DNA-mediated Transformation in Nostoc muscorum, a Nitrogen-fixing Cyanobacterium

Trehan, K. B.; Sinha, Umakant

doi:10.1071/bi9820573

Cited by 7 publications

(7 citation statements)

References 9 publications

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“…Transformation with exogenous DNA, either plasmid, intergeneric or interspecifi c has been successful in a few unicellular species, including Synechococcus sp. PCC 7002 (Stevens and Porter 1980), Synechocystis (Grigorieva and Shestakov 1982), Microcystis aeruginosa PCC 7806 (Dittmann et al, 1997) and Thermosynechococcus elongatus BP-1 (Onai et al, 2004), as well as the heterocystous cyanobacteria Nostoc muscorum and Anabaena tortulosa (Trehan and Sinha 1982) where reciprocal transformation has been exemplifi ed. One of the main diffi culties in establishing a genetic transfer system in cyanobacteria, particularly the fi lamentous strains, is the presence of various restriction-modifi cation systems in these species.…”

Section: Barriers To Transfer Of Foreign Dna Into Cyano-bacteriamentioning

confidence: 99%

Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering

2013

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SUMMARYCyanobacteria display a large diversity of cellular forms ranging from unicellular to complex multicellular fi laments or aggregates. Species in the group present a wide range of metabolic characteristics including the fi xation of atmospheric nitrogen, resistance to extreme environments, production of hydrogen, secondary metabolites and exopolysaccharides. These characteristics led to the growing interest in cyanobacteria across the fi elds of ecology, evolution, cell biology and biotechnology. The number of available cyanobacterial genome sequences has increased considerably in recent years, with more than 140 fully sequenced genomes to date. Genetic engineering of cyanobacteria is widely applied to the model unicellular strains Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942. However the establishment of transformation protocols in many other cyanobacterial strains is challenging. One obstacle to the development of these novel model organisms is that many species have doubling times of 48 h or more, much longer than the bacterial models E. coli or B. subtilis. Furthermore, cyanobacterial defense mechanisms against foreign DNA pose a physical and biochemical barrier to DNA insertion in most strains. Here we review the various barriers to DNA uptake in the context of lateral gene transfer among microbes and the various mechanisms for DNA acquisition within the prokaryotic domain. Understanding the cyanobacterial defense mechanisms is expected to assist in the development and establishment of novel transformation protocols that are specifi cally suitable for this group.

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Section: Barriers To Transfer Of Foreign Dna Into Cyano-bacteriamentioning

confidence: 99%

Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering

2013

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“…Even though NC is known for several single celled cyanobacterial species and one filamentous cyanobacterium, Nostoc muscorum [15], this mechanism was considered as rare trait among cyanobacteria [6,13,14]. The finding of NC in Phormidium lacuna, the first filamentous nonheterocystous cyanobacterium, was therefore unexpected and surprising.…”

Section: Discussionmentioning

confidence: 99%

“…These cyanobacteria have a unicellular lifestyle, and NT was frequently described as a unique feature of few unicellular cyanobacteria [6,13,14]. There is also one report about NT of Nostoc muscorum [15], which belongs to the filamentous cyanobacteria with heterocysts, but no report about other filamentous cyanobacteria.…”

Section: Introductionmentioning

confidence: 99%

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

Nies¹,

Mielke²,

Pochert³

et al. 2020

PLoS ONE

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Research for biotechnological applications of cyanobacteria focuses on synthetic pathways and bioreactor design, while little effort is devoted to introduce new, promising organisms in the field. Applications are most often based on recombinant work, and the establishment of transformation can be a risky, time-consuming procedure. In this work we demonstrate the natural transformation of the filamentous cyanobacterium Phormidium lacuna and insertion of a selection marker into the genome by homologous recombination. This is the first example for natural transformation filamentous non-heterocystous cyanobacterium. We found that Phormidium lacuna is polyploid, each cell has about 20-90 chromosomes. Transformed filaments were resistant against up to 14 mg/ml of kanamycin. Formerly, natural transformation in cyanobacteria has been considered a rare and exclusive feature of a few unicellular species. Our finding suggests that natural competence is more distributed among cyanobacteria than previously thought. This is supported by bioinformatic analyses which show that all protein factors for natural transformation are present in the majority of the analyzed cyanobacteria.

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“…Even though natural competence is known for several single celled cyanobacterial species and one filamentous cyanobacterium, Nostoc muscorum (16), this mechanism was considered as rare trait among cyanobacteria (6, 14, 15). The finding of natural competence in Phormidium lacuna , the first example for the order Oscillatoriales, was therefore unexpected and surprising.…”

Section: Discussionmentioning

confidence: 99%

“…These cyanobacteria have a unicellular lifestyle, and NT was frequently described as a unique feature of few unicellular cyanobacteria (6, 14, 15). We also found one report about NT of the filamentous cyanobacterium Nostoc muscorum (16), but are not aware of other filamentous cyanobacteria.…”

Section: Introductionmentioning

confidence: 89%

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

Nies

Mielke

Pochert

et al. 2019

Preprint

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9Research for biotechnological applications of cyanobacteria focuses on synthetic pathways and 10 bioreactor design, while little effort is devoted to introduce new, promising organisms in the field. 11Applications are most often based on recombinant work, and the establishment of transformation 12 can be a risky, time-consuming procedure. In this work we demonstrate the natural transformation 13 of the filamentous cyanobacterium Phormidium lacuna and insertion of a selection marker into the 14 genome by homologous integration. This is the first example for natural transformation of a member 15 of the order Oscillatoriales. We found that Phormidium lacuna is polyploid, each cell has about 20-16 100 chromosomes. Transformed filaments were resistant against up to 15 mg/ml of kanamycin, and 17 the high resistance feature allowed for rapid segregation into all chromosomes. Formerly, natural 18 transformation in cyanobacteria has been considered a rare and exclusive feature of a few unicellular 19 species. Our finding suggests that natural competence is more distributed among cyanobacteria than 20 previously thought. This is supported by bioinformatic analyses which show that all protein factors 21 for natural transformation are present in the majority of the analyzed cyanobacteria. 22

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DNA-mediated Transformation in Nostoc muscorum, a Nitrogen-fixing Cyanobacterium

Cited by 7 publications

References 9 publications

Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering

Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

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