|本期目录/Table of Contents|

[1]刘 闯 赵 莹** 习慧君 郭康迪 洪坤奇 文才艺**.河南省不同类型土壤可培养放线菌多样性及拮抗活性菌株的筛选*[J].应用与环境生物学报,2019,25(02):1-11.[doi:10.19675/j.cnki.1006-687x.2018.06034]
　LIU Chuang,ZHAO Ying**,XI Huijun,et al.Diversity of culturable actinomycetes in different types of soil in Henan Province and screening of antagonistic strains*[J].Chinese Journal of Applied & Environmental Biology,2019,25(02):1-11.[doi:10.19675/j.cnki.1006-687x.2018.06034]
点击复制

河南省不同类型土壤可培养放线菌多样性及拮抗活性菌株的筛选*()
分享到：

《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年02期
页码:
1-11
栏目:
研究论文
出版日期:
2019-04-25

文章信息/Info

Title:
Diversity of culturable actinomycetes in different types of soil in Henan Province and screening of antagonistic strains*
文章编号:
201806034
作者:
刘 闯 赵 莹** 习慧君 郭康迪 洪坤奇 文才艺**
河南农业大学植物保护学院 郑州 450002
Author(s):
LIU Chuang;  ZHAO Ying**;  XI Huijun;  GUO Kangdi;  HONG Kunqi;  WEN Caiyi**
College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, China
关键词:
放线菌; 多样性; 分离培养; 拮抗活性
Keywords:
actinomycetes ;  diversity;  isolate and culture;  antagonistic activity
DOI:
10.19675/j.cnki.1006-687x.2018.06034
摘要:
分别从河南省濮阳、南阳、信阳、三门峡和商丘等地区采集潮土、黄棕壤、水稻土、褐土、盐碱土等土壤样品45份，采用8种不同的分离培养基对土壤放线菌进行分离和培养；通过16S rRNA基因序列分析对可培养放线菌进行分类鉴定；以7种植物病原菌为靶标，通过对峙培养筛选拮抗活性菌株，并对具有拮抗活性菌株的次级代谢产物合成相关基因进行了初步筛查。结果表明，从5种类型土壤中分离得到放线菌分离物955株，分布于放线菌门的10个目，12个科，17个属，包含626个物种，其中9个分离物为潜在的新种；分离物中链霉菌属菌株751株，占分离菌株的78.64%，为最主要的优势菌属，稀有放线菌204株，占21.36%；不同类型土壤放线菌的分离频率存在一定的差异，其中褐土分离的放线菌数量最多，水稻土的分离数量最少，但是各类型土壤分离的可培养放线菌在属水平上差异不大；筛选获得对植物病原菌有拮抗活性的菌株392株，占总分离物的41%，其中，链霉菌332株，占总拮抗活性菌株的84.7%，稀有放线菌60株，占总拮抗活性菌株的15.3%，且多数活性菌株含有多种生物活性物质合成相关的功能基因。由此可见，河南省土壤放线菌多样性丰富，并蕴藏丰富的稀有放线菌和拮抗活性放线菌资源，具有发掘放线菌新物种和新活性化合物的潜力。图4表6参33
Abstract:
To investigate the diversity of culturable actinomycetes in different types of soil in Henan Province, and screen active actinomycete strains with biocontrol potentiality on plant diseases, 45 soil samples, including fluvo-aquic soil, yellow-brown soil, paddy soil, cinnamon soil, saline-alkali soil, were collected from different districts of Puyang, Nanyang, Xinyang, Sanmenxia and Shangqiu, respectively. Culturable actinomycete strains were isolated by eight differen t separation mediums. B ased on the systematic analysis of 16S rRNA gene sequence, the isolated strains were classified and identified. A ntagonistic activity strains were screened by means of dual culture used s even kinds of plant pathogenic fungi as targets , and preliminary screening of genes related to secondary metabolite synthesis of antagonistic strains were conducted. Totally 955 strains of actinomycete isolates were isolated from 45 soil samples of 5 types of soil, distributed in 10 orders, 12 families and 17 genera, including 626 operational taxonomic units (OTUs), which contained 9 putative species . Among the isolates, 751 strains, accounted for 78.64% belong to the most dominant genus , Streptomyces . 204 rare actinomycetes, were accounted for 21.36%. There existed some differences in screening frequency of actinomycetes among the different types soil , in which t he number of actinomycetes isolated from cinnamon soil were the largest, and that from paddy soil were the least. However, the culturable actinomycetes strains isolated from different types of soil were not s ignificantly different at genus level. 392 strains with antagonistic activity against plant pathogens were obtained, accounted for 41% of total isolates, among witch 332 strains were Streptomyces, accounted for 84.7% of the total antagonistic strains, and the left of 60 strains were rare actinomycetes, accounted for 15.3% , moreover，the vast majority of active strains contained multiple functional genes involved in the synthesis of biologically active substances. Different types of soil in Henan province contain a high diversity of actinomycetes, and the multiple soils are rich in rare actinomycetes and antagonistic actinomycetes resources，which prove to have great potential of novel actinomycetes species and new active compounds.

参考文献/References:

参考文献 [References]
1 Poomthongdee N, Duangmal K, Pathom-Aree W. Acidophilic actinomycetes from rhizosphere soil: diversity and proprieties beneficial to plants[J]. J Antibiot, 2014, 68(2):106–114
2 Genilloud O. Actinomycetes: still a source of novel antibiotics[J]. Nat Prod Rep, 2017, 349(10): 1203-1232
3 Kunova A, Bonaldi M, Saracchi M, Pizzatti C, Chen X, Cortesi P. Selection of Streptomyces against soil borne fungal pathogens by a standardized dual culture assay and evaluation of their effects on seed germination and plant growth[J]. BMC Microbiology, 2016, 16(1):272
4 Sharma M. Actinomycetes: Source, identification, and their applications[J]. Int.J.Curr.Microbiol.App.Sci, 2014, 3(2):801-832
5 Barka EA, Vatsa P, Sanchez L, Gaveau-Vaillant N, Jacquard C, Klenk HP, Clément C, Ouhdouch Y, van Wezel GP. Taxonomy, physiology, and natural products of Actinobacteria[J]. Microbiol Mol Biol R, 2015, 80(1):1-43
6 Koyama R, Matsumoto A, Inahashi Y, Omura S, Takahashi Y. Isolation of actinomycetes from the root of the plant, Ophiopogon japonicus, and proposal of two new species, Actinoallomurus liliacearum sp. nov. and Actinoallomurus vinaceus sp. nov.[J]. J Antibiot, 2012, 65(7):335-340
7 Bonaldi M, Kunova A, Saracchi M, Sardi P, Cortesi P. Streptomycetes as biological control agents against basal drop[J]. Acta horticulturae, 2014, 1044: 313-318
8 李文均, 职晓阳, 唐蜀昆. 我国放线菌系统学研究历史、现状及未来发展趋势[J]. 微生物学通报, 2013, 40 (10): 1860-1873 [Li WJ, Zhi XY, Tang SK. Actinobacterial systematics in China: past, present and future[J]. Microbiology China, 2013, 40 (10): 1860-1873]
9 阮继生. “伯杰氏系统细菌学手册(第二版)”第5卷与我国的放线菌系统学研究[J]. 微生物学报, 2013, 53(6):521-530 [Ruan JS. Bergey’s Manual of Systematic Bacteriology ( 2th edition ) volume 5 and the study of actinomycetes systematic in China[J]. Acta Microbiologica Sinica, 2013, 53(6):521-530]
10 Oren A. 第七届全国微生物资源学术暨国际微生物系统与分类学研讨会：评论和结论[J]. 微生物学通报, 2016, 43(5): 895-898 [Oren A. The 7th National Conference of Microbial Resources&the International Symposium on Microbial Systematics and Taxonomy: comments and conclusions[J]. Microbiology China, 2016, 43(5): 895-898]
11 Bates ST, Clemente JC, Flores GE, Walters WA, Parfrey LW, Knight R, Fierer N. Global biogeography of highly diverse protistan communities in soil [J]. The ISME Journal, 2013,7(3): 652-659
12 李文均, 徐丽华, 姜成林. 云南大学放线菌资源研究历史回顾、发展现状及未来展望[J]. 云南大学学报(自然科学版), 2013, 35(3):428-440 [Li WJ, Xu LH, Jiang CL. Actinobacterial resource and utilization research in Yunnan University: past，present and future[J]. J.?Yuannan Univ. (Nat.?Sci.), 2013, 35(3):428-440]
13 田新朋, 张偲, 李文均. 海洋放线菌研究进展[J]. 微生物学报, 2011, 51(2):161-169 [Tian XP, Zhang S, Li WJ. Advance in marine actinobacterial research - a review [J]. Acta Microbiologica Sinica, 2011, 51(2):161-169]
14 Zhang YQ, Liu HY, Chen J, Yuan LJ, Sun W, Zhang LX, Zhang YQ, Yu LY, Li WJ. Diversity of culturable actinobacteria from Qinghai-Tibet plateau, China. Antonie van Leeuwenhoek, 2010, 98(2): 213–223
15 黄娇, 闫兵法, 黄英. 青藏高原阿里、那曲和海西地区土壤可培养放线菌的多样性[J]. 微生物学报, 2017, 57(09):1342-1351 [Huang J, Yan BF, Huang Y. Diversity of culturable actinobacteria from soils collected in Ali, Naqu and Haixi districts on the Qinghai-Tibet plateau[J]. Acta Microbiologica Sinica, 2017, 57(09):1342-1351]
16 徐丽华, 刘春雷, 常玉梅, 梁利群, 刘金亮, 高国强, 韩启霞. 双标准曲线相对定量PCR试验原理与方法[J]. 生物技术通报, 2011, (01):70-75 [Xu LH, Liu CL, Chang YM, Liang LQ, Liu JL, Gao QG, Han QM. Theory and method of double-standard curves method of relative quantification PCR [J]. Biotech Bull, 2011, (01):70-75]
17 卜春亚, 孙晔, 张天蔚, 王帅, 张英英, 牛薇, 官学琼, 王有年, 师光禄. 一株草莓根腐尖孢镰刀菌拮抗内生细菌的分离鉴定及抑菌特性[J]. 应用与环境生物学报, 2014, 20(02):300-304 [Bu CY, Sun Y, Zhang TW, Wang S, Zhang YY, Niu W, Guan XQ, Wang YN, Shi GL. Isolation and antagonistic character analysis of endophytic bacteria from the strawberry[J]. Chin J Appl Environ Biol, 2014, 20(02):300-304]
18 Wen CY, Zheng DG, Shen SS, Chen JG, Liu WC, Liu T. Streptomyces scabiei subsp. xuchangensis, a novel streptomycete isolate for staurosporine production and a wheat take-all control agent[J]. Int. J. Microbiol. Res., 2012, 4(7):282-289.
19 李小俊, 吴越, 张伟铭, 李静, 刘少伟, 蒋忠科, 黄大林, 孙承航. 河北九莲城淖尔可培养放线菌多样性及抗菌活性筛选[J]. 微生物学通报, 2016, 43(7):1473-1484 [Li XJ, Wu Y, Zhang WM, Li J, Liu SW, Jiang ZK, Huang DL, Sun CH. Biodiversity and antimicrobial activity of culturable actinobacteria isolated from Jiuliancheng Nur in Hebei province[J]. Microbiology China, 2016, 43(7):1473-1484]
20 Ayuso SA, Genilloud O. New PCR primers for the screening of NRPS and PKS-I systems in actinomycetes: detection and distribution of these biosynthetic gene sequences in major taxonomic groups[J]. Microbial Ecology, 2005, 49(1):10-24
21 薛冬, 赵国振, 姚青, 赵海泉, 朱红惠. 肇庆星湖湿地可培养放线菌多样性[J]. 微生物学报, 2015, 55(11):1485-1494 [Xue D, Zhao GZ, Yao Q, Zhao HQ, Zhu HH. Diversity of cultivable actinobacteria in Xinghu wetland sediments[J]. Acta Microbiologica Sinica, 2015, 55(11):1485-1494]
22 Metsä-Ketelä M, Salo V, Halo L, Hautala A, Hakala J, Mäntsälä P, Ylihonko K. An efficient approach for screening minimal PKS genes from Streptomyces[J]. Fems Microbiology Letters, 1999, 180(1):1-6
23 He W, Wu L, Gao Q, Du Y, Wang Y. Identification of AHBA biosynthetic genes related to geldanamycin biosynthesis in Streptomyces hygroscopicus 17997[J]. Curr. Microbiol.,2006,52(3):197-203
24 凡琴. 放线菌产安莎类和环醇类抗生素合成基因资源的挖掘[D]. 厦门大学, 2014 [Fan Q. Gene mining of ansamycin and aminocyclitol antibiotics in Actinomycetes[D]. Xiamen: Xiamen University, 2014]
25 Sigmund JM, Clark DC, Rainey FA, Anderson AS. Detection of eubacterial 3-hydroxy-3-methylglutaryl coenzyme a reductases from natural populations of Actinomycetes.[J]. Microbial Ecology, 2003, 46(1):106-112
26 Chun J, Oren A, Ventosa A,Christensen H, Arahal DR, da Costa MS, Rooney AP, Yi H, Xu XW, De Meyer S, Trujillo ME. Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes[J]. Int J Syst Evol Microbiol, 2018, 68(1):461-466
27 王保军, 刘双江. 环境微生物培养新技术的研究进展[J]. 微生物学通报，2013，40(1)：6-17 [Wang BJ，Liu SJ. Perspectives on the cultivability of environmental microorganisms[J]. Microbiology China，2013,40(1):6-17]
28 谭红妍, 陈宝瑞, 闫瑞瑞, 辛小平, 陶金. 草地土壤微生物特性及其对人为干扰响应的研究进展[J]. 草地学报, 2014, 22(6):1163-1170 [Tan HY, Chen BR, Yan RR, Xin XP, Tao J. Advances on soil microbiological characteristics of grassland ecosystems and its response to human disturbances[J]. Acta Agrectir Sin, 2014, 22(6):1163-1170]
29 朱金山, 张慧, 马连杰, 廖敦秀, 杨星勇, 王龙昌, 王定勇. 不同沼灌年限稻田土壤微生物群落分析[J]. 环境科学, 2018, 39(05):2400-2411 [Zhu JS, Zhang H, Ma LJ, Liao DX, Yang JY, Wang LC, Wang DY. Diversity of the microbial community in rice paddy soil with biogas slurry irrigation analyzed by illumina sequencing technology[J]. Chin.?J.?Envir.?Sci., 2018, 39(05):2400-2411]
30 陈力力, 高必达. 微生物抗植物病毒活性物的研究进展[J]. 中国生物防治学报，2006，22(4)：255-260 [Chen LL, Gao BD. Progress of the research on microbial metabolites against plant virus disease[J]. Chinese J. Biol Control, 2006, 22(4):255-260]
31 Ezziyyani M, Requena ME, Egea-Gilabert C, Candela ME. Biological control of phytophthora root rot of pepper using Trichoderma harzianum and Streptomyces rochei in combination[J]. J. Phytopathol, 2007, 155(6):342-349
32 Srivastava S, Patel JS, Singh HB, Sinha A, Sarma BK. Streptomyces rochei SM3 induces stress tolerance in chickpea against Sclerotinia sclerotiorum and NaCl[J]. J. Phytopathol, 2015, 163(7-8):583-592
33 李威, 肖熙鸥, 李可, 高晓敏, 吕玲玲. 娄彻氏链霉菌XL-6的抑菌活性及对茄子幼苗的防病促生效应[J]. 中国生物防治学报, 2017, 33(4):531-536 [Li W, Xiao XO, Li K, Gao XM, Lv LL. The antibiotic activity of fermentation broth produced from Streptomyces rochei XL-6 and its influence on bacterial wilt control and plant growth in eggplant seedling[J]. Chinese J. Biol Control, 2017, 33(4):531-536]

相似文献/References:

[1]贺达汉,郑哲民.草原不同沙化地段蝗虫与植物群落多样性的变化及相互关系的数值分析[J].应用与环境生物学报,1997,3(01):6.
　He Dahan,Zheng Zhemin.MATHEMATICAL ANALYSIS OF THE CHANGES IN GRASSHOPPER AND PLANT COMMUNITY DIVERSITIES AND THE RELATIONSHIPS BETWEEN HEM IN THE DESERTIFICATION OF STEPPE[J].Chinese Journal of Applied & Environmental Biology,1997,3(02):6.
[2]李成,江建平,刘志君,等.溪洛渡水电站对两栖爬行动物多样性影响的预测与保护对策[J].应用与环境生物学报,2001,7(05):452.
　LI Cheng,et al..Prediction of effects of Xiluodu hydropower Station on the diversity of amphibians and reptiles and their conservation countermeasures[J].Chinese Journal of Applied & Environmental Biology,2001,7(02):452.
[3]周可新,许木启,曹宏,等.活性污泥系统原生动物多样性动态及其与运转效能的关系[J].应用与环境生物学报,2007,13(06):840.
　ZHOU Kexin,et al..Dynamics of Protozoa Diversity and Their Relationship with Performance of Activated Sludge System[J].Chinese Journal of Applied & Environmental Biology,2007,13(02):840.
[4]田耀华,冯玉龙.微生物研究在土壤质量评估中的应用[J].应用与环境生物学报,2008,14(01):132.
　TIAN Yaohua,et al..Application of Microbial Research in Evaluation of Soil Quality[J].Chinese Journal of Applied & Environmental Biology,2008,14(02):132.
[5]贾玉珍 张春雨 赵秀海** 孟庆繁 夏富才.长白山红松阔叶林不同恢复阶段蛾类组成和多样性研究*[J].应用与环境生物学报,2008,14(05):630.
[6]朱新玉,高宝嘉,** 袁胜亮 胡云川.森林-草原交错带土壤节肢动物的群落结构及其季节变化*[J].应用与环境生物学报,2008,14(05):635.
[7]贾玉珍,赵秀海,孟庆繁.温度和降雨对长白山红松阔叶林蝶类组成和多样性的影响[英文][J].应用与环境生物学报,2010,16(01):7.[doi:10.3724/SP.J.1145.2010.00007]
　JIA Yuzhen,ZHAO Xiuhai,MENG Qingfan.Effect of Temperature and Rainfall on Composition and Diversity of Butterfly in Broad-Leaved Korean Pine Forest[J].Chinese Journal of Applied & Environmental Biology,2010,16(02):7.[doi:10.3724/SP.J.1145.2010.00007]
[8]郭建英,万方浩,吴岷.不同棉花品种及种植区域对土壤无脊椎动物群落结构的影响[J].应用与环境生物学报,2010,16(02):205.[doi:10.3724/SP.J.1145.2010.00205]
　GUO Jianying,WAN Fanghao,WU Min.Effects of Cotton Varieties and Planting Areas on Soil Invertebrate Community Structures[J].Chinese Journal of Applied & Environmental Biology,2010,16(02):205.[doi:10.3724/SP.J.1145.2010.00205]
[9]关统伟,赵珂,夏占峰,等.新疆高盐环境土壤放线菌分离培养基比较[J].应用与环境生物学报,2010,16(03):429.[doi:10.3724/SP.J.1145.2010.00429]
　GUAN Tongwei,ZHAO Ke,XIA Zhanfeng,et al.Comparison of Isolation Media for Actinobacteria from Different Saline Environments in Xinjiang, China[J].Chinese Journal of Applied & Environmental Biology,2010,16(02):429.[doi:10.3724/SP.J.1145.2010.00429]
[10]甘慧媚,李靖,谭凤仪,等.深圳福田红树林底栖纤毛虫的群落生态学[J].应用与环境生物学报,2010,16(03):363.[doi:10.3724/SP.J.1145.2010.00363]
　GAN Huimei,LI Jing,TAM NORA Fengyi,et al.Community Ecology of Benthic Ciliates in Futian Mangroves, Shenzhen, China[J].Chinese Journal of Applied & Environmental Biology,2010,16(02):363.[doi:10.3724/SP.J.1145.2010.00363]
[11]邓辉,王成,吕豪豪,等.堆肥过程放线菌演替及其木质纤维素降解研究进展[J].应用与环境生物学报,2013,19(04):581.[doi:10.3724/SP.J.1145.2013.00581]
　DENG Hui,WANG Cheng,LÜ,et al.Research Progress in Succession of Actinomycetal Communities and Their Capacity of Degrading Lignocellulose During Composting Process[J].Chinese Journal of Applied & Environmental Biology,2013,19(02):581.[doi:10.3724/SP.J.1145.2013.00581]

备注/Memo

备注/Memo:
收稿日期: 2018-06-26 接受日期 Accepted: 2018-07-11
*河南省自然科学基金项目（162300410136）和公益性行业（农业）科研专项（201503110）资助?
**通讯作者（E-mail: wencaiyi1965@163.com; nying2009@126.com）
点击摘要页题目后的“PDF”可下载阅读全文；本文为已录用的作者修定稿，尚未经编辑全面修改。
引用本文请注明出处本刊；发表刊期和页码将以正式出版时的安排为准，但DOI确定不变。
更新日期/Last Update: 2018-08-02