两株樟树促生抗病内生细菌的分离、筛选及鉴定

doi:10.3969/j.issn.1000-2006.201802022

国家林草科技领军期刊
中国精品科技期刊
中国高校百佳科技期刊
江苏省新闻出版政府奖期刊奖
RCCSE林学权威期刊（A+）
CSCD核心期刊
Scopus数据库收录期刊
中文核心期刊
SCD核心期刊

作者加群：102861116

微信公众号：南京林业大学学报

高级检索

南京林业大学学报（自然科学版） ›› 2018, Vol. 42 ›› Issue (06): 128-136.doi: 10.3969/j.issn.1000-2006.201802022

两株樟树促生抗病内生细菌的分离、筛选及鉴定

陆蓝翔¹,江明明¹,王焱²,张岳峰²,张洪良²,叶建仁^1*

(1.南京林业大学林学院,南方现代林业协同创新中心,江苏南京 210037; 2.上海市林业总站,上海 200072)

出版日期:2018-11-30 发布日期:2018-11-30
基金资助:
收稿日期:2018-02-24 修回日期:2018-07-19
基金项目:上海市科技兴农重点攻关项目(沪农科攻字(2014)第5-6号); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:陆蓝翔(382433922@qq.com)。*通信作者:叶建仁(jrye@njfu.com.cn),教授。

Isolation, screening and identification of endophytic bacteria from Cinnamomum camphora that promote growth and antagonistic pathogen

LU Lanxiang¹, JIANG Mingming¹,WANG Yan²,ZHANG Yuefeng²,ZHANG Hongliang²,YE Jianren^1*

(1. College of Forestry, Nanjing Forestry University, Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing 210037,China; 2. Shanghai Forestry Station, Shanghai 200072, China)

Online:2018-11-30 Published:2018-11-30

摘要/Abstract

摘要： 【目的】解决城市绿化用樟树黄化病、溃疡病近年来多发的情况。【方法】本研究采集了上海地区健康的行道樟树枝条,分离、筛选并鉴定具有促生抗病作用的樟树内生细菌。通过组织切片法、组织匀浆法从健康樟树组织中分离内生细菌,再通过与2种樟树病害的病原菌进行平皿对抗试验,筛选出对樟树病害具有拮抗作用的菌株,同时还研究了拮抗细菌的解无机磷、解有机磷能力,以及分泌吲哚乙酸(IAA)的能力和耐盐性。在上海嘉定进行了施用优良菌株的田间试验,统计其促进生长情况。【结果】从樟树组织中分离到候选内生细菌共50株,其中有11株菌对樟树病害病原菌具有拮抗效果,4株菌具有解无机磷能力,2株菌具有解有机磷能力,2株菌具有产IAA能力,5株菌有较好的耐盐性。对其中综合性状优秀的2个菌株进行种类鉴定,菌株ZS-1被鉴定为微杆菌(Microbacterium resistens),菌株ZS-3被鉴定为巨大芽孢杆菌(Bacillus megaterium),发现2株菌对樟树扦插苗株高有较好的促生效果。【结论】筛选鉴定了樟树体内的2株优良内生细菌,为樟树促生抗病优良内生菌的研究与应用奠定了良好的基础。

Abstract: 【Objective】Determine the underlying cause of chlorosis in urban greening and canker of Cinnamomum camphora (L.)Presl. 【Method】 We collected the branches and leaves of camphor-tree of Shanghai area, and aimed to isolate endophytic bacterium from camphor-tree. The isolated strains were screened and identified. They could antagonize pathogenic fungi of camphor-tree via phosphorus-solubilizing ability. The endogenous bacteria were isolated from healthy branches and leaves of camphor-tree by tissue slicing method or tissue homogenation method. The endophytic bacteria were screened against the pathogenic fungi in plate for determining the effect of resolved inorganic phosphorus and organic phosphorus by Molybdenum antimony colorimetric method, the effect of indole acetic acid(IAA)production by Salkowski colorimetric method, and its salt resistance.【Result】 Fifty candidate strains were isolated from camphor-tree, of which 11 strains were antagonistic to the pathogens of camphor-tree, 4 strains showed the effect of resolved inorganic phosphate, and 2 strains showed the effect of resolved organic phosphorus. Two strains had the ability of IAA generation, and 5 strains had different salt resistances. Considering the excellent traits, strain ZS-1 was identified as Microbacterium resistens and strain ZS-3 was identified as Bacillus megaterium. Both these strains could increase the height of camphor-tree.【Conclusion】This study reveals the species of endophytic bacteria in camphor-tree and establishes the basis for the biological control and promote the growth of camphor-tree. This study for the first time aimed to determine the underlying causes of diseases in urban greening through the screening of endophytic bacteria from healthy branches and leaves of camphor-tree in Shanghai.

中图分类号:

陆蓝翔,江明明,王焱,等. 两株樟树促生抗病内生细菌的分离、筛选及鉴定[J]. 南京林业大学学报（自然科学版）, 2018, 42(06): 128-136.

LU Lanxiang, JIANG Mingming,WANG Yan,ZHANG Yuefeng,ZHANG Hongliang,YE Jianren. Isolation, screening and identification of endophytic bacteria from Cinnamomum camphora that promote growth and antagonistic pathogen[J].Journal of Nanjing Forestry University (Natural Science Edition), 2018, 42(06): 128-136.DOI: 10.3969/j.issn.1000-2006.201802022.

参考文献

[1] 江明明.几株优良内生菌的分离及鉴定[D].南京:南京林业大学,2016.
JIANG M M.Isolation and identification about several strains of endophytic bacteria[D].Nanjing:Nanjing Forestry University,2016.
[2] 王明生,吴小芹,王焱,等.上海市樟树病害种类调查及病害特征[J].中国森林病虫,2011,30(2):24-28.DOI:10.3969/j.issn.1671-0886.2011.02.008.
WANG M S,WU X Q,WANG Y,et al. Investigation on the species and characteristics of the diseases in Cinnamomum camphora(L.)in Shanghai[J]. Forest Pest and Disease,2011,30(2):24-28.
[3] 袁建康,张英.上海地区香樟黄化病综合防治技术[J].上海农业科技,2006(2):100.DOI:10.3969/j.issn.1001-0106.2006.02.089.
YUAN J K,ZHANG Y.Camphor-tree chlorosis integrated control technology in Shanghai area[J]. Agricultural Science and Technology, 2006(2):100.
[4] 葛建明,张伟,管丽琴,等.香樟炭疽病菌生物学特性及其植物源农药的筛选[J].上海交通大学学报(农业科学版),2005(4):401-405,442. DOI:10.3969/j.issn.1671-9964.2005.04.015.
GE J M, ZHANG W, GUAN L Q, et al. Biological characteristics of Colletotrichum gloeosporioides and its screening of the plant fungicide[J]. Journal of Shanghai Jiaotong University(Agricultural Science),2005(4):401-405,442.
[5] 郭立中,邓先琼,韦石泉.樟树的一种新病害——樟树溃疡病病原菌鉴定[J].植物病理学报,1995(1):28.
GUO L Z,DENG X Q,WEI S Q. Identification on the fungal pathogen of the canker of Camphor-tree[J]. Acta Phytopathologica Sinica,1995(1):28.
[6] 邓先琼,郭立中.樟树溃疡病病原菌再鉴定[J].湖南林业科技,2005(1):39-40,43.DOI:10.3969/j.issn.1003-5710.2005.01.011.
DENG X Q,GUO L Z.Pathogen identification of canker of Cinnamomum camphora(L.)Presl[J].Hunan Forestry Science and Technology,2005(1):39-40,43.
[7] 郭立中,邓先琼.樟树溃疡病研究Ⅲ. 病害发生发展规律[J].湖南农学院学报,1995(2):147-150,160.DOI:10.13331/j.cnki.jhau.1995.02.010.
GUO L Z,DENG X Q. Camphor-tree canker research Ⅲ. the regular pattern[J].Journal of Hunan Agricultural University(Natural Sciences),1995(2):147-150,160.
[8] HARSH N S K, CHANDRA S. A new twig canker disease of Cinnamomum camphora[J]. Indian Journal of Forestry, 2008(2):217-219.
[9] 杨蕾,周国英,梁军,等.防治杨树溃疡病内生菌的分离筛选及鉴定[J].植物保护学报,2014,41(4):438-446. DOI:10.13802/j.cnki.zwbhxb.2014.04.013
YANG L,ZHOU G Y,LIANG J,et al.Isolation.screening and identification of the endogenous microorganisms antagonizing poplar canker[J]. Acta Phytophylacica Sinica,2014,41(4):438-446.
[10] 谢安强.桉树内生菌分离筛选及对宿主生长和抗逆性的影响[D].福州:福建农林大学, 2010.
XIE A Q. Research on photosynthetic characteristics of Eucaly-ptus clones[D]. Fuzhou:Fujian Agrieultural and Forestry University,2010.
[11] 周旋旋.枫杨内生菌抑菌与促生作用初探[D].大连:辽宁师范大学,2008.
ZHOU X X. Primary research into inhibition effect and accelerant of endophytic bacteria and endophytic fungi of Pterocarya steonoptera C. DC[D].Dalian:Liaoning Normal University, 2008.
[12] 唐旭.松枯梢病拮抗细菌的筛选及其抗病机制初探[D].南京:南京林业大学,2017.
TANG X. Screening of antagonistic bacteria against Sphaeropsis sapinea and mechanism of antagomism[D]. Nanjing:Nanjing Forestry University,2017.
[13] BUCHANA R E, GIBBONS N E.伯杰细菌鉴定手册[M]. 8版.北京:科学出版社,1984.
BUCHANA R E, GIBBONS N E. Bergey’s manual of determinative bacteriology[M](eighth ed.). Beijing:Science Press,1984.
[14] 李顺鹏.微生物学实验指导[M]. 北京:中国农业出版社,2015.
LI S P.Microbiological experimental guidance[M].Beijing:China Agriculture Press,2015.
[15] 袁艺.氯乙酰胺除草剂降解菌株Microbacterium sp.T4-7的鉴定及其降解相关酶的酶学特性[D].南京:南京农业大学,2012.
YUAN Y. Identification of chloroacetamide herbicides degrsfating bacterium Microbacterium sp. T4-7 and characterization of its degradation enzyme[D]. Nanjing:Nanjing Agricultural University, 2012.
[16] 魏奇.几株优良内生细菌在上海生态林上的应用研究[D]. 南京:南京林业大学,2017.
WEI Q. Application and research of several excellent endophytic bacteria in Shanghai ecological forest[D]. Nanjing:Nanjing Forestry University,2017.
[17] 付东磊.黄土台塬典型植被恢复措施下土壤胡敏酸降解特性[D].杨凌:西北农林科技大学,2014.
FU D L. Humic acid degradation characteristics under typical vegetation restoration measures of the loess plateau[D].Yangling:Northwest A&F University,2014.
[18] 胡逸晨.细菌Microbacteriumsp.4-7降解聚乙烯[D].武汉:华中农业大学,2015.
HU Y C.Degradation of polyethylene by Microbacterium sp.4-7[D]. Wuhan:Huazhong Agricultural University, 2015.
[19] 韩秋惠,李冲伟,王梅,等.新分离Microbacterium sp.XT11菌产黄原胶降解酶生产条件的优化研究[J].生物技术通报,2006(S1):469-471,475.DOI:10.3969/j.issn.1002-5464.2006.z1.103.
HAN Q H,LI C W,WANG M,et al. Production of Xanthan-degrading enzyme by newly isolated Microbacterium sp. XT11[J].Biotechnology Bulletin,2006(S1):469-471,475.
[20] CHAO W,PRIYANKA S,YEON J K,et al. Characterization and antimicrobial application of biosynthesized gold and silver nanoparticles by using Microbacterium resistens[J].Artificial Cells, Nanomedicine, and Biotechnology, 2015,44(7):1-8.DOI: 10.3109/21691401.2015.1089253
[21] BRENDA ROM’AN-PONCE,DIANA Miryel REZA-V’AZQUE,SONIA G E,et al.Plant growth-promoting traits in Rhizobacteria of heavy metal-resistant plants and their effects on Brassicanigra seed germination [J].Pedosphere,27(3):511-526, 2017.DOI:10.1016/S1002-0160(17)60347-3
[22] 羊宋贞,姚青,孙晓棠,等. 一株番茄青枯菌拮抗菌的鉴定及抗病效果初探[J].微生物学通报,2007(5):859-862.DOI:10.3969/j.issn.0253-2654.2007.05.008.
YANG S Z,YAO Q,SUN X T,et al. Identification of an antagonistic bacterium against Ralstonia solanacearun and primary study of effect on disease resistance[J].Biotechnology Bulletin,2007(5):859-862.
[23] 张新建,黄玉杰,杨合同,等.通过导入几丁质酶基因提高巨大芽孢杆菌的生防效果[J].云南植物研究,2007(6):666-670.DOI:10.3969/j.issn.2095-0845.2007.06.011.
ZHANG X J,HUANG Y J,YANG H T,et al.Improving biocontrol effect of Bacillus megterium(Bacillaceae)on plant disease through genetic modification with Chitinase gene[J]. Acta Botanica Yunnanica,2007(6):666-670.
[24] 赵树民,李晓东,虞方伯,等.巨大芽孢杆菌LY02对黑麦草修复重金属污染土壤的影响[J].水土保持学报,2017,31(5):340-344.DOI:10.13870/i.cnki.stbcxb.2017.05.053
ZHAO S M,LI X D,YU F B,et al. Effect of Bacillus megaterum LY02 on phytoremediation of heavy metal from contaminated soil by Lolium perenne L.[J].Journal of Soil and Water Conservation,2017,31(5):340-344.
[25] 陈超燕. 樟树黄化病发生原因及其致病机理的研究[D].合肥:安徽农业大学,2005.
CHEN C Y. Studies on the causal factors and mechanism of Camphor tree yellows[D]. Hefei:Anhui Agricultural University,2005.

两株樟树促生抗病内生细菌的分离、筛选及鉴定

Isolation, screening and identification of endophytic bacteria from Cinnamomum camphora that promote growth and antagonistic pathogen

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

作者

读者

审稿

[1]	彭萌萌, 吴红渠, 张佳雯, 闫丽琼, 曹传旺, 孙丽丽. 基于RNAi技术解析美国白蛾HcAnk1和HcAnk2基因功能及对HcNPV的敏感性[J]. 南京林业大学学报（自然科学版）, 2024, 48(3): 181-190.
[2]	何乃磊, 张金生, 林文树. 基于深度学习多目标检测技术的林火图像识别研究[J]. 南京林业大学学报（自然科学版）, 2024, 48(3): 207-218.
[3]	李岩松, 杨艳蓉, 张文艺, 张乐英, 黄傲, 张贻荣. 西南地区不同下垫面雷电活动特征与森林雷击火的关系[J]. 南京林业大学学报（自然科学版）, 2024, 48(3): 219-228.
[4]	方静, 张书曼, 严善春, 武帅, 赵佳齐, 孟昭军. 两种丛枝菌根真菌复合接种对青山杨叶片抗美国白蛾的影响[J]. 南京林业大学学报（自然科学版）, 2024, 48(2): 144-154.
[5]	祝艳艳, 贾瑞瑞, 付钰, 常林, 岳远征, 杨秀莲, 王良桂. 不同楸树品种对茎腐病的抗性差异研究[J]. 南京林业大学学报（自然科学版）, 2024, 48(2): 155-165.
[6]	张馨方, 王广鹏, 张树航, 李颖, 郭燕. 不同抗螨性板栗差异次生代谢物筛选与分析[J]. 南京林业大学学报（自然科学版）, 2024, 48(2): 234-240.
[7]	赵亚楠, 孙天骅, 王利峰, 许强, 刘军侠, 高宝嘉, 周国娜. 油松抗性相关激素与代谢物对油松毛虫取食与剪叶刺激的响应[J]. 南京林业大学学报（自然科学版）, 2024, 48(1): 219-226.
[8]	张丞慧, 祖国浩, 王海洋, 薛昊. 蝇克跳小蜂属1中国新记录种(膜翅目:跳小蜂科)[J]. 南京林业大学学报（自然科学版）, 2024, 48(1): 214-218.
[9]	孙凯丽, 贺春玲, 胡俊杰, 方全博, 栾科, 任迎丰, 肖治术. 岩田蜾蠃𧎥在黄喙蜾蠃腹部的寄生习性研究[J]. 南京林业大学学报（自然科学版）, 2023, 47(6): 243-250.
[10]	程方, 孙婷玉, 叶建仁. 抗松针褐斑病湿地松未成熟合子胚胚性愈伤组织的诱导[J]. 南京林业大学学报（自然科学版）, 2023, 47(6): 175-182.
[11]	于赐刚, 郭晓平, 马月, 张振华, 刘燕, 董姗姗, 孙硕. 浙江松阳县鸟类群落结构和多样性分析[J]. 南京林业大学学报（自然科学版）, 2023, 47(5): 231-236.
[12]	刘佳磊, 白润娥, 张锴, 文才艺, 闫凤鸣. 我国桂花树上常见粉虱种类记述[J]. 南京林业大学学报（自然科学版）, 2023, 47(5): 237-244.
[13]	张文文, 王劲, 王秋华, 张曦妍, 曹恒茂, 龙腾腾. 基于MODIS的云南省2001—2020年林火发生时空特征分析[J]. 南京林业大学学报（自然科学版）, 2023, 47(5): 73-79.
[14]	杨乐, 黄晓君, 包玉海, 包刚, 佟斯琴, 苏都毕力格. 无人机航高对落叶松毛虫虫害遥感监测精度的影响[J]. 南京林业大学学报（自然科学版）, 2023, 47(4): 13-22.
[15]	高家军, 张旭, 郭颖, 刘昱坤, 郭安琪, 石蒙蒙, 王鹏, 袁莹. 融合Swin Transformer的虫害图像实例分割优化方法研究[J]. 南京林业大学学报（自然科学版）, 2023, 47(3): 1-10.