春兰根中可分泌吲哚乙酸的内生细菌多样性

doi:10.3724/SP.J.1003.2010.195

生物多样性 ›› 2010, Vol. 18 ›› Issue (2): 182-187. DOI: 10.3724/SP.J.1003.2010.195

春兰根中可分泌吲哚乙酸的内生细菌多样性

刘琳¹, 孙磊¹^,^*(), 张瑞英¹, 姚娜², 李潞滨²^,^*

¹河北大学生命科学学院, 河北省微生物多样性研究与应用实验室, 河北保定 071002
²中国林业科学研究院林业研究所, 国家林业局林木培育重点实验室, 北京 100091

收稿日期:2009-09-15 接受日期:2009-12-16 出版日期:2010-03-20 发布日期:2010-03-20
通讯作者: 孙磊,李潞滨
作者简介:sunlei1018@126.com
*E-mail：lilubin@126.com;
基金资助:
国家“十一五”科技支撑计划课题“主要商品花卉优质高产新品种选育”之专题“优质高产热带兰新品种选育”(2006BAD01A18);“948”项目“花卉新品种创制技术引进与创新”(2009-4-C08);河北省自然科学基金项目(C2009000180);. 河北大学引进人才专项基金(2006-088);海南省重大科技研发专项“热带花卉产业可持续发展关键技术研究”课题(080102)

Diversity of IAA-producing endophytic bacteria isolated from the roots of Cymbidium goeringii

Liu Lin¹, Sun Lei¹^,^*(), Zhang Ruiying¹, Yao Na², Li Lubin²^,^*

¹College of Life Sciences, Hebei University, Key Laboratory of Microbial Diversity Research and Application of Hebei Province, Baoding, Hebei 071002
²Research Institute of Forestry, Chinese Academy of Forestry; Key Laboratory of Tree Breeding and Cultivation, State Forestry Administration, Beijing 100091

Received:2009-09-15 Accepted:2009-12-16 Online:2010-03-20 Published:2010-03-20
Contact: Sun Lei,Li Lubin

摘要/Abstract

摘要：

植物内生细菌可通过分泌吲哚乙酸(Indole-3-acetic acid, IAA)等方式促进植物生长。本研究以温室盆栽春兰(Cymbidium goeringii)为材料, 采用分离培养方法对春兰根中可分泌IAA的内生细菌多样性进行了研究。从春兰根组织中共分离纯化得到了256株内生细菌, 其中57株具有分泌IAA的能力, 占总菌数的22.3%。根据ARDRA(amplified ribosomal DNA restriction analysis)及16S rDNA系统发育分析结果, 将57株内生细菌划分为25个组, 分属于6大类群, 分别为变形菌门的α-变形菌纲(35.1%)、γ-变形菌纲(14.0%)和β-变形菌纲(8.8%)、厚壁菌门(33.3%)、放线菌门(7.0%)及拟杆菌门(1.8%)。其中变形菌门的α-变形菌纲和厚壁菌门为优势类群, 类芽孢杆菌属(Paenibacillus)为优势菌属, 且为高产IAA的主体菌属。另外, 测序结果显示有4个菌株的序列与已知细菌的最高序列相似性低于97.0%, 可能为潜在的新种或新属。研究结果表明春兰根中分泌IAA的内生细菌具有丰富的多样性。这一结果可为研究和开发植物促生细菌提供基础资料。

关键词: Cymbidium goeringii, 植物促生细菌, ARDRA, 16S rDNA

Abstract

Endophytic bacteria can directly stimulate plant growth in several ways, including production of plant hormones such as indole-3-acetic acid (IAA). We investigated the diversity of IAA-producing endophytic bacteria in the roots of Cymbidium goeringii using culture-dependent approaches. A total of 57 IAA-producing strains were obtained from 256 strains isolated from the interior roots of C. goeringii. Based on amplified ribosomal DNA restriction analyses (ARDRA) and 16S rDNA sequences, these 57 strains were divided into 25 groups. The IAA-producing endophytic bacteria were grouped into six phyla, i.e., Alphaproteobacteria (35.1%), Firmicutes (33.3%), Gammaproteobacteria (14.0%), Betaproteobacteria (8.8%), Actinobacteria (7.0%) and Bacteroidetes (1.8%). The dominant groups were Alphaproteobacteria and Firmicutes, and the dominant genus, Paenibacillus, produced relatively high levels of IAA. In addition, four potential novel taxonomic units were found. Our results revealed diverse and abundant communities of IAA-producing endophytic bacteria in the roots of C. goeringii and highlight the microbial resources represented by plant growth-promoting bacteria.

Key words: Cymbidium goeringii, plant growth-promoting bacteria, ARDRA, 16S rDNA

刘琳, 孙磊, 张瑞英, 姚娜, 李潞滨 (2010) 春兰根中可分泌吲哚乙酸的内生细菌多样性. 生物多样性, 18, 182-187. DOI: 10.3724/SP.J.1003.2010.195.

Liu Lin, Sun Lei, Zhang Ruiying, Yao Na, Li Lubin (2010) Diversity of IAA-producing endophytic bacteria isolated from the roots of Cymbidium goeringii. Biodiversity Science, 18, 182-187. DOI: 10.3724/SP.J.1003.2010.195.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2010.195

https://www.biodiversity-science.net/CN/Y2010/V18/I2/182

图/表 2

参考文献 27

[1]	Aagot N, Nybroe O, Nielsen P, Johnsen K (2001) An altered Pseudomonas diversity is recovered from soil by using nutrient-poor Pseudomonas-selective soil extract media . Applied and Environmental Microbiology, 67,5233-5239. DOI URL PMID
[2]	Arshad M, Frankenberger WT (1991) Microbial production of plant hormones. Plant and Soil, 133,1-8.
[3]	Berge O, Guinebretière MH, Achouak W, Normand P, Heulin T (2002) Paenibacillus graminis sp. nov. and Paenibacillus odorifer sp. nov., isolated from plant roots, soil and food . International Journal of Systematic and Evolutionary Microbiology, 52,607-616. DOI URL PMID
[4]	Currah RS, Zelmer CD, Hambleton S, Richardson KA (1997) Fungi from orchid mycorrhizas. In: Orchid Biology: Reviews and Perspectives. VII (eds Arditti J, Pridgeon AM),pp.117-170. Kluwer Academic Publishers, Dordrecht.
[5]	Da Mota FF, Gomes EA, Seldin L (2008) Auxin production and detection of the gene coding for the Auxin Efflux Carrier (AEC) protein in Paenibacillus polymyxa. Journal of Microbiology, 46,257-264.
[6]	Dias ACF, Costa FEC, Andreote FD, Lacava PT, Teixeira MA, Assumpção LC, Araújo WL, João L, Azevedo JL, Melo IS (2008) Isolation of micropropagated strawberry endophytic bacteria and assessment of their potential for plant growth promotion. World Journal of Microbiology and Biotechnology, 25,189-195.
[7]	Du RQ (杜荣骞) (1999) Biostatistics (生物统计学). Higher Education Press, Beijing. (in Chinese)
[8]	Edwards U, Rogall T, Blöker H, Emde M, Böttger EC (1989) Isolation and direct complete nucleotide determination of entire genes: characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Research, 17,7843-7853. DOI URL PMID
[9]	Glickmann E, Dessaux Y (1995) A critical examination of the specificity of the salkowski reagent for indolic compounds produced by phytopathogenic bacteria. Applied and Environmental Microbiology, 2,793-796.
[10]	Kado CI (1991) Plant pathogenic bacteria. In: The Prokaryotes (eds Balows A, Truper HG, Dworkin M, Schleifer KH)pp.659-674. Springer-Verlag, New York.
[11]	Kamneva SV, Muronets EM (1999) Genetic control of the processes of interaction of bacteria with plants in associations. Genetika, 35,1480-1494.
[12]	Kawai M, Matsutera E, Kanda H, Yamaguchi N, Tani K, Nasu M (2002) 16S ribosomal DNA-based analysis of bacterial diversity in purified water used in pharmaceutical manufacturing processes by PCR and denaturing gradient gel electrophoresis. Applied and Environmental Microbiology, 68,699-704. DOI URL PMID
[13]	Kuklinsky-Sobral J, Araujo WL, Mendes R, Geraldi IO, Pizzirani-Kleiner AA, Azevedo JL (2004) Isolation and characterization of soybean-associated bacteria and their potential for plant growth promotion. Environmental Microbiology, 6,1244-1251. DOI URL PMID
[14]	Lane DJ (1991) 16S/23S rRNA sequencing. In: Nucleic Acid Techniques in Bacterial Systematics (eds Stackebrandt E, Goodfellow M),pp.115-175. John Wiley & Sons, Chichester, United Kingdom.
[15]	Li JH, Wang ET, Chen WF, Chen WX (2008) Genetic diversity and potential for promotion of plant growth detected in nodule endophytic bacteria of soybean grown in Heilongjiang Province of China. Soil Biology and Biochemistry, 40,238-246.
[16]	Libbert E, Risch H (1969) Interactions between plants and epiphytic bacteria regarding their auxin metabolism.V. Isolation and identification of the IAA-producing and destroying bacteria from pea plants. Physiologia Plantarum, 22,51-58.
[17]	Lodewyckx C, Vangronsveld J, Porteous F, Moore ERB, Taghavi S, Mezgeay M, van der Lelie D (2002) Endophytic bacteria and their potential application. Critical Reviews in Plant Sciences, 86,583-606.
[18]	Long HH, Schmidt DD, Baldwin IT (2008) Native bacterial endophytes promote host growth in a species-specific manner; phytohormone manipulations do not result in common growth responses. PLoS ONE, 3,e2702. URL PMID
[19]	Mendes R, Pizzirani-Kleiner AA, Araujo WL, Raaijmakers JM (2007) Diversity of cultivated endophytic bacteria from sugarcane: genetic and biochemical characterization of Burkholderia cepacia complex isolates . Applied and Environmental Microbiology, 73,7259-7267. DOI URL PMID
[20]	Rasmussen HN (2002) Recent developments in the study of orchid mycorrhiza. Plant and Soil, 244,149-163.
[21]	Sarwar M, Frankenberger WT (1994) Tryptophan dependent biosynthesis of auxins in soil. Plant and Soil, 160,97-104.
[22]	Selim S, Negrel J, Govaerts C, Gianinazzi S, Tuinen DV (2005) Isolation and partial characterization of antagonistic peptides produced by Paenibacillus sp. strain B2 isolated from the sorghum mycorrhizosphere . Applied and Environmental Microbiology, 71,6501-6507.
[23]	Sun L (孙磊), Bi XB (毕晓宝), Li LB (李潞滨), Liu L (刘琳), Han JG (韩继刚), Yang K (杨凯) (2009) Primary research on the isolation method of root endophytic bacteria of Cymbidium goeringii. Journal of Agricultural University of Hebei (河北农业大学学报), 32,42-46. (in Chinese with English abstract)
[24]	Taghavi B, Garafola C, Monchy S, Newman L, Hoffman A, Weyens N, Barac T, Vangronsveld J,van der Lelie D (2009) Genome survey and characterization of endophytic bacteria exhibiting a beneficial effect on growth and development of poplar trees. Applied and Environmental Microbiology, 75,748-757. DOI URL PMID
[25]	Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24,1596-1599. DOI URL PMID
[26]	Tsavkelova EA, Cherdyntseva TA, Netrusov AI (2005) Auxin production by bacteria associated with orchid roots. Microbiology, 74,46-53.
[27]	Tsavkelova EA, Cherdyntseva TA, Botina SG, Netrusov AI (2007) Bacteria associated with orchid roots and microbial production of auxin. Microbiological Research, 162,69-76. URL PMID

类群 Group	分型测序菌株Isolate	菌株数 No.of strains	IAA分泌量 IAA production (mg·L^-1· (OD₆₀₀)^-1)	最相近菌种(登录号) Nearest strain (accession no.)	序列相似性Similarity (%)
α-变形菌纲Alphaproteobacteria (35.1%)	R209	7	1.0^#-26.3^#	Rhizobium miluonense (EF061096)	100.0
	R128-2	1	5.8^#	R. huautlense (AF025852)	97.1
	R141	1	3.6^#	R. giardinii (U86344)	97.3
	R224-3	4	10.0^#-43.1^#	Skermanella aerolata (DQ672568)	90.1
	R120	2	23.9*-47.1^#	Devosia insulae (EF012357)	97.8
	R90	2	0.5*-1.7^#	Sphingopyxis macrogoltabida (AB372254)	99.8
	R83-1	1	2.3^#	Altererythrobacter epoxidivorans (DQ304436)	96.3
	R178	1	14.9^#	Mesorhizobium opportunistum (AY601515)	97.5
	R191	1	25.8*	Agrobacterium tumefaciens (EU445236)	100.0
β-变形菌纲Betaproteobacteria (8.8%)	R94	2	0.7^#-5.3^#	Variovorax paradoxus (AJ420329)	99.0
	R199	1	1.9^#	V. soli (DQ432053)	98.4
	R163	1	18.9^#	Burkholderia caribensis (Y17009)	99.1
	R235-1	1	13.1*	Herbaspirillum chlorophenolicum (NR_024804)	99.4
γ-变形菌纲 Gammaproteobacteria (14.0%)	R215-2	7	10.1*-14.6^#	Dyella yeojuensis (DQ181549)	100.0
γ-变形菌纲 Gammaproteobacteria (14.0%)	R192	1	3.3*	Pseudoxanthomonas japonensis (AB008507)	99.7
厚壁菌门 Firmicutes (33.33%)	R138	11	1.1*-114.8^#	Paenibacillus agaridevorans (NR_025490)	98.0
	R172	4	37.6^#-44.0^#	P. contaminans (EF626690)	94.6
	R12-2	2	26.3^#-26.4^#	P. alkaliterrae (AY960748)	96.3
	R59	1	7.7^#	Bacillus aerophilus (AJ831844)	100.0
	R95	1	11.6^#	B. niabensis (DQ176422)	99.8
放线菌门 Actinobacteria (7.0%)	R73-1	1	1.7^#	Nocardioides aquiterrae (AF529063)	98.3
	R100	1	15.3^#	Brachybacterium paraconglomeratum (AJ415377)	100.0
	R152	1	9.6^#	Arthrobacter niigatensis (AB248526)	100.0
	R205	1	1.6^#	Streptomyces albaduncus (AY999757)	98.7
拟杆菌门 Bacteroidetes (1.8%)	R156-1	1	2.1*	Chitinophaga terrae (AB267724)	97.4

类群 Group	分型测序菌株Isolate	菌株数 No.of strains	IAA分泌量 IAA production (mg·L^-1· (OD₆₀₀)^-1)	最相近菌种(登录号) Nearest strain (accession no.)	序列相似性Similarity (%)
α-变形菌纲Alphaproteobacteria (35.1%)	R209	7	1.0^#-26.3^#	Rhizobium miluonense (EF061096)	100.0
	R128-2	1	5.8^#	R. huautlense (AF025852)	97.1
	R141	1	3.6^#	R. giardinii (U86344)	97.3
	R224-3	4	10.0^#-43.1^#	Skermanella aerolata (DQ672568)	90.1
	R120	2	23.9*-47.1^#	Devosia insulae (EF012357)	97.8
	R90	2	0.5*-1.7^#	Sphingopyxis macrogoltabida (AB372254)	99.8
	R83-1	1	2.3^#	Altererythrobacter epoxidivorans (DQ304436)	96.3
	R178	1	14.9^#	Mesorhizobium opportunistum (AY601515)	97.5
	R191	1	25.8*	Agrobacterium tumefaciens (EU445236)	100.0
β-变形菌纲Betaproteobacteria (8.8%)	R94	2	0.7^#-5.3^#	Variovorax paradoxus (AJ420329)	99.0
	R199	1	1.9^#	V. soli (DQ432053)	98.4
	R163	1	18.9^#	Burkholderia caribensis (Y17009)	99.1
	R235-1	1	13.1*	Herbaspirillum chlorophenolicum (NR_024804)	99.4
γ-变形菌纲 Gammaproteobacteria (14.0%)	R215-2	7	10.1*-14.6^#	Dyella yeojuensis (DQ181549)	100.0
γ-变形菌纲 Gammaproteobacteria (14.0%)	R192	1	3.3*	Pseudoxanthomonas japonensis (AB008507)	99.7
厚壁菌门 Firmicutes (33.33%)	R138	11	1.1*-114.8^#	Paenibacillus agaridevorans (NR_025490)	98.0
	R172	4	37.6^#-44.0^#	P. contaminans (EF626690)	94.6
	R12-2	2	26.3^#-26.4^#	P. alkaliterrae (AY960748)	96.3
	R59	1	7.7^#	Bacillus aerophilus (AJ831844)	100.0
	R95	1	11.6^#	B. niabensis (DQ176422)	99.8
放线菌门 Actinobacteria (7.0%)	R73-1	1	1.7^#	Nocardioides aquiterrae (AF529063)	98.3
	R100	1	15.3^#	Brachybacterium paraconglomeratum (AJ415377)	100.0
	R152	1	9.6^#	Arthrobacter niigatensis (AB248526)	100.0
	R205	1	1.6^#	Streptomyces albaduncus (AY999757)	98.7
拟杆菌门 Bacteroidetes (1.8%)	R156-1	1	2.1*	Chitinophaga terrae (AB267724)	97.4

春兰根中可分泌吲哚乙酸的内生细菌多样性

Diversity of IAA-producing endophytic bacteria isolated from the roots of Cymbidium goeringii

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