Nannochloropsis have attracted sustained interest from algal biodiesel researchers due to their high biomass accumulation rate and high lipid content. There are six recognized species in the Nannochloropsis genus that are phylogenetically divided into Nannochloropsis gaditana, Nannochloropsis salina, Nannochloropsis granulata, Nannochloropsis limnetica, Nannochloropsis oceanica and Nannochloropsis oculata. In this study, the potential of 9 Nannochloropsis species from the 6 genus for biodiesel production was evaluated by determining their growth rate, biomass accumulation, lipid productivity, lipid composition, fatty acid profiles and biodiesel properties. The results showed that the best strain was N. oceanica IMET1, with lipid productivity of 158.76 ± 13.83 mg L(-1)day(-1), TAG production of 1.67 ± 0.20 g/L, favorable fatty acid profiles of C16-C18 (56.62 ± 1.96%) as well as suitable biodiesel properties of higher cetane number (54.61 ± 0.25), lower iodine number (104.85 ± 2.80 g I2/100g) and relative low cloud point (3.45 ± 0.50°C). N. oceanica IMET1 could be consider as valuable feedstock for microalgal biodiesel production.

Duckweed has been considered as a valuable feedstock for bioethanol production due to its high biomass and starch production. To investigate the effects of light conditions on duckweed biomass and starch production, Lemna aequinoctialis 6000 was cultivated at different photoperiods (12:12, 16:8 and 24:0h) and light intensities (20, 50, 80, 110, 200 and 400μmolm(-2)s(-1)). The results showed that the duckweed biomass and starch production was increased with increasing light intensity and photoperiod except at 200 and 400μmolm(-2)s(-1). Considering the light cost, 110μmolm(-2)s(-1) was optimum light condition for starch accumulation with the highest maximum growth rate, biomass and starch production of 8.90gm(-2)day(-1), 233.25gm(-2) and 98.70gm(-2), respectively. Moreover, the results suggested that high light induction was a promising method for duckweed starch accumulation. This study provides optimized light conditions for future industrial large-scale duckweed cultivation.

Energy crises and environmental pollution have caused considerable concerns; duckweed is considered to be a promising new energy plant that may relieve such problems. Lemna aequinoctialis strain 6000, which has a fast growth rate and the ability to accumulate high levels of starch was grown in both Schenk & Hildebrandt medium (SH) and in sewage water (SW). The maximum growth rates reached 10.0 g DW m⁻² day⁻¹ and 4.3 g DW m⁻² day⁻¹, respectively, for the SH and SW cultures, while the starch content reached 39% (w/w) and 34% (w/w). The nitrogen and phosphorus removal rate reached 80% (SH) and 90% (SW) during cultivation, and heavy metal ions assimilation was observed. About 95% (w/w) of glucose was released from duckweed biomass hydrolysates, and then fermented by Angel yeast with ethanol yield of 0.19 g g⁻¹ (SH) and 0.17 g g⁻¹ (SW). The amylose/amylopectin ratios of the cultures changed as starch content increased, from 0.252 to 0.155 (SH) and from 0.252 to 0.174 (SW). Lemna aequinoctialis strain 6000 could be considered as valuable feedstock for bioethanol production and water resources purification.

Summary: The fast growth, ease of metabolic labelling and potential for feedstock and biofuels production make duckweeds not only an attractive model system for understanding plant biology, but also a potential future crop. However, current duckweed research is constrained by the lack of efficient genetic manipulation tools. Here, we report a case study on genome editing in a duckweed species, Lemna aequinoctialis, using a fast and efficient transformation and CRISPR/Cas9 tool. By optimizing currently available transformation protocols, we reduced the duration time of Agrobacterium‐mediated transformation to 5–6 weeks with a success rate of over 94%. Based on the optimized transformation protocol, we generated 15 (14.3% success rate) biallelic LaPDS mutants that showed albino phenotype using a CRISPR/Cas9 system. Investigations on CRISPR/Cas9‐mediated mutation spectrum among mutated L. aequinoctialis showed that most of mutations were short insertions and deletions. This study presents the first example of CRISPR/Cas9‐mediated genome editing in duckweeds, which will open new research avenues in using duckweeds for both basic and applied research.