Genetic transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) using particle bombardment and glass-bead agitation

Cui, Yulin; Jiang, Peng; Wang, Jinfeng; Li, Fuchao; Chen, Yingjie; Zheng, Guoting; Qin, Song

doi:10.1007/s00343-012-1093-z

Cited by 12 publications

(14 citation statements)

References 16 publications

(12 reference statements)

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“…In the present research, particle bombardment was successfully used to transfer the foreign plasmid into the P. subcordiformis chloroplast. However, this method was less efficient for nuclear transformation of P. subcordiformis than glass-bead agitation [18]. This difference may be related to the fact that P. subcordiformis has a single large cup-shaped chloroplast surrounding the nucleolus, which may facilitate particles entering the former while blocking them from the latter.…”

Section: Discussionmentioning

confidence: 96%

“…For chloroplast transformation, the commonly used primary markers are the aadA gene [14] – [15] and the neo gene [16] , which confer resistance to spectinomycin–streptomycin and kanamycin, respectively; these antibiotics inhibit protein synthesis by the plastid ribosome [17] . We previously found [18] that P. subcordiformis was not sensitive to spectinomycin, streptomycin, or kanamycin but was very sensitive to the herbicide Basta. Basta contains a tripeptide of two Ala residues and an analog of Glu called phosphinothricin (PPT).…”

Section: Introductionmentioning

confidence: 99%

“…The bar gene encoding phosphinothricin acetyltransferase (PAT) confers tolerance to PPT. The interaction of PPT, GS, and PAT can be used to select positive colonies in nuclear transformations of higher plants and algae [18] , [20] – [23] . In tobacco chloroplast transformation, the foreign bar gene could confer resistance to PPT in transformed plants but failed in direct selection for positive colonies [24] .…”

Section: Introductionmentioning

confidence: 99%

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Chloroplast Transformation of Platymonas (Tetraselmis) subcordiformis with the bar Gene as Selectable Marker

2014

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The objective of this research was to establish a chloroplast transformation technique for Platymonas (Tetraselmis) subcordiformis. Employing the gfp gene as a reporter and the bar gene as a selectable marker, transformation vectors of P. subcordiformis chloroplast were constructed with endogenous fragments rrn16S–trnI (left) and trnA–rrn23S (right) as a recombination site of the chloroplast genome. The plasmids were transferred into P. subcordiformis via particle bombardment. Confocal laser scanning microscopy indicated that the green fluorescence protein was localized in the chloroplast of P. subcordiformis, confirming the activity of the Chlamydomonas reinhardtii promoter. Cells transformed with the bar gene were selected using the herbicide Basta. Resistant colonies were analyzed by PCR and Southern blotting, and the results indicated that the bar gene was successfully integrated into the chloroplast genome via homologous recombination. The technique will improve genetic engineering of this alga.

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Section: Discussionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Chloroplast Transformation of Platymonas (Tetraselmis) subcordiformis with the bar Gene as Selectable Marker

2014

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“…C. reinhardtii, Dunaliella salina, Tetraselmis sp. are among microalgae with reported protocols for transformation with this method (68)(69)(70). Cell wall-deficient strains such as C. reinhardtii mutants or cells that do not possess a thick cell wall like D. salina have higher efficiencies of transformation than their wt-cell wall or thick cell wall relatives (71).…”

Section: Transformation Methodsmentioning

confidence: 99%

“…However, some antibiotics such as streptomycin and kanamycin, cannot be used for selection even in low salt concentrations (112)(113)(114)(115). For Marine microalgae species, selectable markers/ resistance genes (Table 1) are BAR/phosphinothricin, BLE/zeocin, CAT/chloramphenicol, PDS(L504R)/ norflurazon, NAT/nourseothricin (70,74,(116)(117)(118).…”

Section: Selection Of Transformantsmentioning

confidence: 99%

Genetic manipulation of microalgae for the production of bioproducts

et al. 2018

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Microalgae have been used during the past four decades in the Bio-industries for the production of high added value products and development of useful approaches with environmental applications. The fast growing rate, simple growth requirements and using sunlight as the major source of energy are the key factors for usage of algae. In the past 15 years, a considerable progress has been made regarding the use of microalgae for production of proteins, nutraceuticals, food supplements, molecular tags for diagnostics and fixation of greenhouse gases. Nevertheless, genetic manipulation of microalgae still remains a fairly un-explored area which could boost the production of bioproducts. It is anticipated that in the near future use of microalgae will revolutionize its applications in diverse industries. The aim of this work is to present a critical review on potential of microalgae for the production of high-added value molecules, their practical applications, and the role of genetic engineering in its utilization as a unique niche in industry. In addition, current challenges within synthetic biology approaches are discussed.

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