Natural transformation of the filamentous cyanobacterium <i>Phormidium lacuna</i>

Nies, Fabian; Mielke, Marion; Pochert, Janko; Lamparter, Tilman

doi:10.1101/870006

Cited by 2 publications

(4 citation statements)

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“…TP1 was adapted by a previously described natural transformation protocol for the filamentous, non-heterocystous cyanobacterium Phormidium lacuna HE10DO (9). 25 ml Chlorogloeopsis cultures grown in BG11 at 37 °C were sonicated for min at maximum intensity using Elmasonic S 60 ultrasound unit (Elma Schmidbauser GmbH, Singen, Germany) to lose cell aggregates and generate a homogenous cell suspension.…”

Section: Natural Transformation Of Cyanobacteriamentioning

confidence: 99%

“…The role of the here-listed genes in natural competence was recently validated in Synechococcus elongatus PCC (17). The distribution of genes essential for natural competence in cyanobacterial genomes suggest that natural competence is a prevalent trait within the phylum (9,17,18).…”

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confidence: 91%

“…To validate the computational prediction we tested for the presence of natural competence in Chlorogloeopsis using a previously developed natural transformation protocol (9). Given the low homologous recombination efficiency in Chlorogloeopsis spp.…”

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Natural competence inChlorogloeopsis fritschiiPCC 6912 and other ramified cyanobacteria

Springstein

Nies

2020

Preprint

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17Lateral DNA transfer plays an important role in the evolution of genetic diversity in 18 prokaryotes. DNA acquisition via transformation involves the uptake of DNA from the 19 environment. The ability of recipient cells to actively transport DNA into the cytoplasm -20 termed natural competencedepends on the presence of type IV pili and competence 21 proteins. Natural competence has been described in cyanobacteria for several organisms 22 including unicellular and filamentous species. However, the presence of natural competence 23 in ramified cyanobacteria, which are considered the peak of cyanobacterial morphological 24 complexity, remains unknown. Here we show that ramified cyanobacteria harbour the genes 25 essential for natural competence and experimentally demonstrate natural competence in the 26 ramified cyanobacterium Chlorogloeopsis fritschii PCC 6912 (hereafter Chlorogloeopsis). 27Searching for homologs to known natural competence genes in ramified cyanobacteria 28 showed that these genes are conserved in the majority of tested isolates. Experimental 29 validation of natural competence using several alternative protocols demonstrates that 30Chlorogloeopsis could be naturally transformed with a replicative plasmid. Our results show 31 that natural competence is a common trait in ramified cyanobacteria and that natural 32 transformation is likely to play an important role in cyanobacteria evolution. 33 34 Importance Cyanobacteria are crucial players in the global biogeochemical cycles where 35 they contribute to CO 2 -and N 2 -fixation. Their main ecological significance is the oxygen-36 producing photosynthetic apparatus that contributes to contemporary food chains. Ramified 37 cyanobacteria form true-branching and multiseriate cell filament structures that represent a 38 peak of prokaryotic multicellularity. Species in that group inhabit fresh and marine water 39 habitats, thermal springs, arid environments, as well as endolithic and epiphytic habitats. 40Here we show that ramified cyanobacteria harbor the mechanisms required for DNA 41 acquisition via natural transformation. The prevalence of mechanisms for natural uptake of 42 DNA has implications for the role of DNA acquisition in the evolution of cyanobacteria. 43Furthermore, presence of mechanisms for natural transformation in ramified cyanobacteria 44 opens up new possibilities for genetic modification of ramified cyanobacteria. 45 46

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Section: Natural Transformation Of Cyanobacteriamentioning

confidence: 99%

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Natural competence inChlorogloeopsis fritschiiPCC 6912 and other ramified cyanobacteria

Springstein

Nies

2020

Preprint

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“…Others, like Caulobacter crescentus are monoploid irrespective to the growth rate (9). Alternatively, some bacteria maintain multiple chromosome copies per cell (polyploidy) irrespective of growth rate as observed in many cyanobacteria (10)(11)(12)(13). Ploidy of cyanobacteria is highly variable, it is influenced by growth phase, chemical and physical parameters and stress conditions (10,12,14,15).…”

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confidence: 99%

The Genome Copy Number of the Thermophilic Cyanobacterium Thermosynechococcus elongatus E542 Is Controlled by Growth Phase and Nutrient Availability

Riaz

Xiao

Chen

et al. 2021

Appl Environ Microbiol

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Recently isolated thermophilic cyanobacterium Thermosynechococcus E542 is a promising strain for the fundamental and applied research. Here, we used several improved ploidy estimation approaches, which include qPCR, spectrofluorometry and flow cytometry to precisely determine the ploidy level in Thermosynechococcus E542 across different growth stages, nutritional, and stress conditions. The distribution of genome copies per cell among the populations of Thermosynechococcus E542 was also analysed. The strain tends to maintain 3 to 4 genome copies per cell in lag phase, early growth phase or stationary phase under standard conditions. Increased ploidy (5.5 ± 0.3) was observed in exponential phase; hence ploidy level is growth phase regulated. Nearly no monoploid cells were detected in all growth phases, and prolonged stationary phase could not yield ploidy level lower than 3 under standard conditions. During the late growth phase, significantly higher ploidy level was observed in the presence of bicarbonate (7.6 ±0.7) and high phosphate (6.9 ±0.2) at the expense of the reduced percentage of di- and triploid cells. Meanwhile, the reduction in phosphates decreased the average ploidy level by increasing the percentage of mono- and diploid cells. In contrast, temperature and antibiotic stress reduced the percentage of mono-, di and triploid cells, yet maintaining average ploidy. The results indicate a possible causality between growth rate, stress and genome copy number across the conditions tested, but the exact mechanism is yet to be elucidated. Furthermore, the spectrofluorometric approach presented here is a quick and straightforward ploidy estimation method with reasonable accuracy. Importance Present study revealed the genome copy number (ploidy) status in thermophilic cyanobacterium Thermosynechococcus E542 is regulated by growth phase and various environmental parameters to give us a window into understanding the role of polyploidy. Increased ploidy level is found associated with the higher metabolic activity and increased vigor by acting as backup genetic information to compensate for damage to the other chromosomal copies. Several improved ploidy estimation approaches are presented in this work that may upgrade the ploidy estimation procedure for cyanobacteria in the future. Furthermore, a new spectrofluorometric method presented here is a rapid and straightforward method of ploidy estimation with reasonable accuracy compared to other laborious methods.

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Natural transformation of the filamentous cyanobacterium Phormidium lacuna

Cited by 2 publications

References 49 publications

Natural competence inChlorogloeopsis fritschiiPCC 6912 and other ramified cyanobacteria

Natural competence inChlorogloeopsis fritschiiPCC 6912 and other ramified cyanobacteria

The Genome Copy Number of the Thermophilic Cyanobacterium Thermosynechococcus elongatus E542 Is Controlled by Growth Phase and Nutrient Availability

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