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Rhizosphere Inhibition of Cucumber Fusarium Wilt by Different Surfactinexcreting Strains of Bacillus subtilis

  • Jia, Ke;Gao, Yu-Han;Huang, Xiao-Qin;Guo, Rong-Jun;Li, Shi-Dong
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.140-151
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    • 2015
  • Bacillus subtilis B006 strain effectively suppresses the cucumber fusarium wilt caused by Fusarium oxysporum f. sp. cucumerinum (Foc). The population dynamics of Foc, strain B006 and its surfactin over-producing mutant B841 and surfactin-deficient mutant B1020, in the rhizosphere were determined under greenhouse conditions to elucidate the importance of the lipopeptides excreted by these strains in suppressing Foc. Results showed that B. subtilis strain B006 effectively suppressed the disease in natural soil by 42.9%, five weeks after transplanting, whereas B841 and B1020 suppressed the disease by only 22.6% and 7.1%, respectively. Quantitative PCR assays showed that effective colonization of strain B006 in the rhizosphere suppressed Foc propagation by more than 10 times both in nursery substrate and in field-infected soil. Reduction of Foc population at the cucumber stems in a range of $0.96log_{10}ng/g$ to $2.39log_{10}ng/g$ was attained at the third and the fifth weeks of B006 treatment in nursery substrate. In field-infected soil, all three treatments with B. subtilis suppressed Foc infection, indicated by the reduction of Foc population at a range of $2.91log_{10}ng/g$ to $3.36log_{10}ng/g$ at the stem base, one week after transplanting. This study reveals that the suppression of fusarium wilt disease is affected by the effective colonization of the surfactin-producing B. subtilis strain in the rhizosphere. These results improved our understanding of the biocontrol mechanism of the B. subtilis strain B006 in the natural soil and facilitate its application as biocontrol agent in the field.

Biosynthesis of rare 20(R)-protopanaxadiol/protopanaxatriol type ginsenosides through Escherichia coli engineered with uridine diphosphate glycosyltransferase genes

  • Yu, Lu;Chen, Yuan;Shi, Jie;Wang, Rufeng;Yang, Yingbo;Yang, Li;Zhao, Shujuan;Wang, Zhengtao
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.116-124
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    • 2019
  • Background: Ginsenosides are known as the principal pharmacological active constituents in Panax medicinal plants such as Asian ginseng, American ginseng, and Notoginseng. Some ginsenosides, especially the 20(R) isomers, are found in trace amounts in natural sources and are difficult to chemically synthesize. The present study provides an approach to produce such trace ginsenosides applying biotransformation through Escherichia coli modified with relevant genes. Methods: Seven uridine diphosphate glycosyltransferase (UGT) genes originating from Panax notoginseng, Medicago sativa, and Bacillus subtilis were synthesized or cloned and constructed into pETM6, an ePathBrick vector, which were then introduced into E. coli BL21star (DE3) separately. 20(R)-Protopanaxadiol (PPD), 20(R)-protopanaxatriol (PPT), and 20(R)-type ginsenosides were used as substrates for biotransformation with recombinant E. coli modified with those UGT genes. Results: E. coli engineered with $GT95^{syn}$ selectively transfers a glucose moiety to the C20 hydroxyl of 20(R)-PPD and 20(R)-PPT to produce 20(R)-CK and 20(R)-F1, respectively. GTK1- and GTC1-modified E. coli glycosylated the C3-OH of 20(R)-PPD to form 20(R)-Rh2. Moreover, E. coli containing $p2GT95^{syn}K1$, a recreated two-step glycosylation pathway via the ePathBrich, implemented the successive glycosylation at C20-OH and C3-OH of 20(R)-PPD and yielded 20(R)-F2 in the biotransformation broth. Conclusion: This study demonstrates that rare 20(R)-ginsenosides can be produced through E. coli engineered with UTG genes.

Phenolic acids in Panax ginseng inhibit melanin production through bidirectional regulation of melanin synthase transcription via different signaling pathways

  • Jianzeng Liu ;Xiaohao Xu ;Jingyuan Zhou;Guang Sun ;Zhenzhuo Li;Lu Zhai ;Jing Wang ;Rui Ma ;Daqing Zhao;Rui Jiang ;Liwei Sun
    • Journal of Ginseng Research
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    • v.47 no.6
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    • pp.714-725
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    • 2023
  • Background: Our previous investigation indicated that the preparation of Panax ginseng Meyer (P. ginseng) inhibited melanogenesis. It comprised salicylic acid (SA), protocatechuic acid (PA), p-coumaric acid (p-CA), vanillic acid (VA), and caffeic acid (CA). In this investigation, the regulatory effects of P. ginseng phenolic acid monomers on melanin production were assessed. Methods: In vitro and in vivo impact of phenolic acid monomers were assessed. Results: SA, PA, p-CA and VA inhibited tyrosinase (TYR) to reduce melanin production, whereas CA had the opposite effects. SA, PA, p-CA and VA significantly downregulated the melanocortin 1 receptor (MC1R), cycle AMP (cAMP), protein kinase A (PKA), cycle AMP-response element-binding protein (CREB), microphthalmia-associated transcription factor (MITF) pathway, reducing mRNA and protein levels of TYR, tyrosinase-related protein 1 (TYRP1), and TYRP2. Moreover, CA treatment enhanced the cAMP, PKA, and CREB pathways to promote MITF mRNA level and phosphorylation. It also alleviated MITF protein level in α-MSH-stimulated B16F10 cells, comparable to untreated B16F10, increasing the expression of phosphorylation glycogen synthase kinase 3β (p-GSK3β), β-catenin, p-ERK/ERK, and p-p38/p38. Furthermore, the GSK3β inhibitor promoted p-GSK3β and p-MITF expression, as observed in CA-treated cells. Moreover, p38 and ERK inhibitors inhibited CA-stimulated p-p38/p38, p-ERK/ERK, and p-MITF increase, which had negative binding energies with MC1R, as depicted by molecular docking. Conclusion: P. ginseng roots' phenolic acid monomers can safely inhibit melanin production by bidirectionally regulating melanin synthase transcription. Furthermore, they reduced MITF expression via MC1R/cAMP/PKA signaling pathway and enhanced MITF post-translational modification via Wnt/mitogen-activated protein kinase signaling pathway.

Evaluation of Coarsely Ground Wheat as a Replacement for Ground Corn in the Diets of Lactating Dairy Cows

  • Guo, Y.Q.;Zou, Y.;Cao, Z.J.;Xu, X.F.;Yang, Z.S.;Li, Shengli
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.7
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    • pp.961-970
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    • 2013
  • Eight multiparous Holstein cows ($569{\pm}47$ kg of BW; $84{\pm}17$ DIM) were used to evaluate the effects of different levels of coarsely ground wheat (CGW) as replacements for ground corn (GC) in diets on feed intake and digestion, ruminal fermentation, lactation performance, and plasma metabolites profiles in dairy cows. The cows were settled in a replicated $4{\times}4$ Latin square design with 3-wk treatment periods; four cows in one of the replicates were fitted with rumen cannulas. The four diets contained 0, 9.6, 19.2, and 28.8% CGW and 27.9, 19.2, 9.6, and 0% GC on dry matter (DM) basis, respectively. Increasing dietary levels of CGW, daily DM intake tended to increase quadratically (p = 0.07); however, apparent digestibility of neutral detergent fiber (NDF) and acid detergent fiber (ADF) were significantly decreased (p<0.01) in cows fed the 28.8% CGW diets. Ruminal pH remained in the normal physiological range for all dietary treatments at all times, except for the 28.8% CGW diets at 6 h after feeding; moreover, increasing dietary levels of CGW, the daily mean ruminal pH decreased linearly (p = 0.01). Increasing the dietary levels of CGW resulted in a linear increase in ruminal propionate (p<0.01) and ammonia nitrogen ($NH_3$-N) (p = 0.06) concentration, while ruminal acetate: propionate decreased linearly (p = 0.03) in cows fed the 28.8% CGW diets. Milk production was not affected by diets; however, percentage and yield of milk fat decreased linearly (p = 0.02) when the level of CGW was increased. With increasing levels of dietary CGW, concentrations of plasma beta-hydroxybutyric acid (BHBA) (p = 0.07) and cholesterol (p<0.01) decreased linearly, whereas plasma glucose (p = 0.08), insulin (p = 0.02) and urea nitrogen (p = 0.02) increased linearly at 6 h after the morning feeding. Our results indicate that CGW is a suitable substitute for GC in the diets of dairy cows and that it may be included up to a level of 19.2% of DM without adverse effects on feed intake and digestion, ruminal fermentation, lactation performance, and plasma metabolites if the cows are fed fiber-sufficient diets.

Photon Energy Dependence of the Sensitivity of LiF TLDs Loaded with Thin Material (얇은 박막을 얹은 TLD 반응감도의 광자 에너지에 대한 의존성)

  • Min Byongim J;Kim Sookil;Loh John J.K;Cho Young Kap
    • Radiation Oncology Journal
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    • v.17 no.3
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    • pp.256-260
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    • 1999
  • Purpose : An investigation has been carried out on the factors which affect the response reading of thermoluminescent dosimeters (TLD-100) loaded with thin material in high energy Photon. The aim of the study was to assess the energy response of TLD-100 to the therapeutic ranges of photon beam. Materials and Methods : In this technique, TLD-100 (abbreviated as TLD) chips and three different thin material (Tin, Gold, and Tissue equivalent plastic plate) which mounted on the TLD chip were used in the clinical photon beam. The thickness of each metal plates was 0.1 mm and TE plastic plate was 1 mm thick. These compared with the photon energy dependence of the sensitivities of TLD (normal chip), TLD loaded with Tin or Gold plate, for the photon energy range 6 MV to 15 MV, which was of interest in radiotherapy. Results : The enhancement of surface dose in the TLD with metal plate was clearly detected. The TLD chips with a Gold plate was found to larger response by a factor of 1.83 in 10 MV photon beam with respect to normal chip. The sensitivity of TLD loaded with Tin was less than that for normal TLD and TLD loaded with Gold. The relative sensitivity of TLD loaded with metal has little energy dependence. Conclusion : The good stability and linearity with respect to monitor units of TLD loaded with metal were demonstrated by relative measurements in high energy Photon ($6\~15$ MV) beams. The TLD laminated with metals embedded system in solid water phantom is a suitable detector for relative dose measurements in a small beam size and surface dose.

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Effects of Adding Super Dose Phytase to the Phosphorus-deficient Diets of Young Pigs on Growth Performance, Bone Quality, Minerals and Amino Acids Digestibilities

  • Zeng, Z.K.;Wang, D.;Piao, X.S.;Li, P.F.;Zhang, H.Y.;Shi, C.X.;Yu, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.2
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    • pp.237-246
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    • 2014
  • Two experiments were conducted to evaluate the efficacy of feeding an Escherichia coli (E. coli) derived phytase to pigs fed P deficient, corn-soybean meal diets. In Exp. 1, one hundred and twenty crossbred piglets ($9.53{\pm}0.84$ kg) were allocated to one of five treatments which consisted of four low P diets (0.61% Ca, 0.46% total P and 0.24% non-phytate P) supplemented with 0, 500, 1,000, or 20,000 FTU/kg E. coli phytase as well as a positive control formulated to be adequate in all nutrients (0.77% Ca, 0.62% total P and 0.42% non-phytate P). The treatments were applied to six pens with four pigs per pen for 28 days. In Exp. 2, ten crossbred pigs ($19.66{\pm}1.16$ kg) fitted with ileal T-cannula were used in a nutrient balance study. The pigs were assigned to treatments similar to those used in Exp. 1 in a doubly replicated $5{\pm}4$ incomplete Latin square design (5 diets with 4 periods). Each period consisted of a 5-d adjustment period followed by a 3-d total collection of feces and urine and then a 2-d collection of ileal digesta. Supplementation with phytase linearly increased (p<0.05) weight gain, feed intake, feed efficiency, bone breaking strength and fat-free dry and ash bone weight. There were linear increases (p<0.01) in the apparent ileal digestibility (AID) of DM, GE, CP, Ca, total P, inositol hexaphosphate ($IP_6$) and some AA with increasing dose of E. coli phytase. Pigs fed 20,000 FTU/kg had a greater (p<0.05) AID of IP6 (80% vs 59% or 64%, respectively) than pigs fed diets with 500 or 1,000 FTU/kg phytase. There were linear increases (p<0.05) in the total tract digestibility of Ca, total P, Na, K, Mg, and Zn as well as in the retention of Mg and Zn with increased phytase dose. The retention and utilization of Cu, and the total tract digestibility of CP and Cu quadratic increased (p<0.05) with increased phytase dose. In conclusion, supplementation of 500 FTU of phytase/kg and above effectively hydrolyzed phytate in low-P corn-soybean diets for pigs. In addition, a super dose of phytase (20,000 FTU/kg) hydrolyzed most of the IP6 and consequently further improved mineral use, protein utilization and performance.

Prediction of Digestible and Metabolizable Energy Content and Standardized Ileal Amino Acid Digestibility in Wheat Shorts and Red Dog for Growing Pigs

  • Huang, Q.;Piao, X.S.;Ren, P.;Li, D.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.12
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    • pp.1748-1758
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    • 2012
  • Two experiments were conducted to evaluate the effects of chemical composition of wheat shorts and red dog on energy and amino acid digestibility in growing pigs and to establish prediction models to estimate their digestible (DE) and metabolizable (ME) energy content and as well as their standardized ileal digestible (SID) amino acid content. For Exp. 1, sixteen diets were fed to thirty-two growing pigs according to a completely randomized design during three successive periods. The basal diet was based on corn and soybean meal while the other fifteen diets contained 28.8% wheat shorts (N = 7) or red dog (N = 8), added at the expense of corn and soybean meal. Over the three periods, each diet was fed to six pigs with each diet being fed to two pigs during each period. The apparent total tract digestibility (ATTD) of energy in wheat shorts and red dog averaged 75.1 and 87.9%. The DE values of wheat shorts and red dog averaged 13.8 MJ/kg (range 13.1 to 15.0 MJ/kg) and 15.1 MJ/kg (range 13.3 to 16.6 MJ/kg) of dry matter, respectively. For Exp. 2, twelve growing pigs were allotted to two $6{\times}6$ Latin Square Designs with six periods. Ten of the diets were formulated based on 60% wheat shorts or red dog and the remaining two diets were nitrogen-free diets based on cornstarch and sucrose. Chromic oxide (0.3%) was used as an indigestible marker in all diets. There were no differences (p>0.05) in SID values for the amino acids in wheat shorts and red dog except for lysine and methionine. Apparent ileal digestibility (AID) and SID values for lysine in different sources of wheat shorts or red dog, which averaged 78.1 and 87.8%, showed more variation than either methionine or tryptophan. A stepwise regression was performed to establish DE, ME and amino acid digestibility prediction models. Data indicated that fiber content and amino acid concentrations were good indicators to predict energy values and amino acid digestibility, respectively. The present study confirms the large variation in the energy content and amino acid digestibility in wheat shorts and red dog, and describes the factors that influence this variation and presents equations based on chemical composition that could probably be used to predict the DE and ME values as well as the amino acid digestibility of wheat shorts and red dog.

Identification of genes involved in inbreeding depression of reproduction in Langshan chickens

  • Xue, Qian;Li, Guohui;Cao, Yuxia;Yin, Jianmei;Zhu, Yunfen;Zhang, Huiyong;Zhou, Chenghao;Shen, Haiyu;Dou, Xinhong;Su, Yijun;Wang, Kehua;Zou, Jianmin;Han, Wei
    • Animal Bioscience
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    • v.34 no.6
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    • pp.975-984
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    • 2021
  • Objective: Inbreeding depression of reproduction is a major concern in the conservation of native chicken genetic resources. Here, based on the successful development of strongly inbred (Sinb) and weakly inbred (Winb) Langshan chickens, we aimed to evaluate inbreeding effects on reproductive traits and identify candidate genes involved in inbreeding depression of reproduction in Langshan chickens. Methods: A two-sample t-test was performed to estimate the differences in phenotypic values of reproductive traits between Sinb and Winb chicken groups. Three healthy chickens with reproductive trait values around the group mean values were selected from each of the groups. Differences in ovarian and hypothalamus transcriptomes between the two groups of chickens were analyzed by RNA sequencing (RNA-Seq). Results: The Sinb chicken group showed an obvious inbreeding depression in reproduction, especially for traits of age at the first egg and egg number at 300 days (p<0.01). Furthermore, 68 and 618 differentially expressed genes (DEGs) were obtained in the hypothalamus and ovary between the two chicken groups, respectively. In the hypothalamus, DEGs were mainly enriched in the pathways related to vitamin metabolism, signal transduction and development of the reproductive system, such as the riboflavin metabolism, Wnt signaling pathway, extracellular matrix-receptor interaction and focal adhesion pathways, including stimulated by retinoic acid 6, serpin family F member 1, secreted frizzled related protein 2, Wnt family member 6, and frizzled class receptor 4 genes. In the ovary, DEGs were significantly enriched in pathways associated with basic metabolism, including amino acid metabolism, oxidative phosphorylation, and glycosaminoglycan degradation. A series of key DEGs involved in folate biosynthesis (gamma-glutamyl hydrolase, guanosine triphosphate cyclohydrolase 1), oocyte meiosis and ovarian function (cytoplasmic polyadenylation element binding protein 1, structural maintenance of chromosomes 1B, and speedy/RINGO cell cycle regulator family member A), spermatogenesis and male fertility (prostaglandin D2 synthase 21 kDa), Mov10 RISC complex RNA helicase like 1, and deuterosome assembly protein 1) were identified, and these may play important roles in inbreeding depression in reproduction. Conclusion: The results improve our understanding of the regulatory mechanisms underlying inbreeding depression in chicken reproduction and provide a theoretical basis for the conservation of species resources.

Effects of protein content and the inclusion of protein sources with different amino acid release dynamics on the nitrogen utilization of weaned piglets

  • Hu, Nianzhi;Shen, Zhiwen;Pan, Li;Qin, Guixin;Zhao, Yuan;Bao, Nan
    • Animal Bioscience
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    • v.35 no.2
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    • pp.260-271
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    • 2022
  • Objective: We aimed to investigate the effect of the differing amino acid (AA) release dynamics of two protein sources on the growth performance, nitrogen deposition, plasma biochemical parameters, and muscle synthesis and degradation of piglets when included in their diets at normal and low concentrations. Methods: Forty-eight piglets (Duroc×Landrace×Large White) with initial body weight of 7.45±0.58 kg were assigned to six groups and fed one of 6 diets. The 6 dietary treatments were arranged by 3×2 factorial with 3 protein sources and 2 dietary protein levels. They are NCAS (a normal protein content with casein), NBlend (a normal protein content with blend of casein and corn gluten meal), NCGM (a normal protein content with corn gluten meal), LCAS (a low protein content with casein), LBlend (a low protein content with blend of casein and corn gluten meal), LCGM (a low protein content with corn gluten meal). The release dynamics of AA in these diets were determined by in vitro digestion. The digestibility, utilization and biological value of nitrogen in piglets were determined by micro Kjeldahl method. Plasma insulin was measured by enzyme-linked immunosorbent assay kits. The protein expression of mediators of muscle synthesis and degradation was determined by western blotting. Results: Although the consumption of a low-protein diet supplemented with crystalline AA was associated with greater nitrogen digestion and utilization (p<0.05), the final body weight, growth performance, nitrogen deposition, and phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein 1 in the muscle of pigs in the low-protein diet-fed groups were lower than those of the normal-protein diet-fed groups (p<0.05) because of the absence of non-essential AA. Because of the more balanced release of AA, the casein (CAS) and Blend-fed groups showed superior growth performance, final body weight and nitrogen deposition, and lower expression of muscle ring finger 1 and muscle atrophy F-box than the CGM-fed groups (p<0.05). Conclusion: We conclude that the balanced release of AA from CAS containing diets and mixed diets could reduce muscle degradation, favor nitrogen retention, % intake and improve growth performance in pigs consuming either a normal- or low-protein diet.

Analysis of soil coarse pore fraction by major factors for evaluation of water conservation function potential in forest soil (산림토양의 수원함양기능 잠재력 평가를 위한 주요 인자별 토양 조공극률 분석)

  • Li, Qiwen;Lim, Hong-Geun;Moon, Hae-Won;Nam, Soo-Youn;Kim, Jae-Hoon;Choi, Hyung-Tae
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.25 no.6
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    • pp.35-50
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    • 2022
  • As the water shortage has become a noticeable issue due to climate change, forests play an importance role as the provider of water supply service. There is, however, little information about the relationships between the factors used in the estimation of water supply service and coarse pore fraction of forest soil which determines the potential of water supply. To find out whether there would be an amelioration in the scoring system of water supply service estimation, we examined all factors except meteorological one and additionally, analyzed 4 extra factors that might be related with coarse pore fraction of soil. A total of 2,214 soil samples were collected throughout South Korea to measure coarse pore fractions from 2015 to 2020. First, the result of average coarse pore fraction of all samples showed 32.98±6.59% which was consistent with previous studies. And the results of non-parametric analysis of variance indicated that only two of eleven factors that was used in the scoring system matched the results of coarse pore fraction of forest soils. Tree canopy coverage showed no difference among categories, and slope also showed no significance at level of 0.05 in the linear regression analysis. Additionally, the applicability of 4 extra factors were confirmed, as the result of coarse pore fractions of soil samples were different for various categories of each factor. Therefore, the scoring system of water supply service of forest should be revised to improve accuracy.