Based on CiteSpace Insights into Illicium verum Hook. f. Current Hotspots and Emerging Trends and China Resources Distribution.

Illicium verum Hook. f. is a globally significant spice, which is recognized in China as a food-medicine homolog and extensively utilized across the pharmaceutical, food, and spice industries. China boasts the world's leading resources of I. verum, yet its comprehensive utilization remains relatively underexplored. Through a resource survey of I. verum and the application of bibliometric visualization using CiteSpace, this study analyzed 324 papers published in the Web of Science Core Collection (WOSCC) from 1962 to 2023 and 353 core documents from China's three major databases (CNKI, Wanfang Database, and VIP Database). I. verum from Guangxi province towards various southern provinces in China, with autumn fruits exhibited superior quality and market value over their spring fruits. Literature in WOSCC emerged earlier, with a research emphasis on food science technology and pharmacology pharmacy domains. WOSCC research on I. verum could be divided into two phases: an embryonic period (1962-2001) and a growth period (2002-2023), showing an overall upward trend in publication. The three major Chinese databases contain a larger number of publications, with a focus on the food sector, which could be categorized into three stages: an embryonic period (1990-1999), a growth period (2000-2010), and a stable period (2011-2023), with an overall downward trend in publication. Both Chinese and international research hotspots converge on the medical applications of I. verum, with antioxidant bioactivity research emerging as a prevailing trend. This study delineated the resource distribution of I. verum across China and identified the research hotspots and trends both in China and internationally. The findings are beneficial for guiding researchers in swiftly establishing their research focus and furnishing decision-makers with a comprehensive reference for industry information.

Illicium verum L. (Star Anise) Essential Oil: GC/MS Profile, Molecular Docking Study, In Silico ADME Profiling, Quorum Sensing, and Biofilm-Inhibiting Effect on Foodborne Bacteria.

Illicium verum, or star anise, has many uses ranging from culinary to religious. It has been used in the food industry since ancient times. The main purpose of this study was to determine the chemical composition, antibacterial, antibiofilm, and anti-quorum sensing activities of the essential oil (EO) obtained via hydro-distillation of the aerial parts of Illicium verum. Twenty-four components were identified representing 92.55% of the analyzed essential oil. (E)-anethole (83.68%), limonene (3.19%), and α-pinene (0.71%) were the main constituents of I. verum EO. The results show that the obtained EO was effective against eight bacterial strains to different degrees. Concerning the antibiofilm activity, trans-anethole was more effective against biofilm formation than the essential oil when tested using sub-inhibitory concentrations. The results of anti-swarming activity tested against P. aeruginosa PAO1 revealed that I. verum EO possesses more potent inhibitory effects on the swarming behavior of PAO1 when compared to trans-anethole, with the percentage reaching 38% at a concentration of 100 µg/mL. The ADME profiling of the identified phytocompounds confirmed their important pharmacokinetic and drug-likeness properties. The in silico study using a molecular docking approach revealed a high binding score between the identified compounds with known target enzymes involved in antibacterial and anti-quorum sensing (QS) activities. Overall, the obtained results suggest I. verum EO to be a potentially good antimicrobial agent to prevent food contamination with foodborne pathogenic bacteria.

A Comprehensive Review of the Pharmacology, Chemistry, Traditional Uses and Quality Control of Star Anise (Illicium verum Hook. F.): An Aromatic Medicinal Plant.

Illicium verum Hook. F., also known as star anise, is one of the most important plants of the genus Anise in the family Magnoliaceae. I. verum not only has the functions of warming Yang, dispersing cold, regulating Qi and relieving pain but can also be used as a condiment to increase flavor as well as reconcile and remove fish smells. Currently, 201 chemical constituents have been identified from star anise; among these, star anise oil and shikimic acid are the two most widely used and studied chemical components in star anise, with the oil accounting for a large proportion of the total. This review integrates, classifies and updates studies related to the botany, pharmacology, phytochemistry, traditional and modern uses and quality control of star anise, with a special reference to its phytochemical composition and pharmacological activity. It will provide a reference for further research on this important medicinal plant. In addition, the broad applications and research profiles of star anise essential oil and shikimic acid are highlighted. Our review indicates that the research prospects regarding star anise are very broad and worthy of further investigation.

Safety and efficacy of a feed additive consisting of a fraction of the essential oil from the fruit and leaves of Illicium verum Hook.f. (star anise terpenes) for use in all animal species (FEFANA asbl).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of a fraction of the essential oil from the fruit and leaves of Illicium verum Hook.f. (star anise terpenes), when used as a feed additive for all animal species. The additive contains up to 25% estragole by specification and is obtained by a manufacturing process which results in the enrichment of this genotoxic carcinogen. This is not in line with the principles outlined in the general approach to assess the safety for the target species of botanical preparations which contain compounds that are genotoxic and/or carcinogenic when used as feed additives. Therefore, the FEEDAP Panel considered it was inappropriate to perform an assessment of the safety and efficacy of star anise terpenes for its use as a feed additive.

Antidiabetic Effect of Star Anise (Illicium verum) in Streptozotocin-induced Diabetic Rats.

AIM: The current study aimed to evaluate the antidiabetic potential of Illicium verum fruits. BACKGROUND: Illicium verum fruits are frequently used by the Moroccan population in the treatment of diabetes. METHODS: The antihyperglycemic effect of the aqueous extract of Illicium verum fruits (AEIVF) in rats was assessed. The effects of AEIVF (20 mg/kg) on glycemia and lipid profile as well as its phytochemical and antioxidant properties were evaluated. RESULTS: In normal and diabetic rats, AEIVF reduced blood glucose levels 6 hours after administration. Furthermore, after 7 days of treatment, glycemia was lowered in diabetic rats, and this extract exhibited an antioxidant activity. CONCLUSION: The study shows that Illicium verum possesses a potent antidiabetic activity. In addition, the toxicity of AEIVF was evaluated and the LD50 value was found to be greater than the 2 g/kg dose.

Safety and efficacy of a feed additive consisting of an essential oil derived from the fruit of Illicium verum Hook.f. (star anise oil) for use in all animal species (FEFANA asbl).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of star anise oil from the fruit (without or with the presence of plant leaves) of Illicium verum Hook.f., when used as a sensory additive in feed and water for drinking for all animal species. For long-living and reproductive animals, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) considered of low concern the use of the additive in complete feed at 0.6 mg/kg for laying hens and rabbits, 1.0 mg/kg for sows and dairy cows, 1.6 mg/kg for sheep/goats, horses and cats, 1.9 mg/kg for dogs and 6.5 mg/kg for ornamental fish. For short-living animals, the Panel had no safety concern when the additive is used at 83.3 mg/kg for veal calves, 73.3 mg/kg for sheep/goats, cattle for fattening and horses for meat production, 83.8 mg/kg for salmonids, 24.8 mg/kg for turkeys for fattening, 18.5 mg/kg chickens for fattening, 33.3 mg/kg for piglets, 40 mg/kg for pigs for fattening and 29.3 mg/kg for rabbits for meat production. These conclusions were extrapolated to other physiologically related species. For any other species, the additive was considered of low concern at 0.6 mg/kg. The use of star anise oil in animal feed is expected to be of no concern for consumers and for the environment. The additive under assessment should be considered as an irritant to skin and eyes, and as a dermal and respiratory sensitiser. Due to the high concentration of estragole (≥ 1%), the additive is classified as suspected of causing genetic defects and of causing cancer and should be handled accordingly. Since the fruit of I. verum and its preparations are recognised to flavour food and their function in feed would be the same, no further demonstration of efficacy was necessary.

Root rot Caused by Setophoma terrestris on Illicium verum in Yunnan Province, China.

Star anise (Illicium verum Hook. f.), a genus of star anise in the family Magnoliaceae, is an important cash crop of "medicinal and food" origin, mainly from China. In August 2021, root rot of I. verum was first observed on more than 80% of the plants grown within a 500 hectares area in Wenshan city, Yunnan Province. At the early stage of the disease, the phloem of the root was dark yellow-brown, and the leaves turn yellow. With further disease development, the whole root became black (Fig. 1a, 1b), and the leaves gradually fall off, affecting the growth, yield and eventually caused death of the whole plant. A total of 20 root samples were collected from typical symptomatic plant roots with 20 years old in Wenshan City (23°18'12″N, 103°56'98″E) and were cut into 2 × 2 mm pieces at the junction of infected and healthy tissue. Each sample was surface-sterilized with 3% NaClO and 75% alcohol for 60 s before rinsing three times with distilled water. The sterile filter paper (5×5 cm) was used to dry the tissue, and samples were cultured on potato dextrose agar (PDA) amended with streptomycin sulfate (50 µg/ml). Plates were incubated at 25°C in the dark in the incubator. From 9 isolates obtained in culture, 7 exhibited the morphology described by Boerema et al. (Boerema et al. 2004) for Setophoma sp. The hyphae were hyaline and septate (Fig.1c). After 14 days of culture on V8 juice agar, white round colonies are formed, but there is no groove in the middle of the colonies (Fig.1d), and transparent, oval, or cylindrical conidia were produced, 6.0-8.0 x 2.5 to 4.0 um (Fig.1e). DNA was extracted from a representative isolate BJGF-04 for molecular identification using a fungal genomic DNA extraction kit (Solarbio, Beijing, China). Polymerase chain reactions (PCRs) were performed with primers ITS1/ITS4 for the internal transcribed spacer (ITS) region (White et al. 1990) and primers T1/ß-Sandy-R for the ß-tubulin gene (TUB) region (Yang et al. 2017) and primers NL3/ LR5 for 28S large subunit rDNA (LSU) region (Hu et al. 2021) and NS1/ NS4 for 5.8S large subunit rDNA (SSU) region (Mahesha et al. 2021). Newly generated representative sequences were deposited in GenBank: ITS sequence (ON645256), TUB sequence (ON854484), and LSU sequence (ON644445), SSU sequence (ON644451). were sequenced and blasted, showing 99 to 100% sequence homology with known S. terrestris. Pathogenicity was performed using one-year asymptomatic plants of I. verum. A conidial suspension (1 x 106 conidia/ml) collected from V8 juice cultures with 0.05% Tween buffer was poured at a volume of 10 ml/plant. Three individual seedlings were used as replicates for each treatment, and sterile water was used as the negative control. All plants were placed in an artificial climate incubator at 25°C under 90% relative humidity. After 20 days, all inoculated plants showed symptoms identical to those described above, whereas controls remained healthy. Setophoma terrestris was reisolated from the infected roots, which was confirmed by morphological and molecular identification, which completed Koch's postulates. To our knowledge, this is the first report of S. terrestris as a causal agent of root rot on I. verum in China.

First report of anthracnose caused by Colletotrichum siamense on Illicium verum in China.

Aniseed (Illicium verum) is a woody spice tree that has been grown in China for a long time. Anthracnose is an important disease of aniseed, which can cause severe leaf drop. In Sep. 2020, severe anthracnose was observed in Shanglin (23°35'5"N, 108°19'51"E), Nanning, Guangxi in China, and the incidence was 85%. The symptoms at the early stage were small, round and watery, then became larger and gradually turned brown. The acervuli would appear at the later stage, and contain many conidia. Leaves with disease were randomly sampled from 10 plants, and were cut into small rectangular pieces of 0.5×1 cm, and disinfected with 75% alcohol 1 min, with 0.1% HgCl2 3 min. After washing with sterile water 3 times, they were placed onto potato dextrose agar (PDA) medium and incubated at 25°C for 5 days. The average colony growth rate was 11.85 mm/d in 7 days. The colony was white or light gray in the initial stage, with dense aerial mycelium, and the central mycelium of the colony was dark grey in the later stage. Conidia were colorless, single spore, smooth, cylindrical, both ends obtuse, with an average size of 14.95 ± 0.97 µm × 5.46 ± 0.44 µm (n = 100). The conidial appressorium was oval or club-shaped, brown, margin intact, with an average size of 7.83 ± 1.21 µm × 5.82 ± 0.58µm (n = 50). Three strains GXNN02, GXNN03 and GXNN05 were selected for further study. Primer pairs T1/ßt2b, ACT512/ACT783, GDF/GDR, CHS1-79F / CHS1-354R and ITS1/ITS4 (Weir et al. 2012) were used to amplify tubulin (TUB), actin (ACT), 3-phosphate glyceraldehyde dehydrogenase (GAPDH), chitinase (CHS1) and the internal transcribed spacers of rDNA (ITS) respectively. BLASTn searches showed that the TUB (ON619861-63 ), ACT (ON619852-54), GAPDH (ON619855-57), CHS1 (ON619858-60) and ITS (ON573028-30) sequences had the highest similarity to Colletotrichum siamense with up to 99% (699/702, 676/679, 699/702) identity for TUB (JX010404.1); 99% (281/282, 253/254, 249/250) identity for ACT (JX009518.1); 99% (275/277, 275/277, 239/241) identity for GAPDH (JX009924.1); 99% (296/299, 296/299, 259/262) identity for CHS1 (JX009865.1); up to 99% (527/530, 485/487, 527/530) identity for ITS (JX010171.1) of ex-type ICMP 18578. A ML tree was constructed by combining 5 sequenced loci, and three isolates clustered in the C. siamense clade with 94% bootstrap support. Therefore, combined with the morphological characteristics, the pathogens were identified as C. siamense. In a pathogenicity test, these three isolates were tested on 9 healthy aniseed seedlings with at least 10 leaves, and 3 seedlings as control. The leaves were surface disinfected with 75% alcohol, and then wiped with sterilized water three times. Holes were made near the edge of the leaves and were sprayed with conidial solution (6×106 spores/mL) in test groups, and use sterilized water as control. Then the leaves were sealed inside a plastic bag for 48 h to retain moisture. Brown spot and black acervuli, similar to the symptoms in the field, were observed on the leaves in test groups within 10-15 days. No symptoms were observed on the negative control leaves. The pathogens were reisolated from the treated infected leaves and were identified as C. siamense, thus fulfilling Koch's postulates. The pathogenicity test was confirmed by repeating in triplicate. The isolation frequency of C. siamense in our samples was 82.50%. To our knowledge, this is the first report of C. siamense in China. Further research on the occurrence of the disease will help prevent the spread of the disease.

Toxicological and molecular adverse effect of Illicium verum fruit constituents toward Bradysia procera.

BACKGROUND: Bradysia procera, a ginseng stem fungus gnat, is one of the most serious insect pests of Korean ginseng (Panax ginseng), causing significant damage to plant growth. The goal of this study was to determine the toxicity and mechanism of action of phenylpropanoids (trans-anethole and estragole) isolated from the methanol extract and hydrodistillate of Illicium verum fruit against third-instar larvae and eggs of Bradysia procera. RESULTS: The filter-paper mortality bioassay revealed that estragole [median lethal concentration (LC50 ) = 4.68 g/cm2 ] has a significant fumigant effect, followed by trans-anethole (LC50 = 43.92 g/cm2 ). However, estragole had the lowest toxic effect when compared to commercially available insecticides. After 7 days, estragole and trans-anethole at 75 g/cm2 inhibited egg hatchability up to 97% and 93%, respectively. At 0.09 g/cm2 , insecticides had an inhibitory effect on egg-hatching ability ranging from 88% to 94%. Furthermore, in both closed and open containers, these active constituents were able to consistently induce vapor-phased toxicity. Both estragole and trans-anethole have the ability to inhibit acetylcholinesterase (AChE), which is involved in neurotransmitter function. However, the active constituent estragole from I. verum fruit acted as a potent AChE inhibitor and had a slightly lower effect on cyclic adenosine monophosphate (AMP) than octopamine alone. CONCLUSION: This finding suggests that estragole may influence Bradysia procera neurotransmitter function via both the AChE and octopaminergic receptors. More research is needed to demonstrate the potential applications of I. verum fruit-derived products as potential larvicides and ovicides for Bradysia procera population control. © 2022 Society of Chemical Industry.

Identification of a promising inhibitor from Illicium verum (star anise) against the main protease of SARS-CoV-2: insights from the computational study.

SARS-CoV-2, the causing agent of coronavirus disease (COVID-19), first broke out in Wuhan and rapidly spread worldwide, resulting in a global health emergency. The lack of specific drugs against the coronavirus has made its spread challenging to control. The main protease (Mpro) is a key enzyme of SARS-CoV-2 used as a key target in drug discovery against the coronavirus. Medicines derived from plant phytoconstituents have been widely exploited to treat various diseases. The present study has evaluated the potential of Illicium verum (star anise) phytoconstituents against Mpro by implementing a computational approach. We performed molecular docking and molecular dynamics simulation study with a set of 60 compounds to identify their potential to inhibit the main protease (Mpro) of SARS-CoV-2. DFT study and post dynamics free energy calculations were also performed to strengthen the findings. The identified four compounds by docking study exhibited the highest potential compared to other selected phytoconstituents. Further, density functional theory (DFT) calculation, molecular dynamics simulation and post dynamics MM-GBSA energy calculation predicted Verimol-G as a potential compound, which formed stable interactions through the catalytic dyad residues. The HOMO orbital energy (-0.250038) from DFT and the post dynamics binding free energy calculation (-73.33 Kcal/mol) correlate, suggesting Verimol-G is the best inhibitor compared to the other phytoconstituents. This compound also complies with the ADME properties of drug likeliness. Thus, based on a computational study, we suggest that Verimol G may be developed as a potential inhibitor against the main protease to combat COVID-19.Communicated by Ramaswamy H. Sarma.

The Biological Activity of Illicium verum (Star Anise) on Lernaea cyprinacea-Infested Carassius auratus (Goldfish): In Vivo Study.

Lernaea cyprinacea infestation is considered a serious economic problem in the fish market. An assessment to control this parasite is needed to manage this problem. The Illicium verum oil extract has considerable antioxidant activity and scavenges 96.22% of free radicals; the high antioxidant activity refers to the phenolic content presence. The extract contains minerals, especially K, fibers, and dry matter. So, the Illicium verum ingredients were tested against this copepod for in vitro and in vivo investigation with the assessment of the treatment trial using a scanning electron microscope and evaluating the change in different immunological genes in goldfish. Female parasitic L. cyprinacea worms were blackish and hairy. The in vitro study on L. cyprinacea adults using star anise revealed that the LC50 was 12.5 and 25 µg/mL for 2 and 1 h exposure periods, respectively. Interleukin (IL-1ß) and IL-6 were grossly upregulated in C. auratus-infested skin by L. cyprinacea after treatment by 1 week, then declined after 3 weeks. In contrast, TNF-α was 18 folds upregulated in the first week after treatment, with a decline after 3 weeks. In conclusion, star anise is recommended as a safe and economical agent for controlling L. cyprinacea infestation in fish.

Safety and efficacy of a feed additive consisting of a tincture derived from the fruit of Illicium verum Hook f. (star anise tincture) for use in all animal species (FEFANA asbl).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of a tincture from the fruit (pericarps and seeds) of Illicium verum Hook f. (star anise tincture) when used as a sensory feed additive for all animal species. The product is a ■■■■■ solution, with a dry matter content of approximately 1.86%. The product contained on average 0.2588% polyphenols (of which 0.0229% were flavonoids, including 0.0036% rutin), anethole (0.018%) and estragole (0.00039%). The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the star anise tincture is safe at the maximum proposed use levels of 200 mg/kg complete feed for horses and 50 mg/kg complete feed for all other animal species. The FEEDAP Panel considered that the use in water for drinking is safe provided that the total daily intake of the additive does not exceed the daily amount which is considered safe when consumed via feed. No safety concern would arise for the consumer from the use of star anise tincture up to the maximum proposed use levels in feed. Star anise tincture should be considered as irritant to skin and eyes, and as a dermal and respiratory sensitiser. When handling the tincture, exposure of unprotected users to estragole cannot be excluded. Therefore, to reduce the risk, the exposure of the users should be minimised. The use of star anise tincture as a flavour in animal feed was not expected to pose a risk for the environment. Since the fruit of I. verum and its preparations are recognised to provide flavour in food and their function in feed would be essentially the same, no demonstration of efficacy was considered necessary.

Preparation and antibacterial and antioxidant ability of ß-cyclodextrin complexes of vaporized Illicium verum essential oil.

Compared with traditional liquid-liquid embedding method and solid-liquid embedding method of inclusion complexes of ß-cyclodextrin (ß-CD) inclusion of essential oil to form stable properties, the gas-liquid embedding method was applied to encapsulate vaporized illicium verum essential oil (IvEO), with ß-CD as wall materials so that core and wall materials molecules are in active state during complexing process. At optimal conditions with a mass ratio of 1:10, temperature of 80°C, time of 1 h, the ß-CD-IvEO inclusion complexes (ß-CD-IvEO-ICs) had an encapsulation efficiency (EE) of 84.55 ± 2.31%. Fourier transform infrared spectroscopy (FTIR) revealed the encapsulation of IvEO into inclusion complexes, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) demonstrated the enhanced thermal stability of IvEO after embedding. Furthermore, the reducing power and 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO)-scavenging capacity displayed certain capacity of antioxidation in a short time but stronger antioxidative activities as reaction time was extended. The diameter of growth zone (DGZ) indicated stronger antibacterial activity of ß-CD-IvEO-ICs against Escherichia coli, Bacillus subtilis, Staphylococcus epidermidis, and Staphylococcus aureus. Moreover, the ß-CD-IvEO-ICs could induce the bacteria producing more reactive oxygen species (ROS) than IvEO, resulting in bacterial death.

Effects of Illicium verum Hook. f. (Chinese herb) enriched diet on growth performance, immune response and disease resistance in Catla catla [Hamilton] fingerlings against Aeromonashydrophila.

Our present study investigates the effects of Illicium verum enriched diet on growth performance, immune response and disease resistance in Catla catla fingerlings against Aeromonas hydrophila. Experimental groups, i.e., infected C. catla were fed with 5 g/kg, 10 g/kg and 15 g/kg of I. verum diet for a period of 30 days showed a significant increase in their final body weight (FBW) over control. There was a significant increase (p < 0.05) in total weight gain (TWG), specific growth rate (SGR), feed conversion rate (FCR), protein efficiency ratio (PER), condition factor (CF) and survival rate (SR) in 10 g/kg and 15 g/kg groups compared to control; also from 31 to 60 days these groups showed an enhanced (p < 0.05) complement activity, phagocytic activity, respiratory burst activity and lysosome activity. The experimental group fed with 10 g/kg of I. verum diet showed the least cumulative mortality rate (5%) compared to 5 g/kg (30%) and 15 g/kg (15%) experimental groups and in control (10%) and negative control (75%). Our present study highlights that challenged fish fed with the mid dosage of 10 g/kg showed a significant increase in overall growth performance, non susceptibility to diseases and tolerance against A. hydrophila in C. catla compared to other experimental groups including the group given with high dosage of 15 g/kg. Thus our results suggest 10 g/kg as the optimal dosage of I. verum that can be used to enrich the fish diet for a better performance and effective immunostimulation in commercially important C. catla for its sustainable culture and is reported for the first time.

Combinations of plant essential oils and their major compositions inducing mortality and morphological abnormality of Aedes aegypti and Aedes albopictus.

Extensive uses of synthetic insecticides to control mosquito's populations have induced the insects to develop resistance against them, rendering them ineffective today. Moreover, they cause serious impacts on human health and the ecosystem. Therefore, safe and effective natural alternatives are needed. This study evaluated the larvicidal and pupicidal activities of essential oils (EOs) from Illicium verum and Zanthoxylum limonella and the major constituents against Aedes aegypti and Aedes albopictus mosquitoes as well as recorded their morphological aberrations at death. The GC-MS analysis showed that trans-anethole was the major constituent of I. verum EO, and limonene was the major constituent of Z. limonella EO. Both were more effective against the larvae and pupae of Ae. aegypti than those of Ae. albopictus. A 2.5% I. verum EO + 2.5% trans-anethole combination showed the highest larvicidal and pupicidal effects against Ae. aegypti and Ae. albopictus with an LT50 ranging from 0.2-6.9 h. Between the two tested constituents, trans-anethole exhibited stronger larvicidal and pupicidal activities (LC50 ranging 2.4-3.4%) against the two tested mosquito species than d-limonene (LC50 ranging 2.5-3.7%). Most importantly, 5% trans-anethole, 5% d-limonene, and 2.5% I. verum EO + 2.5% trans-anethole were more effective (LT50 ranging 0.1-0.3 h) than 1% (w/w) temephos (LT50 ranging 2.9-3.1 h). Morphological aberrations at death observed were such as color pigment and thorax shape abnormalities. To conclude, trans-anethole, d-limonene, and a combination of I. verum EO + trans-anethole, are natural compounds that not only are as effective as temephos at the time of this study, but should be also be much safer to human health.

The first report of Lasiodiplodia theobromae causing leaf spot on Illicium verum in China.

Star anise (Illicium verum) production is an important industry in parts of southern China (Wang et al. 2011). The production of star anise (cultivar Dahong) was seriously affected by a leaf spot disease during the summer of 2020 in Rong County, Guangxi province, China. Approximately 20% to 30% of the trees (n = 200, 15-16 years old) had obvious leaf spots on more than 80% of the leaves. Local growers had to cut down the seriously diseased trees. Symptomatic samples were collected and sent to us at the end of August 2021. A single brown-edged round gray spot appeared on each leaf. The spot was ~20 mm in diameter. The margin was cut into 5 mm pieces, then disinfested with 3% NaOCl for 30 s, 75% ethanol for 30 s and sterile deionized water for 1 min before dried and placed on potato dextrose agar (PDA) medium at 25°C in the dark. A total of 25 fungal isolates were obtained (isolation rate 82%). Genomic DNAs was extracted from the mycelia of these isolates and three diagnostic regions including ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999) and Bt2a/Bt2b (Glass and Donaldson 1995) were amplified. The colony morphology on PDA and the sequences of the five isolates BJ20-1, BJ20-4, BJ20-5, BJ20-7, BJ20-8 were identical. Fungal colonies had light gray mycelium and black pigment on PDA. The average colony growth rate was 4.25±0.31 cm per day and no spores were produced. Sequences of the representative isolate BJ20-1 were deposited in GenBank (Accession nos. OK483326, OL547596 and OL547597). BLASTn search indicated high identity 99.6%, 98.58% and 100% to ITS (AY640255), EF1-α (AY640258), and ß-tubulin (KU887532) of Lasiodiplodia theobromae, respectively. Combined phylogenetic analysis using MEGAX (Kumar et al. 2018) clustered BJ20-1 and L. theobromae CBS164.96 in one clade. To test pathogenicity, 2-years-old healthy I. verum trees (cultivar Dahong) maintained in a greenhouse were inoculated. Leaves were surface sterilized with 70% ethanol. One PDA plug (5mm in diameter) was placed on each wound acupunctured with a sterile needle pick. Ten PDA plugs with mycelial of BJ20-1 growing 7 days on it were inoculated on five leaves. Four sterile PDA plugs placed on two leaves served as controls. All the plugs were removed from the leaves the day after inoculation. The experiment was repeated three times. At 1 day post-inoculation (DPI), brown expanding lesions were observed on the inoculated leaves. At 7 DPI, a mature ellipse of necrosis formed with 18±4 mm in diameter with black pycnidia in the center. Conidia were observed in the pycnidia. The immature conidia were thick-walled, hyaline, aseptate and ellipsoid, measuring 20 - 25.2 × 11 - 13 µm (n = 25). The mature conidia were dark brown with one central septum, measuring 24.3 - 27 × 13 - 14 µm (n = 25). At 10 DPI, the control leaves remained asymptomatic. Re-isolation was successful from the spot on the inoculated leaves. The colony morphology and molecular identification of the re-isolations were all the same as that of BJ20-1. In conclusion, the morphological and molecular evidence consistently indicated these isolates were L. theobromae. Koch's postulates were fulfilled that L. theobromae was pathogenic on star anise. Although L. theobromae has been reported to cause leaf spot disease on Camellia sinensis (Bao et al, 2021), Kadsura longipedunculata (Fan et al, 2020) and Broussonetia papyrifera (Luo et al, 2020), etc., this is the first report of L. theobromae causing leaf spot on I. verum in China. Due to the leaf spot disease resulting in serious yield reduction on star anise, accurate pathogen identification in this study would significantly improve the control of the leaf spot disease on star anise.

Co-Treatment with the Herbal Medicine SIP3 and Donepezil Improves Memory and Depression in the Mouse Model of Alzheimer's Disease.

BACKGROUND: Alzheimer's disease (AD) is a lethal, progressive neurodegenerative disorder that has been linked to a deficiency of the neurotransmitter acetylcholine. Currently, many acetylcholinesterase inhibitors, such as donepezil, are widely used for the treatment of AD. On the other hand, the efficacy of long-term donepezil use is limited. SIP3, a mixture of three herbal extracts from Santalum album, Illicium verum, and Polygala tenuifolia, is a new formula derived from traditional Korean herbal medicine. OBJECTIVE: We assessed the synergistic effect of SIP3 and donepezil co-treatment on symptoms of AD using APP/PS1 transgenic mice. METHODS: In this study, a Drosophila AD model and SH-SY5Y clles were used to assess the toxicity of SIP3, and APPswe/PS1dE9 (APP/PS1) transgenic mice were used to evaluate the cognitive-behavioral and depression-like behavior effect of SIP3 and donepezil co-treatment on symptoms of AD. The cerebral cortex or hippocampus transcriptomes were analyzed by RNA sequencing and miRNA to investigate the molecular and cellular mechanisms underlying the positive effects of SIP3 on AD. RESULTS: In the passive avoidance test (PAT) and Morris water maze (MWM) test, the combination of SIP3 and donepezil improved the learning capabilities and memory of APP/PS1 mice in the mid-stage of AD compared to the group treated with donepezil only. In addition, co-administration of SIP3 and donepezil effectively reduced the depression-like behavior in the forced swimming and tail suspension tests. Furthermore, RNA sequencing of the cerebral cortex transcriptome and miRNA of the hippocampus showed that the gene expression profiles after a low dose SIP3 co-treatment were more similar to those of the normal phenotype mice than those obtained after the donepezil treatment alone. The Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, showed that differentially expressed genes were involved in the locomotor behavior and neuroactive ligand-receptor interactions. These results suggest that a co-treatment of low dose SIP3 and donepezil improves impaired learning, memory, and depression in the mid-stage of AD in mice. CONCLUSION: Co-treatment of low dose SIP3 and donepezil improves impaired learning, memory, and depression in the mid-stage of AD in mice.

Anti-glycation properties of Illicium verum Hook. f. fruit in-vitro and in a diabetic rat model.

BACKGROUND: Chronic hyperglycemic triggers the non-enzymatic glycation of biomolecules, resulting in the production of advanced glycation endproducts, that lead to several micro- and macrovascular complications. Therefore, the discovery of new, effective, and safe anti-glycation agents is an important need. One of the best choices for the management of diabetes is to use complementary and alternative medicinal therapies. Therefore, the present study was designed to evaluate the anti-glycation activity of ethanolic extract of Illicium verum Hook. f. (Star anise, a frequently used spice and medicinally important herb). METHODS: The anti-glycation activity of ethanolic extract of Illicium verum Hook. f. was determined by using both in-vitro and in-vivo assays. HSA-fructose glycation model was employed to assess the in-vitro inhibition of protein glycation, additionally cross-linked AGEs (formed by incubating lysozyme with fructose) were assessed by SDS polyacrylamide gel electrophoresis. Dual inhibitory mechanisms, i.e., antioxidant and metal chelating activities, were also evaluated by using DPPH, ABTS, and Fe (II)-chelation assays. Acute toxicity of I. verum extract was also performed (by administrating different doses i.e. 2,000, 1,500, 1,000, and 500 mg/kg of body weight). Finally, in-vivo anti-glycation potential was evaluated by 7 weeks of administration of I. verum extract in streptozotocin-induced diabetic rats. RESULTS: In HSA-fructose glycation model, extract of I. verum showed a good inhibitory activity with IC50 value of 0.11±0.001 mg/mL, as compared to the standard inhibitor, rutin (IC50 = 0.02±0.01 mg/mL). Extract of I. verum showed inhibitory activity in DPPH, and ABTS radical scavenging assays with IC50 values of 130±1.0, and 57±2.0 µg/mL, respectively, while it was found to be inactive in the Fe+2-chelation assay. The extract was found to be non-toxic, and reduce the elevated blood glucose, urea, lipid, liver function parameters, and renal AGEs levels in streptozotocin-induced diabetic rats. CONCLUSIONS: These results suggest that I. verum supplementation might help to reduce the burden of AGEs, and may have potential in preventing diabetes-associated complications.

Antiviral and Antioxidant Components from the Fruits of Illicium verum Hook.f. (Chinese Star Anise).

Illicium verum Hook.f. (Chinese star anise), a known Chinese traditional spice, is commonly applied in Chinese cuisine and cooking in Southeast Asia. As a kind of medicinal and edible resource, the fruit of I. verum has attracted great attention for its chemical constituents and physiological activities. In this work, the phytochemical study of the fruits of I. verum led to the isolation and identification of 20 compounds, including 6 new lignans and phenylpropanoids (1-6) and 14 known ones (7-20). Their structures were characterized by extensive analysis of spectroscopic data (IR, UV, high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), one-dimensional (1D) and two-dimensional (1D) NMR), electronic circular dichroism (ECD) calculation, and by comparison with literature data. Meanwhile, all compounds (1-20) were evaluated for their antiviral and antioxidant activities. Especially, compound 7 [(-)-bornyl p-coumarate] showed strong antiviral activities against influenza virus A/Puerto Rico/8/34 H1N1 (PR8) with an IC50 value of 1.74 ± 0.47 µM, which is much better than those of Tamiflu (IC50 = 10.01 ± 0.92 µM) and ribavirin (IC50 = 10.76 ± 1.60 µM). The antiviral activity against PR8 of compound 7 was reported for the first time, which was sufficiently confirmed by cell counting kit 8 (CCK-8), cytopathic effect (CPE) reduction, and immunofluorescence assays. In this study, the discovery of antiviral and antioxidant components from the fruits of I. verum could benefit the further development and utilization of this plant.