Marine Microalgae 5.4 Genetic Engineering of Microalgae 55
Part A | 5.4
Table 5.2 Microalgal strains achieved for the stable transformation
Phylum species
Organelle
Transformation methods
Gene knock-down References
Diatom
Cyclotella cryptica
Nucleus
Biolistic
[5.36]
Cylindrotheca fusiformis
Nucleus
Biolistic
[5.37]
Chaetoceros sp.
Nucleus
Biolistic
[5.38]
Navicula saprophila
Nucleus
Biolistic
[5.36]
Phaeodactylum tricornutum
Nucleus
Biolistic
X
[5.39, 40]
Thalassiosira pseudonana
Nucleus
Biolistic
[5.41]
Fistulifera sp.
Nucleus
Biolistic
X
[5.42]
Chlorophyta
Chlamydomonas reinhardtii
Nucleus
Biolistic, Electroporation, Glass beads,
Agrobacterium
X
[5.43–48]
Chloroplast
Biolistic
[5.9]
Mitochondoria Biolistic
[5.33]
Chlorella spp.
Nucleus
Biolistic, Electroporation, Agrobacterium
[5.49–53]
Dunaliella spp.
Nucleus
Biolistic, Electroporation , Glass beads
X
[5.54–58]
Haematococcus pluvialis
Nucleus
Biolistic, Agrobacterium
[5.59, 60]
Volvox carteri
Nucleus
Biolistic
[5.61]
Dinoflagellate
Amphidinium sp.
Nucleus
Glass beads
[5.62]
Symbiodinium microadriaticum Nucleus
Glass beads
[5.62]
Rhodophyta
Cyanidioschyzon merolae
Nucleus
Glass beads
X
[5.63, 64]
Porphyridium spp.
Chloroplast
Biolistic, Agrobacterium
[5.65, 66]
Euglenophyta
Euglena gracilis
Chloroplast
Biolistic
[5.67]
Eustigmatophyte
Nannochloropsis spp.
Nucleus
Electroporation
[5.35, 68]
the optimization of the gene transformation method for
each specific species turns out to be important. Depending on the physiological characteristics of microalgal
cells, electroporation, glass beads-mediated transformation, agrobacterium-mediated transformation, and
biolistics have frequently been used. Moreover, the
level of target protein varied due to multiple insertion,
random integration, and (or) gene silencing [5.80]. Stable transformants that have already been reported are
summarized in Table 5.2.
Biolistics, also referred to as a gene gun that was
originally designed for the delivery of nucleic acid
through the cell wall of intact plant cells, has been
mostly applied for microalgae gene transformation. The
payload in this system is a plasmid DNA-coated tungsten particle (particle size: 0:61:6 m), which can be
shot with helium gas. After bombardment, the tungsten
particles were shot down to the plant organism or the
cell culture on the petri dish. Some cells that are not
disrupted by the firing may envelope the DNA-coated
tungsten particles and the DNA can then migrate to and
integrate into the plant chromosome [5.81]. The transformation efficiency of this methodology is not related
to the physical property of the host cell but is highly
controlled by the gas pressure at the point of firing.
Therefore, theoretically, despite the hard cell wall and
frustules, gene transformation can be achieved when the
gas pressure is high enough.
Electroporation is a phenomenon when the electrical conductivity and permeability of the cell membrane
increase by the externally applied electrical field. If the
host cells and plasmids are mixed together, the plasmids can be transferred into the host cells through
the transient holes in the cell membrane generated by
the electronic shock. Electroporation-based gene transformation methodology has been commonly used to
Part A | 5.4
Table 5.2 Microalgal strains achieved for the stable transformation
Phylum species
Organelle
Transformation methods
Gene knock-down References
Diatom
Cyclotella cryptica
Nucleus
Biolistic
[5.36]
Cylindrotheca fusiformis
Nucleus
Biolistic
[5.37]
Chaetoceros sp.
Nucleus
Biolistic
[5.38]
Navicula saprophila
Nucleus
Biolistic
[5.36]
Phaeodactylum tricornutum
Nucleus
Biolistic
X
[5.39, 40]
Thalassiosira pseudonana
Nucleus
Biolistic
[5.41]
Fistulifera sp.
Nucleus
Biolistic
X
[5.42]
Chlorophyta
Chlamydomonas reinhardtii
Nucleus
Biolistic, Electroporation, Glass beads,
Agrobacterium
X
[5.43–48]
Chloroplast
Biolistic
[5.9]
Mitochondoria Biolistic
[5.33]
Chlorella spp.
Nucleus
Biolistic, Electroporation, Agrobacterium
[5.49–53]
Dunaliella spp.
Nucleus
Biolistic, Electroporation , Glass beads
X
[5.54–58]
Haematococcus pluvialis
Nucleus
Biolistic, Agrobacterium
[5.59, 60]
Volvox carteri
Nucleus
Biolistic
[5.61]
Dinoflagellate
Amphidinium sp.
Nucleus
Glass beads
[5.62]
Symbiodinium microadriaticum Nucleus
Glass beads
[5.62]
Rhodophyta
Cyanidioschyzon merolae
Nucleus
Glass beads
X
[5.63, 64]
Porphyridium spp.
Chloroplast
Biolistic, Agrobacterium
[5.65, 66]
Euglenophyta
Euglena gracilis
Chloroplast
Biolistic
[5.67]
Eustigmatophyte
Nannochloropsis spp.
Nucleus
Electroporation
[5.35, 68]
the optimization of the gene transformation method for
each specific species turns out to be important. Depending on the physiological characteristics of microalgal
cells, electroporation, glass beads-mediated transformation, agrobacterium-mediated transformation, and
biolistics have frequently been used. Moreover, the
level of target protein varied due to multiple insertion,
random integration, and (or) gene silencing [5.80]. Stable transformants that have already been reported are
summarized in Table 5.2.
Biolistics, also referred to as a gene gun that was
originally designed for the delivery of nucleic acid
through the cell wall of intact plant cells, has been
mostly applied for microalgae gene transformation. The
payload in this system is a plasmid DNA-coated tungsten particle (particle size: 0:61:6 m), which can be
shot with helium gas. After bombardment, the tungsten
particles were shot down to the plant organism or the
cell culture on the petri dish. Some cells that are not
disrupted by the firing may envelope the DNA-coated
tungsten particles and the DNA can then migrate to and
integrate into the plant chromosome [5.81]. The transformation efficiency of this methodology is not related
to the physical property of the host cell but is highly
controlled by the gas pressure at the point of firing.
Therefore, theoretically, despite the hard cell wall and
frustules, gene transformation can be achieved when the
gas pressure is high enough.
Electroporation is a phenomenon when the electrical conductivity and permeability of the cell membrane
increase by the externally applied electrical field. If the
host cells and plasmids are mixed together, the plasmids can be transferred into the host cells through
the transient holes in the cell membrane generated by
the electronic shock. Electroporation-based gene transformation methodology has been commonly used to