Effect of an extraction solvent on the antioxidant quality of Pinus densiflora needle extract / 药物分析学报

Pinus densiflora needle extract (PDNE) is widely reported to have many pharmacological activities including antioxidant potential. However, the solvent system used for extraction greatly affects its antioxidant quality. Hence, in the present study, we investigated the effect of a different ratio (vol/vol) of ethanol to water (0-100％) in the extraction of PDNE with potent antioxidant capacity. The chemical assays, 2,2-diphenyl-1 picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), were conducted to assess the antioxidant potential of PDNE. Subsequently, the cytoprotective effect of PDNE was determined using tert-butyl hydroperoxide (TBHP)-challenged HepG2 cellular model. The needle extracts from 40％ ethanol (PDNE-40) showed greater radical scavenging activity followed by 60％, 20％, 80％, 0％ and 100％ ethanol extracts. EC50 value of the most active extract, PDNE-40, was 8.56 ± 0.51 μg/mL, relative to 1.34 ± 0.28 μg/mL of the standard trolox (for ABTS radical), and 75.96 ± 11.60 μg/mL, relative to 4.83 ± 0.26 μg/mL of the standard trolox (for DPPH radical). Either PDNE-20 or PDNE-40 pretreatment remarkably decreased the levels of reactive oxygen species (ROS), lipid peroxides and protein carbonyls in TBHP-challenged HepG2 cells. In addition, both PDNE-20 and PDNE-40 significantly reversed the decreased ratio of reduced (GSH) to oxidized (GSSG) glutathione. Moreover, these two extracts showed a significant inhibitory effect on TBHP-induced nuclear damage and loss of cell viability. In summary, the inclusion of 40％ ethanol in water for extraction of Pinus densiflora needle greatly increases the antioxidant quality of the extract.

Ectomycorrhizal Fungal Communities of Red Pine (Pinus densiflora) Seedlings in Disturbed Sites and Undisturbed Old Forest Sites

This study aimed to investigate differences in ectomycorrhizal (ECM) fungal communities between disturbed sites and undisturbed old forest sites. ECM root tips of Pinus densiflora were collected from 4 sites disturbed by human activities and 3 undisturbed old forest sites adjacent to the disturbed sites. Results in this study showed that the number of ECM root tips, species diversity, and number of species were significantly higher in the disturbed sites than in the undisturbed sites, suggesting that the ECM fungal community structure was affected by the degree of disturbance.

Molecular Identification of Endophytic Fungi Isolated from Needle Leaves of Conifers in Bohyeon Mountain, Korea

Fungal endophytes are microfungi that live in plants without causing apparent symptoms of infection. This study was conducted to identify endophytic fungi isolated from leaves of coniferous trees in Bohyeon Mountain of Korea. We collected leaves of two species of coniferous trees, Pinus densiflora and Pinus koraiensis, from 11 sites in the study area. A total 58 isolates were obtained and identified using molecular and morphological characteristics. Four species of endophytic fungi were isolated from P. densiflora: Lophodermium conigenum, Leotiomycetes sp., Septoria pini-thunbergii, and Polyporales sp., while two fungal species were isolated from P. koraiensis: Eurotiomycetes sp. and Rhytismataceae sp. The most frequently isolated species were L. conigenum and S. pini-thunbergii.

Effect of Soil Ameliorators on Ectomycorrhizal Fungal Communities that Colonize Seedlings of Pinus densiflora in Abandoned Coal Mine Spoils

In this study, the effect of soil ameliorators on ectomycorrhizal (ECM) fungal communities in coal mine spoils was investigated. Organic fertilizers and slaked lime were applied as soil ameliorators in 3 abandoned coal mine spoils. One year after the initial treatment, roots of Pinus densiflora seedlings were collected and the number of ECM species, colonization rate, and species diversity were assessed. The results showed that the soil ameliorators significantly increased ECM colonization on the roots of P. densiflora. The results suggest that soil ameliorators can have a positive effect on ECM fungi in terms of growth of host plants and show the potential use of soil ameliorator treatment for revegetation with ECM-colonized pine seedlings in the coal mine spoils.

Growth response of Pinus densiflora seedlings inoculated with three indigenous ectomycorrhizal fungi in combination

Pinus densiflora seedlings were inoculated with three indigenous ectomycorrhizal fungi (Cenococcum geophilum, Rhizopogon roseolus and Russula densifolia) in single-, two-, and three-species treatments. After 8 months, the colonization rates of each ectomycorrhizal species, seedling growth and the nutrition were assessed in each treatment. P. densiflora seedlings inoculated with different ECM species composition showed an increase in height and basal diameter and improved seedling root and shoot nutrition concentrations compared to control treatment. Generally, combined inoculation had a more positive influence on the seedlings than the single inoculation. The three-species inoculation presented the highest growth and basal diameter and concentration of most nutrients except potassium. In conclusion, the results provided strong evidence for benefits of combined inoculation with the indigenous ectomycorrhizal fungi on P. densiflora seedlings under controlled conditions.

Spore Dispersion of Tricholoma matsutake at a Pinus densiflora Stand in Korea

The spore of Tricholoma matsutake is considered to be the starting point of the mushroom growth cycle, but the mechanism of mycelial development from the spore stage is not yet clarified. In this study, we tried to measure how far the spores of T. matsutake disperse from a fruiting body located at a Pinus densiflora stand in Korea. We established 16 slide glasses coated with glycerin near a fruiting body in four directions separated by four different distance intervals within a mushroom productive stand after removing all other fruiting bodies from three plots. The number of dispersed spores increased with time from the first day (475 spores/cm2) to the fourth day (836 spores/cm2) after the pileus opened. The number of spores dispersed downward was about 1.5 times greater than that dispersed toward the ridge. The number of dispersed spores decreased exponentially as the distance from each fruiting body increased. More than 95% of the spores dropped within a meter from the fruiting body, with 75% dropping within 0.5 m. Even so, the number of spores dispersed over 5 m from the fruiting body was more than 50 million when considering the total number of spores produced by a fruiting body is about 5 billion.

Effects of Ectomycorrhizal Fungi on Growth of Seedlings of Pinus densiflora

This study was conducted to investigate the different effects of ectomycorrhizal fungal (ECMF) species on the growth of seedlings of Pinus densiflora, and the effects of ECMF diversity on plant productivity. A total of five species of ECMF were isolated from root tips of pine seedlings collected from Mt. Songni and used as inocula. Pots containing pine seedlings were inoculated with either a single ECMF species or a mixture of five ECMF species. All of the seedlings formed ECM on their roots except for the control plants. The pine seedlings' growth responses varied by the different ECMF species. Also, pine seedlings inoculated with a mixture of five ECMF species showed the highest growth response. The results of the study suggest that the colonization of diverse species of ECMF will increase plant productivity, and the selection of suitable ECMF species could be an important factor for plant growth.

Effects of Pine Needle Butanol Fraction on Acetylcholine (ACh) and Its Related Enzymes in Brain of Rats

This study was designed to investigate the effects of buthanol (BuOH) fraction of pine (Pinus densiflora Sieb et Zucc) needle on cholesterol and lipofuscin (LF) accumulations, acetylcholine (ACh) and its related enzyme activities such as choline acetyltransferase (CAhT) and acetylcholinesterase (AChE), and monoamone oxidase-B (MAO-B) activity, which destroyed the catecholamine-related neurotransmitters in brain membranes of Sprague-Dawley (SD) rats. Male SD rats were fed basic diets (control group) or experimental diets (BuOH-25, BuOH-50 and BuOH-100) for 45 days. Cholesterol accumulations in mitochondria and microsomes were significantly inhibited (about 14 - 17% and 23 - 34%, respectvely) in BuOH-50 and BuOH-100 groups, whereas LF levels were significantly inhibited (about 10 - 14%) in BuOH-50 and BuOH-100 groups compared with control group. ACh levels and ChAT activities were significantly increased (about 11 - 17% and 11 - 23%, respectively) in membranes of BuOH-50 and BuOH-100 groups compared with control group. AChE activities were significantly increased (about 14 - 17%) in membranes of BuOH-50 and BuOH-100 groups. There was no significant difference in MAO-B activities between control and experimental diet groups. The results suggest that butanol fraction of pine needle may play an effective role in an antiaging effect and improving a learning and memory impairments.

Effects of Pine Needle Ethyl Acetate Fraction on Acetylcholine(ACh) and Its Related Enzymes in Brain of Rats

This study was designed to investigate the effects of ethyl acetate (EtOAc) fraction of pine (Pinus densiflora Sieb et Zucc) needle on cholesterol and lipofuscin (LF) accumulations, acetylcholine (ACh) and its related enzyme activities such as choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), and monoamine oxidase-B (MAO-B) activity, which destroyed the catecholamine related neurotransmitters in brain membranes of Sprague- Dawley (SD) rats. Male SD rats were fed basic diets (control group) and experimental diets (EtOAc-25, EtOAc-50 and EtOAc-100) for 45 days. Cholesterol accumulations in mitochondria and microsomes were significantly inhibited (11.8 - 12.1% and 9.6 - 13.0%, respectvely) in EtOAc-50 and EtOAc-100 groups. ACh levels and ChAT activities were significantly increased about 10% in membranes of EtOAc-100 group compared with control group. AChE activities were significantly increased about 8 - 12% in membranes of EtOAc-50 and EtOAc-100 groups compared with control group. MAO-B activities were significantly inhibited about 10% in membrane of EtOAc-100 group compared with control group. These results suggest that ethyl acetate fraction of pine needle may play an effective role in inhibiting cholesterol and improving a membrane fluidity, and learning and memory impairments.

Fungi Colonizing Sapwood of Japanese Red Pine Logs in Storage

The Korean sawmills have recently recognized the importance of prevention of fungal discoloration due to increased losses in revenue. Before establishing integrated control strategies of fungal discoloration, more complete knowledge about causal organisms is needed. As a first step, we initiated a through survey of fungi colonizing commercially important softwood (Pinus densiflora, Pinus koraiensis, and Pinus radiata) logs and lumber in Korea. In this paper we report results obtained from Japanese red pine (Pinus densiflora) log study. In summer 2000, fungi were isolated from Japanese red pine logs in storage, and identified based on their cultural and morphological characteristics. A total of 595 fungi were isolated, representing 21 genera and 30 species. Mold fungi, mostly Trichoderma species, were the most frequently isolating fungi, representing more than half of all isolates. Dematiaceous fungi represented approximately one fifth of the isolates, and Rhinocladiella atorvirens was the most abundant in all samples. Opiostoma species represented 7% of all isolates from cores planted on malt extract agar (MEA) and the incidence of these species doubled with the addition of streptomycin and cycloheximide to MEA. The results indicate that Japanese red pine sapwood is susceptible to colonization by a variety of fungal species. As a result, control strategies that concentrate on one fungus may have limited success because of interference from competing flora.