生物多样性 ›› 2015, Vol. 23 ›› Issue (1): 61-67. DOI: 10.17520/biods.2014189
王盼盼, 武永秀, 宋彤彤, 马春玲, 赵文, 王瑛, 孙磊*()
收稿日期:
2014-09-15
接受日期:
2014-12-30
出版日期:
2015-01-20
发布日期:
2015-05-04
通讯作者:
孙磊
作者简介:
E-mail: sunlei1018@126.com基金资助:
Panpan Wang, Yongxiu Wu, Tongtong Song, Chunling Ma, Wen Zhao, Ying Wang, Lei Sun*()
Received:
2014-09-15
Accepted:
2014-12-30
Online:
2015-01-20
Published:
2015-05-04
Contact:
Lei Sun
摘要:
惠兰(Cymbidium faberi)是中国兰属代表种之一, 具有很高的观赏价值和经济价值, 对其内生细菌进行研究不仅可以丰富植物内生细菌资源, 还可以为探讨兰花与微生物之间的相互作用关系提供基础数据。本研究采用分离培养方法及16S rRNA基因序列测定对天目山野生蕙兰、在温室培养1年后的蕙兰根内生细菌遗传多样性进行了研究。结果表明: 从野生蕙兰根内分离得到的97株细菌分属于变形菌门的α-变形菌纲、β-变形菌纲、γ-变形菌纲及厚壁菌门的13个属, 其最优势类群为γ-变形菌纲(86.60%), Lelliottia (26.80%)为最优势菌属。从温室盆栽蕙兰根内分离得到的52株细菌分属于变形菌门的α-变形菌纲、β-变形菌纲、γ-变形菌纲及放线菌门的9个属, 优势类群为β-变形菌纲(48.08%), 优势菌属为草螺菌属(Herbaspirillum) (34.62%), 其中菌株ehR17为潜在的新种。这些结果表明天目山野生蕙兰可培养根内生细菌多样性较其在温室培养1年后更为丰富, 同时也说明植物内生细菌的群落结构与生长环境密切相关。
王盼盼, 武永秀, 宋彤彤, 马春玲, 赵文, 王瑛, 孙磊 (2015) 野生及温室盆栽蕙兰可培养根内生细菌的遗传多样性. 生物多样性, 23, 61-67. DOI: 10.17520/biods.2014189.
Panpan Wang, Yongxiu Wu, Tongtong Song, Chunling Ma, Wen Zhao, Ying Wang, Lei Sun (2015) Genetic diversity of culturable endophytic bacteria in the roots of wild and greenhouse Cymbidium faberi. Biodiversity Science, 23, 61-67. DOI: 10.17520/biods.2014189.
类群 Group | 菌株数 Strain numbers | 所占比例 % | 代表菌株 Representative strains | 最相近菌种(登录号) Nearest strain (accession no.) | 相似度 Similarity (%) |
---|---|---|---|---|---|
野生蕙兰 Wild C. faberi | |||||
α-变形菌纲 Alphaproteobacteria (2.06%) | 2 | ||||
1 | 1.03 | 6hN10 | Agrobacterium tumefaciens (AJ389904) | 100.00 | |
1 | 1.03 | 6hR78 | Devosia riboflavin (AJ549086) | 99.85 | |
β-变形菌纲 Betaproteobacteria (4.12%) | 4 | ||||
4 | 4.12 | 6hR5 | Burkholderia stabilis (AF148554) | 99.67 | |
γ-变形菌纲 Gammaproteobacteria (86.60%) | 84 | ||||
13 | 13.40 | 5hR3 | Cedecea neteri (AB086230) | 98.58 | |
1 | 1.03 | 6hR30 | Enterobacter asburiae LF7a (CP003026) | 99.09 | |
4 | 4.12 | 5hR6 | Erwinia rhapontici (ERU80206) | 98.51 | |
9 | 9.28 | 6hN2 | Klebsiella michiganensis (JQ070300) | 99.30 | |
21 | 21.65 | 6hR10 | Kluyvera cryocrescens (AF310218) | 99.34 | |
26 | 26.80 | 6hR36 | Lelliottia amnigena (AB004749) | 99.10 | |
2 | 2.06 | 6hR56 | Pantoea septic (EU216734) | 98.86 | |
8 | 8.25 | 6hN21 | Serratia plymuthica (AJ233433) | 98.49 | |
厚壁菌门 Firmicutes (7.22%) | 7 | ||||
1 | 1.03 | 6hR3 | Bacillus luciferensis (AJ419629) | 98.31 | |
6 | 6.19 | 6hR75 | Paenibacillus kribbensis (AF391123) | 97.62 | |
温室盆栽蕙兰 C. faberi in the greenhouse | |||||
α-变形菌纲 Alphaproteobacteria (34.62%) | 18 | ||||
2 | 3.85 | ehN1 | Novosphingobium rosa (D13945) | 98.26 | |
6 | 11.54 | ehN5 | Rhizobium mayense (JX855172) | 100.00 | |
9 | 17.31 | ehR13 | Rhizobium multihospitium (EF035074) | 100.00 | |
1 | 1.92 | ehR17 | Sphingomonas polyaromaticivorans (EF467848) | 96.52 | |
β-变形菌纲 Betaproteobacteria (48.08%) | 25 | ||||
6 | 11.54 | ehR33 | Burkholderia lata (CP000150) | 99.71 | |
18 | 34.62 | ehK14 | Herbaspirillum chlorophenolicum (AB094401) | 99.18 | |
1 | 1.92 | ehR14 | Variovorax paradoxus (D88006) | 99.56 | |
γ-变形菌纲 Gammaproteobacteria (15.38%) | 8 | ||||
7 | 13.46 | ehR5 | Dyella koreensis (AY884571) | 99.43 | |
1 | 1.92 | ehK21 | Moraxella osloensis (X74897) | 99.86 | |
放线菌门 Actinobacteria (1.92%) | 1 | ||||
1 | 1.92 | ehT15 | Microbacterium oxydans (Y17227) | 100.00 |
表1 野生及温室盆栽蕙兰根内生细菌16S rRNA基因序列相似性分析
Table 1 Similarity analysis of the 16S rRNA gene partial sequences of endophytic bacteria from the roots of wild Cymbidium faberi and C. faberi in the greenhouse
类群 Group | 菌株数 Strain numbers | 所占比例 % | 代表菌株 Representative strains | 最相近菌种(登录号) Nearest strain (accession no.) | 相似度 Similarity (%) |
---|---|---|---|---|---|
野生蕙兰 Wild C. faberi | |||||
α-变形菌纲 Alphaproteobacteria (2.06%) | 2 | ||||
1 | 1.03 | 6hN10 | Agrobacterium tumefaciens (AJ389904) | 100.00 | |
1 | 1.03 | 6hR78 | Devosia riboflavin (AJ549086) | 99.85 | |
β-变形菌纲 Betaproteobacteria (4.12%) | 4 | ||||
4 | 4.12 | 6hR5 | Burkholderia stabilis (AF148554) | 99.67 | |
γ-变形菌纲 Gammaproteobacteria (86.60%) | 84 | ||||
13 | 13.40 | 5hR3 | Cedecea neteri (AB086230) | 98.58 | |
1 | 1.03 | 6hR30 | Enterobacter asburiae LF7a (CP003026) | 99.09 | |
4 | 4.12 | 5hR6 | Erwinia rhapontici (ERU80206) | 98.51 | |
9 | 9.28 | 6hN2 | Klebsiella michiganensis (JQ070300) | 99.30 | |
21 | 21.65 | 6hR10 | Kluyvera cryocrescens (AF310218) | 99.34 | |
26 | 26.80 | 6hR36 | Lelliottia amnigena (AB004749) | 99.10 | |
2 | 2.06 | 6hR56 | Pantoea septic (EU216734) | 98.86 | |
8 | 8.25 | 6hN21 | Serratia plymuthica (AJ233433) | 98.49 | |
厚壁菌门 Firmicutes (7.22%) | 7 | ||||
1 | 1.03 | 6hR3 | Bacillus luciferensis (AJ419629) | 98.31 | |
6 | 6.19 | 6hR75 | Paenibacillus kribbensis (AF391123) | 97.62 | |
温室盆栽蕙兰 C. faberi in the greenhouse | |||||
α-变形菌纲 Alphaproteobacteria (34.62%) | 18 | ||||
2 | 3.85 | ehN1 | Novosphingobium rosa (D13945) | 98.26 | |
6 | 11.54 | ehN5 | Rhizobium mayense (JX855172) | 100.00 | |
9 | 17.31 | ehR13 | Rhizobium multihospitium (EF035074) | 100.00 | |
1 | 1.92 | ehR17 | Sphingomonas polyaromaticivorans (EF467848) | 96.52 | |
β-变形菌纲 Betaproteobacteria (48.08%) | 25 | ||||
6 | 11.54 | ehR33 | Burkholderia lata (CP000150) | 99.71 | |
18 | 34.62 | ehK14 | Herbaspirillum chlorophenolicum (AB094401) | 99.18 | |
1 | 1.92 | ehR14 | Variovorax paradoxus (D88006) | 99.56 | |
γ-变形菌纲 Gammaproteobacteria (15.38%) | 8 | ||||
7 | 13.46 | ehR5 | Dyella koreensis (AY884571) | 99.43 | |
1 | 1.92 | ehK21 | Moraxella osloensis (X74897) | 99.86 | |
放线菌门 Actinobacteria (1.92%) | 1 | ||||
1 | 1.92 | ehT15 | Microbacterium oxydans (Y17227) | 100.00 |
图1 蕙兰根内生细菌16S rRNA基因序列系统发育树。(A)野生蕙兰; (B)温室盆栽蕙兰。
Fig. 1 A dendrogram based on the 16S rRNA gene partial sequences of endophytic bacteria. GenBank accession numbers are given in parentheses. Numbers at the nodes indicate the bootstrap values (>50%) based on 1,000 replicates. (A) A dendrogram of roots endophytic bacteria of wilid Cymbidium faberi. (B) A dendrogram of roots endophytic bacteria of C. faberi in the greenhouse.
类群 Group | 属 Genera | |
---|---|---|
野生蕙兰 Wild | 温室盆栽蕙兰 In the greenhouse | |
Alphaproteobacteria | Agrobacterium | Novosphingobium |
Devosia | Rhizobium | |
- | Sphingomonas | |
Betaproteobacteria | Burkholderia | Burkholderia |
- | Herbaspirillum | |
- | Variovorax | |
Gammaproteobacteria | Cedecea | Dyella |
Enterobacter | Moraxella | |
Erwinia | - | |
Klebsiella | - | |
Kluyvera | - | |
Lelliottia | - | |
Pantoea | - | |
Serratia | - | |
Firmicutes | Bacillus | - |
Paenibacillus | - | |
Actinobacteria | - | Microbacterium |
表2 野生蕙兰和温室盆栽蕙兰根内生细菌群落结构比较
Table 2 Comparison of endophytic bacteria community struc- tures in the roots from wild Cymbidium faberi and C. faberi in the greenhouse
类群 Group | 属 Genera | |
---|---|---|
野生蕙兰 Wild | 温室盆栽蕙兰 In the greenhouse | |
Alphaproteobacteria | Agrobacterium | Novosphingobium |
Devosia | Rhizobium | |
- | Sphingomonas | |
Betaproteobacteria | Burkholderia | Burkholderia |
- | Herbaspirillum | |
- | Variovorax | |
Gammaproteobacteria | Cedecea | Dyella |
Enterobacter | Moraxella | |
Erwinia | - | |
Klebsiella | - | |
Kluyvera | - | |
Lelliottia | - | |
Pantoea | - | |
Serratia | - | |
Firmicutes | Bacillus | - |
Paenibacillus | - | |
Actinobacteria | - | Microbacterium |
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