Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sediment
Abstract In the classical microbial isolation technique, the isolation process inevitably destroys all microbial interactions and thus makes it difficult to culture the many microorganisms that rely on these interactions for survival. In this study, we designed a simple coculture technique named the...
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| Format: | Article |
| Language: | English |
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Wiley
2024-03-01
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| Series: | mLife |
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| Online Access: | https://doi.org/10.1002/mlf2.12093 |
| _version_ | 1827301983565905920 |
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| author | Jing Zhang Qi‐Yun Liang Da‐Shuai Mu Fengbai Lian Ya Gong Mengqi Ye Guan‐Jun Chen Yuqi Ye Zong‐Jun Du |
| author_facet | Jing Zhang Qi‐Yun Liang Da‐Shuai Mu Fengbai Lian Ya Gong Mengqi Ye Guan‐Jun Chen Yuqi Ye Zong‐Jun Du |
| author_sort | Jing Zhang |
| collection | DOAJ |
| description | Abstract In the classical microbial isolation technique, the isolation process inevitably destroys all microbial interactions and thus makes it difficult to culture the many microorganisms that rely on these interactions for survival. In this study, we designed a simple coculture technique named the “sandwich agar plate method,” which maintains microbial interactions throughout the isolation and pure culture processes. The total yield of uncultured species in sandwich agar plates based on eight helper strains was almost 10‐fold that of the control group. Many uncultured species displayed commensal lifestyles. Further study found that heme was the growth‐promoting factor of some marine commensal bacteria. Subsequent genomic analysis revealed that heme auxotrophies were common in various biotopes and prevalent in many uncultured microbial taxa. Moreover, our study supported that the survival strategies of heme auxotrophy in different habitats varied considerably. These findings highlight that cocultivation based on the “sandwich agar plate method” could be developed and used to isolate more uncultured bacteria. |
| first_indexed | 2024-04-24T16:36:31Z |
| format | Article |
| id | doaj.art-e51a92da758b46b797d456810a32ca00 |
| institution | Directory Open Access Journal |
| issn | 2770-100X |
| language | English |
| last_indexed | 2024-04-24T16:36:31Z |
| publishDate | 2024-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | mLife |
| spelling | doaj.art-e51a92da758b46b797d456810a32ca002024-03-29T16:09:55ZengWileymLife2770-100X2024-03-013114315510.1002/mlf2.12093Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sedimentJing Zhang0Qi‐Yun Liang1Da‐Shuai Mu2Fengbai Lian3Ya Gong4Mengqi Ye5Guan‐Jun Chen6Yuqi Ye7Zong‐Jun Du8State Key Laboratory of Microbial Technology, Institute of Microbial Technology Shandong University Qingdao ChinaMarine College Shandong University Weihai ChinaState Key Laboratory of Microbial Technology, Institute of Microbial Technology Shandong University Qingdao ChinaMarine College Shandong University Weihai ChinaState Key Laboratory of Microbial Technology, Institute of Microbial Technology Shandong University Qingdao ChinaState Key Laboratory of Microbial Technology, Institute of Microbial Technology Shandong University Qingdao ChinaState Key Laboratory of Microbial Technology, Institute of Microbial Technology Shandong University Qingdao ChinaMarine College Shandong University Weihai ChinaState Key Laboratory of Microbial Technology, Institute of Microbial Technology Shandong University Qingdao ChinaAbstract In the classical microbial isolation technique, the isolation process inevitably destroys all microbial interactions and thus makes it difficult to culture the many microorganisms that rely on these interactions for survival. In this study, we designed a simple coculture technique named the “sandwich agar plate method,” which maintains microbial interactions throughout the isolation and pure culture processes. The total yield of uncultured species in sandwich agar plates based on eight helper strains was almost 10‐fold that of the control group. Many uncultured species displayed commensal lifestyles. Further study found that heme was the growth‐promoting factor of some marine commensal bacteria. Subsequent genomic analysis revealed that heme auxotrophies were common in various biotopes and prevalent in many uncultured microbial taxa. Moreover, our study supported that the survival strategies of heme auxotrophy in different habitats varied considerably. These findings highlight that cocultivation based on the “sandwich agar plate method” could be developed and used to isolate more uncultured bacteria.https://doi.org/10.1002/mlf2.12093cultivationgrowth factorshemesandwich agar plateuncultured bacteria |
| spellingShingle | Jing Zhang Qi‐Yun Liang Da‐Shuai Mu Fengbai Lian Ya Gong Mengqi Ye Guan‐Jun Chen Yuqi Ye Zong‐Jun Du Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sediment mLife cultivation growth factors heme sandwich agar plate uncultured bacteria |
| title | Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sediment |
| title_full | Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sediment |
| title_fullStr | Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sediment |
| title_full_unstemmed | Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sediment |
| title_short | Cultivating the uncultured: Harnessing the “sandwich agar plate” approach to isolate heme‐dependent bacteria from marine sediment |
| title_sort | cultivating the uncultured harnessing the sandwich agar plate approach to isolate heme dependent bacteria from marine sediment |
| topic | cultivation growth factors heme sandwich agar plate uncultured bacteria |
| url | https://doi.org/10.1002/mlf2.12093 |
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