients in autotetraploid of Dioscorea zingibiernsis].

OBJECTIVE: To improve the yield and quality of Dioscrea zingibiernsis. METHODS: The Biochemical indices and the diosgenin's content were analysed in the autotetraploid lines. RESULTS: The results showed that the activities of APX, SOD and POD in most of autotetraploid lines were higher than that in diploid line or close to it, there was also difference between autotetraploid lines and control lines in the SDS-PAGE of soluble proteins. The content of diosgenin in most autotetraploid plantlets were higher than that in the control. CONCLUSION: There were difference between autotetraploid and control lines in the content of diosgenin and Biochemical indices, therefore, inducing autotetraploid could be an effective way to breeding the superior varieties.

[Effect of N, P, K on yield and quality of Rabdosia rubescens].

OBJECTIVE: To research the three fertilizer of N, P, K impacting Rabdosia rubescens on the yield and quality. METHODS: The optimum design of three factors with D-saturation design plan of N, P, K was adopted in the pot experiment. RESULTS: The fertilizers of N, P, K made an obvious improvement in the growth, yield and quality of Rabdosia rubescens. CONCLUSIONS: The N fertilizer plays an important role in the growth of plants. The K fertilizer makes a great impact on the accumulating of oridonin. The N, P, K fertilizers applied reasonably can make the Rabdosia rubescens high yields and improve its quality.

3,5-Dihydr-oxy-2-methyl-4H-pyran-4-one.

In the title compound, C(6)H(6)O(4), inter- and intra-molecular hydrogen bonds are observed which help to establish the crystal structure. There are weak π-π interactions between pyran rings separated by 3.5692 (9) Å.

[A survey of Cornus officinalis varieties].

According to the survey of Cornus officinalis in the main planting areas of Henan, Zhejiang and Shanxi province, the fruit shape can be divided into seven types, cylindricform, ellipticform, long pear-shape, short pear-shape, long cylindricform, short cylindricform and spindlefrom. These fruit types, growth habit and dried pulp productivity can provide scientific basis for breeding cultivation of Cornus officinalis varietis.