新疆伊犁马铃薯根际芽胞杆菌纯培养多样性

doi:10.17520/biods.2016348

摘要/Abstract

摘要：

了解马铃薯根际土壤中芽胞杆菌种类多样性, 可为挖掘芽胞杆菌新资源提供基础。从新疆伊犁州9个地点采集了30份马铃薯根际土壤样品, 采用可培养法从中分离芽胞杆菌, 通过16S rRNA基因同源性鉴定了分离菌株的分类地位。共获得芽胞杆菌349株, 基于菌落形态特征和16S rRNA基因序列确定了66个代表菌株, 其中14株与近缘种模式菌株的16S rRNA基因序列相似性介于93.0-98.5%之间, 为潜在芽胞杆菌新种。349株芽胞杆菌属于9属66种, 分别为芽胞杆菌属(Bacillus)的41个种(41/66, 62.1%), 为最优势属; 类芽胞杆菌属(Paenibacillus)有9种(9/66, 13.6%), 赖氨酸芽胞杆菌属(Lysinibacillus)共6种(6/66, 9.1%), 嗜冷芽胞杆菌属(Pscychrobacillus)共4种(4/66, 6.1%), 虚构芽胞杆菌属(Fictibacillus)有2种(2/66, 3.0%), 短芽胞杆菌属(Brevibacillus)、大洋芽胞杆菌属(Oceanobacillus)、鲁梅尔芽胞杆菌属(Rummelibacillus)和解硫胺素芽胞杆菌属(Aneurinibacillus)皆为1种。新疆9个地点马铃薯根际土壤中芽胞杆菌含量为2.20-8.86 × 10⁴ cfu/g, 其中特克斯蒙古乡马铃薯根际土壤中芽胞杆菌的菌落含量和种类最多, 分别为8.86 × 10⁴ cfu/g和37种; 尼勒克县马场马铃薯根际土壤中的芽胞杆菌含量最少, 特克斯去昭苏路上和阜康西沟村的芽胞杆菌种类最少, 仅9种。但每个地点的优势种均相同, 均为简单芽胞杆菌(Bacillus simplex)和阿氏芽胞杆菌(B. aryabhattai)。芽胞杆菌种类可划分为高频度分布型(简单芽胞杆菌等4种)、中频度分布型(植物内生芽胞杆菌等12种)和低频度分布型(其余51种芽胞杆菌, 如蕈状芽胞杆菌)。新疆9个地点芽胞杆菌种类分布可分为高含量高丰度型(特克斯蒙古乡和农技推广园)和低含量低丰度型(如尼勒克县马场等地)。相关性分析发现, 马铃薯根际芽胞杆菌种类分布与海拔高度无显著相关性。新疆马铃薯根际土壤蕴含种类丰富的芽胞杆菌资源, 可为芽胞杆菌功能菌株开发提供丰富的菌种来源。

关键词: 芽胞杆菌, 马铃薯根际土壤, 多样性

Abstract:

This study aims to understand the diversity of Bacillus-like species found in potato rhizosphere soils in Xinjiang, and provide a basis for mining new resources. Bacillus-like bacteria were isolated from potato rhizosphere soils using the cultivable method and preliminarily identified based on the 16S rRNA gene sequences. The spatial distribution pattern of the Bacillus-like bacteria was analyzed using frequency distribution and number. A total of 349 Bacillus-like isolates were obtained from 30 soil samples, and identified as 66 species belonging to 9 genera and 2 families (Bacillaceae and Paenibacillaceae) within Bacilli by 16S rRNA gene sequences. Sixty six selective strains were chosen to be further analyzed based on colony morphology and 16S rRNA gene sequences, and 14 strains were potential novel species based on the lower similarities found between 93.0-98.5% in their closed references type strains. The species numbers of each genus were 41 for Bacillus (41/66, 62.1%), 9 for Paenibacillus (9/66, 13.6%), 6 for Lysinibacillus (6/66, 9.1%), 4 for Pscychrobacillus (4/66, 6.1%), 2 for Fictibacillus (2/66, 3.0%), and 1 for Brevibacillus, Oceanibacillus, Rummelibacillus and Aneurinibacillus, respectively. In the potato rhizosphere soils, the content range of Bacillus-like species were 2.20-8.86×10⁴ cfu/g. The highest colony content and Bacillus-like species, e.g. 8.86×10⁴ cfu/g and 37 species, respectively, occurred in the Mongolian township of Tekes, while the lowest colony content was found in a stud-farm in Nilka County and the lowest Bacillus-like species (9 species) were found in Xigou Village of Fukang City and on the way from Tekes to Zhaosu County. There was large difference in the distribution of Bacillus-like species in the potato rhizosphere soils, but each site was characterized by the same dominant species, Bacillus simplex and Bacillus aryabhattai. Based on their occurrence frequency, the Bacillus-like species could be discriminated as high frequency species including Bacillus simplex, middle frequency species including Bacillus endophyticus, and low frequency species including Bacillus mycoides. Based on the number of Bacillus-like species, 9 collecting sites were divided into two groups, namely high content and richness and low content and richness. Moreover, there was no significant correlation between the distribution of Bacillus-like species and altitude. Thus, there were many Bacillus-like species in potato rhizosphere soils, which provide rich resources for exploring functional strains.

Key words: Bacillus species, potato rhizosphere soil, diversity

刘国红, 刘波, 车建美, 陈倩倩, 林乃铨, 崔卫东. 新疆伊犁马铃薯根际芽胞杆菌纯培养多样性[J]. 生物多样性, 2017, 25(8): 856-863.

Guohong Liu, Bo Liu, Jianmei Che, Qianqian Chen, Naiquan Lin, Weidong Cui. Diversity of Bacillus-like species isolated from potato rhizosphere soils in Yili, Xinjiang[J]. Biodiv Sci, 2017, 25(8): 856-863.

图/表 4

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编号 No.	采集地点 Collection site	海拔 Altitude (m)	经纬度 Location
S1	尼勒克县尼勒克镇公社种子队 Seed Team of Nilka Township, Nilka County	1,138	43°46°28? N, 82°35°01? E
S2	阜康市上户沟乡西沟村 Xigou Village, Shanghugou Township, Fukang City	1,261	44°00°17? N, 88°33°46? E
S3	尼勒克县马场 Machang, Nilka County	1,139	43°45°58? N, 82°34°43? E
S4	尼勒克县沙勒漫乡 Shaleman Township, Nilka County	1,110	43°47°08? N, 82°32°15? E
S5	昭苏县农业局试验场 Testing Ground of Agricultural Bureau, Zhaosu County	1,856	44°09'46? N, 81°09°14? E
S6	特克斯蒙古乡(施药) Mongolia Township, Tekes County (pesticide applied)	1,171	43°13°25? N, 81°54°27? E
S7	特克斯蒙古乡 Mongolia Township, Tekes County	1,181	43°13°17? N, 81°52°33? E
S8	特克斯去昭苏路上 On the way from Tekes to Zhaosu County	1,808	-
S9	特克斯县农业技术推广园 Agro Technical Extension Garden, Tekes County	1,181	43°13°17? N, 81°52°33? E

编号 No.	采集地点 Collection site	海拔 Altitude (m)	经纬度 Location
S1	尼勒克县尼勒克镇公社种子队 Seed Team of Nilka Township, Nilka County	1,138	43°46°28? N, 82°35°01? E
S2	阜康市上户沟乡西沟村 Xigou Village, Shanghugou Township, Fukang City	1,261	44°00°17? N, 88°33°46? E
S3	尼勒克县马场 Machang, Nilka County	1,139	43°45°58? N, 82°34°43? E
S4	尼勒克县沙勒漫乡 Shaleman Township, Nilka County	1,110	43°47°08? N, 82°32°15? E
S5	昭苏县农业局试验场 Testing Ground of Agricultural Bureau, Zhaosu County	1,856	44°09'46? N, 81°09°14? E
S6	特克斯蒙古乡(施药) Mongolia Township, Tekes County (pesticide applied)	1,171	43°13°25? N, 81°54°27? E
S7	特克斯蒙古乡 Mongolia Township, Tekes County	1,181	43°13°17? N, 81°52°33? E
S8	特克斯去昭苏路上 On the way from Tekes to Zhaosu County	1,808	-
S9	特克斯县农业技术推广园 Agro Technical Extension Garden, Tekes County	1,181	43°13°17? N, 81°52°33? E

样本编号 Sample no.	芽胞杆菌含量 Bacillus colonies content (×10⁴ cfu/g)	种类数 Species number
S1	3.25	19
S2	5.00	9
S3	2.20	11
S4	6.60	15
S5	3.97	15
S6	7.70	15
S7	8.86	37
S8	4.30	9
S9	3.76	31

样本编号 Sample no.	芽胞杆菌含量 Bacillus colonies content (×10⁴ cfu/g)	种类数 Species number
S1	3.25	19
S2	5.00	9
S3	2.20	11
S4	6.60	15
S5	3.97	15
S6	7.70	15
S7	8.86	37
S8	4.30	9
S9	3.76	31