Purification and characterization of selenium-glutathione peroxidase from hamster liver

Hamster liver glutathione peroxidase was purified to homogeneity in three chromatographic steps and with 30% yield. The purified enzyme had a specific activity of approximately 500 μmol cumene hydroperoxide reduced/min/mg of protein at 37 °C, pH 7.6, and 0.25 mm GSH. The enzyme was shown to be a tetramer of indistinguishable subunits, the molecular weight of which was approximately 23,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A single isoelectric point of 5.0 was attributed to the active enzyme. Amino acid analysis determined that selenocysteine, identified as its carboxymethyl derivative, was the only form of selenium. One residue of cysteine was found to be present in each glutathione peroxidase subunit. The presence of tryptophan was colorimetrically determined. Pseudo-first-order kinetics of inactivation of the enzyme by iodoacetate was observed at neutral pH with GSH as the only reducing agent. An optimal pH of 8.0 at 37 °C and an activation energy of 3 kcal/mol at pH 7.6 were found. A ter-uni-ping-pong mechanism was shown by the use of an integrated-rate equation. At pH 7.6, the apparent second-order rate constants for reaction of glutathione peroxidase with hydroperoxides were as follows: k₁ (t-butyl hydroperoxide), 7.06 × 10⁵ mm min^?1; k₁ (cumene hydroperoxide), 1.04 × 10⁶ mm^?1 min^?1; k₁ (p-menthane hydroperoxide), 1.2 × 10⁶ mm^?1 min^?1; k₁ (diisopropylbenzene hydroperoxide), 1.7 × 10⁶ mm^?1 min^?1; k₁ (linoleic acid hydroperoxide), 2.36 × 10⁶ mm^?1 min^?1; k₁ (ethyl hydroperoxide), 2.5 × 10⁶ mm^?1 min^?1; and k₁ (hydrogen peroxide), 2.98 × 10⁶ mm^?1 min^?1. It is concluded that for bulky hydroperoxides, the more hydrophobic the substrate, the faster its reduction by glutathione peroxidase.     

Selenium as Inducer of Glutathione Peroxidase in low-CO(2)-Grown Chlamydomonas reinhardtii

Culture of the green alga Chlamydomonas reinhardtii in the medium containing sodium selenite caused the activity of ascorbate peroxidase to disappear and the appearance of glutathione peroxidase. The induced maximum activity of glutathione peroxidase reached 350 micromole (milligram chlorophyll hour)⁻¹ under assay conditions used. The enzymic properties of the selenite-induced glutathione peroxidase closely resembled those of animal glutathione peroxidase that contains selenium.     

Role of salicylic acid in regulating ultraviolet radiation-induced oxidative stress in pepper leaves

The effect of salicylic acid (SA) counteracting the UV-A, UV-B, and UV-C-induced action on pepper (Capsicum annuum L.) plants was studied. For this purpose, the activities of antioxidant enzymes (peroxidase, polyphenol oxidase, ascorbate peroxidase, catalase, and glutathione reductase) were measured. Plants were sprayed with SA and treated with UV-A (320–390 nm), UV-B (312 nm), and UV-C (254 nm) radiation with a density of 6.1, 5.8, and 5.7 W/m². The activities of antioxidant enzymes were enhanced in leaves in response to UV-B and UV-C radiation. SA treatment moderated an increase in the activities of some antioxidant enzymes (peroxidase, ascorbate peroxidase, catalase, and glutathione reductase) in plants that were treated with UV radiation. The activity of antioxidant enzyme polyphenol oxidase in plants that were treated with UV-B, UV-C, and SA was significantly increased. The aim of the present study was to investigate the possible protective effect of SA treatment on UV-A, UV-B, and UV-C stress.     

Stand density and species richness affect carbon storage and net primary productivity in early and late successional temperate forests differently

How stand density and species richness affect carbon (C) storage and net primary productivity (NPP) changes with forest succession is poorly understood. We quantified the C storage of trees and the aboveground NPP in an early successional secondary birch forest (birch forest) and a late successional mixed broadleaf-Korean pine (Pinus koraiensis) forest (mixed forest) in northeastern China. We found that: 1) tree C storage in the mixed forest (120.3 Mg C ha^?1) was significantly higher than that in the birch forest (78.5 Mg C ha^?1), whereas the aboveground NPP was not different between the two forest types; and 2) only stand density had a positive linear relationship with tree C storage and aboveground NPP in the birch forest. In the mixed forest, both tree C storage and aboveground NPP were significantly affected by the combination of the stand density and species richness. The tree C storage to stand density and species richness relationships were hump-shaped. The aboveground NPP increased with increasing stand density, but its relationship to species richness was hump-shaped. We conclude that the effect of stand density and species richness on tree C storage and aboveground NPP was influenced by forest stand succession, and such effects should be considered in studying stand density- and species richness- ecosystem function (e.g., C storage and NPP) relationships in temperate forest ecosystems.     

Regeneration of a Coastal Pine (Pinus thunbergii Parl.) Forest 11 Years after Thinning,Niigata, Japan

To examine the effects of thinning intensity on wind vulnerability and regeneration in a coastal pine (Pinus thunbergii) forest, thinning with intensities of 20%, 30% and 50% was conducted in December 1997; there was an unthinned treatment as the control (total 8 stands). We re-measured the permanent sites to assess the regeneration characteristics 11 years after thinning. In the 50% thinned stand, seedlings aged from 2 to 10 years exhibited the highest pine seedling density and growth. The age composition ranged from 1–3 years with densities of 9.9 and 5.1 seedlings m⁻² in 30% and 20% thinned stands; only 1-year-old seedlings with a density of 6.1 seedlings m⁻² in the unthinned stand. Similar trends were found for the regeneration of broadleaved species such as Robinia pseudoacacia and Prunus serrulata. We speculate that the canopy openness and moss coverage contributed to the regeneration success in the 50% thinned stand, while the higher litter depth and lack of soil moisture induced the regeneration failure in the unthinned stand. The stands thinned at 20% or 30% were less favourable for pine regeneration than the stands thinned at 50%. Therefore, thinning with less than 30% canopy openness (20% and 30% thinned stands) should be avoided, and thinning at higher than 30% canopy openness (50% thinned stand, approximately 1500 stems ha⁻¹ at ages 40–50 years) is suggested for increasing regeneration in the coastal pine forest. The implications of thinning-based silviculture in the coastal pine forest management are also discussed. The ongoing development of the broadleaved seedlings calls for further observations.     

六盘山典型森林伴随降水的总有机碳(TOC)通量变化特征

在六盘山香水河小流域,选择6种典型森林样地,测定了2011年生长季的大气降水、穿透水、干流、枯落物渗漏水和主根系层(0—30 cm深)土壤渗漏水的总有机碳(TOC)浓度及其相应的通量变化。结果表明,在降水转化为由穿透雨和干流组成的林下降水中,所有样地的TOC浓度都不同程度地增大;虽然林冠截持使林下降水减小,但因雨水淋洗和与林冠发生碳交换,各样地林下降水携带的生长季TOC通量(kg/hm2)(华北落叶松人工林132.28、华山松次生林106.56、油松人工林94.10、灌木林79.49、桦木林66.52、辽东栎次生林63.01)都比林外降水(53.17)不同程度地明显增大,整体看来,林冠的TOC淋出作用在针叶林很大,在阔叶林较弱。在6种森林样地的枯落物层渗漏水中,其TOC浓度彼此相差不大,平均为24.51 mg/L,高于林冠穿透水的TOC浓度;受枯落物截持部分降水及与枯落物TOC交换的影响,4个样地枯落物渗漏水的TOC通量(kg/hm2)(桦木次生林84.35、野李子灌丛129.35、辽东栎次生林79.21、油松人工林114.93)都比其林下降水TOC通量增加了,但华北落叶松人工林和华山松次生林的TOC通量分别降至90.76和104.90 kg/hm2。在测定的华北落叶松人工林和华山松次生林的主根系层(0—30 cm)土壤渗漏水中,TOC浓度均低于枯落物渗漏水;由于水量减小和与土壤发生碳交换,土壤渗漏水的TOC通量均显著低于枯落物渗漏水,两个林分样地分别降至43.04和66.33 kg/hm2。整体来看,林外降水携带的TOC输入通量在林地TOC输入中占有重要地位,林冠的TOC淋洗使其程度不同地增加TOC通量,枯落物层具有增加或减少TOC通量的作用,但主根系层土壤会显著减少TOC输出通量,所以是固定TOC的重要场所。     

Specificity of an HPETE peroxidase from rat PMN

The 15,000xg supernatant of sonicated rat PMN contains 5-lipoxygenase that converts arachidonic acid to 5-hydroperoxyeicosatetraenoic acid (5-HPETE) and leukotriene A₄ and an HPETE peroxidase that catalyzes reduction of the 5-HPETE. The specificity of this HPETE peroxidase for peroxides, reducing agents, and inhibitors has been characterized to distinguish this enzyme from other peroxidase activities. In addition to 5-HPETE, the HPETE peroxidase will catalyze reduction of 15-hydroperoxyeicosatetraenoic acid, 13-hydroperoxyoctadecadienoic acid, and 15-hydroperoxy-8,11,13-eicosatrienoic acid, but not cumene or t-butylhydroperoxides. The HPETE peroxidase accepted 5 of 11 thiols tested as reducing agents. However, glutathione is >15 times more effective than any other thiol tested. Other reducing agents, ascorbate, NADH, NADPH, phenol, p-cresol, and homovanillic acid, were not accepted by HPETE peroxidase. This enzyme is not inhibited by 10 mM KCN, 2 mM aspirin, 2 mM salicylic acid, or 0.5 mM indomethacin. When 5-[¹⁴C]HPETE is generated from [¹⁴C]arachidonic acid in the presence of unlabeled 5-HPETE and the HPETE peroxidase, the 5-[¹⁴C]HETE produced is of much lower specific activity than the [¹⁴C]arachidonic acid. This indicates that the 5-[¹⁴C]HPETE leaves the active site of 5-lipoxygenase and mixes with the unlabeled 5-HPETE in solution prior to reduction and is a kinetic demonstration that 5-lipoxygenase has no peroxidase activity. Specificity for peroxides, reducing agents, and inhibitors differentiates HPETE peroxidase from glutathione peroxidase, phospholipid-hydroperoxide glutathione peroxidase, a 12-HPETE peroxidase, and heme peroxidases. The HPETE peroxidase could be a glutathione S-transferase selective for fatty acid hydroperoxides.     

In vitro synthesis of glutathione peroxidase from selenite Translational incorporation of selenocysteine

The synthesis of glutathione peroxidase from [⁷⁵Se]selenite was studied in slices and cell-free extracts from rat liver. The incorporation of [⁷⁵Se]selenocysteine at the active site was detected by carboxymethylation and hydrolysis of partially purified glutathione peroxidase (glutathione:hydrogen peroxide oxidoreductase, EC 1.11.1.9) in the presence of [³H]selenocysteine and subsequent amino acid analysis. The synthesis of glutathione peroxidase in slices was inhibited by cycloheximide or puromycin and ⁷⁵Se was incorporated from [⁷⁵Se]selenite into free selenocysteine and selenocysteyl tRNA. Increasing concentrations of selenocystine caused a progressive dilution of the ⁷⁵Se and a corresponding decrease in glutathione peroxidase labeling. In cell-free systems, [⁷⁵Se]selenocysteyl tRNA was the best substrate for glutathione peroxidase synthesis. These results indicate the existence in rat liver of the de novo synthesis of free selenocysteine and a translational pathway of selenocysteine incorporation into glutathione peroxidase     

Chloroplast superoxide and hydrogen peroxide scavenging systems from pea leaves: Seasonal variations

Levels of chloroplast antioxidants and enzymes that scavenge oxygen racidals were followed in the leaves of pea plants (Pisum sativum L. cv. Meteor) grown under glasshouse conditions between April 1984 and May 1985. While little variation in pigment levels or superoxide dismutase activity was detected during this period, plants grown in early summer (May–June) contained appreciably higher levels of ascorbate, ascorbate peroxidase and glutathione reductase than plants grown in winter (Dec–Jan.). The role of light intensity in regulating levels of chloroplast antioxidants was examined further using pea plants grown in a constant environment chamber under 100 or 400 μmol m⁻² s ⁻¹ photon flux density. Chloroplasts isolated from plants grown at the higher light intensity contained significantly higher levels of ascorbate, ascorbate peroxidase, glutathione reductase and dehydroascorbate reductase. These data suggest that light intensity may have an important influence on the level and activity of chloroplast antioxidants and oxygen radical scavenger enzymes.     

The Protective Effect of N-Acetylcysteine on Oxidative Stress in the Brain Caused by the Long-Term Intake of Aspartame by Rats

Long-term intake of aspartame at the acceptable daily dose causes oxidative stress in rodent brain mainly due to the dysregulation of glutathione (GSH) homeostasis. N-Acetylcysteine provides the cysteine that is required for the production of GSH, being effective in treating disorders associated with oxidative stress. We investigated the effects of N-acetylcysteine treatment (150 mg kg^?1, i.p.) on oxidative stress biomarkers in rat brain after chronic aspartame administration by gavage (40 mg kg^?1). N-Acetylcysteine led to a reduction in the thiobarbituric acid reactive substances, lipid hydroperoxides, and carbonyl protein levels, which were increased due to aspartame administration. N-Acetylcysteine also resulted in an elevation of superoxide dismutase, glutathione peroxidase, glutathione reductase activities, as well as non-protein thiols, and total reactive antioxidant potential levels, which were decreased after aspartame exposure. However, N-acetylcysteine was unable to reduce serum glucose levels, which were increased as a result of aspartame administration. Furthermore, catalase and glutathione S-transferase, whose activities were reduced due to aspartame treatment, remained decreased even after N-acetylcysteine exposure. In conclusion, N-acetylcysteine treatment may exert a protective effect against the oxidative damage in the brain, which was caused by the long-term consumption of the acceptable daily dose of aspartame by rats.     

Some characteristics of 75Se-P,a selenoprotein found in rat liver and plasma,and comparison of it with selenoglutathione peroxidase

The selenoenzyme glutathione peroxidase cannot account for all the physiological effects of selenium in rat liver. Therefore, a study was carried out with the ultimate aim of identifying selenoproteins other than glutathione peroxidase. The incorporation of ⁷⁵Se, given as ⁷⁵SeO₃^2?, into centrifugally separated fractions of selenium-deficient and control rat livers was determined. In selenium-deficient liver much less ⁷⁵Se was incorporated into the 105,000g supernatant fraction than in controls, so this fraction was studied further by gel filtration, ion-exchange, and hydroxylapatite chromatography. Selenoglutathione peroxidase and another selenoprotein, called ⁷⁵Se-P, were separated and identified. Both these selenoproteins were also found in plasma. Selenium deficiency had opposite effects on incorporation of ⁷⁵Se by these proteins. It decreased ⁷⁵Se incorporation by glutathione peroxidase at 3 and 72 h after ⁷⁵Se injection but increased ⁷⁵Se incorporation by ⁷⁵Se-P. This suggests that ⁷⁵Se-P competes for available selenium better than does glutathione peroxidase when the element is in short supply. Apparent molecular weights of ⁷⁵Se-P from liver and plasma determined by gel filtration were, respectively, 83,000 and 79,000, which indicate proteins smaller than glutathione peroxidase. Cycloheximide pretreatment of the rat blocked ⁷⁵Se incorporation into plasma ⁷⁵Se-P. These experiments establish the existence of a selenoprotein, ⁷⁵Se-P, in rat liver and plasma which is chromatographically distinct from glutathione peroxidase and which incorporates ⁷⁵Se differently from glutathione peroxidase. ⁷⁵Se-P may account for some of the physiological effects of selenium.     

Cd~(2+)胁迫对小桐子幼苗叶片抗氧化系统的影响

以小桐子幼苗为材料,设置不同浓度CdCl_2处理,测定Cd~(2+)胁迫对小桐子幼苗叶片中可溶性蛋白、丙二醛(MDA)含量,以及5种抗氧化酶活性和2种抗氧化剂含量的变化,探讨镉胁迫对小桐子幼苗抗氧化系统的影响。结果表明:(1)Cd~(2+)胁迫导致小桐子幼苗叶片中可溶性蛋白含量降低、MDA含量增加;(2)随着镉胁迫时间的延长,幼苗叶片中愈创木酚过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、抗坏血酸专一性过氧化酶(APX)、谷胱甘肽还原酶(GR)等抗氧化酶活性表现出先升高然后降低的变化趋势;(3)幼苗叶片中还原型抗坏血酸(ASA)和还原型谷胱甘肽(GSH)含量随着胁迫时间延长而降低,但其中氧化型抗坏血酸(DHA)和氧化型谷胱甘肽(GSSG)含量则升高。研究表明,镉胁迫初期能诱导小桐子幼苗抗氧化系统活性显著增强,提高其抗氧化能力,但随着胁迫时间的延长,致使其抗氧化酶的活性和抗氧物质含量下降,植株遭受明显氧化胁迫,幼苗生长受到镉的严重毒害。     

Lipid peroxidation, H2O2 content, and antioxidants during acclimatization of Abrus precatorius to ex vitro conditions

An efficient, rapid, and reproducible plant regeneration protocol was successfully developed for Abrus precatorius L. using mature nodal explants excised from a 5-year-old field grown plant. The highest shoot regeneration frequency (87 %) with maximum number of multiple shoots (15.0) and shoot length (4.8 cm) were recorded on Murashige and Skoog (MS) medium amended with 2.5 μM thidiazuron, 120 mg dm^?3 polyvinylpyrrolidone, and 0.5 μM α-naphthalene acetic acid. The best treatment for maximum root (4.0) induction was half strength MS medium supplemented with 1.5 μM indole-3-butyric acid. The in vitro plantlets with well-developed shoots and roots were successfully transferred into plastic cups with Soilrite and acclimatized in a culture room under photon flux density (PFD) of 150 μmol m^?2 s^?1, thereafter transferred to a greenhouse with PFD of 300 μmol m^?2 s^?1, and finally to a field with 70 % survival rate. During the acclimatization period (0–49 d), leaf chlorophyll and carotenoid content increased whereas malondialdehyde and H₂O₂ content decreased probably due to increasing activities of antioxidant enzymes (catalase, superoxide dismutase, glutathione reductase, and ascorbate peroxidase). Our work suggests that micropropagated plants developed an antioxidant enzymatic protective system to avoid oxidative stress during establishment under ex vitro environment.     

Patterns of Biomass and Carbon Distribution across a Chronosequence of Chinese Pine (Pinus tabulaeformis) Forests

Patterns of biomass and carbon (C) storage distribution across Chinese pine (Pinus tabulaeformis) natural secondary forests are poorly documented. The objectives of this study were to examine the biomass and C pools of the major ecosystem components in a replicated age sequence of P. tabulaeformis secondary forest stands in Northern China. Within each stand, biomass of above- and belowground tree, understory (shrub and herb), and forest floor were determined from plot-level investigation and destructive sampling. Allometric equations using the diameter at breast height (DBH) were developed to quantify plant biomass. C stocks in the tree and understory biomass, forest floor, and mineral soil (0–100 cm) were estimated by analyzing the C concentration of each component. The results showed that the tree biomass of P. tabulaeformis stands was ranged from 123.8 Mg·ha^–1 for the young stand to 344.8 Mg·ha^–1 for the mature stand. The understory biomass ranged from 1.8 Mg·ha^–1 in the middle-aged stand to 3.5 Mg·ha^–1 in the young stand. Forest floor biomass increased steady with stand age, ranging from 14.9 to 23.0 Mg·ha^–1. The highest mean C concentration across the chronosequence was found in tree branch while the lowest mean C concentration was found in forest floor. The observed C stock of the aboveground tree, shrub, forest floor, and mineral soil increased with increasing stand age, whereas the herb C stock showed a decreasing trend with a sigmoid pattern. The C stock of forest ecosystem in young, middle-aged, immature, and mature stands were 178.1, 236.3, 297.7, and 359.8 Mg C ha^–1, respectively, greater than those under similar aged P. tabulaeformis forests in China. These results are likely to be integrated into further forest management plans and generalized in other contexts to evaluate C stocks at the regional scale.     

Antioxidants rescue photoreceptors in rd1 mice: Relationship with thiol metabolism

We have previously shown that the use of a combination of antioxidants delayed the degeneration process in rd1 mouse retina. In an effort to understand the mechanism of action of these substances (zeaxanthin, lutein, α-lipoic acid, glutathione, and Lycium barbarum extract) the changes in the levels of several proteins and oxidative stress markers in the rd1 retina have been studied. The treatment increased glutathione peroxidase activity and glutathione levels and decreased cystine concentrations in rd1 retinas. Considering all the results obtained from treated and untreated animals, a high correlation was present between glutathione concentration and glutathione peroxidase activity, and there was a negative correlation between glutathione retinal concentration and number of TUNEL-positive cells. No difference was observed between the numbers of nNOS- and NADPH-diaphorase-positive cells in treated and untreated rd1 mice. Thiol contents and thiol-dependent peroxide metabolism seem to be directly related to the survival of photoreceptors in rd1 mouse retina.     

24-Epibrassinolide reduces stress in nematode-infected tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis> L.) plants cultured <Emphasis Type="Italic">in vitro</Emphasis>

24-Epibrassinolide (EBL) is considered the most probable brassinosteroid (BR) candidate that could be used for practical application in agriculture. EBL-induced stress-protective properties were evaluated in in vitro-grown tomato (Solanum lycopersicum L.) varieties Pusa Ruby (susceptible to nematodes) and PNR-7 (resistant to nematodes) during nematode pathogenesis. Sterilized tomato seeds treated with 10^?11, 10^?9, or 10^?7 M EBL and germinated in vitro were inoculated with second-stage juveniles of Meloidogyne incognita [(Kofoid and White) Chitwood]. Whole plant analyses of morphological and biochemical parameters 7 d after inoculation showed significant improvements in plant growth and development for both varieties and a highly significant reduction in the number of galls in the susceptible variety. Increased specific activities of antioxidative enzymes (catalase, ascorbate peroxidase, glutathione reductase, glutathione peroxidase, guaiacol peroxidase, and superoxide dismutase) were observed in EBL-treated seedlings of both varieties, but increases were higher in the resistant variety. A highly significant increase in antioxidants (ascorbic acid content, total flavonoid content, total glutathione content, and total phenolic content) was observed in EBL-treated Pusa Ruby seedlings, whereas in PNR-7, significant increases were found except for total flavonoid content, which increased non-significantly. Confocal microscopic images showed amelioration of stress in roots of EBL-treated seedlings as indicated by the decrease in level of green fluorescence in them as compared to untreated and nematode-inoculated roots.     

桂东不同林龄马尾松人工林的生物量及其分配特征

根据5a、15a、21a、32a、60a生的5个不同林龄的15块1 000m2样地(3次重复)调查资料,利用21株不同年龄和径阶的马尾松样木数据,建立以胸径(D)为单变量的生物量回归方程.采用样木回归分析法(乔木层)和样方收获法(灌木层、草本层、地上凋落物)获取不同林龄马尾松人工林的生物量,并分析了其组成、分配特征及不同林龄生物量的变化趋势.结果表明:(1)林分的总生物量随林龄而增加,5a、15a、21a、32a和60a生马尾松人工林生物量分别为15.03、125.93、183.51、191.53、405.31 Mg/hm2,其中活体植物占75.01％～94.19％,地上凋落物占0.86％～24.99％.(2)层次分配方面乔木层占绝对优势,占90.20％～98.35％,且随林龄的增加而增大,其次为地上凋落物,占0.86％～24.99％;草本层和灌木层生物量较小,分别占0.47％～34.85％和0.32％～27.00％,均随林龄的增加呈递减趋势.(3)乔木层器官分配以干所占比例最高,占49.93％～83.10％,且随林龄而增加;根相对比较稳定,占6.97％～12.82％;枝、叶分别占11.75％～14.83％、1.33％～23.65％,均随林龄增大而下降.灌木层器官分配除幼龄林为根＞枝＞叶,其余的均呈枝＞根＞叶的趋势.草本层中龄林和近熟林生物量地下＞地上,其他林龄生物量地上＞地下.(4)各林龄凋落物生物量在3.48～6.68Mg/hm2,规律性不强.(5)马尾松人工林乔木层各器官及林分生物量具有良好的优化增长模型,其32a生林分生物量高于同林龄的楠木人工林,低于热带雨林,是一种速生丰产、固碳潜力大的优良造林树种.     

Biomass Expansion Factors of Natural Japanese Red Pine (<Emphasis Type="Italic">Pinus densiflora</Emphasis>) Forests in Korea

Biomass expansion factors, which convert the timber volume (or dry weight) to biomass, are used to estimate the forest biomass and account for the carbon budget at the national or regional level. This study estimated the biomass conversion and expansion factors (BCEF), root to shoot ratio (R), biomass expansion factors (BEF) of natural Japanese Red Pine (Pinus densiflora Sieb. et Zucc.) forests based on direct field measurements and publications in Korea. This study attempted to fit the non-linear relationships between the biomass expansion factors (BCEF and BEF) and main stand factors [stand age, tree height, and diameter at breast height (DBH)]. The relationship between BEF and each main stand factor was expressed as a simple logarithmical equation. The BCEF was also expressed as a logarithmical equation of the tree height, DBH, and stand volume, whereas there was no significant relationship between BCEF and stand age. The mean value for BCEF, BEF, and R was 0.5821 Mg m⁻³ (n = 22, SD = 0.1196), 1.4465 (n = 22, SD = 0.2905), and 0.2220 (n = 17, SD = 0.0687), respectively. The values of the biomass expansion factors in this study may indicate much representativeness to estimate forest biomass in natural Japanese Red Pine forests of Korea than the default values given by the IPCC (2003, 2006).