Cinnamic acid, coumarin and vanillin: Alternative phenolic compounds for efficient Agrobacterium-mediated transformation of the unicellular green alga, Nannochloropsis sp.

San, Thye; Chen, Chin-Fong; Yee, Willy; Aziz, Azimon Abdul; Loh, Saw-Hong

doi:10.1016/j.mimet.2011.01.005

Cited by 97 publications

(52 citation statements)

References 22 publications

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“…Transient transformation of Nannochloropsis sp. using Agrobacterium has been described (33). Transformation of N. oculata after enzymatic digestion of the cell wall has also been reported (34), but the lack of a selectable marker gene in the transformation construct necessitated PCR screening of hundreds of colonies to identify a few stable transformants.…”

Section: Discussionmentioning

confidence: 99%

High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp.

Kilian¹,

Benemann²,

Niyogi³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

409

301

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Algae have reemerged as potential next-generation feedstocks for biofuels, but strain improvement and progress in algal biology research have been limited by the lack of advanced molecular tools for most eukaryotic microalgae. Here we describe the development of an efficient transformation method for Nannochloropsis sp., a fast-growing, unicellular alga capable of accumulating large amounts of oil. Moreover, we provide additional evidence that Nannochloropsis is haploid, and we demonstrate that insertion of transformation constructs into the nuclear genome can occur by high-efficiency homologous recombination. As examples, we generated knockouts of the genes encoding nitrate reductase and nitrite reductase, resulting in strains that were unable to grow on nitrate and nitrate/nitrite, respectively. The application of homologous recombination in this industrially relevant alga has the potential to rapidly advance algal functional genomics and biotechnology.

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Section: Discussionmentioning

confidence: 99%

High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp.

Kilian¹,

Benemann²,

Niyogi³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

409

301

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show abstract

“…Agrobacterium has been used extensively to 315 transfer DNA to eukaryotic algae in the green algal lineage with subsequent chromosomal integration (29)(30)(31)(32). In contrast, to our knowledge, the only other example beyond our results of using A. tumefaciens to deliver DNA maintained as an episome in the recipient was with yeast (14).…”

Section: Discussionmentioning

confidence: 87%

Additional conjugation systems for inter-domain plasmid transfer to the diatom Phaeodactylum tricornutum

Kang

Bielinski

Bolt

et al. 2017

Preprint

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Bacterial conjugation utilizes a type IV secretion system and a DNA transfer mechanism to deliver DNA from one cell to another. Conjugative partners are conventionally confined to the prokaryotic domain. In a prominent exception, Agrobacterium tumefaciens type IV secretion-mediated transfer of DNA to plant cells can result in subsequent chromosomal integration. Recently, we demonstrated interdomain conjugation from Escherichia coli to the diatom Phaeodactylum tricornutum with the subsequent maintenance of an episome at chromosomal copy numbers if it contains diatom centromeres or centromere-like elements. The genes involved in the conjugation process can be separated into those encoding the type IV secretion system, also called the mating pair formation (MPF) genes, and genes involved in DNA processing called the mobilization (MOB) genes. Various protein families compose each class of conjugation genes, including common MOB types F, P, and Q and MPF types F, P, and T. The conjugative transfer from E. coli to P. tricornutum was demonstrated with a vector expressing MOBP and MTFP. Here we show that the MOBPsystem can be deleted and complemented with a MOBQ system in E.coli-diatom conjugations with subsequent episomal maintenaince. Utilization of both MOBP and MOBQ systems results in substantially higher efficiencies in E. coli-diatom conjugation. Finally, we demonstrate conjugative gene transfer between P. tricornutum and A. tumefaciens expressing a MPFT, the first demonstration of this system in diatoms,resulting in episomal maintainance or chromosomal integration, depending on the ex-conjugant. The promiscuity of MOB and MTF systems permitting prokaryote to diatom conjugative DNA transfer suggest major environmental and evolutionary importance of this process. The increased efficiency of dual MOB systems immediately improves genetic engineering in diatoms and has interesting basic cellular biology implications.

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“…The transformation systems in the marine microalgae Nannochloropsis sp. and Dunaliella bardawil were also recently established with this method (Anila et al, 2011;Cha et al, 2011). In the Dunaliella study, the transformation frequency obtained (41.0 ± 4 cfu per 10 6 cells) was no higher than those reported for glass beads transformation and electroporation, but the transformants obtained were found to be stable for 18 months (Anila et al, 2011).…”

Section: Agrobacterium Tumefaciens-mediated Genetic Transformationmentioning

confidence: 93%

“…It was found that cinnamic acid, vanillin and coumarin produced higher percentages of GUS positive cells in Nannochloropsis sp. transformations compared to acetosyringone, providing possible alternative Agrobacterium vir gene inducers that are more potent than the commonly used acetosyringone (Cha et al, 2011).…”

Section: Agrobacterium Tumefaciens-mediated Genetic Transformationmentioning

confidence: 99%

Advances in genetic engineering of marine algae

Qin

Lin

Jiang

2012

Biotechnology Advances

140

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Cinnamic acid, coumarin and vanillin: Alternative phenolic compounds for efficient Agrobacterium-mediated transformation of the unicellular green alga, Nannochloropsis sp.

Cited by 97 publications

References 22 publications

High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp.

High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp.

Additional conjugation systems for inter-domain plasmid transfer to the diatom Phaeodactylum tricornutum

Advances in genetic engineering of marine algae

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