生物多样性 ›› 2016, Vol. 24 ›› Issue (10): 1154-1163. DOI: 10.17520/biods.2016121
刘国红, 刘波*(), 朱育菁, 车建美, 葛慈斌, 苏明星, 唐建阳
收稿日期:
2016-05-04
接受日期:
2016-10-24
出版日期:
2016-10-20
发布日期:
2016-11-10
通讯作者:
刘波
基金资助:
Guohong Liu, Bo Liu*(), Yujing Zhu, Jianmei Che, Cibin Ge, Mingxing Su, Jianyang Tang
Received:
2016-05-04
Accepted:
2016-10-24
Online:
2016-10-20
Published:
2016-11-10
Contact:
Liu Bo
摘要:
了解芽胞杆菌资源多样性, 可为芽胞杆菌功能资源挖掘和菌剂开发提供基础。从台湾地区8个市(县)采集土壤样本, 从20份土壤样品中分离获得了136株芽胞杆菌, 采用16S rRNA基因同源性将其鉴定为芽胞杆菌科的2个属、20个种。分别属于芽胞杆菌属(Bacillus)的16个种和赖氨酸芽胞杆菌属(Lysinibacillus)的4个种。根据分离频度分析得知, 台湾地区土壤中的芽胞杆菌优势菌群为阿氏芽胞杆菌(Bacillus aryabhattai)、苏云金芽胞杆菌(B. thuringiensis)和蜡样芽胞杆菌(B. cereus), 其他种类分布极其不均匀。芽胞杆菌Shannon多样性指数为1.2925-2.5850, 最高的为台中市和嘉义市(2.5850), 最低的为桃园县(1.2925)。根据分离频度对芽胞杆菌种类的聚类分析显示, 当欧式距离λ = 20时, 芽胞杆菌种类可分为高频度分布类型如阿氏芽胞杆菌(B. aryabhattai), 低频度分布类型如简单芽胞杆菌(B. simplex)。依据分离频度对8个采样点间的聚类分析未发现采样点间的芽胞杆菌种类分布的相关性。本研究认为台湾地区土壤中蕴藏着丰富芽胞杆菌种类多样性高, 具有很大的开发潜力。
刘国红, 刘波, 朱育菁, 车建美, 葛慈斌, 苏明星, 唐建阳 (2016) 台湾地区芽胞杆菌物种多样性. 生物多样性, 24, 1154-1163. DOI: 10.17520/biods.2016121.
Guohong Liu, Bo Liu, Yujing Zhu, Jianmei Che, Cibin Ge, Mingxing Su, Jianyang Tang (2016) Diversity of Bacillus-like species in Taiwan. Biodiversity Science, 24, 1154-1163. DOI: 10.17520/biods.2016121.
序号 No | 土样编号 Soil no. | 市/县 City/county | 采集地点 Location | 生境类型 Habitat type | 采集日期 Collecting date |
---|---|---|---|---|---|
1 | 4570 | 台北市 Taipei City | 阳明山花钟 Flower-clock, Yangmingshan | 草地 Grassland | 2011-08-28 |
2 | 4569 | 阳明山蒋介石雕像 Statue of Chaing Kai-shek, Yangmingshan | 草地 Grassland | 2011-08-28 | |
3 | 4568 | 士林官邸 Chaing Kai-shek Shilin Residence | 芭乐树下 Rhizosphere soils of a Guava tree | 2011-08-28 | |
4 | 4566 | 士林官邸 Chaing Kai-shek Shilin Residence | 白千层树下 Rhizosphere soils of Melaleuca alternifolia | 2011-08-28 | |
5 | 4567 | 士林官邸 Chaing Kai-shek Shilin Residence | 草地 Grassland | 2011-08-28 | |
6 | 4559 | 桃园县 Taoyuan County | 慈湖石像 Stone in Tzuhu | 裸土 Bare soil | 2011-08-27 |
7 | 4561 | 慈湖蒋寝 Chiang Kai-shek Bedroom in Tzuhu | 裸土 Bare soil | 2011-08-27 | |
8 | 4564 | 桃园大溪 Ta-Hsi town, Taoyuan | 九里香根际 Rhizosphere soils of Murraya paniculata | 2011-08-27 | |
9 | 4562 | 慈湖 Tzuhu | 黄花槐 Rhizosphere soils of Sophora xanthantha | 2011-08-27 | |
10 | 4575 | 中坜高速休息区 Highway resting area, Chungli, Taoyuan | 裸土 Bare soil | 2011-08-29 | |
11 | 4574 | 新竹县 Hsinchu County | 新竹食品研究所 Institute of Food Research, Hsinchu | 草地 Grassland | 2011-08-29 |
12 | 4576 | 苗栗县 Miaoli County | 西湖高速休息区 Highway resting area, Xizi Bay Spot, Kaohsiung | 裸土 Bare soil | 2011-08-30 |
13 | 4572 | 台中市 Taichung City | 台湾大学农学院 College of Agriculture, Taiwan University | 草地 Grassland | 2011-08-29 |
14 | 4582 | 南投县 Nantou County | 中台寺 Chung Tai Template | 林地 Forest land | 2011-08-27 |
15 | 4577 | 日月潭 Sun-Moon Lake | 草地 Grassland | 2011-08-30 | |
16 | 4579 | 日月潭宏观寺 Soils at Hongguan Temple, Sun-Moon Lake | 草地 Grassland | 2011-08-30 | |
17 | 4583 | 台一农场 Tai Yi Farm, Nantou | 草地 Grassland | 2011-08-31 | |
18 | 4580 | 嘉义市 Chiayi City | 嘉义大学 Chiayi University | 草地 Grassland | 2011-08-31 |
19 | 4587 | 高雄市 Kaohsiung City | 高雄农友种业 Kaohsiung Nongyou Seeding | 草地 Grassland | 2011-09-01 |
20 | 4585 | 高雄农友种业 Kaohsiung Nongyou Seeding | 草地 Grassland | 2011-09-01 |
表1 台湾土壤样本信息
Table 1 Information of soil samples from Taiwan
序号 No | 土样编号 Soil no. | 市/县 City/county | 采集地点 Location | 生境类型 Habitat type | 采集日期 Collecting date |
---|---|---|---|---|---|
1 | 4570 | 台北市 Taipei City | 阳明山花钟 Flower-clock, Yangmingshan | 草地 Grassland | 2011-08-28 |
2 | 4569 | 阳明山蒋介石雕像 Statue of Chaing Kai-shek, Yangmingshan | 草地 Grassland | 2011-08-28 | |
3 | 4568 | 士林官邸 Chaing Kai-shek Shilin Residence | 芭乐树下 Rhizosphere soils of a Guava tree | 2011-08-28 | |
4 | 4566 | 士林官邸 Chaing Kai-shek Shilin Residence | 白千层树下 Rhizosphere soils of Melaleuca alternifolia | 2011-08-28 | |
5 | 4567 | 士林官邸 Chaing Kai-shek Shilin Residence | 草地 Grassland | 2011-08-28 | |
6 | 4559 | 桃园县 Taoyuan County | 慈湖石像 Stone in Tzuhu | 裸土 Bare soil | 2011-08-27 |
7 | 4561 | 慈湖蒋寝 Chiang Kai-shek Bedroom in Tzuhu | 裸土 Bare soil | 2011-08-27 | |
8 | 4564 | 桃园大溪 Ta-Hsi town, Taoyuan | 九里香根际 Rhizosphere soils of Murraya paniculata | 2011-08-27 | |
9 | 4562 | 慈湖 Tzuhu | 黄花槐 Rhizosphere soils of Sophora xanthantha | 2011-08-27 | |
10 | 4575 | 中坜高速休息区 Highway resting area, Chungli, Taoyuan | 裸土 Bare soil | 2011-08-29 | |
11 | 4574 | 新竹县 Hsinchu County | 新竹食品研究所 Institute of Food Research, Hsinchu | 草地 Grassland | 2011-08-29 |
12 | 4576 | 苗栗县 Miaoli County | 西湖高速休息区 Highway resting area, Xizi Bay Spot, Kaohsiung | 裸土 Bare soil | 2011-08-30 |
13 | 4572 | 台中市 Taichung City | 台湾大学农学院 College of Agriculture, Taiwan University | 草地 Grassland | 2011-08-29 |
14 | 4582 | 南投县 Nantou County | 中台寺 Chung Tai Template | 林地 Forest land | 2011-08-27 |
15 | 4577 | 日月潭 Sun-Moon Lake | 草地 Grassland | 2011-08-30 | |
16 | 4579 | 日月潭宏观寺 Soils at Hongguan Temple, Sun-Moon Lake | 草地 Grassland | 2011-08-30 | |
17 | 4583 | 台一农场 Tai Yi Farm, Nantou | 草地 Grassland | 2011-08-31 | |
18 | 4580 | 嘉义市 Chiayi City | 嘉义大学 Chiayi University | 草地 Grassland | 2011-08-31 |
19 | 4587 | 高雄市 Kaohsiung City | 高雄农友种业 Kaohsiung Nongyou Seeding | 草地 Grassland | 2011-09-01 |
20 | 4585 | 高雄农友种业 Kaohsiung Nongyou Seeding | 草地 Grassland | 2011-09-01 |
代表菌株 Strain | 最相近菌种 Closed match | 16S rRNA登录号 16S rRNA accession no. | 相似性 Similarity (%) |
---|---|---|---|
FJAT-14427 | 假蕈状芽胞杆菌 Bacillus pseudomycoides | KC013911 | 100 |
FJAT-14428 | 黄海芽胞杆菌 Bacillus marisflavi | KC013912 | 99.71 |
FJAT-14429 | 朝日芽胞杆菌 Bacillus asahii | KC013913 | 99.87 |
FJAT-14435 | 简单芽胞杆菌 Bacillus simplex | KC013914 | 100 |
FJAT-14438 | 解木糖赖氨酸芽胞杆菌 Lysinibacillus xylanilyticus | KC013915 | 99.86 |
FJAT-14439 | 阿氏芽胞杆菌 Bacillus aryabhattai | KC013916 | 100 |
FJAT-14454 | 枯草芽胞杆菌 Bacillus subtilis | KC013931 | 99.89 |
FJAT-14481 | 蕈状芽胞杆菌 Bacillus mycoides | KC013917 | 97.98 |
FJAT-14485 | 长赖氨酸芽胞杆菌 Lysinibacillus macroides | KC013918 | 99.37 |
FJAT-14491 | 嗜气芽胞杆菌 Bacillus aerophilus | KC013919 | 100 |
FJAT-14498 | 壁芽胞杆菌 Bacillus muralis | KC013920 | 100 |
FJAT-14510 | 纺锤形赖氨酸芽胞杆菌 Lysinibacillus fusiformis | KC013921 | 100 |
FJAT-14535 | 土壤芽胞杆菌 Bacillus soli | KC013922 | 98.51 |
FJAT-14559 | 婴儿芽胞杆菌 Bacillus infantis | KC013923 | 99.81 |
FJAT-14564 | 球形赖氨酸芽胞杆菌 Lysinibacillus sphaericus | KC013925 | 98.81 |
FJAT-14586 | 苏云金芽胞杆菌 Bacillus thuringiensis | KC013926 | 100 |
FJAT-14602 | 坚强芽胞杆菌 Bacillus firmus | KC013927 | 98.86 |
FJAT-14603 | 海洋沉积芽胞杆菌 Bacillus oceanisediminis | KC013928 | 98.95 |
FJAT-14607 | 病研所芽胞杆菌 Bacillus idriensis | KC013929 | 100 |
FJAT-14613 | 蜡样芽胞杆菌 Bacillus cereus | KC013930 | 100 |
表2 台湾地区土壤中芽胞杆菌的16S rRNA鉴定
Table 2 Identification of Bacillus-like species by 16S rRNA sequencing
代表菌株 Strain | 最相近菌种 Closed match | 16S rRNA登录号 16S rRNA accession no. | 相似性 Similarity (%) |
---|---|---|---|
FJAT-14427 | 假蕈状芽胞杆菌 Bacillus pseudomycoides | KC013911 | 100 |
FJAT-14428 | 黄海芽胞杆菌 Bacillus marisflavi | KC013912 | 99.71 |
FJAT-14429 | 朝日芽胞杆菌 Bacillus asahii | KC013913 | 99.87 |
FJAT-14435 | 简单芽胞杆菌 Bacillus simplex | KC013914 | 100 |
FJAT-14438 | 解木糖赖氨酸芽胞杆菌 Lysinibacillus xylanilyticus | KC013915 | 99.86 |
FJAT-14439 | 阿氏芽胞杆菌 Bacillus aryabhattai | KC013916 | 100 |
FJAT-14454 | 枯草芽胞杆菌 Bacillus subtilis | KC013931 | 99.89 |
FJAT-14481 | 蕈状芽胞杆菌 Bacillus mycoides | KC013917 | 97.98 |
FJAT-14485 | 长赖氨酸芽胞杆菌 Lysinibacillus macroides | KC013918 | 99.37 |
FJAT-14491 | 嗜气芽胞杆菌 Bacillus aerophilus | KC013919 | 100 |
FJAT-14498 | 壁芽胞杆菌 Bacillus muralis | KC013920 | 100 |
FJAT-14510 | 纺锤形赖氨酸芽胞杆菌 Lysinibacillus fusiformis | KC013921 | 100 |
FJAT-14535 | 土壤芽胞杆菌 Bacillus soli | KC013922 | 98.51 |
FJAT-14559 | 婴儿芽胞杆菌 Bacillus infantis | KC013923 | 99.81 |
FJAT-14564 | 球形赖氨酸芽胞杆菌 Lysinibacillus sphaericus | KC013925 | 98.81 |
FJAT-14586 | 苏云金芽胞杆菌 Bacillus thuringiensis | KC013926 | 100 |
FJAT-14602 | 坚强芽胞杆菌 Bacillus firmus | KC013927 | 98.86 |
FJAT-14603 | 海洋沉积芽胞杆菌 Bacillus oceanisediminis | KC013928 | 98.95 |
FJAT-14607 | 病研所芽胞杆菌 Bacillus idriensis | KC013929 | 100 |
FJAT-14613 | 蜡样芽胞杆菌 Bacillus cereus | KC013930 | 100 |
图1 基于16S rRNA基因序列构建的芽胞杆菌的NJ树。数字表示各节点的自展支持率数值(> 50%)
Fig. 1 Neighbor-joining (NJ) phylogenetic tree of Bacillus-like species based on 16S rRNA gene sequences. Numbers indicate bootstrap values (> 50%) of NJ.
种名 Species | 台北市 Taipei City | 桃园县 Taoyuan County | 新竹县 Hsinchu County | 苗栗县 Miaoli County | 南投县 Nantou County | 台中市 Taichung City | 嘉义市 Chiayi City | 高雄市 Kaohsiung City | 总体 Total |
---|---|---|---|---|---|---|---|---|---|
阿氏芽胞杆菌 Bacillus aryabhattai | 100 | 100 | 0 | 0 | 75 | 100 | 0 | 100 | 85 |
苏云金芽胞杆菌 B. thuringiensis | 80 | 80 | 100 | 100 | 75 | 100 | 100 | 50 | 75 |
蜡样芽胞杆菌 B. cereus | 20 | 80 | 0 | 0 | 25 | 100 | 100 | 0 | 55 |
假蕈状芽胞杆菌 B. pseudomycoides | 60 | 20 | 0 | 0 | 0 | 0 | 100 | 100 | 35 |
解木糖赖氨酸芽胞杆菌 Lysinibacillus xylanilyticus | 20 | 40 | 0 | 0 | 25 | 0 | 0 | 0 | 25 |
纺锤形赖氨酸芽胞杆菌 L. fusiformis | 0 | 20 | 100 | 0 | 0 | 100 | 0 | 50 | 15 |
壁芽胞杆菌 B. muralis | 0 | 20 | 100 | 0 | 0 | 0 | 0 | 0 | 10 |
黄海芽胞杆菌 B. marisflavi | 20 | 0 | 100 | 0 | 0 | 0 | 0 | 0 | 10 |
坚强芽胞杆菌 B. firmus | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 0 | 10 |
简单芽胞杆菌 B. simplex | 40 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 10 |
土壤芽胞杆菌 B. soli | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 0 | 10 |
蕈状芽胞杆菌 B. mycoides | 0 | 0 | 0 | 0 | 25 | 0 | 0 | 0 | 5 |
朝日芽胞杆菌 B. asahii | 20 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 5 |
枯草芽胞杆菌 B. subtilis | 20 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 5 |
嗜气芽胞杆菌 B. aerophilus | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 0 | 5 |
L. macroides | 0 | 0 | 0 | 100 | 0 | 0 | 0 | 0 | 5 |
婴儿芽胞杆菌 B. infantis | 0 | 0 | 0 | 100 | 0 | 0 | 0 | 0 | 5 |
球形赖氨酸芽胞杆菌 L. sphaericus | 0 | 0 | 0 | 0 | 25 | 0 | 0 | 0 | 5 |
病研所芽胞杆菌 B. idriensis | 0 | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 5 |
海洋沉积芽胞杆菌 B. oceanisediminis | 0 | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 5 |
表3 台湾各县市芽胞杆菌种类分离频度(%)
Table 3 The frequency (%) of Bacillus-like species isolated from soils in Taiwan
种名 Species | 台北市 Taipei City | 桃园县 Taoyuan County | 新竹县 Hsinchu County | 苗栗县 Miaoli County | 南投县 Nantou County | 台中市 Taichung City | 嘉义市 Chiayi City | 高雄市 Kaohsiung City | 总体 Total |
---|---|---|---|---|---|---|---|---|---|
阿氏芽胞杆菌 Bacillus aryabhattai | 100 | 100 | 0 | 0 | 75 | 100 | 0 | 100 | 85 |
苏云金芽胞杆菌 B. thuringiensis | 80 | 80 | 100 | 100 | 75 | 100 | 100 | 50 | 75 |
蜡样芽胞杆菌 B. cereus | 20 | 80 | 0 | 0 | 25 | 100 | 100 | 0 | 55 |
假蕈状芽胞杆菌 B. pseudomycoides | 60 | 20 | 0 | 0 | 0 | 0 | 100 | 100 | 35 |
解木糖赖氨酸芽胞杆菌 Lysinibacillus xylanilyticus | 20 | 40 | 0 | 0 | 25 | 0 | 0 | 0 | 25 |
纺锤形赖氨酸芽胞杆菌 L. fusiformis | 0 | 20 | 100 | 0 | 0 | 100 | 0 | 50 | 15 |
壁芽胞杆菌 B. muralis | 0 | 20 | 100 | 0 | 0 | 0 | 0 | 0 | 10 |
黄海芽胞杆菌 B. marisflavi | 20 | 0 | 100 | 0 | 0 | 0 | 0 | 0 | 10 |
坚强芽胞杆菌 B. firmus | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 0 | 10 |
简单芽胞杆菌 B. simplex | 40 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 10 |
土壤芽胞杆菌 B. soli | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 0 | 10 |
蕈状芽胞杆菌 B. mycoides | 0 | 0 | 0 | 0 | 25 | 0 | 0 | 0 | 5 |
朝日芽胞杆菌 B. asahii | 20 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 5 |
枯草芽胞杆菌 B. subtilis | 20 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 5 |
嗜气芽胞杆菌 B. aerophilus | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 0 | 5 |
L. macroides | 0 | 0 | 0 | 100 | 0 | 0 | 0 | 0 | 5 |
婴儿芽胞杆菌 B. infantis | 0 | 0 | 0 | 100 | 0 | 0 | 0 | 0 | 5 |
球形赖氨酸芽胞杆菌 L. sphaericus | 0 | 0 | 0 | 0 | 25 | 0 | 0 | 0 | 5 |
病研所芽胞杆菌 B. idriensis | 0 | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 5 |
海洋沉积芽胞杆菌 B. oceanisediminis | 0 | 0 | 0 | 0 | 0 | 0 | 100 | 0 | 5 |
样本编号 Sample no. | 样本来源 Source | 菌种数 Strain number | Shannon指数 Shannon index (H') | Shannon指数平均值 Average of Shannon index |
---|---|---|---|---|
4570 | 台北市 Taipei City | 6 | 2.5850 | 2.0850 |
4569 | 5 | 2.3220 | ||
4568 | 4 | 2.0000 | ||
4566 | 5 | 2.3220 | ||
4567 | 3 | 1.5850 | ||
4559 | 桃园县 Taoyuan County | 2 | 1.0000 | 1.2925 |
4561 | 5 | 2.3220 | ||
4564 | 4 | 2.0000 | ||
4562 | 5 | 2.3220 | ||
4575 | 3 | 1.5850 | ||
4574 | 新竹县 Hsinchu County | 4 | 2.0000 | 2.0000 |
4576 | 苗栗县 Miaoli County | 5 | 2.3220 | 2.3220 |
4572 | 台中市 Taichung City | 6 | 2.5850 | 2.5850 |
4582 | 南投县 Nantou County | 3 | 1.5850 | 1.5850 |
4577 | 3 | 1.5850 | ||
4579 | 4 | 2.0000 | ||
4583 | 3 | 1.5850 | ||
4580 | 嘉义市 Chiayi City | 6 | 2.5850 | 2.5850 |
4585 | 高雄市 Kaohsiung City | 3 | 1.5850 | 1.7925 |
4587 | 4 | 2.0000 |
表4 台湾各县市样本芽胞杆菌种类多样性
Table 4 The diversity of Bacillus-like species of the soil samples of counties and cities in Taiwan
样本编号 Sample no. | 样本来源 Source | 菌种数 Strain number | Shannon指数 Shannon index (H') | Shannon指数平均值 Average of Shannon index |
---|---|---|---|---|
4570 | 台北市 Taipei City | 6 | 2.5850 | 2.0850 |
4569 | 5 | 2.3220 | ||
4568 | 4 | 2.0000 | ||
4566 | 5 | 2.3220 | ||
4567 | 3 | 1.5850 | ||
4559 | 桃园县 Taoyuan County | 2 | 1.0000 | 1.2925 |
4561 | 5 | 2.3220 | ||
4564 | 4 | 2.0000 | ||
4562 | 5 | 2.3220 | ||
4575 | 3 | 1.5850 | ||
4574 | 新竹县 Hsinchu County | 4 | 2.0000 | 2.0000 |
4576 | 苗栗县 Miaoli County | 5 | 2.3220 | 2.3220 |
4572 | 台中市 Taichung City | 6 | 2.5850 | 2.5850 |
4582 | 南投县 Nantou County | 3 | 1.5850 | 1.5850 |
4577 | 3 | 1.5850 | ||
4579 | 4 | 2.0000 | ||
4583 | 3 | 1.5850 | ||
4580 | 嘉义市 Chiayi City | 6 | 2.5850 | 2.5850 |
4585 | 高雄市 Kaohsiung City | 3 | 1.5850 | 1.7925 |
4587 | 4 | 2.0000 |
图2 基于芽胞杆菌分离频度采用欧氏距离模型和Ward’s法构建的台湾地区芽胞杆菌种类的聚类树
Fig. 2 A dendrogram of Bacillus-like species isolated from Taiwan based on frequency of Bacillus-like species. The Ward’s method and Euclidean distance model were used to construct dendrogram.
图3 基于芽胞杆菌分离频度采用欧氏距离模型和Ward’s法构建的台湾地区土样采集点聚类树
Fig. 3 A dendrogram of collection sites in Taiwan based on frequency of Bacillus-like species. The Ward’s method and Euclidean distance model were used to construct dendrogramt.
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