08.08.2015   Biomass and Biofuels from Microalgae

Genetic Engineering in Other Algae

Apart from C. reinhardtii, few algal species have been subjected to extensive genomic manipulation. As it seems unlikely that C. reinhardtii will be used for commercial biofuel applications, this needs to be remedied. Because of the phy­logenetic and structural diversity of algae, methods established for C. reinhardtii cannot necessarily be easily transferred to other species and may require major adaptations. Therefore, recent efforts have been made to develop molecular toolkits to increase the range of other more suitable algal species for commercial production scenarios.

A number of algae species have been transformed successfully, and an overview is given in Table 11.2. For example, Euglena gracilis was transformed with an antibiotic resistance marker (Doetsch et al. 2001) and Porphyridium spp. using a herbicide resistance cassette (Lapidot et al. 2002) RNAi has also been used to engineer nuclear genes in the chlorophyte Dunaliella salina (Sun et al. 2008) and in the diatom Phaeodactylum tricornutum (De Riso et al. 2009). Applicable genetic modifications of green algae for industry are the transformation of Haematococcus pluvialis (Steinbrenner and Sandmann 2006; Teng et al. 2002), an important

Table 11.2 Amenable strains and transformation systems

Compartment

Phylum

Class

Species

Method

References

Nuclear

Chlorophyta

Chlorophyceae

Chlamydomonas

reinhardtii

A, B, E, G, S

Debuchy et al. (1989)

Dunahay (1993) and Kindle et al. (1989)

Kindle (1990), Kumar et al. (2004) and Mayfield and Kindle (1990)

Rochaix and Vandillewijn (1982), Shimogawara et al. (1998) and Fernandez et al. (1989)

Molnar et al. (2009) and Stevens et al. (1996)

Berthold et al. (2002) and Sizova et al. (2001)

Cerutti et al. (1997a)

Goldschmidt-Clermont

(1991)

Schroda et al. (2000)

Dunaliella salina

B, E, G

Feng et al. (2009) and Geng et al. (2004)

Sun et al. (2005) and Wang et al. (2007),

Tan et al. (2005) and Li et al. (2007)

Eudorina elegans

B

Lerche and Hallmann (2013)

Gonium

pectorale

B

Lerche and Hallmann (2009)

Haematococcus

pluvialis

A, B

Teng et al. (2002)

Kathiresan et al. (2009) and Steinbrenner and Sandmann (2006)

Volvox carteri

Hallmann and Rappel (1999) and Hallmann and Sumper (1994)

Hallmann and Sumper (1996) and Jakobiak et al. (2004)

(Schiedlmeier et al. 1994)

Trebouxiophyceae

Chlorella

ellipsoidea

PT-E

Jarvis and Brown (1991) and Kim et al. (2002)

Bai et al. (2013) and Liu et al. (2013)

Chlorella

saccharophila

PT-E

Maruyama et al. (1994)

Chlorella

sorokiniana

B

Dawson et al. (1997) and Hawkins and Nakamura

(1999)

Chlorella

vulgaris

B

Hawkins and Nakamura

(1999)

Dinoflagellate

Dinophyceae

Amphidinium

spp.

S

ten Lohuis and Miller

(1998)

Symbiodinium

microadriaticum

S

ten Lohuis and Miller

(1998)

Heterokontophyta

Bacillariophyceae (diatoms)

Chaetoceros

salsugineum

B

Miyagawa-Yamaguchi et al. (2011)

(continued)

Table 11.2 (continued)

Compartment

Phylum

Class

Species

Method

References

Chaetoceros

debilis

B

Miyagawa-Yamaguchi et al. (2011)

Chaetoceros

setoensis

B

Miyagawa-Yamaguchi et al. (2011)

Chaetoceros

tenuissimus

B

Miyagawa-Yamaguchi et al. (2011)

Cyclotella

cryptica

B

Dunahay et al. (1995)

Cylindrotheca

fusiformis

B

Fischer et al. (1999) and Poulsen and Kroger (2005)

Navicula

saprophila

B

Dunahay et al. (1995)

Phaeodactylum

tricornutum

B

Apt et al. (1996), De Riso et al. (2009), Falciatore et al. (1999) and Zaslavskaia et al. (2000)

Miyagawa et al. (2009), Sakaguchi et al. (2011) and Zaslavskaia et al. (2001)

Thalassiosira

weissflogii

B

Falciatore et al. (1999)

Thalassiosira

pseudonana

B

Poulsen et al. (2006)

Eustigmatophyceae

Nannochloropsis

sp.

A, E

Cha et al. (2011) and Kilian et al. (2011)

Nannochloropsis

gaditana

E

Li et al. (2014) and Radakovits et al. (2012)

Nannochloropsis

granulata

E

Li et al. (2014)

Nannochloropsis

oculata

PT-E

Chen et al. (2008), Li et al. (2014) and Li and Tsai (2009)

Nannochloropsis

oceanica

E

Vieler et al. (2012) and Li et al. (2014)

Nannochloropsis

salina

E

Li et al. (2014)

Rhodophyta

Cyanidiophyceae

Cyanidioschyzon

merolae

E, PEG

Fujiwara et al. (2013), Minoda et al. (2004) and Ohnuma et al. (2008), (2009)

Chloroplast

Chlorophyta

Chlorophyceae

Chlamydomonas

reinhardtii

B, G

Boynton et al. (1988) and O’Neill et al. (2012)

Haematococcus

pluvialis

B

Gutierrez et al. (2012)

Dunaliella sp

B

Purton et al. (2013)

Scenedesmus sp

B

Purton et al. (2013)

Prasinophyceae

Platymonas

subcordiformis

B

Cui et al. (2014)

Euglenophyta

Euglenoidea

Euglena gracilis

B

Doetsch et al. (2001)

Porphyridiophyta

Porphyridiophyceae

Porphyridium sp

B

Lapidot et al. (2002)

Mitochondria

Chlorophyta

Chlorophyceae

Chlamydomonas

reinhardtii

B

Randolph-Anderson et al. (1993), Remacle et al. (2006) and Yamasaki et al. (2005)

Methods A Agrobacterium, B biolistic bombardment, E electroporation, G glass bead agitation, S silicon carbide whiskers, PT protoplast transformation, and PEG with polyethylene glycol

producer of astaxanthin, and Dunaliella salina (Feng et al. 2009; Geng et al. 2004; Sun et al. 2005; Tan et al. 2005) used for P-carotene production. Diatoms are also important commercial sources for aquaculture feedstock, specialty oils such as omega-3 fatty acids, and are used in nanotechnology due to their unique silica frustules. There has been one report of a nuclear transformation of dinoflagellates (ten Lohuis and Miller 1998). Red algae have been used for both chloroplast transformation (Lapidot et al. 2002) and nuclear transformation (Cheney et al. 2001; Minoda et al. 2004). A human growth hormone (hGH) has been successfully expressed in the nucleus of Chlorella vulgaris (Hawkins and Nakamura 1999) and a fish growth hormone (GH) in Nannochloropsis oculata (Chen et al. 2008). Transformation techniques using a cellulolytic enzyme to weaken the cell walls and make the cells more competent for the uptake of foreign DNA have been suc­cessfully applied to the green algae Chlorella ellipsoidea (Liu et al. 2013) and may be applicable for the transformation of other algal species with tough cell walls in future. A synthetic biology approach to engineer complex photosynthetic traits from diverse algae into a more controllable production strains has been shown using an ex vivo genome assembly to transfer genes for core photosystem subunits from Scenedesmus into multiple loci in the Chlamydomonas plastid genome (O’Neill et al. 2012).

Given the recent expansion of interest in microalgae, a broader repertoire of genome sequences and analytical and molecular engineering tools are being reported and will provide the foundation for a broad range of biofuel applications, some of which are covered in the following section.

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