Development of Loop-Mediated Isothermal Amplification Assays for the Rapid and Accurate Diagnosis of Exserohilum turcicum for Field Applications.

Northern corn leaf blight (NCLB), caused by Exserohilum turcicum, is one of the most devastating foliar diseases of maize. Rapid and accurate diagnosis for this disease is urgently needed but still limited. Here, we establish a field-deployable diagnostic method to detect E. turcicum based on loop-mediated isothermal amplification (LAMP) assays. A software application called K-mer Elimination by Cross-reference was used to search for the specific sequences belonging to E. turcicum by comparing the whole genome sequence between E. turcicum and other known maize pathogens. Five LAMP primer sets were designed based on specific and single-copy fragments of E. turcicum. Post-LAMP analyses indicated that only the primer set, Et9468_set1, was the most suitable, producing a ladder-like amplification pattern in the agarose gel electrophoresis and a strong fluorescence signal in the presence of SYBR Green I. The LAMP assay using Et9468_set1 primers demonstrated a high level of specificity in distinguishing E. turcicum from six other common fungal pathogens of maize, as well as 12 more fungal and oomycete strains including the epiphytic fungi from maize leaves and other crop pathogens. Moreover, it exhibited remarkable sensitivity by detecting five copies per reaction, which was approximately 104 times more sensitive compared with conventional PCR. The LAMP assay successfully detected E. turcicum in field maize leaves without DNA extraction, demonstrating its suitability for rapid on-spot detection of NCLB. Our study provides a direct LAMP diagnostic method to detect E. turcicum, which enables on-site pathogen detection in the field and the development of preventive strategies for NCLB management.

Genome Assembly and Comparative Analysis of the Egg Parasitoid Wasp Trichogramma dendrolimi Shed Light on the Composition and Evolution of Olfactory Receptors and Venoms.

Trichogramma dendrolimi is one of the most successfully industrialized Trichogramma species used to control agricultural and forestry pests in China. However, the molecular mechanisms underlying its host recognition and parasitism remain largely unknown, partially due to the limited genome information of this parasitoid wasp. Here, we present a high-quality de novo assembly of T. dendrolimi through a combination of Illumina and PacBio sequencing technologies. The final assembly had a length of 215.2 Mb and contains 316 scaffolds with a scaffold N50 size of 1.41 Mb. Repetitive sequences with a length of 63.4 Mb and 12,785 protein-coding genes were identified. Significantly expanded gene families were identified to be involved in the development and regulatory processes, while remarkably contracted gene families were involved in the transport processes in T. dendrolimi. The olfactory and venom-associated genes were identified in T. dendrolimi and 24 other hymenopteran species, using uniform methods combining BLAST and HMM profiling. The identified venom genes of T. dendrolimi were enriched in antioxidant activity, tricarboxylic acid cycle, response to oxidative stress and cell redox homeostasis. Our study provides an important resource for comparative genomics and functional studies to interpret the molecular mechanisms underlying host recognition and parasitism of Trichogramma species.

Gradually Increasing the Temperature Reduces the Diapause Termination Time of Trichogramma dendrolimi While Increasing Parasitoid Performance.

Trichogramma dendrolimi Matsumura is widely used as a biological control agent of many lepidopteran pests. Diapause has been used as an effective method to preserve the Trichogramma products during mass rearing production. However, it currently takes at least 70 days to break diapause, and we tested whether gradually increasing the temperature instead of using constant temperature could reduce the time of diapause termination and offer a higher flexibility to Trichogramma producers. The diapause termination rates of individuals kept at different conditions were measured, and five groups for which diapause termination rate reached the 95% were selected to test five biological parameters, including the number of eggs parasitized, the parasitism and emergence rates, the female sex ratio, the wing deformation rate, and the parasitoid longevity. Compared to the currently used procedure (70 days at 3 °C), treatments with at least two different temperatures resulted in higher parasitism and emergence rates while keeping the other parameters constant. The treatment that consisted of at least two different temperatures preceded by only 55 days of induction period had the highest population trend index, meaning that the population under these conditions grows more rapidly. Our results demonstrate that gradually increasing temperature allows T. dendrolimi to complete diapause earlier than at present while increasing its potential pest control capacity and providing additional flexibility in mass production of T. dendrolimi.

A new species of Hemiptarsenus Westwood (Hymenoptera, Eulophidae) from China, with a key to Chinese species.

A new species, Hemiptarsenus jilinus Tao, sp. nov., is described and illustrated. All the type specimens were reared from Chromatomyia horticola (Goureau) (Diptera: Agromyzidae), a leafminer attacking the plants Ixeris polycephala Cass. and Pterocypsela indica (L.) Shih, in Jilin Province, north-eastern China. A key to Chinese species of the genus is provided.

High-throughput profiling of diapause regulated genes from Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae).

BACKGROUND: The parasitoid wasp, Trichogramma dendrolimi, can enter diapause at the prepupal stage. Thus, diapause is an efficient preservation method during the mass production of T. dendrolimi. Previous studies on diapause have mainly focused on ecological characteristics, so the molecular basis of diapause in T. dendrolimi is unknown. We compared transcriptomes of diapause and non-diapause T. dendrolimi to identify key genes and pathways involved in diapause development. RESULTS: Transcriptome sequencing was performed on diapause prepupae, pupae after diapause, non-diapause prepupae, and pupae. Analysis yielded a total of 87,022 transcripts with an average length of 1604 bp. By removing redundant sequences and those without significant BLAST hits, a non-redundant dataset was generated, containing 7593 sequences with an average length of 3351 bp. Among them, 5702 genes were differentially expressed. The result of Gene Ontology (GO) enrichment analysis revealed that regulation of transcription, DNA-templated, oxidation-reduction process, and signal transduction were significantly affected. Ten genes were selected for validation using quantitative real-time PCR (qPCR). The changes showed the same trend as between the qPCR and RNA-Seq results. Several genes were identified as involved in diapause, including ribosomal proteins, zinc finger proteins, homeobox proteins, forkhead box proteins, UDP-glucuronosyltransferase, Glutathione-S-transferase, p53, and DNA damage-regulated gene 1 (pdrg1). Genes related to lipid metabolism were also included. CONCLUSIONS: We generated a large amount of transcriptome data from T. dendrolimi, providing a resource for future gene function research. The diapause-related genes identified help reveal the molecular mechanisms of diapause, in T. dendrolimi, and other insect species.

Remarkable impact of amino acids on ginsenoside transformation from fresh ginseng to red ginseng.

BACKGROUND: Amino acids are one of the major constituents in Panax ginseng, including neutral amino acid, acidic amino acid, and basic amino acid. However, whether these amino acids play a role in ginsenoside conversion during the steaming process has not yet been elucidated. METHODS: In the present study, to elucidate the role of amino acids in ginsenoside transformation from fresh ginseng to red ginseng, an amino acids impregnation pretreatment was applied during the steaming process at 120°C. Acidic glutamic acid and basic arginine were used for the acid impregnation treatment during the root steaming. The ginsenosides contents, pH, browning intensity, and free amino acids contents in untreated and amino acid-treated P. ginseng samples were determined. RESULTS: After 2 h of steaming, the concentration of less polar ginsenosides in glutamic acid-treated P. ginseng was significantly higher than that in untreated P. ginseng during the steaming process. However, the less polar ginsenosides in arginine-treated P. ginseng increased slightly. Meanwhile, free amino acids contents in fresh P. ginseng, glutamic acid-treated P. ginseng, and arginine-treated P. ginseng significantly decreased during steaming from 0 to 2h. The pH also decreased in P. ginseng samples at high temperatures. The pH decrease in red ginseng was closely related to the decrease in basic amino acids levels during the steaming process. CONCLUSION: Amino acids can remarkably affect the acidity of P. ginseng sample by altering the pH value. They were the main influential factors for the ginsenoside transformation. These results are useful in elucidating why and how steaming induces the structural change of ginsenoside in P. ginseng and also provides an effective and green approach to regulate the ginsenoside conversion using amino acids during the steaming process.

Identification of Serratia marcescens isolated from Antheraea pernyi eggs and determination of bacterial pathogenicity and transmission pathway.

The Chinese oak silkworm, Antheraea pernyi, is an important alternate host for mass production of Trichogramma parasitoids, which play a vital role in the biological control of lepidopterous pests in China. The quality of A. pernyi eggs is particularly important for mass production of these parasitoids. In this study, a pathogenic bacterium, isolated and purified from A. pernyi eggs that had turned gray in color, was identified as Serratia marcescens. We used morphology, biochemistry and 16S rDNA analysis to characterize the strain, which was named "APE strain". Serratia marcescens APE strain was determined to be the causal bacterium associated with the disease in the eggs, verified by a test based on Koch's Postulates. We tested the pathogenicity of S. marcescens APE strain on A. pernyi eggs; the percentage of diseased (gray) eggs reached 57.78% when uninfected eggs were exposed to a concentration of 1 × 109 cfu/mL bacterial suspension for 7 h. S. marcescens was transmitted mechanically by Trichogramma parasitoids. The transmission rate was 25.56%. In a horizontal transmission test, the highest percentage of uninfected eggs that developed infections was 51.43% after being treated with contents of diseased eggs for 12 h. In a vertical transmission test, the number of infected eggs per treated adult female was 63.8-92.3 after treatment with different S. marcescens concentrations, significantly lower than the 304.3 eggs per female in the control group. Furthermore, the percentage of infected (gray) eggs produced by adult moths exposed to bacteria in the larval stage was 80.5-85.3%.

Microbial Conversion of Protopanaxadiol-Type Ginsenosides by the Edible and Medicinal Mushroom Schizophyllum commune: A Green Biotransformation Strategy.

Previous studies have shown that many kinds of microorganisms, including bacteria, yeasts, and filamentous fungi, can convert parent ginsenosides into minor ginsenosides. However, most microorganisms used for ginsenoside transformations may not be safe for food consumption and drug development. In this study, 24 edible and medicinal mushrooms were screened by high-performance liquid chromatography analyses for their ability to microbiologically transform protopanaxadiol (PPD)-type ginsenosides. We observed that the degradation of ginsenosides by Schizophyllum commune was inhibited by high concentrations of sugar in the culture medium. However, the inhibition was avoided by maintaining sugar concentration below 15 g L-1. S. commune showed a strong ability to convert PPD-type ginsenosides (Rb1, Rc, Rb2, and Rd) into minor ginsenosides (F2, C-O, C-Y, C-Mc1, C-Mc, and C-K). The production and bioconversion rates of minor ginsenosides were significantly higher than those previously reported by food microorganisms. The fermentation process is efficient, nontoxic, eco-friendly, and economical, and the required biotransformation systems are readily available. This is the first report about the biotransformation of major ginsenosides into minor ginsenosides through fermentation by edible and medicinal mushrooms. Our results provide a green biodegradation strategy in transformation of ginsenosides using edible and medicinal mushrooms.

Selectivity of novel and traditional insecticides used for management of whiteflies on the parasitoid Encarsia formosa.

BACKGROUND: Encarsia formosa Gahan is the most important parasitoid exploited for the control of whitefly pests of vegetable crops. However, the non-target effects of recently developed insecticides for controlling whiteflies toward this biocontrol agent is little documented. Here we evaluated the susceptibility of E. formosa adults and juveniles to eight commonly used insecticides against whiteflies. RESULTS: Residual toxicity tests on glass showed that E. formosa adults were susceptible (over 98% mortality after 6 h treated) to field concentrations of the tested neonicotinoids (imidacloprid, acetamiprid, nitenpyram and thiamethoxam), abamectin, cyantraniliprole, and sulfoxaflor at their field concentrations. Pyriproxyfen was harmless to both adult and pupae of E. formosa. A risk quotient analysis showed that toxicity of pyriproxyfen to E. formosa adults was the lowest, followed by acetamiprid and cyantraniliprole. In the leaf residues test, mortality of E. formosa adults under all insecticides decreased when the residue age increased. Interestingly, E. formosa experiencing contact with the residues of most insecticides, except pyriproxyfen and abamectin, parasitized significantly more hosts 5-days after treatment, suggesting a hormetic effect on parasitism. CONCLUSION: The risk assessment of insecticides indicated that pyriproxyfen and cyantraniliprole were the least toxic to E. formosa. Our results suggest that the inclusion of the insecticides in sustainable integrated pest management programs of whiteflies should be carefully evaluated on a case-by-case basis. © 2019 Society of Chemical Industry.

Advantages of diapause in Trichogramma dendrolimi mass production on eggs of the Chinese silkworm, Antheraea pernyi.

BACKGROUND: Trichogramma dendrolimi Matsumura is widely used as a biological control agent for control of lepidopteran agricultural pests in China and can be successfully reared using the large eggs of the Chinese silkworm, Antheraea pernyi. In this study, biological parameters of diapaused, non-diapaused, and cold-stored T. dendrolimi were investigated for two generations on host eggs of A. pernyi under laboratory conditions. RESULTS: The cold-stored T. dendrolimi performed more poorly than diapaused and non-diapaused T. dendrolimi based on biological parameters. In the F1 generation, the non-diapaused T. dendrolimi had a higher emergence rate, longer longevity, and a lower proportion of deformed individuals than diapaused T. dendrolimi. In the F2 generation, the diapaused T. dendrolimi had a lower proportion of unemerged parasitoids than non-diapaused T. dendrolimi. However, the diapaused T. dendrolimi had a higher number of parasitized hosts than non-diapaused T. dendrolimi in both generations. In the F1 generation, the index of population trend (I) was found to be similar for diapaused (140.792) and non-diapaused (141.542) T. dendrolimi and was approximately 3.4 times that of cold-stored T. dendrolimi (41.698). In the F2 generation, the diapaused T. dendrolimi showed the largest I (146.791), followed by non-diapaused (136.859) and cold-stored T. dendrolimi (59.607). CONCLUSION: In a 3-year augmentative field release from 2014 to 2016, diapaused T. dendrolimi showed effective parasitism on eggs of the Asian corn borer, Ostrinia furnacalis. These results indicated that diapaused T. dendrolimi can be an efficient alternative method for mass rearing of T. dendrolimi for long-term cold storage. © 2017 Society of Chemical Industry.

Dynamic Changes in Neutral and Acidic Ginsenosides with Different Cultivation Ages and Harvest Seasons: Identification of Chemical Characteristics for Panax ginseng Quality Control.

In this study, dynamic changes in ginsenoside content and ratios in the Panax ginseng root were investigated with different cultivation ages and different collection months, using high-performance liquid chromatography (HPLC). Our data indicate that changes in ginsenoside Ro and malonyl ginsenosides content were dependent on the ginseng cultivation age (p < 0.05); especially, the Ro content varied from 0.16 to 4.91 mg/g, with a difference about 30-fold. Further, we found that the samples of 5 and 6-year-old P. ginseng had high Ro/Re ratio, whereas two and three-year-old P. ginseng possessed low Ro/Re ratio. Thus, the Ro/Re ratio can be used as a characteristic marker for differentiating the age of the root. The relative content of ginsenosides Rg1 and Re were affected by the ginseng's harvest season. The Re content was higher than the Rg1 content in May and June, but lower than the Rg1 content from August to October. Thus, the Rg1/Re ratio can be used as a characteristic marker for differentiating the ginseng's harvest seasons. These results indicate that the chemical characteristics of P. ginseng at different cultivation ages and harvest seasons are clearly different, which may cause differences in pharmacological activities and therapeutic effects. In addition, we developed HPLC coupled with hierarchical cluster analysis and principal component analysis methods to identify the cultivation age and harvest season of P. ginseng using characteristic ginsenosides. Our results showed that this method can be used to discriminate the cultivation age and harvest season of P. ginseng.

Geographic Variation of Diapause Induction Rates in Trichogramma drendrolimi (Hymenoptera: Trichogrammatidae) in China.

Diapause variation is a key factor affecting the development and success of parasitoids employed as biological control agents. Trichogramma dendrolimi Matsumura is mass-reared for biocontrol purposes in several countries around the world. Here we investigated the rate of diapause induction in three populations of T. dendrolimi from different regions in China: Heilongjiang (44° 55' N, 128° 26' E; HLJ), Liaoning (40° 18' N, 123° 22' E; LN), and Jiangsu province (32° 30' N, 120° 09' E; JS). We measured the rates of diapause induction for populations from each region when exposed to temperatures of 8, 10, 12, and 14 °C, for periods ranging from 5 to 35 d. Results showed that both the temperature and exposure duration influenced diapause induction in the three populations of T. dendrolimi. The HLJ and LN populations showed the highest percentage of diapause under 12 and 10 °C, regardless of time period tested. After 20 d at 12 and 10 °C, 97.42% and 95.94% of individuals of HLJ and LN, respectively, entered diapause. After treatment for 20-35 d under 8 °C, or 15-25 d under 10 °C, the diapause rate of LN was higher than those of the other two populations. In contrast, the higher temperatures (12-14 °C) induced diapause rapidly and steadily in HLJ. The diapause rate of JS was low for all temperatures and time periods. After 30 d at 12 °C, the JS population reached its highest diapause rate (29.56%). Our results showed significant variance in diapause rates among geographically distinct populations of T. dendrolimi in response to various abiotic conditions.

Remarkable Impact of Acidic Ginsenosides and Organic Acids on Ginsenoside Transformation from Fresh Ginseng to Red Ginseng.

Panax ginseng contains many chemical components, including acidic ginsenosides and organic acids. However, whether these acidic substances play a role in ginsenoside transformation during steaming treatment has not yet been explored. In this paper, the content of neutral ginsenosides, acidic ginsenosides, and their degradation products in unsteamed and steamed P. ginseng were simultaneously quantified by high-performance liquid chromatography. We observed that neutral ginsenosides were converted to rare ginsenosides during the root steaming but not during the individual ginsenoside steaming. In contrast, acidic malonyl ginsenosides released malonic acid and acetic acid through demalonylation, decarboxylation, deacetylation reactions during the steaming at 120 °C. These malonyl ginsenosides not only were converted to rare ginsenosides but also promoted the degradation of neutral ginsenosides. Further studies indicated that a low concentration of organic acid was the determining factor for the ginsenoside conversion. The related mechanisms were deduced to be mainly acidic hydrolysis and dehydration. In summary, acidic ginsenosides and organic acids remarkably affected ginsenoside transformation during the steaming process. Our results provide useful information for precisely understanding the ginsenoside conversion pathways and mechanisms underlying the steaming process.

Parasitism and Suitability of Fertilized and Nonfertilized Eggs of the Rice Striped Stem Borer, Chilo suppressalis (Lepidoptera: Crambidae), for Trichogramma Parasitoids.

Trichogramma spp. are egg parasitoids largely used worldwide for biological control of lepidopteran pests. Host quality (related to host size and age, host species, etc.) can influence parasitism preference and fitness of the parasitoid offspring. However, few studies have documented the performance of Trichogramma parasitoids when they parasitize nonfertilized eggs of their lepidopteran hosts. In the present study, we investigated the performance of three indigenous Trichogramma species ( Trichogramma japonicum Ashmead, Trichogramma dendrolimi Matsumura, and Trichogramma chilonis Ishii) on fertilized and nonfertilized eggs of their native host, Chilo suppressalis (Walker). The results showed that the three Trichogramma species tested could recognize fertilized and nonfertilized eggs of C. suppressalis . The different fertilization status eggs of C. suppressalis were all accepted by T. japonicum , T. chilonis , and T. dendrolimi with a clear tendency that they preferred to parasitize fertilized eggs whether under no-choice or choice conditions. All Trichogramma species successfully completed development in parasitized eggs of C. suppressalis regardless if the host eggs were fertilized or not. There was similar adult emergence rate, development time, and female progeny between fertilized and nonfertilized eggs for all Trichogramma species with an exception of development for T. chilonis . Trichogramma chilonis had shorter developmental time in fertilized eggs than in nonfertilized ones. Whether in fertilized or nonfertilized eggs, T. dendrolimi had longer development time than other two Trichogramma species. However, T. dendrolimi had the largest percentage of female progeny on fertilized eggs, followed by T. japonicum , and T. chilonis had the least percentage of female progeny. The present study indicates that Trichogramma parasitoids (i) can distinguish fertilized vs. nonfertilized host eggs, and (ii) could use nonfertilized host eggs for successful offspring development (despite showing lower preference for such eggs) [corrected].

Parasitism and Suitability of Different Egg Ages of the Leguminivora glycinivorella (Lepidoptera: Tortricidae) for Three Indigenous Trichogramma Species.

Pod borers are economically important soybean pests in temperate and tropical regions. However, the biological control of these pests using their natural insect enemies has been poorly studied to date. Indigenous natural populations of three Trichogramma (Hymenoptera: Trichogrammatidae) species, Trichogramma chilonis Ishii, Trichogramma ostriniae Pang & Chen, and Trichogramma leucaniae Pang & Chen, were collected from Leguminivora glycinivorella (Matsumura) (Lepidoptera: Tortricidae) eggs in soybean fields in China. In this study, we compare the parasitic capacities and suitabilities of three indigenous Trichogramma species on L. glycinivorella eggs at various ages. Host eggs of all ages were accepted by T. chilonis, T. ostriniae, and T. leucaniae. T. chilonis tended to parasitize 0-2-d-old eggs more than 3-4-d-old eggs. There were no significant differences in parasitism between the 0-2-d-old eggs and the 1-4-d-old eggs for T. ostriniae and T. leucaniae. For eggs at various ages, T. chilonis parasitized the smallest number of eggs, while T. leucaniae and T. ostriniae exhibited similar parasitic capacities. With 0-d-old host eggs, T. ostriniae developed over the longest time period (8.7 d), and T. leucaniae produced the most female progeny (87.9%). Both T. leucaniae and T. ostriniae had similar developmental times, survival rates and percentages of female progeny with 1-3-d-old eggs. These results show that T. leucaniae can parasitize host eggs at varying ages with the best development and suggest that it may be a valuable biological control agent for soybean pod borers.

Effects of rearing host species on the host-feeding capacity and parasitism of the whitefly parasitoid Encarsia formosa.

Parasitoids of the Encarsia genus (Hymenoptera: Aphelinidae) are important biological control agents against whiteflies. Some of the species in this genus not only parasitize their hosts, but also kill them through host feeding. The whitefly parasitoid, Encarsia formosa Gahan, was examined to determine whether the rearing host species affects its subsequent host-feeding capacity and parasitism. E. formosa wasps were reared on Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) and Bemisia tabaci (Gennadius) 'Q', and their subsequent host-feeding capacity and parasitism of T. vaporariorum and B. tabaci were examined. E. formosa reared on T. vaporariorum were significantly larger in body size than those reared on B. tabaci, but these wasps killed a similar number of whitefly nymphs by host feeding when they attacked the same host species on which they were reared. Regardless of the species on which it was reared, E. formosa fed significantly more on the B. tabaci nymphs than on the T. vaporariorum nymphs. The number of whitefly nymphs parasitized by E. formosa differed between the wasps reared on T. vaporariorum and those reared on B. tabaci depending on which whitefly species was offered as a host. In addition, the wasps reared on T. vaporariorum parasitized significantly more on T. vaporariorum than those reared on B. tabaci. The wasps reared on B. tabaci, however, parasitized similar numbers of whiteflies of both host species. The results indicated that the host-feeding capacity of E. formosa was affected more by the host species attacked than by the rearing host species, but the parasitism was affected by the host species attacked and the rearing host species. Generally, E. formosa reared on T. vaporariorum killed more T. vaporariorum nymphs by parasitism and host feeding than those reared on B. tabaci. Additionally, a similar number of B. tabaci nymphs were killed by parasitism and host feeding regardless of the rearing host species. Currently coexistence of B. tabaci and T. vaporariorum on vegetable crops usually occurs in some areas; our results may provide helpful information on using mass-reared parasitoids against mixed whitefly infestations in biological control programs.

Antidiabetic effects of malonyl ginsenosides from Panax ginseng on type 2 diabetic rats induced by high-fat diet and streptozotocin.

ETHNOPHARMACOLOGY RELEVANCE: Ginseng (Panax ginseng C. A. Meyer) has been recorded to treat 'Xiao-ke' (emaciation and thirst) symptom in many ancient Chinese medical literatures (such as 'Shen Nong Ben Cao Jing') for thousands of years. 'Xiao-ke' symptom, in general, indicates diabetes mellitus. AIM OF THE STUDY: Malonyl ginsenosides (MGR) are natural ginsenosides which exist in both fresh and air-dried ginseng. The objective of this study is to determine the antidiabetic function of MGR on type 2 diabetes. MATERIALS AND METHODS: High fat diet-fed and streptozotocin-induced diabetic rats were treated with 50 and 100mg/kg/d of MGR or vehicle for 3 weeks. The effects of MGR on fasting blood glucose (FBG), intraperitoneal glucose tolerance test (IPGTT), serum insulin (SI), insulin tolerance test (ITT), body weight, total cholesterol (TC), and triglyceride (TG) levels in type 2 diabetic rats were measured. RESULTS: After 3 weeks of treatment, MGR administration showed significantly lower FBG levels compared to the diabetic control group. In glucose tolerance test, IPGTT data showed that both MGR 50 and 100mg/kg groups significantly increased the glucose disposal after glucose load. The ITT also showed improvement of insulin sensitivity during 120 min of insulin treatment. In addition, MGR reduced TG and TC contents while showed no effect on body weight in diabetic rats. CONCLUSION: The findings from this study suggest that MGR can alleviate hyperglycemia, hyperlipemia and insulin resistance of type 2 diabetes.

The effects of dynamic changes of malonyl ginsenosides on evaluation and quality control of Panax ginseng C.A. Meyer.

To clarify the effects of malonyl ginsenosides (MGR) on evaluation and quality control of Panax ginseng, the contents of neutral and malonyl ginsenosides from P. ginseng were examined by high-performance liquid chromatography equipped with UV-VIS detector (HPLC-UV) during extraction, processing and storage. Several solvents, including water, ethanol, methanol, and n-butanol were used in the cold-soaked extraction (CSE). Among the four extraction solvents, methanol was found to be the most efficient. CSE was compared with other extraction methods such as Soxhlet extraction (SE), heat reflux extraction (HRE), ultrasonic-assisted extraction (UAE), and microwave-assisted extraction (MAE). The content of MGR showed significant differences, higher in CSE and UAE; lower in MAE and HRE; no MGR could be detected after SE. However, the total contents of neutral and malonyl ginsenosides were not different. Meanwhile, white ginseng, stored at 25°C in air of low humidity, showed a marked decrease in the concentration of MGR from 1.19% to 0.63% but with an increase in the neutral ginsenosides from 1.12% to 1.53% after 0-9-month storage. The results indicated that MGR changed dynamically in P. ginseng with different extraction solvents, extraction methods and increasing storage time. The total ginsenosides was not only underestimated but also determined imprecisely by ignoring malonyl ginsenosides. On the basis of our results, we suggest that malonyl ginsenosides should be transformed into the corresponding neutral ginsenosides during sample preparation for quality control and evaluation of P. ginseng. Then the content of six neutral ginsenosides in samples was used as the true level of total ginsenosides. The results reported here might provide useful information for accurate evaluation and quality control of P. ginseng.

Isolation and characterization of a new ginsenoside from the fresh root of Panax Ginseng.

A new saponin, malonylginsenoside Ra3, was isolated from the fresh root of Panax ginseng, along with four known ginsenosides. The new compound was identified as (20S)-protopanaxadiol-3-O-(6-O-malonyl-beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranoside-20-O-beta-D-xylopyranosyl(1-->3)-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranoside on the basis of extensive 1D and 2D NMR as well as HRESI-MS spectroscopic data analysis.

Biotransformation of ginsenoside Rf to Rh1 by recombinant beta-glucosidase.

An Aspergillus niger strain was isolated from the soil around ginseng fruit. In vitro enzyme assays showed that this strain had the ability to transform total ginsenosides (TGS) into several new products. In a further biochemical study, a beta-glucosidase gene isolated from this strain, bgl1, was expressed in Saccharomyces cerevisiae. His-tagged BGL1 protein (approximately 170 kD) showed the ability to transform ginsenoside Rf into Rh(1).