生物多样性 ›› 2019, Vol. 27 ›› Issue (8): 903-910.doi: 10.17520/biods.2019211

• • 上一篇    下一篇

大小兴安岭可培养细菌的资源多样性

刘君1, 王宁2, 崔岱宗2, 卢磊2, 赵敏2, *()   

  1. 1. 内蒙古大兴安岭林业科学技术研究所, 内蒙古牙克石 022150
    2. 东北林业大学生命科学学院, 哈尔滨 150040
  • 收稿日期:2019-07-01 接受日期:2019-09-03 出版日期:2019-08-20
  • 通讯作者: 赵敏 E-mail:82191513@163.com
  • 基金项目:
    科技部科技基础性工作专项(2014FY210400)

Diversity of bacterial resources in the Greater and Lesser Khinggan Mountains

Jun Liu1, Ning Wang2, Daizong Cui2, Lei Lu2, Min Zhao2, *()   

  1. 1. Inner Mongolia Daxing’anling Forestry Science and Technology Research Institute, Yakeshi, Inner Mongolia 022150;
    2. College of Life Sciences, Northeast Forestry University, Harbin 150040
  • Received:2019-07-01 Accepted:2019-09-03 Online:2019-08-20
  • Contact: Zhao Min E-mail:82191513@163.com

细菌作为微生物中的重要组成部分, 在有机质的形成与分解、维持生态系统平衡、促进动植物发育等多方面都有着重要作用。2014-2017年, 我们采用常见培养基分离纯培养法及16S rRNA基因序列分析方法对大小兴安岭地区土壤可培养细菌的群落结构和多样性进行了调查研究。结果表明: 从大小兴安岭地区的17个自然保护区内不同生境的土壤中分离获得3,180个菌株, 隶属于24属120种。其中, 芽孢杆菌属(Bacillus)的种数和株数最多, 分别为38种和2,419株, 是大小兴安岭地区可培养细菌的绝对优势类群(占总株数的76.1%); 其次是短杆菌属(Brevibacterium)(13.0%)。大兴安岭地区的物种数、Simpson多样性指数和Shannon-Wiener指数高于小兴安岭地区。优势类群芽孢杆菌属的枯草芽孢杆菌(B. subtilis)、苏云金芽孢杆菌(B. thuringiensis)、巨大芽孢杆菌(B. megaterium)等有重要的生产及科学研究价值。

关键词: 大兴安岭, 小兴安岭, 细菌多样性, 纯培养, 16S rRNA, 芽孢杆菌属

As an important part of all microorganisms, bacteria play a vital role in the formation and decomposition of organic matter, the maintenance of ecosystem balance and the promotion of both animal and plant development. Here, the community structure and population diversity of culturable soil bacteria in the Greater and Lesser Khinggan Mountains were studied from 2014 to 2017 using the culture-dependent 16S rRNA sequencing method. Three thousand one hundred eighty bacterial strains were isolated from different habitats across 17 nature reserves in the Greater and Lesser Khinggan Mountains. Twenty-four genera and 120 species were identified from the specimens. Among them, Bacillus had the largest number of species and strains, 38 and 2,419, respectively. Bacillus was the dominant genus (76.1%) for this culture of bacteria from the Greater and Lesser Khinggan Mountains, followed by Brevisbasillus (13.0%). The species number, Simpson diversity index and Shannon-Wiener index were higher in the Greater Khinggan Mountains than the Lesser Khinggan Mountains. The subsequent analysis of the dominant genus of Bacillus showed that B. subtilis, B. thuringiensis and B. megaterium were important for the local economy and future scientific research.

Key words: the Great Khinggan Mountains, the Lesser Khinggan Mountains, bacterial diversity, pure culture, 16S rRNA, Bacillus

表1

大小兴安岭地区样地概况"

保护区 Nature reserve 位置 Location 样地类型 Plot type
大兴安岭 The Greater Khinggan Mountains
呼中国家级自然保护区
Huzhong National Nature Reserve
122°96′ E
51°94′ N
白桦、百里香、杜香、花楸、黑豆树、接骨木、落叶松、鹿蹄草、毛赤杨、山杨、小叶章、钻天柳等
Betula platyphylla, Thymus mongolicus, Ledum palustre, Sorbus pohuashanensis, Vaccinium uliginosum, Sambucus williamsii, Larix gmelinii, Pyrola calliantha, Alnus sibirica, Populus davidiana, Deyeuxia angustifolia, Chosenia arbutifolia, etc.
汗马国家级自然保护区
Hanma National Nature Reserve
122°40′ E
51°61′ N
稗草、赤杨、杜香、风毛菊、黑豆树、鹿蹄草、落叶松、云杉、偃松、榆树、钻天柳、珍珠梅等
Echinochloa crusgalli, Alnus japonica, Ledum palustre, Saussurea japonica, Vaccinium uliginosum, Pyrola incarnate, Larix gmelinii, Picea asperata, Pinus pumila, Ulmus pumila, Chosenia arbutifolia, Sorbaria sorbifolia, etc.
红花尔基樟子松国家级自然保护区
Honghuaerji Pinus sylvestris National Nature Reserve
119°98′ E
48°25′ N
草木樨、刺儿菜、旱柳、黄芩、龙牙菜、鹿蹄草、千屈菜、绣线菊、樟子松等
Melilotus officinalis, Cirsium setosum, Salix matsudana, Scutellaria baicalensis, Agrimonia pilosa, Pyrola calliantha, Lythrum salicaria, Spiraea salicifolia, Pinus sylvestris var. mongolica, etc.
赛罕乌拉国家级自然保护区
Saihanwula National Nature Reserve
118°69′ E
44°22′ N
翠雀、地榆、黑桦、柳兰、柳穿鱼、柳叶风毛菊、狼牙委陵菜、蒙古栎、牛蒡、蛇床、山马兰、山芍药、羊蹄、野火球等
Delphinium grandiflorum, Sanguisorba officinalis, Betula dahurica, Epilobium angustifolium, Linaria vulgaris, Saussurea salicifolia, Potentilla cryptotaeniae, Quercus mongolica, Arctium lappa, Cnidium monnieri, Kalimeris lautureana, Paeonia obovata, Rumex japonicus, Trifolium lupinaster, etc.
达里诺尔国家级自然保护区
Dalinoer National Nature Reserve
116°71′ E
43°37′ N
白叶蒿、干枝梅、芦苇、野豌豆、野韭菜、问荆等
Artemisia leucophylla, Armeniaca mume, Phragmites australis, Vicia sepium, Acorus tatarinowii, Equisetum arvense, etc.
双河国家级自然保护区
Shuanghe National Nature Reserve
125°32′ E
52°46′ N
白桦、百里香、草甸、春榆、稠李、杜香、风毛菊、落叶松、柳兰、鹿蹄草、毛赤杨、暖木条荚蒾、越桔、小叶章、钻天柳、珍珠梅、樟子松等
Betula platyphylla, Thymus mongolicus, Meadow, Ulmus davidiana, Padus racemosa, Ledum palustre,
Saussurea japonica, Larix gmelinii, Epilobium angustifolium, Pyrola calliantha, Alnus sibirica, Viburnum burejaeticum, Vaccinium vitis-idaea, Deyeuxia angustifolia, Chosenia arbutifolia, Sorbaria sorbifolia, Pinus sylvestris var. mongolica, etc.
南瓮河国家级自然保护区
Nanwenghe National Nature Reserve
125°11′ E
51°29′ N
白桦、胡枝子、柳兰、落叶松、毛赤杨、毛榛、土庄绣线菊、蚊子草、玉竹等
Betula platyphylla, Lespedeza bicolor, Epilobium angustifolium, Larix gmelinii, Alnus sibirica, Corylus mandshurica, Spiraea pubescens, Filipendula palmata, Polygonatum odoratum, etc.
科尔沁国家级自然保护区
Horchin National Nature Reserve
121°62′ E
44°95′ N
草问荆、大果榆、黄芩、合掌消、旋覆花等
Equisetum pratense, Ulmus macrocarpa, Scutellaria baicalensis, Cynanchum amplexicaule, Inula japonica, etc.
图牧吉国家级自然保护区
Tumuji National Nature Reserve
123°04′ E
46°24′ N
白叶蒿、灰绿藜、狗尾草、山杏、野豌豆、野韭菜、问荆等
Artemisia leucophylla, Chenopodium glaucum, Setaria viridis, Armeniaca sibirica, Vicia sepium, Acorus tatarinowii, Equisetum arvense, etc.
辉河国家级自然保护区
Huihe National Nature Reserve
118°52′ E
48°36′ N
查干山湿地、稠李、沙地樟子松林、特莫呼珠季节性湿地等
Chagan Mountain Wetland, Padus racemosa, Pinus sylvestris var. mongolica in sandy land, Temohuzhu seasonal wetland, etc.
小兴安岭 The Lesser Khinggan Mountains
逊别拉河自然保护区
Xunbiela River Nature Reserve
127°70′ E
49°40′ N
白桦、落叶松、蒙古栎、硕桦、榆树、樟子松、钻天柳等
Betula platyphylla, Larix gmelinii, Quercus mongolica, Betula costata, Ulmus pumila, Pinus sylvestris var. mongolica, Chosenia arbutifolia, etc.
五大连池国家级自然保护区
Wudalianchi National Nature Reserve
126°07′ E
48°38′ N
白桦、黑桦、落叶松、蒙古栎、青杨、硕桦、樟子松等
Betula platyphylla, B. dahurica, Larix gmelinii, Quercus mongolica, Populus cathayana, Betula costata, Pinus sylvestris var. mongolica, etc.
胜山国家级自然保护区
Shengshan National Nature Reserve
127°74′ E
49°53′ N
白桦暴马丁香、落叶松、毛赤杨、青杨、山梨、硕桦、樟子松等
Betula platyphylla, Syringa reticulata var. amurensis, Larix gmelinii, Alnus sibirica, Populus cathayana, Pyrus ussuriensis, Betula costata, Pinus sylvestris var. mongolica, etc.
乌伊岭国家级自然保护区
Wuyiling National Nature Reserve
129°26′ E
48°36′ N
白桦、赤杨、红松、冷杉、落叶松、湿地、山槐、云杉等
Betula platyphylla, Alnus japonica, Pinus koraiensis, Abies fabri, Larix gmelinii, Wetland, Albizia kalkora, Picea asperata, etc.
丰林国家级自然保护区
Fenglin National Nature Reserve
129°11′ E
48°06′ N
白桦、冷杉、落叶松、红松、忍冬、硕桦、山桃、色木槭、云杉等
Betula platyphylla, Abies fabri, Larix gmelinii, Pinus koraiensis, Lonicera japonica, Betula costata, Amygdalus davidiana, Acer mono, Picea asperata, etc.
凉水国家级自然保护区
Liangshui National Nature Reserve
128°90′ E
47°18′ N
白桦、阔叶混交林、云杉、针叶混交林等
Betula platyphylla, Broadleaf mixed forest, Picea asperata, Coniferous mixed forest, etc.
大亮子河国家森林公园
Daliangzihe National Forest Park
129°39′ E
47°00′ N
白桦、草地、黑桦、红松、落叶松、蒙古栎、硕桦、毛榛、毛赤杨、榆树、针阔混交林等
Betula platyphylla, Grassland, Betula dahurica, Pinus koraiensis, Larix gmelinii, Quercus mongolica, Betula costata, Corylus mandshurica, Alnus sibirica, Ulmus pumila, Coniferous-broadleaf mixed forest, etc.

表2

小兴安岭地区细菌多样性"

样地
Sample area
物种数
No. of species
Simpson 指数
Simpson index
Shannon-Wiener指数
Shannon-Wiener index
Pielou指数
Pielou index
逊别拉河自然保护区 Xunbielahe Nature Reserve 19 0.382 0.629 0.454
五大连池国家级自然保护区 Wudalianchi National Nature Reserve 18 0.288 0.650 0.404
胜山国家级自然保护区 Shengshan National Nature Reserve 15 0.405 0.694 0.431
乌伊岭国家级自然保护区 Wuyiling National Nature Reserve 15 0.322 0.601 0.547
丰林国家级自然保护区 Fenglin National Nature Reserve 39 0.251 0.652 0.297
凉水国家级自然保护区 Liangshui National Nature Reserve 21 0.626 1.282 0.659
大亮子河国家森林公园 Da Liangzihe National Forest Park 26 0.220 0.537 0.258
小兴安岭地区 The Lesser Khinggan Mountains 61 0.326 0.833 0.300

表3

大兴安岭地区细菌多样性"

样地
Sample area
物种数
Species
Simpson指数
Simpson index
Shannon-Wiener指数
Shannon-Wiener index
Pielou指数
Pielou index
达里诺尔国家级自然保护区 Dalinor National Nature Reserve 16 0.310 0.559 0.403
汗马国家级自然保护区 Hanma National Nature Reserve 18 0.558 1.101 0.566
红花尔基樟子松林国家级自然保护区
Honghuaerji Pinus sylvestris National Nature Reserve
14 0.219 0.486 0.302
呼中国家级自然保护区 Huzhong National Nature Reserve 11 0.504 0.805 0.732
科尔沁国家级自然保护区 Horqin National Nature Reserve 38 0.413 0.865 0.416
南瓮河国家级自然保护区 Nanwenghe National Nature Reserve 28 0.540 1.200 0.521
赛罕乌拉国家级自然保护区 Saihanwula National Nature Reserve 9 0.318 0.552 0.502
双河国家级自然保护区 Shuanghe National Nature Reserve 24 0.427 0.958 0.492
图牧吉国家级自然保护区 Tumuji National Nature Reserve 25 0.395 0.638 0.460
辉河国家级自然保护区 Huihe National Nature Reserve 33 0.518 0.972 0.467
大兴安岭地区 The Greater Khinggan Mountains 92 0.441 0.967 0.323

表4

大小兴安岭芽孢杆菌菌株鉴定结果"

代表菌株 Strain 中文名称 Chinese name 相似序列 Similar sequences 相似度 Similarity 登录号 Login number
5-BH16 枯草芽孢杆菌 Bacillus subtilis (NR_102783.1) 100% MH484225
5-L3 平流层芽孢杆菌 Bacillus stratosphericus (NR_118441.1) 99% MH484226
CJ-21 胍胶降解菌 Bacillus paralicheniformis (NR_137421.1) 100% MH484227
CL-2 堀越氏芽孢杆菌 Bacillus horikoshii (NR_040852.1) 99% MH484228
D-GZ8 坚强芽抱杆菌 Bacillus firmus (NR_112635.1) 99% MH484229
J-GW6 莫海威芽孢杆菌 Bacillus mojavensis (NR_118290.1) 99% MH484230
J-GW18 解淀粉芽孢杆菌 Bacillus amyloliquefaciens (NR_117946.1) 99% MH484231
J-ZH13 假蕈状芽孢杆菌 Bacillus pseudomycoides (NR_113991.1) 99% MH484232
K-CQ18 病研所芽孢杆菌 Bacillus idriensis (NR_043268.1) 100% MH484233
K-DY17 盐敏芽孢杆菌 Bacillus halmapalus (NR_026144.1) 98% MH484234
K-SX3 蚯蚓芽孢杆菌 Bacillus eiseniae (NR_108906.1) 98% MH484235
K-HZ4 农研所芽孢杆菌 Bacillus niabensis (NR_043334.1) 99% MH484236
K-RB10 植物内芽孢杆菌 Bacillus endophyticus (NR_025122.1) 99% MH484237
L-FH4 抗根结线虫芽孢杆菌 Bacillus toyonensis (NR_121761.1) 100% MH484238
L-FH15 炭疽芽孢杆菌 Bacillus anthracis (NR_074453.1) 100% MH484239
N-BH9 短小芽孢杆菌 Bacillus pumilus (NR_074977.1) 99% MH484240
N-BH14 Bacillus luciferensis (NR_025511.1) 99% MH484241
N-CD5 特基拉芽孢杆菌 Bacillus tequilensis (NR_104919.1) 99% MH484242
R-BH3 萎缩芽孢杆菌 Bacillus atrophaeus (NR_024689.1) 99% MH484243
T-HH14 蕈状芽孢杆菌 Bacillus mycoides (NR_113990.1) 99% MH484244
T-HS1 蜡样芽孢杆菌 Bacillus cereus (NR_074540.1) 99% MH484245
T-HS9 巨大芽孢杆菌 Bacillus megaterium (NR_117473.1) 99% MH484246
TJ-4 嗜硼芽孢杆菌 Bacillus boroniphilus (NR_041275.1) 99% MH484247
TJ-7 咸海鲜芽孢杆菌 Bacillus jeotgali (NR_025060.1) 99% MH484248
TJ-12 漳州芽孢杆菌 Bacillus zhangzhouensis (NR_148786.1) 99% MH484249
TJ-18 黄海芽孢杆菌 Bacillus marisflavi (NR_118437.1) 99% MH484250
T-KH1 阿耶波多氏芽孢杆菌 Bacillus aryabhattai (NR_115953.1) 100% MH484251
TS-12 钻特省芽孢杆菌 Bacillus drentensis (NR_114085.1) 99% MH484252
T-YS3 简单芽孢杆菌 Bacillus simplex (NR_114919.1) 99% MH484253
T-YS29 Bacillus cytotoxicus (NR_074914.1) 98% MH484254
V-CY6 海泥芽孢杆菌 Bacillus oceanisediminis (NR_117285.1) 99% MH484255
W-LSH2 死谷芽孢杆菌 Bacillus vallismortis (KF419122.1) 99% MH484256
WY-BH37 沙福芽孢杆菌 Bacillus safensis (NR_113945.1) 99% MH484257
WY-YS35 空气芽孢杆菌 Bacillus aerius (NR_118439.1) 99% MH484258
X-FH3 烟酸芽孢杆菌 Bacillus niacini (NR_113777.1) 99% MH484259
X-L14 酸快生芽孢杆菌 Bacillus acidiceler (NR_043774.1) 99% MH484260
X-YS8 苏云金芽孢杆菌 Bacillus thuringiensis (NR_102506.1) 100% MH484261
Z-MCY1 地衣芽孢杆菌 Bacillus licheniformis (NR_118996.1) 99% MH484262
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(in Chinese with English abstract) [ 赵雅慧, 张舒琳, 吴家法, 黄新静, 吴霜, 张政, 杨立芳, 龙寒, 禤金彩, 姜明国 ( 2018) 山口红树林根际土壤可培养细菌多样性及其活性筛选. 海洋学报, 40, 138-151.]
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