Next Article in Journal
Polychaete Diversity and Functional Trait Composition in Subtropical Mangrove Ecosystems
Previous Article in Journal
Effect of Long-Term Burning and Mowing Regimes on Ant Communities in a Mesic Grassland
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Diversity
Volume 15
Issue 9
10.3390/d15090997
diversity-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Diversity and Distribution Characteristics of Culturable Bacteria in Burqin Glacier No. 18, Altay Mountains, China

by
Mao Tian
1,2,3,
Puchao Jia
1,2,4,
Yujie Wu
1,2,4,
Xue Yu
2,3,
Shiyu Wu
1,2,3,
Ling Yang
5,
Binglin Zhang
1,2,3,
Feiteng Wang
1,2,
Guangxiu Liu
1,2,4,
Tuo Chen
1,2,3,* and
Wei Zhang
1,2,4,*
1
State Key Laboratory of Cryospheric Sciences, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China
2
University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, China
3
Key Laboratory of Extreme Environmental Microbial Resources and Engineering, Lanzhou 730000, China
4
Key Laboratory of Desert and Desertification, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China
5
School of Biological and Pharmaceutical Engineering, Lanzhou Jiaotong University, Lanzhou 730070, China
*
Authors to whom correspondence should be addressed.
Diversity 2023, 15(9), 997; https://doi.org/10.3390/d15090997
Submission received: 30 July 2023 / Revised: 27 August 2023 / Accepted: 5 September 2023 / Published: 7 September 2023
Browse Figures
Review Reports Versions Notes

Abstract

:
Ecosystems away from human disturbance provide an ideal paradigm for microbial ecology research. The Burqin glacier No. 18 in the Chinese Altay Mountains is such an ecosystem; however, there are no prior studies on the microbiology in the area. Here, we isolated 902 bacterial strains on the Burqin glacier No. 18 to determine the diversity and distribution characteristics of microorganisms. Isolated strains belonged to six phyla (in the order of dominance: Proteobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, and Deinococcus-Thermus) and 90 genera. Our results also demonstrated the presence of a high proportion of potential new species (43%) in the Burqin glacier No. 18, and 67% of the potential new species were isolated at 25 °C. Species diversity varied among habitats, with the lowest diversity in surface ice and the highest diversity in the soil farthest from the glacier terminus. The pigmented colonies made up 52.7% of all isolates, with yellow-colored colonies being the most abundant (18.8%). This study indicates that the Burqin glacier No. 18 hosts rich bacterial strain diversity, and may represent a significant potential source of new functional and pigmented bacteria for the development of critical pharmaceuticals.

1. Introduction

Mountain glaciers are widely distributed around the world, forming where snowfall accumulation exceeds seasonal melting [1]. Mountain glacier ecosystems are extreme environments characterized by low temperatures, limited oxygen, and minimal nutrition [2]. Existing research indicates that all microorganisms can survive in glacier environments, including bacteria, fungi, archaea, and viruses [3,4].
Glacial microorganisms participate in important biogeochemical cycles, have a significant impact on the global carbon cycle, and play an important ecological role in their habitats [2]. Microorganisms released during glacier melting with climate change will have a profound impact on the microbial diversity and ecological functions of surrounding and downstream ecosystems [5].
Mountain glaciers are ecosystems with diverse habitats and high habitat heterogeneity; habitats colonized by microorganisms include ice, snow, meltwater runoff, and soil [1]. Based on the vertical stratification characteristics, spatial location, environmental characteristics, and nutrient types of the colonizing microorganisms, mountain glacier ecosystems are divided into five ecological zones: sunlight-penetrated supraglacial zone, englacial zone, subglacial zone, proglacial streams, and glacier foreland [4,6].
Previous studies on culturable bacteria based on 16S rRNA showed that soil microorganisms in the foreland of Tianshan No. 1 glacier [7] belonged to Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Actinobacteria, Bacteroides, and Deinococcus-Thermus. Glacier microorganisms isolated from the Antarctic [8], Alaska [9], and Qinghai-Tibet Plateau [10,11] belonged to Betaproteobacteria, Gammaproteobacteria, Alphaproteobacteria, Actinobacteria, and Baceroidetes. Current studies on glacier microorganisms are mostly focused on a single ecological zone [12,13], and few studies have compared microbial communities from different ecological zones and different habitats. However, such studies will help to better understand microbial life and succession processes, and their ecological effects in extreme environments.
Existing studies of glacier microorganisms in China have focused primarily on the Tianshan Mountains [7], Qilian Mountains [14], and the Tibetan Plateau [15,16]. To the best of our knowledge, there is no prior research on glacier microorganisms in the Chinese Altay Mountains. In order to fill this knowledge gap, we determined the community structure and distribution characteristics of culturable microorganisms in different habitats of different ecological zones of the Burqin glacier No. 18. This study provides a comprehensive overview of bacterial diversity in the Burqin glacier No. 18, a large number of new and pigmented strain sources, and lays the foundation for further studies of microbial function and natural pigment isolation.

2. Materials and Methods

2.1. Sample Collection

Sampling was conducted on the Burqin glacier No. 18 (49°4′41″ N, 87°27′28″ E), which is a northeast-oriented cirque glacier located on the southern side of Altay Mountains, Xinjiang Province, China (Figure 1). The glacier covers an area of 1.17 km2 and extends in elevation from 2589 to 3273 m [17]. All samples were collected in July 2021. The distance between the three sampling sites was approximately 50–60 m. In each locale, three independent samples (1–2 m distance) with 5–6 L volume from each sampling site were collected. Surface snow (BS), surface ice (BI), deep ice (BDI), and surface meltwater (BW) samples were collected at altitudes of 2750, 2680, 2680, and 2600 m, respectively. BS and BI samples were collected from 0–10 cm below the surface of the glacier, and BDI samples were collected at 2 m below ice surface with a sterile ice pick; these samples were packed into sterile Whirl-Pak bags (Nasco, Salida, CA, USA) after collection. BW samples were collected using 500 mL sterile plastic sampling bottles. Three types of soil were collected in the glacier foreland: 30 m from the glacier terminus (BFS1), 130 m from the glacier terminus (BFS2), and 230 m from the glacier terminus (BFS3); surface soil samples (0–5 cm depth) were placed into separate sterile Whirl-Pak bags (Nasco, Salida, CA, USA). All samples were collected in three glacier ecological zones; BS, BI, and BW were collected in the supraglacial zone, BDI was from the englacial zone, and BFS1, BFS2, and BFS3 were located in the glacier foreland zone. The different types of samples from Burqin glacier No. 18 corresponded to different habitats. As noted, they were transported frozen in insulated containers to the State Key Laboratory of Cryosphere Science in Lanzhou, China, for storage at −20 °C prior to laboratory analyses. The samples were slowly thawed at 4 °C on an ultraclean bench. Sterile gloves, laboratory coats, and hair covers were worn during all handling procedures.

2.2. Physicochemical Analysis of Samples

All samples were analyzed for pH, electrical conductivity (EC), total organic carbon (TOC), inorganic carbon (IC), total nitrogen (TN), and K+, Ca2+, Na+, Mg2+, Cl−, and SO42−. The pH was measured using a pH meter (PT-10, Sartorius, Göttingen, Germany) and EC was measured using a conductivity meter (DDSJ-308A, Leici, Shanghai, China) in glacial foreland soil samples using fresh soil and water at a ratio of 1:5 (w/v). The contents of TOC, IC, and TN were measured with an element analyzer (Elementar Vario-EL, Langenselbold, Hessen, Germany). The ion concentration was determined with atomic absorption spectrometry (AAS) (Thermo Fisher Scientific, Waltham, MA, USA).

2.3. Bacteria Isolation and Cultivation

Five grams of glacier foreland soil samples was mixed in a 50 mL sterile plastic flask with 25 mL sterile saline solution and sterile glass beads. Flasks with samples were first fully shaken and mixed with an oscillator, and then shaken in an orbital shaker at 200 rpm at 25 °C. Then, serial dilutions of the suspensions were prepared, and 200 µL of the 10−1–10−4 dilutions was placed on Petri dishes containing Reasoner’s 2A (R2A), peptone–yeast–glucose–vitamin agar (PYGV), Trypticase Soy Broth agar (TSB), 0.1 × Trypticase Soy Broth agar (0.1 × TSB), Luria–Bertani agar (LB), and 0.1 × Luria–Bertani agar (0.1 × LB) medium for the isolation and enumeration of culturable bacteria. The formulation of the media is shown in Table S1. R2A, PYGV, 0.1 × TSB, and 0.1 × LB were oligotrophic media; TSB and LB were eutrophic media.
Surface ice, surface snow, deep ice, and glacier meltwater do not need dilution and were placed directly on the medium. Each sample was cultured at least three times to meet the needs of statistical analysis. The inoculated Petri dishes were sealed with parafilm membrane, and the uninoculated Petri dishes were used as negative blanks. The cultures were inverted and placed at 10 and 25 °C in an incubator for 3–4 weeks. After bacterial colonies were formed, the colonies on each Petri dish were counted and purified with the quadrant streak plate method. Purified strains were stored in a −80 °C freezer and supplemented with 20% (v/v) glycerol.

2.4. 16S rRNA Gene Analysis

The genomic DNA of the strain was extracted using the Bacterial DNA Extraction Kit (Omega Bio-Tek, Norcross, GA, USA) according to the manufacturer’s instructions. 16S rRNA gene was amplified using bacterial universal primers 27F and 1492R (10 µM) [18]. PCR was carried out in a final volume of 25 µL using 12.5 µL Green Taq Mix (Vazyme), 10.5 µL double-distilled water, 1 µL of each primer, and 1 µL template DNA. The PCR reaction cycling conditions were as follows: initial denaturation at 94 °C for 5 min, followed by 30 cycles of denaturation at 95 °C for 30 s, annealing at 55 °C for 30 s, elongation at 72 °C for 45 s, and final elongation step at 72 °C for 10 min. The PCR was confirmed successful by using 2% agarose gel electrophoresis and PCR products were sent to Beijing Qinke Biotechnology Co., Ltd. Xi’an Branch (Xi’an, China), for sequencing with an Applied Biosystems 3730 XL sequencer. The bacterial 16S rRNA gene sequence obtained by sequencing was determined with an EzTaxon-e server [19]. The thresholds of 98.7 and 97% for 16S rRNA gene similarity were used to assess whether the isolates were potential new species [20].

2.5. Data Analysis and Visualization

The map of sampling sites on the Burqin glacier No. 18 was drawn using ArcGIS version 10.5. One-way analysis of variance (ANOVA) was performed using SPSS 27.0. Venn diagrams of shared and exclusive taxa were drawn by TBtools version 1.108 [21]. Origin 2021 was used for drawing other graphics.

2.6. Nucleotide Sequence Accession Numbers

All sequence data in this study were submitted to the GenBank database, and the sequence accession number is OQ646887-OQ647788.

3. Results

3.1. Physicochemical Properties Associated with Different Glacier Habitats

The physicochemical characteristics of the different habitats varied significantly (Table S2). The pH ranged from 5.86 to 6.43 across habitats, indicating slight acidity, with BFS1 exhibiting the highest pH. Conductivity varied widely among habitats, ranging from 1.55 to 14.41 µS/cm, with BDI exhibiting the lowest conductivity. Concentration of K+ ions was lowest in samples collected from BW (14.42 μg/L), and highest in samples from other habitats at 38.8 to 1391 μg/L. Concentration of Cl− ions differed significantly among habitats. Concentration of TOC ranged from 0.99 to 1337.37 ppm. Concentrations of TN exhibited no significant differences between BS, BI, BW, and BDI.

3.2. Bacterial Numbers

The number of culturable bacteria ranged among different habitats from 3.48 × 102 to 6.83 × 105 CFU mL−1/g−1 at 25 °C and 2.25 × 102 to 1.65 × 105 CFU mL−1/g−1 at 10 °C (Figure 2). The number of culturable bacteria in BFS in the glacier foreland zone was greater than that in BS, BI, and BW in the supraglacial zone and BDI in the englacial zone. In R2A and 0.1 × TSB media, the number of culturable bacteria in the glacier foreland was significantly different from that in the englacial zone at both culture temperatures, but the numbers did not differ significantly among supraglacial zone habitats.

3.3. Bacterial Community Analysis of Different Culture Temperatures

The 902 isolates were classified into 6 phyla, 11 classes, 30 orders, 48 families, and 90 genera. The similarity range of 16S rRNA sequence was 82.3 to 100% (Table S3). The strains were affiliated with Proteobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, and Deinococcus-Thermus. At the genus level, Pseudomonas, Sphingomonas, Streptomyces, Massilia, Janthinobacterium, and Rhodococcus were the dominant genera. In addition, 3.2% of the genera had only one strain.
Differences in the composition of culturable bacteria among the habitats at the phylum level at 10 °C are shown in Figure 3a. Proteobacteria was the most abundant phylum (49.3%), followed by Actinobacteria (44.3%), Bacteroidetes (3.6%), Cyanobacteria (2.2%), and Deinococcus-Thermus (0.3%) and Firmicutes (0.28%). Bacteria abundance varied greatly and significantly among the zones. The glacier foreland zone contained all six phyla, and Actinobacteria was dominant (58.5–62.3%). The supraglacial and englacial zones contained only Proteobacteria, Actinobacteria, and Bacteroidetes, and the dominant phylum was Proteobacteria (70.2–96.4%). Among the less abundant phyla, Cyanobacteria was only present in BFS1; Firmicutes, and Deinococcus-Thermus were only present in BFS3, and Bacteroidetes was present in all zones except surface snow.
Differences in the composition of culturable bacteria among the habitats at the phylum level at 25 °C are shown in Figure 3b. Actinobacteria was the most abundant phylum (48.1%), followed by Proteobacteria (44.8%), Bacteroidetes (3.0%), Firmicutes (1.5%), Deinococcus-Thermus (1.5%), and Cyanobacteria (1.3%). The predominant phylum was Actinobacteria (37.4–64.6%) in the glacier foreland zone and Proteobacteria in the supraglacial and englacial zones (66.7–95.5%); these results were similar to those at 10 °C. Furthermore, only Proteobacteria and Bacteroidetes were present in BS and BI. In contrast to the results at 10 °C, Cyanobacteria, Deinococcus-Thermus, and Firmicutes were present in BFS1 and BFS2, and only BFS1 and BFS2 contained all six phyla. Actinobacteria was the dominant bacteria in the glacier foreland zone, and Proteobacteria was the dominant phylum in the supraglacial and englacial zones at 10 and 25 °C.
Composition of culturable bacteria at the genus level at 10 °C revealed that the isolates belonged to 46 genera (Figure 4a). Cryobacterium was the dominant genus (10.6%), followed by Pseudomonas (9.8%), Sphingomonas (8.9%), Streptomyces (7.2%), and Massilia (7.0%). Additionally, richness varied greatly among the habitats, with BFS3 exhibiting the highest species richness and BI the lowest; species richness of deep ice was higher than that of surface ice. In addition, dominant genera differed in almost every habitat, with Streptomyces (23.5%) in BFS3, Sphingomonas (23.1%) in BFS2, Cryobacterium (35.1%) in BFS1, Pseudomonas (29.8%) in BW, Massilia (40.9%) in BDI, Pseudomonas in BI (68.2%), and Janthinobacterium in BS (42.9%).
Culturable bacteria composition at 25 °C revealed that a total of 543 isolates belonged to 77 genera (Figure 4b). Pseudomonas was a dominant genus in all habitats (7.2%), followed by Massilia (5.3%), Mycolicibacterium (5.2%), and Streptomyces (5.2%). In addition, dominant genera on the surface of the supraglacial and englacial zones were consistent with those at 10 °C, with Pseudomonas being the dominant genus in BI (68.2%) and BW (23.3%), Janthinobacterium (61.1%) in BS, and Massilia (52.4%) in BDI. Dominant genera of the glacier foreland zone habitats were completely different at 25 °C than at 10 °C, with Mycolicibacterium (8.2%) and Nocardioides (8.2%) as the dominant genera in BFS3, and Streptomyces (9.9%) and Noviherbaspirillum (15.7%), respectively, in BFS2 and BFS1.

3.4. Potential New Species

The distribution of isolated strains differed among habitats at different temperatures (Figure 5). Among all isolates, those from BFS3 were the most abundant, followed by those from BFS2, BFS1, and BW. In contrast, the total number of isolates from BS, BI, and BDI was slightly more than that from BW, and these differences were small. The number of potential new species in isolates from BS and BDI was greater, while that from other habitats was lower at 10 °C than at 25 °C.
There were 389 strains with 16S rRNA gene sequence similarity of less than 98.7% and 55 strains with 16S rRNA gene sequence similarity of less than 97%, respectively, accounting for 43.1 and 6.1% of the total isolates (Figure 5). Among them, the 16S rRNA gene sequence similarity of potential new species from BDI and BFS2 was more than 50%. Strains with 16S rRNA gene sequence similarity of less than 97% originated mainly from BFS1, BFS2, and BFS3, that is, soil in the glacier foreland zone.

3.5. Venn Diagram of Shared and Exclusive Taxa

The co-isolation of genera in different habitats was compared via Venn diagram representations (Figure 6 and Table S4). BW, BFS1, BFS2, and BFS3 had 4, 8, 7, and 29 unique genera, respectively, while BS, BI, and BDI had no unique genera. In addition, there were 11 genera that existed in at least four habitats, namely, Arthrobacter, Cryobacterium, Janthinobacterium, Massilia, Mucilaginibacter, Nocardioides, Pseudarthrobacter, Pseudomonas, Sphingomonas, Subtercola, and Variovorax; it is worth noting that Massilia was obtained from all seven habitats. Moreover, the genera shared by three glacial zones were Cryobacterium, Massilia, Mucilaginibacter, Janthinobacterium, and Subtercola, indicating that they remain even after glacial melting. Seventy-one genera (78.9%) originated from the glacier foreland zone alone, indicating that the glacier foreland zone is a rich microbial resource zone in the glacier ecosystem.

3.6. Isolation Efficiency of Bacterial Taxa with Different Conditions

A total of 543 isolates (60.20%) were obtained from the six media used in this study at 25 °C, while only 359 isolates were obtained at 10 °C. The number of isolates at 25 °C was greater than that at 10 °C in all but the TSB medium and the number of isolates in the R2A medium at both culture temperatures exceeded 100 (Figure S1). There were 33 shared genera between both incubation temperatures, 13 unique genera at 10 °C and 44 unique genera at 25 °C (Table S5).
The diversity and abundance of genera isolated from oligotrophic media were greater than those isolated from eutrophic media (Figure 7). Eleven genera were isolated from the six media: Agreia, Conyzicola, Cryobacterium, Janthinobacterium, Mucilaginibacter, Pseudarthrobacter, Pseudomonas, Rhodococcus, Sphingomona, Streptomyces, and Subtercol. Pseudomonas was dominant in TSB and PYGV, while Massilia (8.1%) was dominant in R2A, Rhodococcus (10.1%) in 0.1 × TSB, Sphingomonas (8.6%) in 0.1 × LB, and Streptomyces (19.4%) in LB.
The diversity of bacteria was analyzed for the culture medium for isolation of the genus; 61.1% of the genera were isolated from at least two media, while 10 genera were isolated from only one of the media (Table S6). In addition, the oligotrophic media R2A, PYGV, 0.1 × TSB, and 0.1 × LB co-cultured 12 genera of isolates, and 17 isolates were unique to the oligotrophic medium R2A, indicating that the oligotrophic medium was relatively better at cultivating the glacier microorganisms (Table S6).

3.7. Variation of Pigmented Colonies in Different Habitats

Based on their color, pigmented strains were found in 52.7%, and these strains were divided into eight groups (yellow, black, white, purple, pink, green, orange, and brown) (Figure 8). Among all pigmented isolates, yellow colonies were dominant and made up 18.8% of all colonies, followed by orange (17.0%), pink (12.2%), brown (2.3%), green (1.7%), black (0.6%), and purple (0.1%). The percentage of pigmented strains was lowest in surface ice (BS), and much higher in deep ice (BDI). The only purple-pigmented strain, Janthinobacterium rivuli, was isolated from surface snow (BS). There were at least five types of pigmented strains in glacier foreland soil, and the farther from the glacier terminus, the lower the percentage of pigment production.

4. Discussion

In this study, we separated and identified 902 isolates from the Burqin glacier No. 18, and classified them into six phyla and 90 genera. Proteobacteria (46.7%) and Actinobacteria (46.6%) accounted for 93.3% of the isolates. Previous studies have shown that the bacteria isolated from glacier ecosystems worldwide belonged to the phyla Actinobacteria, Proteobacteria, Firmicutes, Bacteroidetes, and Deinococcus-Thermus [10,22]. We were able to isolate the above five phyla from the glacier environment, and, additionally, autotrophic cyanobacteria.
The Burqin glacier No. 18 has fostered a large source of potential new species of microorganisms. In this study, 43.1% of the isolates had 16S rRNA sequence similarity values of less than 98.7%, and 6.1% of less than 97%. In a previous study in the Gurbantunggut Desert [23], potential novel species with the 16S rRNA gene sequence similarity values of <98.7% accounted for 25.2%, and those with similarity values of <97% accounted for 5.5%. In comparison, the proportions of potential new species with 16S rRNA similarity value less than 98.7 and 97% were 10.6 and 5.5%, respectively, in Europe’s Tabernas Desert [24]. This highlights the fact that the Burqin glacier No. 18 is a treasure trove of potential new species.
Thirty-four of the ninety identified genera, representing 37.8% of the diversity at the genus level, were found in one medium alone. Furthermore, the percentage of genera isolated exclusively from the other six media used ranged from 1 to 19%. This result indicates that diversity is highly dependent on the use of multiple media, which supports the idea that media combinations and diluting can increase the discovery of different and new microbial taxa [24]. The isolation efficiency of the oligonutrient medium was higher than that of eutrophic medium in this study, indicating that glacial microorganisms were more adapted to a low nutrient environment.
Habitats in different ecological zones of the glacier exhibited differences in species abundance. The phylum Proteobacteria was dominant in the supraglacial and the englacial zones, while Actinobacteria was dominant in the glacier foreland zone. The dominant genera were Pseudomonas and Janthinobacterium in the supraglacial zone, Massilia and Janthinobacterium in the englacial zone, and Streptomyces and Sphingomonas in the glacier foreland zone. The dominant bacterial phyla isolated from the Burqin glacier No. 18 foreland were similar to those from the Tianshan No. 1 glacier foreland [7], Dongkemadi glacier foreland [25], and Himalayan Pindari glacier foreland [26], indicating that culturable bacteria in glacial foreland soil may be less affected by geographical and environmental factors. The number of culturable strains in glacier foreland habitats increased with the distance from the glacier terminus, and this result was consistent with the study by Cazzolla [27] and due to the strong ecosystem developmental dynamics of glacier foreland soils with the characteristics of primary succession [28].
The bacterial dominant phyla isolated from the supraglacial and englacial zone were different from those found in Pakistan’s Siachen Glacier [29], Yuzhufeng Glacier ice core [11], and Qinghai–Tibet Plateau ice core [10], indicating that the culturable bacteria in glaciers may be greatly affected by local environments and bacteria sources [11]. Microbial diversity in deep ice in this study was higher than that in surface ice, which may be due to differences in microbial strategies for fulfilling their energy requirements in different habitats [4]. The supraglacial zone microorganisms were mainly derived from aeolian deposition, in which the microorganisms originating from aerosols, dust, and precipitation events directly determined composition and quantity of ice and snow microorganisms [30]. In addition, harsh environments such as strong radiation, low temperature, and low oxygen concentration and nutrients can also affect the supraglacial bacterial community structure and diversity [31]. Microorganisms within the englacial zone were less connected to atmospheric processes, and nutrients they need are transported through liquid water in ice cracks [4]. A total of 90 genera were isolated in this study from different habitats in the Burqin glacier No. 18, more than the number of genera in other extreme environments [23,24]; this indicates that the culturomics method is critical in glacial microbial research.
In this study, cyanobacteria were isolated only from the glacier foreland zone. Although cyanobacteria constitute a relatively small part of microbial communities in glacial ecosystems, they are an integral part [32,33]. Cyanobacteria are major primary producers in polar and mountain glacier ecosystems and contribute significantly to the cryosphere’s nitrogen and carbon cycles [34]. These autotrophic microorganisms sustain their own life activities through photosynthesis while the chemoautotrophic and heterotrophic microorganisms utilize the accumulated carbon [4,35]. Some cyanobacteria can tolerate and survive in low-temperature, arid, and UV-exposed environments [36]; cyanobacteria isolated from the Burqin glacier No. 18 provided an excellent resource for future research on low-temperature adaptability, drought resistance, and radiation resistance of Cyanobacteria.
Actinobacteria are abundant in extreme biospheres and exhibit an excellent ability to produce various enzymes and secondary metabolites to combat harsh conditions [37,38]. Actinobacteria are a rich source of novel important bioactive metabolites for pharmaceutical utilization, and the discovery of beneficial bioactive compounds from extreme-environment Actinobacteria can help address antibiotic resistance [38]. Surprisingly, two rare strains of Actinobacteria, Microbacterium sp. and Kribbella sp., were identified from the glacier foreland of the Burqin glacier No. 18 in this study, and Kribbella sp. was a potential new species. In particular, Microbacterium may be important in the modulation of communities through inhibition of competitors growth [28,39]. Notably, our isolated strain Streptomyces avidinii BFS3LP15 was capable of producing transparent droplets, and droplet production was used as a criterion for the description of species [40]. In addition, we also identified nine strains of four species of the Deinococcus genus in the Deinococcus-Thermus phylum, all of which were potential new species; Deinococcus exhibits strong radiation resistance. Deinococcus-Thermus phylum includes many species of thermophiles, along with those resistant to extreme radiation [41]. Discovering Deinococcus-Thermus strains in a glacier environment is conducive to further study of its UV resistance and cold adaptation mechanisms.
Concerning previous studies, the percentage of pigmented bacteria in ice cores of Zadang Glacier, Mengdagangri Glacier, and Yuzhufeng were 61, 55, and 89% [11,42]. The percentage of pigment-producing bacteria in this study was lower than in previous studies, but still dominant in the Burqin glacier No. 18. The proportion of pigmented bacteria in the supraglacial zone was much lower than that in the englacial zone, probably because bacteria in the supraglacial zone have not had sufficient time to undergo environmental selection and evolutionary mutation, and bacteria in the englacial zone have been deposited in the glacier for much longer than those in the surface layer; these results support the hypothesis that pigments play an important role in bacterial adaptation to the cold environment [43].
Purple-colored heterotrophic bacteria and autotrophic cyanobacteria were among the many pigmented bacteria isolated from the Burqin glacier No. 18. Our isolated strain, Janthinobacterium rivuli, produced a purple pigment, identified as violacein, which could be used as an anticancer or sensitizer for cisplatin drugs in cervical cancer [44]. Violacein also has anti-inflammatory and anti-viral activities and is involved in the tolerance of bacteria to UV irradiation and low temperature [45].
Cyanobacteria are a rich source of many natural pigments [46]. Cyanobacterial pigments are natural colorants with strong antioxidant and anti-mutagenic properties that play an important role not only in diseases including cancer, diabetes, and arthritis, but also as natural food colors [47,48]. Cyanobacterial pigments have promising applications in dietary supplements and cosmetics, and in pharmaceutical and textile industries [46]. Therefore, isolation and culture of pigmented microorganisms from glacial environments is of great importance for the development and application of natural pigments.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/d15090997/s1, Figure S1: Number of isolates under different culture temperatures and media; Table S1: Media used for isolation in this study; Table S2: Physicochemical properties of different glacial habitats; Table S3: Stain taxonomic information based on 16S rRNA gene; Table S4: List of coincident genera comparing all seven habitats; Table S5: List of coincident genera comparing two temperatures; Table S6: List of coincident genera comparing all six media.

Author Contributions

Conceptualization, M.T., T.C. and W.Z.; methodology, M.T. and L.Y.; formal analysis, Y.W., X.Y. and S.W.; writing—original draft preparation, M.T., P.J. and G.L.; writing—review and editing, M.T., B.Z. and F.W.; funding acquisition, B.Z. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the National Science Foundation of China, grant number 42371156, and the Third Comprehensive Scientific Expedition of Xinjiang Uyghur Autonmous Region (2022xikk0802).

Data Availability Statement

All data in this study are within the manuscript and the Supplemental Materials.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Hotaling, S.; Hood, E.; Hamilton, T.L. Microbial Ecology of Mountain Glacier Ecosystems: Biodiversity, Ecological Connections and Implications of a Warming Climate. Environ. Microbiol. 2017, 19, 2935–2948. [Google Scholar] [CrossRef]
  2. Margesin, R.; Collins, T. Microbial Ecology of the Cryosphere (Glacial and Permafrost Habitats): Current Knowledge. Appl. Microbiol. Biotechnol. 2019, 103, 2537–2549. [Google Scholar] [CrossRef]
  3. Anesio, A.M.; Laybourn-Parry, J. Glaciers and Ice Sheets as a Biome. Trends Ecol. Evol. 2012, 27, 219–225. [Google Scholar] [CrossRef]
  4. Boetius, A.; Anesio, A.M.; Deming, J.W.; Mikucki, J.A.; Rapp, J.Z. Microbial Ecology of the Cryosphere: Sea Ice and Glacial Habitats. Nat. Rev. Microbiol. 2015, 13, 677–690. [Google Scholar] [CrossRef]
  5. Cameron, K.A.; Müller, O.; Stibal, M.; Edwards, A.; Jacobsen, C.S. Glacial Microbiota Are Hydrologically Connected and Temporally Variable. Environ. Microbiol. 2020, 22, 3172–3187. [Google Scholar] [CrossRef]
  6. Rime, T.; Hartmann, M.; Frey, B. Potential Sources of Microbial Colonizers in an Initial Soil Ecosystem after Retreat of an Alpine Glacier. ISME J. 2016, 10, 1625–1641. [Google Scholar] [CrossRef]
  7. Wu, X.; Zhang, G.; Zhang, W.; Liu, G.; Chen, T.; Wang, Y.; Long, H.; Tai, X.; Zhang, B.; Li, Z. Variations in Culturable Bacterial Communities and Biochemical Properties in the Foreland of the Retreating Tianshan No. 1 Glacier. Braz. J. Microbiol. 2018, 49, 443–451. [Google Scholar] [CrossRef]
  8. Zdanowski, M.K.; Żmuda-Baranowska, M.J.; Borsuk, P.; Świątecki, A.; Górniak, D.; Wolicka, D.; Jankowska, K.M.; Grzesiak, J. Culturable Bacteria Community Development in Postglacial Soils of Ecology Glacier, King George Island, Antarctica. Polar Biol. 2013, 36, 511–527. [Google Scholar] [CrossRef]
  9. Segata, N.; Izard, J.; Waldron, L.; Gevers, D.; Miropolsky, L.; Garrett, W.S.; Huttenhower, C. Metagenomic Biomarker Discovery and Explanation. Genome Biol. 2011, 12, R60. [Google Scholar] [CrossRef]
  10. Liu, Y.; Priscu, J.C.; Yao, T.; Vick-Majors, T.J.; Michaud, A.B.; Sheng, L. Culturable Bacteria Isolated from Seven High-Altitude Ice Cores on the Tibetan Plateau. J. Glaciol. 2019, 65, 29–38. [Google Scholar] [CrossRef]
  11. Shen, L.; Liu, Y.; Wang, N.; Jiao, N.; Xu, B.; Liu, X. Variation with Depth of the Abundance, Diversity and Pigmentation of Culturable Bacteria in a Deep Ice Core from the Yuzhufeng Glacier, Tibetan Plateau. Extremophiles 2018, 22, 29–38. [Google Scholar] [CrossRef] [PubMed]
  12. Sampaio, D.S.; Almeida, J.R.B.; de Jesus, H.E.; Rosado, A.S.; Seldin, L.; Jurelevicius, D. Distribution of Anaerobic Hydrocarbon-Degrading Bacteria in Soils from King George Island, Maritime Antarctica. Microb. Ecol. 2017, 74, 810–820. [Google Scholar] [CrossRef] [PubMed]
  13. Krauze, P.; Wagner, D.; Yang, S.; Spinola, D.; Kühn, P. Influence of Prokaryotic Microorganisms on Initial Soil Formation along a Glacier Forefield on King George Island, Maritime Antarctica. Sci. Rep. 2021, 11, 13135. [Google Scholar] [CrossRef] [PubMed]
  14. Zhang, B.; Chen, T.; Guo, J.; Wu, M.; Yang, R.; Chen, X.; Wu, X.; Zhang, W.; Kang, S.; Liu, G.; et al. Microbial Mercury Methylation Profile in Terminus of a High-Elevation Glacier on the Northern Boundary of the Tibetan Plateau. Sci. Total Environ. 2020, 708, 135226. [Google Scholar] [CrossRef]
  15. Liu, Y.; Ji, M.; Yu, T.; Zaugg, J.; Anesio, A.M.; Zhang, Z.; Hu, S.; Hugenholtz, P.; Liu, K.; Liu, P.; et al. A Genome and Gene Catalog of Glacier Microbiomes. Nat. Biotechnol. 2022, 40, 1341–1348. [Google Scholar] [CrossRef]
  16. Zhong, Z.-P.; Tian, F.; Roux, S.; Gazitúa, M.C.; Solonenko, N.E.; Li, Y.-F.; Davis, M.E.; Van Etten, J.L.; Mosley-Thompson, E.; Rich, V.I.; et al. Glacier Ice Archives Nearly 15,000-Year-Old Microbes and Phages. Microbiome 2021, 9, 160. [Google Scholar] [CrossRef]
  17. Jin, S.; Li, Z.; Wang, Z.; Wang, F.; Xu, C.; Ai, S. Ice Thickness Distribution and Volume Estimation of Burqin Glacier No. 18 in the Chinese Altay Mountains. J. Arid. Land. 2020, 12, 905–916. [Google Scholar] [CrossRef]
  18. Lane, D.J. 16S/23S RRNA Sequencing. In Nucleic Acid Techniques in Bacterial Systematics; Stackebrandt, E., Goodfellow, M., Eds.; John Wiley and Sons: New York, NY, USA, 1991; pp. 125–175. [Google Scholar]
  19. Yoon, S.H.; Ha, S.-M.; Kwon, S.; Lim, J.; Kim, Y.; Seo, H.; Chun, J. Introducing EzBioCloud: A Taxonomically United Database of 16S RRNA Gene Sequences and Whole-Genome Assemblies. Int. J. Syst. Evol. Microbiol. 2017, 67, 1613–1617. [Google Scholar] [CrossRef]
  20. Chun, J.; Oren, A.; Ventosa, A.; Christensen, H.; Arahal, D.R.; da Costa, M.S.; Rooney, A.P.; Yi, H.; Xu, X.-W.; De Meyer, S.; et al. Proposed Minimal Standards for the Use of Genome Data for the Taxonomy of Prokaryotes. Int. J. Syst. Evol. Microbiol. 2018, 68, 461–466. [Google Scholar] [CrossRef]
  21. Chen, C.; Chen, H.; Zhang, Y.; Thomas, H.R.; Frank, M.H.; He, Y.; Xia, R. TBtools: An Integrative Toolkit Developed for Interactive Analyses of Big Biological Data. Mol. Plant 2020, 13, 1194–1202. [Google Scholar] [CrossRef]
  22. Franzetti, A.; Tagliaferri, I.; Gandolfi, I.; Bestetti, G.; Minora, U.; Mayer, C.; Azzoni, R.S.; Diolaiuti, G.; Smiraglia, C.; Ambrosini, R. Light-Dependent Microbial Metabolisms Drive Carbon Fluxes on Glacier Surfaces. ISME J. 2016, 10, 2984–2988. [Google Scholar] [CrossRef] [PubMed]
  23. Li, S.; Dong, L.; Lian, W.H.; Lin, Z.L.; Lu, C.-Y.; Xu, L.; Li, L.; Hozzein, W.N.; Li, W.J. Exploring Untapped Potential of Streptomyces Spp. in Gurbantunggut Desert by Use of Highly Selective Culture Strategy. Sci. Total Environ. 2021, 790, 148235. [Google Scholar] [CrossRef] [PubMed]
  24. Molina-Menor, E.; Gimeno-Valero, H.; Pascual, J.; Peretó, J.; Porcar, M. High Culturable Bacterial Diversity From a European Desert: The Tabernas Desert. Front. Microbiol. 2021, 11, 583120. [Google Scholar] [CrossRef]
  25. Liu, G.X.; Hu, P.; Zhang, W.; Wu, X.; Yang, X.; Chen, T.; Zhang, M.; Li, S.W. Variations in Soil Culturable Bacteria Communities and Biochemical Characteristics in the Dongkemadi Glacier Forefield along a Chronosequence. Folia Microbiol. 2012, 10, 485–494. [Google Scholar] [CrossRef]
  26. Shivaji, S.; Pratibha, M.S.; Sailaja, B.; Hara Kishore, K.; Singh, A.K.; Begum, Z.; Anarasi, U.; Prabagaran, S.R.; Reddy, G.S.N.; Srinivas, T.N.R. Bacterial Diversity of Soil in the Vicinity of Pindari Glacier, Himalayan Mountain Ranges, India, Using Culturable Bacteria and Soil 16S rRNA Gene Clones. Extremophiles 2011, 15, 1–22. [Google Scholar] [CrossRef] [PubMed]
  27. Cazzolla Gatti, R.; Dudko, A.; Lim, A.; Velichevskaya, A.I.; Lushchaeva, I.V.; Pivovarova, A.V.; Ventura, S.; Lumini, E.; Berruti, A.; Volkov, I.V. The Last 50 Years of Climate-induced Melting of the Maliy Aktru Glacier (Altai Mountains, Russia) Revealed in a Primary Ecological Succession. Ecol. Evol. 2018, 8, 7401–7420. [Google Scholar] [CrossRef]
  28. Yang, G.L.; Hou, S.G.; Le Baoge, R.; Li, Z.G.; Xu, H.; Liu, Y.P.; Du, W.T.; Liu, Y.Q. Differences in Bacterial Diversity and Communities Between Glacial Snow and Glacial Soil on the Chongce Ice Cap, West Kunlun Mountains. Sci. Rep. 2016, 6, 36548. [Google Scholar] [CrossRef]
  29. Rafiq, M.; Hayat, M.; Anesio, A.M.; Jamil, S.U.U.; Hassan, N.; Shah, A.A.; Hasan, F. Recovery of Metallo-Tolerant and Antibiotic Resistant Psychrophilic Bacteria from Siachen Glacier, Pakistan. PLoS ONE 2017, 12, e0178180. [Google Scholar] [CrossRef]
  30. Xiang, S.-R.; Shang, T.-C.; Chen, Y.; Yao, T.-D. Deposition and Postdeposition Mechanisms as Possible Drivers of Microbial Population Variability in Glacier Ice. FEMS Microbiol. Ecol. 2009, 70, 165–176. [Google Scholar] [CrossRef]
  31. Liu, Y.; Yao, T.; Jiao, N.; Kang, S.; Zeng, Y.; Huang, S. Microbial Community Structure in Moraine Lakes and Glacial Meltwaters, Mount Everest: Microbial Diversity in Lakes of Mount Everest. FEMS Microbiol. Lett. 2006, 265, 98–105. [Google Scholar] [CrossRef]
  32. Jungblut, A.D.; Lovejoy, C.; Vincent, W.F. Global Distribution of Cyanobacterial Ecotypes in the Cold Biosphere. ISME J. 2010, 4, 191–202. [Google Scholar] [CrossRef] [PubMed]
  33. Lutz, S.; Anesio, A.M.; Raiswell, R.; Edwards, A.; Newton, R.J.; Gill, F.; Benning, L.G. The Biogeography of Red Snow Microbiomes and Their Role in Melting Arctic Glaciers. Nat. Commun. 2016, 7, 11968. [Google Scholar] [CrossRef] [PubMed]
  34. Chrismas, N.A.M.; Anesio, A.M.; Sánchez-Baracaldo, P. Multiple Adaptations to Polar and Alpine Environments within Cyanobacteria: A Phylogenomic and Bayesian Approach. Front. Microbiol. 2015, 6, 1070. [Google Scholar] [CrossRef] [PubMed]
  35. Møller, A.K.; Søborg, D.A.; Al-Soud, W.A.; Sørensen, S.J.; Kroer, N. Bacterial Community Structure in High-Arctic Snow and Freshwater as Revealed by Pyrosequencing of 16S RRNA Genes and Cultivation. Polar Res. 2013, 32, 17390. [Google Scholar] [CrossRef]
  36. Clarke, A.; Morris, G.J.; Fonseca, F.; Murray, B.J.; Acton, E.; Price, H.C. A Low Temperature Limit for Life on Earth. PLoS ONE 2013, 8, e66207. [Google Scholar] [CrossRef]
  37. Mohammadipanah, F.; Wink, J. Actinobacteria from Arid and Desert Habitats: Diversity and Biological Activity. Front. Microbiol. 2016, 6, 1541. [Google Scholar] [CrossRef]
  38. Hui, M.L.-Y.; Tan, L.T.-H.; Letchumanan, V.; He, Y.-W.; Fang, C.-M.; Chan, K.-G.; Law, J.W.-F.; Lee, L.-H. The Extremophilic Actinobacteria: From Microbes to Medicine. Antibiotics 2021, 10, 682. [Google Scholar] [CrossRef]
  39. Zhao, P.; Xue, Y.; Gao, W.; Li, J.; Zu, X.; Fu, D.; Feng, S.; Bai, X.; Zuo, Y.; Li, P. Actinobacteria –Derived Peptide Antibiotics since 2000. Peptides 2018, 103, 48–59. [Google Scholar] [CrossRef]
  40. Schmidt, K.; Spiteller, D. Ammonia Released by Streptomyces aburaviensis Induces Droplet Formation in Streptomyces violaceoruber. J. Chem. Ecol. 2017, 43, 806–816. [Google Scholar] [CrossRef]
  41. Ho, J.; Adeolu, M.; Khadka, B.; Gupta, R.S. Identification of Distinctive Molecular Traits That Are Characteristic of the Phylum “Deinococcus-Thermus” and Distinguish Its Main Constituent Groups. Syst. Appl. Microbiol. 2016, 39, 453–463. [Google Scholar] [CrossRef]
  42. Shen, L.; Yao, T.; Liu, Y.; Jiao, N.; Kang, S.; Xu, B.; Zhang, S.; Liu, X. Downward-Shifting Temperature Range for the Growth of Snow-Bacteria on Glaciers of the Tibetan Plateau. Geomicrobiol. J. 2014, 31, 779–787. [Google Scholar] [CrossRef]
  43. Liu, Y.; Yao, T.; Jiao, N.; Kang, S.; Huang, S.; Li, Q.; Wang, K.; Liu, X. Culturable Bacteria in Glacial Meltwater at 6350 m on the East Rongbuk Glacier, Mount Everest. Extremophiles 2009, 13, 89–99. [Google Scholar] [CrossRef] [PubMed]
  44. Alem, D.; Marizcurrena, J.J.; Saravia, V.; Davyt, D.; Martinez-Lopez, W.; Castro-Sowinski, S. Production and Antiproliferative Effect of Violacein, a Purple Pigment Produced by an Antarctic Bacterial Isolate. World J. Microbiol. Biotechnol. 2020, 36, 120. [Google Scholar] [CrossRef] [PubMed]
  45. Choi, S.Y.; Yoon, K.; Lee, J.I.; Mitchell, R.J. Violacein: Properties and Production of a Versatile Bacterial Pigment. BioMed Res. Int. 2015, 2015, 1–8. [Google Scholar] [CrossRef] [PubMed]
  46. Sandybayeva, S.K.; Kossalbayev, B.D.; Zayadan, B.K.; Sadvakasova, A.K.; Bolatkhan, K.; Zadneprovskaya, E.V.; Kakimov, A.B.; Alwasel, S.; Leong, Y.K.; Allakhverdiev, S.I.; et al. Prospects of Cyanobacterial Pigment Production: Biotechnological Potential and Optimization Strategies. Biochem. Eng. J. 2022, 187, 108640. [Google Scholar] [CrossRef]
  47. Rastogi, R.P.; Singh, S.P.; Häder, D.-P.; Sinha, R.P. Detection of Reactive Oxygen Species (ROS) by the Oxidant-Sensing Probe 2′,7′-Dichlorodihydrofluorescein Diacetate in the Cyanobacterium Anabaena Variabilis PCC 7937. Biochem. Biophys. Res. Commun. 2010, 397, 603–607. [Google Scholar] [CrossRef]
  48. Manfellotto, F.; Stella, G.R.; Falciatore, A.; Brunet, C.; Ferrante, M.I. Engineering the Unicellular Alga Phaeodactylum Tricornutum for Enhancing Carotenoid Production. Antioxidants 2020, 9, 757. [Google Scholar] [CrossRef]
Diversity 15 00997 g001
Figure 1. Map showing the location of the sampling site on the Burqin glacier No. 18.
Figure 1. Map showing the location of the sampling site on the Burqin glacier No. 18.
Diversity 15 00997 g001
Diversity 15 00997 g002
Figure 2. Culturable bacteria numbers in seven glacial habitats: (a), R2A; (b), PYGV; (c), TSB; (d), 0.1 × TSB; (e), LB; (f), 0.1 × LB. Different letters indicate significant differences at different temperatures (p < 0.05).
Figure 2. Culturable bacteria numbers in seven glacial habitats: (a), R2A; (b), PYGV; (c), TSB; (d), 0.1 × TSB; (e), LB; (f), 0.1 × LB. Different letters indicate significant differences at different temperatures (p < 0.05).
Diversity 15 00997 g002
Diversity 15 00997 g003
Figure 3. Relative abundance of culturable bacteria at the phylum level in seven glacial habitats: (a) culture temperature 10 °C, (b) culture temperature 25 °C.
Figure 3. Relative abundance of culturable bacteria at the phylum level in seven glacial habitats: (a) culture temperature 10 °C, (b) culture temperature 25 °C.
Diversity 15 00997 g003
Diversity 15 00997 g004
Figure 4. Species abundance of culturable bacteria at the genus level in seven glacial habitats: (a) culture temperature 10 °C, (b) culture temperature 25 °C.
Figure 4. Species abundance of culturable bacteria at the genus level in seven glacial habitats: (a) culture temperature 10 °C, (b) culture temperature 25 °C.
Diversity 15 00997 g004
Diversity 15 00997 g005
Figure 5. Number of potential new species under different culture temperatures and habitats. 98.7% and 97% were the thresholds for 16S rRNA gene similarity.
Figure 5. Number of potential new species under different culture temperatures and habitats. 98.7% and 97% were the thresholds for 16S rRNA gene similarity.
Diversity 15 00997 g005
Diversity 15 00997 g006
Figure 6. Venn diagram of shared and exclusive number of the genus in seven glacial habitats. Black dots indicate the presence in the corresponding habitat, and gray dots indicate the absence.
Figure 6. Venn diagram of shared and exclusive number of the genus in seven glacial habitats. Black dots indicate the presence in the corresponding habitat, and gray dots indicate the absence.
Diversity 15 00997 g006
Diversity 15 00997 g007
Figure 7. Heatmap showing the abundances of 30 most abundant genera.
Figure 7. Heatmap showing the abundances of 30 most abundant genera.
Diversity 15 00997 g007
Diversity 15 00997 g008
Figure 8. Variation in proportions of pigmented colonies across glacial habitats.
Figure 8. Variation in proportions of pigmented colonies across glacial habitats.
Diversity 15 00997 g008
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Tian, M.; Jia, P.; Wu, Y.; Yu, X.; Wu, S.; Yang, L.; Zhang, B.; Wang, F.; Liu, G.; Chen, T.; et al. Diversity and Distribution Characteristics of Culturable Bacteria in Burqin Glacier No. 18, Altay Mountains, China. Diversity 2023, 15, 997. https://doi.org/10.3390/d15090997

AMA Style

Tian M, Jia P, Wu Y, Yu X, Wu S, Yang L, Zhang B, Wang F, Liu G, Chen T, et al. Diversity and Distribution Characteristics of Culturable Bacteria in Burqin Glacier No. 18, Altay Mountains, China. Diversity. 2023; 15(9):997. https://doi.org/10.3390/d15090997

Chicago/Turabian Style

Tian, Mao, Puchao Jia, Yujie Wu, Xue Yu, Shiyu Wu, Ling Yang, Binglin Zhang, Feiteng Wang, Guangxiu Liu, Tuo Chen, and et al. 2023. "Diversity and Distribution Characteristics of Culturable Bacteria in Burqin Glacier No. 18, Altay Mountains, China" Diversity 15, no. 9: 997. https://doi.org/10.3390/d15090997

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop