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八角炭疽病拮抗放线菌RX2-2的分离鉴定及其生物活性评价
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广西高校中青年教师基础能力提升项目(2021KY0582);玉林师范学院大学生创新创业训练计划(202010606165);玉林师范学院高层次人才科研启动基金(G2019ZK49)


Isolation,identification,and bioactivity evaluation of an actinomycetes strain RX2-2 against star anise anthracnose
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    摘要:

    【背景】八角炭疽病主要是由哈锐炭疽菌(Colletotrichum horii)引起的真菌性病害,给八角产业带来严重的经济损失。【目的】从八角根系土壤中分离筛选对哈锐炭疽菌具有拮抗作用的放线菌菌株,并对其进行种属鉴定及抗菌活性评价。【方法】采用稀释涂布平板法分离放线菌,并以哈锐炭疽菌作为指示菌,利用平板对峙法筛选具有高拮抗活性的菌株;基于形态特征、培养特性、生理生化特性和16S rRNA基因序列分析对该菌株进行种属鉴定;采用对峙生长法及菌 丝生长速率法对该菌株的拮抗活性进行评价,并采用离体叶片法测定其无菌发酵液的室内防效。【结果】筛选得到一株对哈锐炭疽菌具有较强拮抗活性的菌株RX2-2,其皿内抑制活性达95.48%,该菌对其他7种植物病原真菌和5种常见细菌均具有较好的拮抗作用,抗菌谱较广。发酵液中的抗菌活性物质具有良好的热稳定性。无菌发酵液对感染哈锐炭疽菌的离体八角叶片防控效果高达47.62%。根据该菌的形态特征、生理生化特征及16S rRNA基因序列分析,初步鉴定菌株RX2-2为Streptomyces lunalinharesii。【结论】菌株RX2-2对八角炭疽病有较好的防治效果,具有潜在的应用价值。

    Abstract:

    [Background] Star anise anthracnose is a fungal disease caused by Colletotrichum horii,which has caused serious economic losses to star anise industry in China. [Objective]To isolate and screen out the actinomycetes strains with inhibitory effect on C. horii from the rhizosphere soil of star anise trees.[Methods]Actinomycetes strains were isolated by dilution-plating method,and a plate confrontation experiment was conducted to screen out the strains with antagonistic effects on C. horii.The antagonistic strain was identified based on morphological,physiological,and biochemical characteristics combined with 16S rRNA gene sequence analysis.The antagonistic activity was evaluated based on the mycelial growth rate.The biocontrol effect of the cell-free fermentation broth was tested with detached star anise leaves.[Results]An actinomycetes strain RX2-2 was isolated from the rhizosphere soil of star anise trees,which had a strong antagonistic effect against C. horii, with the inhibition rate of 95.48%.RX2-2 had a broad antimicrobial spectrum,with strong inhibitory effects on other seven plant pathogenic fungal species and five common bacterial species.The antibacterial substances in the fermentation broth of RX2-2 have good thermal stability.The cell-free fermentation broth showed the biocontrol effect up to 47.62% on detached star anise leaves infected with C. horii.RX2-2 was identified as Streptomyces lunalinharesii based on the morphological,physiological,and biochemical characteristics in combination with 16S rRNA gene sequence.[Conclusion]RX2-2 has a promising prospect in the biological control of star anise anthracnose and other plant diseases.

    参考文献
    [1] 钟明顺.八角主要病虫害及科学防治措施探讨[J].绿色科技, 2018(21):121-122 Zhong MS. Discussion on the main pests and diseases of star anise and scientific control measures[J]. Journal of Green Science and Technology, 2018(21):121-122(in Chinese)
    [2] Wang Y, Li MF, Wang L, Tan J, Li R, Jiang ZT, Tang SH, Li TT. Improvement of the stabilities and antioxidant activities of polyphenols from the leaves of Chinese star anise (Illicium verum Hook. f.) using β-cyclodextrin-based metal-organic frameworks[J]. Journal of the Science of Food and Agriculture, 2021, 101(1):287-296
    [3] 梁婵.广西八角人工控制授粉技术研究[J].园艺与种苗, 2021, 41(1):1-4, 15 Liang C. Study on artificial control pollination technology of Illicium verum in Guangxi[J]. Horticulture& Seed, 2021, 41(1):1-4, 15(in Chinese)
    [4] 戴晓萧.广西藤县古龙镇八角产业现状及发展对策研究[D].南宁:广西大学硕士学位论文, 2019 Dai XX. Research on the present situation and development strategy of star anise industry in gulong town Teng County of Guangxi[D]. Nanning:Master's Thesis of Guangxi University, 2019(in Chinese)
    [5] 李慧兰,韦明,杨熔.广西发展八角的优势及前景[J].中国热带农业, 2008(4):18-19 Li HL, Wei M, Yang R. Advantages and prospects of developing star anise in Guangxi[J]. China Tropical Agriculture, 2008(4):18-19(in Chinese)
    [6] 马锦林,曾祥艳,李开祥,邓力,黄开顺.广西八角产业现状及发展战略[J].广西林业科学, 2011, 40(4):336-339 Ma JL, Zeng XY, Li KX, Deng L, Huang KS. Current situation and development strategy of star anise industry in Guangxi[J]. Guangxi Forestry Science, 2011, 40(4):336-339(in Chinese)
    [7] 蒋晓萍,莫小刚,周婵,阙衍文,黄乃秀.广西八角炭疽病年发病节律观测与防治适期分析[J].中国野生植物资源, 2017, 36(6):68-70, 79 Jiang XP, Mo XG, Zhou C, Que YW, Huang NX. Preliminary study of the annual dynamic of Colletotrichum coccdes and control period in Guangxi[J]. Chinese Wild Plant Resources, 2017, 36(6):68-70, 79(in Chinese)
    [8] Alhajj MS, Qasem MA, Al-Mufarrej SI. Inhibitory activity of Illicium verum extracts against avian viruses[J]. Advances in Virology, 2020, 2020:4594635
    [9] 刘瑞新.六万林场八角林病虫害为害结构调查初报[J].广西农学报, 2015, 30(6):50-53 Liu RX. Experimental report on the damage structure of disease and pest of Illicium verum forest in Liuwan Forest Farm[J]. Journal of Guangxi Agriculture, 2015, 30(6):50-53(in Chinese)
    [10] 苏晓琳,蒋晓萍,刘瑞新,莫小刚,何水秋,郭丽娟.八角主要病虫害发生规律研究[J].安徽农业科学, 2019, 47(2):132-134 Su XL, Jiang XP, Liu RX, Mo XG, He SQ, Guo LJ. The regularity of the main insect pests and diseases of Illicium verum[J]. Journal of Anhui Agricultural Sciences, 2019, 47(2):132-134(in Chinese)
    [11] 陈军.百色市右江区八角主要病虫害及其综合防治措施[J].吉林农业, 2014(20):81-82 Chen J. The main pests and diseases of star anise and its integrated control measures in Youjiang District of Baise[J]. Agricultural of Jilin, 2014(20):81-82(in Chinese)
    [12] 廖旺姣,邹东霞,黄乃秀,吴耀军,覃世杰,蒋晓萍.一种八角炭疽病新病原鉴定[J].西南农业学报, 2017, 30(10):2242-2245 Liao WJ, Zou DX, Huang NX, Wu YJ, Qin SJ, Jiang XP. Pathogen identification of new anthracnose of Illicium verum[J]. Southwest China Journal of Agricultural Sciences, 2017, 30(10):2242-2245(in Chinese)
    [13] 李柏秀.藤县八角主要病虫害及防治措施研究[J].大众科技, 2013, 15(3):95-96 Li BX. Star anise in Tengxian main pests and diseases and prevention measures[J]. Popular Science& Technology, 2013, 15(3):95-96(in Chinese)
    [14] Togni PHB, Venzon M, Lagôa ACG, Sujii ER. Brazilian legislation leaning towards fast registration of biological control agents to benefit organic agriculture[J]. Neotropical Entomology, 2019, 48(2):175-185
    [15] 罗姗姗.采后芒果炭疽病拮抗酵母菌的筛选鉴定及生防效果评价[D].海口:海南大学硕士学位论文, 2015 Luo SS. Isolation, screening and biocontrol efficacy assessment of antagonism yeast against postharvest mango anthracnose[D]. Haikou:Masterʼs Thesis of Hainan University, 2015(in Chinese)
    [16] 焦敬华.山药炭疽病生防菌30702的生防特性及分类鉴定[D].海口:海南大学硕士学位论文, 2016 Jiao JH. Yam anthracnose biocontrol 30702 biocontrol characteristics and identification[D]. Haikou:Master's Thesis of Hainan University, 2016(in Chinese)
    [17] 李晓宇.解淀粉芽孢杆菌LXY-6-2筛选、鉴定、诱变与发酵条件优化及其对辣椒根腐病的生物防治研究[D].杨凌:西北农林科技大学硕士学位论文, 2018 Li XY. Screening, identification, mutation and optimization of fermentation conditions of Bacillus amyloliguefaciens LXY-6-2 and its biocontrol effects against Fusarium solani[D]. Yangling:Master's Thesis of Northwest A& F University, 2018(in Chinese)
    [18] 欧阳慧.抗猕猴桃果腐病西姆芽孢杆菌的分离鉴定及抑菌活性的初步研究[D].南昌:江西农业大学硕士学位论文, 2017 Ouyang H. Identifieation of Bacillus siamensis antagonistic to fruit ripe rot of kiwifruit and the tentative study of its antimicrobial substances[D]. Nanchang:Master's Thesis of Jiangxi Agricultural University, 2017(in Chinese)
    [19] 何有为.芽孢杆菌对油菜根肿病和稻瘟病的生防潜能研究[D].武汉:华中农业大学硕士学位论文, 2018 He YW. Biocontrol potential of Bacillus strains against canola clubroot and rice blast[D]. Wuhan:Master's Thesis of Huazhong Agricultural University, 2018(in Chinese)
    [20] 张月静. 7种植物内生和根际放线菌的分离鉴定及抗菌活性研究[D].哈尔滨:东北农业大学博士学位论文, 2015 Zhang YJ. Identification and antibacterial activity of the endophytic and rhizospheric actinomycetes from seven plants[D]. Harbin:Doctoral Dissertation of Northeast Agricultural University, 2015(in Chinese)
    [21] 李庆蒙,王世强,李昆太,黄国强,涂国全,魏赛金.拮抗放线菌JD211的抑菌活性及其鉴定[J].生物灾害科学, 2013, 36(4):394-398 Li QM, Wang SQ, Li KT, Huang GQ, Tu GQ, Wei SJ. Identification of antagonistic actinomycetes strain JD211 and its antimicrobial activity[J]. Biological Disaster Science, 2013, 36(4):394-398(in Chinese)
    [22] 张志斌,邓映明,熊瑶瑶,汪涯,颜日明,朱笃.东乡野生稻内生放线菌分离及菌株S123次级代谢产物分析[J].微生物学通报, 2015, 42(9):1662-1670 Zhang ZB, Deng YM, Xiong YY, Wang Y, Yan RM, Zhu D. Isolation of endophytic actinomycetes from Dongxiang wild rice (Oryza rufipogon) and analysis of secondary metabolite of active strain S123[J]. Microbiology China, 2015, 42(9):1662-1670(in Chinese)
    [23] 沈玥.玉米内生放线菌的筛选、鉴定及抑制玉米大斑病菌机理研究[D].哈尔滨:东北农业大学博士学位论文, 2014 Shen Y. Sreening and identification of endophytic actinomycetes from maize and research on the mechanism of antigungal activity against Exserohilum turcicum[D]. Harbin:Doctoral Dissertation of Northeast Agricultural University, 2014(in Chinese)
    [24] Shirling EB, Gottlieb D. Methods for characterization of Streptomyces species[J]. International Journal of Systematic Bacteriology, 1966, 16(3):313-340
    [25] Williams ST, Goodfellow M, Alderson G, Wellington EM, Sneath PH, Sackin MJ. Numerical classification of Streptomyces and related genera[J]. Journal of General Microbiology, 1983, 129(6):1743-1813
    [26] 辜柳霜,张月凤,张凯,李晓宇,柳志强.胶孢炭疽菌生防放线菌SD-29的鉴定及抗菌活性评价[J].微生物学通报, 2019, 46(4):770-779 Gu LS, Zhang YF, Zhang K, Li XY, Liu ZQ. Identification and antifungal activity of a biocontrol actinomycete strain SD-29 against Colletotrichum gloeosporioides[J]. Microbiology China, 2019, 46(4):770-779(in Chinese)
    [27] 王彦,牛世全,郑豆豆,周璇,朱学泰,孔维宝,韩建山,张爱梅.黄瓜枯萎病拮抗放线菌的筛选、鉴定及发酵条件优化[J].微生物学通报, 2019, 46(5):1062-1073 Wang Y, Niu SQ, Zheng DD, Zhou X, Zhu XT, Kong WB, Han JS, Zhang AM. Screening, identification and optimization of fermentation conditions of an antagonistic actinomycetes to cucumber Fusarium wilt[J]. Microbiology China, 2019, 46(5):1062-1073(in Chinese)
    [28] 潘洁明,张荣意,邓加艾,谭志琼.原始热带雨林鹦歌岭土壤抗MRSA放线菌的分离与筛选[J].生物技术通报, 2018, 34(6):128-133 Pan JM, Zhang RY, Deng JA, Tan ZQ. Isolation and screening of anti-MRSA actinomycetes in the soil from the primitive tropical rainforest yingge ridge[J]. Biotechnology Bulletin, 2018, 34(6):128-133(in Chinese)
    [29] 薛应钰,范万泽,张树武,徐秉良.苹果树腐烂病菌拮抗放线菌的筛选、鉴定及防效[J].应用生态学报, 2016, 27(10):3379-3386 Xue YY, Fan WZ, Zhang SW, Xu BL. Screening, identification and biocontrol effect of antagonistic actinomycetes against the pathogen of Cytospora sp. apple tree[J]. Chinese Journal of Applied Ecology, 2016, 27(10):3379-3386(in Chinese)
    [30] 阎逊初.放线菌的分类和鉴定[M].北京:科学出版社, 1992 Yan XC. Classification and Identification of Actinomycetes[M]. Beijing:Science Press, 1992(in Chinese)
    [31] 中国科学院微生物研究所放线菌分类组.链霉菌鉴定手册[M].北京:科学出版社, 1975 Actinomycete Classification Group. Institute of Microbiology, Chinese Academy of Sciences. Handbook of Streptomyces Identification[M]. Beijing:Science Press, 1975(in Chinese)
    [32] 于杰.一株抗MRSA放线菌的鉴定及其抗菌活性物质的初步分析[D].海口:海南大学硕士学位论文, 2016 Yu J. Identification of an anti-MRSA actinomycetes and preliminary analysis of antibacterial metabolite[D]. Haikou:Master's Thesis of Hainan University, 2016(in Chinese)
    [33] 武哲,张俊,孙蕾,刘彦彦,葛蓓孛,张克诚.放线菌菌株1706的鉴定及其对草莓白粉病的防治效果[J].中国生物防治学报, 2013, 29(3):401-405 Wu Z, Zhang J, Sun L, Liu YY, Ge BB, Zhang KC. Identification of antagonistic strain 1706 and its effect on strawberry powdery mildew[J]. Chinese Journal of Biological Control, 2013, 29(3):401-405(in Chinese)
    [34] 徐大勇,李志栋,李峰,徐顶泉,吴旺丽.番茄灰霉病菌拮抗放线菌的筛选、鉴定及其活性评价[J].中国生物防治学报, 2012, 28(2):298-302 Xu DY, Li ZD, Li F, Xu DQ, Wu WL. Screening, identification and inhibitory activity of an actinomycete strain and against Botrytis cinerea[J]. Chinese Journal of Biological Control, 2012, 28(2):298-302(in Chinese)
    [35] Himaman W, Thamchaipenet A, Pathom-Aree W, Duangmal K. Actinomycetes from Eucalyptus and their biological activities for controlling Eucalyptus leaf and shoot blight[J]. Microbiological Research, 2016, 188/189:42-52
    [36] Subramani R, Aalbersberg W. Marine actinomycetes:an ongoing source of novel bioactive metabolites[J]. Microbiological Research, 2012, 167(10):571-580
    [37] 周凯强,李志洪,覃书漫,巫新月,汤千千,王紫莹,陈安均,张志清,申光辉.余甘子采后软腐病拮抗菌的筛选、鉴定及其防效[J].微生物学通报, 2020, 47(2):459-468 Zhou KQ, Li ZH, Qin SM, Wu XY, Tang QQ, Wang ZY, Chen AJ, Zhang ZQ, Shen GH. Identification and biocontrol effect of antagonistic Streptomyces against postharvest penicillium fruit rot of Phyllanthus emblica L.[J]. Microbiology China, 2020, 47(2):459-468(in Chinese)
    [38] 郭春燕.拮抗放线菌FX-H-2的鉴定、发酵条件优化及生防效果评价[D].泰安:山东农业大学硕士学位论文, 2018 Guo CY. Identification and fermentation condition optimization of antagonistic actinomycetes FX-H-2 and evaluation of its biocontrol effect[D]. Taiʼan:Masterʼs Thesis of Shandong Agricultural University, 2018(in Chinese)
    [39] 陈淑琴,王生荣.放线菌DX23对马铃薯炭疽病菌的拮抗作用及其鉴定[J].甘肃农业大学学报, 2015, 50(2):95-99 Chen SQ, Wang SR. Antagonism of actinomycete DX23 to Colletotrichum coccodes of potato and its identification[J]. Journal of Gansu Agricultural University, 2015, 50(2):95-99(in Chinese)
    [40] 张丽,纪明山,于志国.娄彻氏链霉菌YL-2代谢产物对稻瘟病菌的抑制活性及其稳定性[J].沈阳农业大学学报, 2014, 45(2):143-146 Zhang L, Ji MS, Yu ZG. Antifungal activities and stability of metabolites from Streptomyces rochei YL-2 against Pyricularia oryzae[J]. Journal of Shenyang Agricultural University, 2014, 45(2):143-146(in Chinese)
    [41] Boukaew S, Prasertsan P. Suppression of rice sheath blight disease using a heat stable culture filtrate from Streptomyces philanthi RM-1-138[J]. Crop Protection, 2014, 61:1-10
    [42] 赖宝春,吴振强,戴瑞卿,林明辉,李丰,林德锋,王家瑞.琯溪蜜柚炭疽病拮抗放线菌的筛选和鉴定[J].热带作物学报, 2020, 41(9):1876-1882 Lai BC, Wu ZQ, Dai RQ, Lin MH, Li F, Lin DF, Wang JR. Screening and identification of antagonistic actinomycete against anthracnose disease of Guanxi honey pomelo cultivar[J]. Chinese Journal of Tropical Crops, 2020, 41(9):1876-1882(in Chinese)
    [43] 姚锦爱,黄鹏,陈峰,黄敏敏,余德亿.建兰炭疽病拮抗放线菌的筛选及防治效果[J].中国生物防治学报, 2019, 35(5):805-812 Yao JA, Huang P, Chen F, Huang MM, Yu DY. Screening and effect of antagonistic actinomycetes against anthracnose of Cymbidium ensifolium[J]. Chinese Journal of Biological Control, 2019, 35(5):805-812(in Chinese)
    [44] 张惠茜,王尉,周登博,云天艳,陈宇丰,谢江辉.香蕉枯萎病拮抗放线菌的分离鉴定及抑菌机理分析[J].中国南方果树, 2021, 50(3):1-8 Zhang HX, Wang W, Zhou DB, Yun TY, Chen YF, Xie JH. Isolation and identification of actinomycetes JRGG-11 and its antagonistic mechanism against banana Fusarium wilt[J]. South China Fruits, 2021, 50(3):1-8(in Chinese)
    [45] 赵娟,刘霆,刘伟成,刘德文,张殿朋,卢彩鸽.番茄灰霉病生防链霉菌筛选及鉴定[J].微生物学通报, 2019, 46(10):2548-2558 Zhao J, Liu T, Liu WC, Liu DW, Zhang DP, Lu CG. Screening and identification of the biocontrol Streptomyces against tomato Botrytis cinerea[J]. Microbiology China, 2019, 46(10):2548-2558(in Chinese)
    [46] 卢彩鸽,董红平,张殿朋,王俊丽,刘德文,刘伟成.解淀粉芽胞杆菌MH71摇瓶发酵培养基及发酵条件优化[J].中国生物防治学报, 2015, 31(3):369-377 Lu CG, Dong HP, Zhang DP, Wang JL, Liu DW, Liu WC. Optimization of fermentation medium components and cultural conditions for Bacillus amyloliquefaciens MH71 in flask[J]. Chinese Journal of Biological Control, 2015, 31(3):369-377(in Chinese)
    [47] 瞿佳,孙晓宇,陈锐,门欣,赵玲侠,宁硕瀛.核桃黑斑病拮抗放线菌WMF106发酵条件优化和抑菌物质稳定性[J].微生物学通报, 2022, 49(1):88-100 Qu J, Sun XY, Chen R, Men X, Zhao LX, Ning SY. Optimization of fermentation conditions and stability evaluation of antimicrobial active substance for antagonistic actinomycete WMF106 against walnut blight[J]. Microbiology China, 2022, 49(1):88-100(in Chinese)
    [48] Vela Gurovic MS, Olivera NL. Antibacterial producing actinomycetes from Extra Andean Patagonia[J]. Journal of Arid Environments, 2017, 144:216-219
    [49] 刘一贤,施玉萍,戴利铭,李岚岚,蔡志英.橡胶褐根病拮抗放线菌17-7的筛选、鉴定及发酵条件优化[J].微生物学通报, 2020, 47(1):118-129 Liu YX, Shi YP, Dai LM, Li LL, Cai ZY. Screening, identification and fermentation optimization of an antimicrobial actinomycete strain 17-7 to Phellinus noxius[J]. Microbiology China, 2020, 47(1):118-129(in Chinese)
    [50] Yang YC, Guan ZB, Ding YR, Liao XR, Cai YJ. Fermentation optimization, cloning and sequence analysis of the laccase gene from Shiraia sp. SUPER-H168[J]. Annals of Microbiology, 2015, 65(1):575-583
    [51] 王游游,蒋继志,王雪宁,李明,吴艳清.致病疫霉拮抗细菌WL2鉴定及其发酵液稳定性分析[J].河北农业大学学报, 2017, 40(4):90-95 Wang YY, Jiang JZ, Wang XN, Li M, Wu YQ. Identification of antagonistic bacteria WL2 against Phytophthora infestans and stability analysis of its fermentation broth[J]. Journal of Agricultural University of Hebei, 2017, 40(4):90-95(in Chinese)
    [52] 朱丹璐,刘忠玄,刘雪峰,刘朝霞,王斌.拮抗旱柳枯萎病菌YQT13菌株发酵条件和抑菌稳定性[J].东北林业大学学报, 2017, 45(7):94-97 Zhu DL, Liu ZX, Liu XF, Liu ZX, Wang B. Fermentation conditions and antibacterial stability of Salix matsudana wilt disease antagonistic fungi YQT13[J]. Journal of Northeast Forestry University, 2017, 45(7):94-97(in Chinese)
    [53] 周丽娜,王莉莉,张永娜,杜春梅. 2株放线菌的抗菌活性及分类学地位[J].中国农学通报, 2015, 31(11):182-189 Zhou LN, Wang LL, Zhang YN, Du CM. Antifungal activity and taxonomic status of two actinomycetes[J]. Chinese Agricultural Science Bulletin, 2015, 31(11):182-189(in Chinese)
    [54] Liu R, Deng ZX, Liu TG. Streptomyces species:ideal chassis for natural product discovery and overproduction[J]. Metabolic Engineering, 2018, 50:74-84
    [55] 张超群,戴建荣.放线菌的研究现况与展望[J].中国病原生物学杂志, 2019, 14(1):110-113, 122 Zhang CQ, Dai JR. Status of and prospects for research on actinomycetes[J]. Journal of Pathogen Biology, 2019, 14(1):110-113, 122(in Chinese)
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潘洁明,陈雪玉,贝永建,张雨晴,韦爱香,莫莎,梁紫丽,赖洁玲. 八角炭疽病拮抗放线菌RX2-2的分离鉴定及其生物活性评价[J]. 微生物学通报, 2022, 49(3): 991-1003

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  • 收稿日期:2021-07-23
  • 录用日期:2021-10-27
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