High efficiency of protoplast preparation for artificially cultured Ulva prolifera (Ulvophyceae, Chlorophyta)

Wu, Chunhui; Jiang, Peng; Zhao, Jianmin; Fu, Huihui

doi:10.1007/s00343-018-7058-0

Cited by 10 publications

(9 citation statements)

References 20 publications

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“…We found numerous highly moving bacteria when observing the thalli of U. prolifera by optical microscope (see the Supplementary Video 1 ). In addition, the existence of highly moving bacteria was observed in the protoplast of U. prolifera using enzymatic digestion (Wu et al, accepted ) as well by optical microscope (see the Supplementary Video 2 ). This is the first time the potential endophytic bacteria of U. prolifera have been visualized.…”

Section: Resultsmentioning

confidence: 97%

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Comparative Genomics of Pseudomonas sp. Strain SI-3 Associated With Macroalga Ulva prolifera, the Causative Species for Green Tide in the Yellow Sea

Jiang

Zhao

et al. 2018

Front. Microbiol.

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Algae-bacteria associations occurred widely in marine habitats, however, contributions of bacteria to macroalgal blooming were almost unknown. In this study, a potential endophytic strain SI-3 was isolated from Ulva prolifera, the causative species for the world's largest green tide in the Yellow Sea, following a strict bleaching treatment to eliminate epiphytes. The genomic sequence of SI-3 was determined in size of 4.8 Mb and SI-3 was found to be mostly closed to Pseudomonas stutzeri. To evaluate the characteristics of SI-3 as a potential endophyte, the genomes of SI-3 and other 20 P. stutzeri strains were compared. We found that SI-3 had more strain-specific genes than most of the 20 P. stutzeri strains. Clusters of Orthologous Groups (COGs) analysis revealed that SI-3 had a higher proportion of genes assigned to transcriptional regulation and signal transduction compared with the 20 P. stutzeri strains, including four rhizosphere bacteria, indicating a complicated interaction network between SI-3 and its host. P. stutzeri is renowned for its metabolic versatility in aromatic compounds degradation. However, significant gene loss was observed in several aromatic compounds degradation pathways in SI-3, which may be an evolutional adaptation that developed upon association with its host. KEGG analysis revealed that dissimilatory nitrate reduction to ammonium (DNRA) and denitrification, two competing dissimilatory nitrate reduction pathways, co-occurred in the genome of SI-3, like most of the other 20 P. stutzeri strains. We speculated that DNRA of SI-3 may contribute a competitive advantage in nitrogen acquisition of U. prolifera by conserving nitrogen in NH4+ form, as in the case of microalgae bloom. Collectively, these data suggest that Pseudomonas sp. strain SI-3 was a suitable candidate for investigation of the algae-bacteria interaction with U. prolifera and the ecological impacts on algal blooming.

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Section: Resultsmentioning

confidence: 97%

“…The protoplast of U. prolifera was prepared according to Wu et al ( accepted ). Complete cells and protoplast of U. prolifera were observed by optical microscope to determine if endophytic bacteria exist.…”

Section: Methodsmentioning

confidence: 99%

Comparative Genomics of Pseudomonas sp. Strain SI-3 Associated With Macroalga Ulva prolifera, the Causative Species for Green Tide in the Yellow Sea

Jiang

Zhao

et al. 2018

Front. Microbiol.

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“…Green macroalgae may be in the forefront of protoplast mediated propagation technologies. Protoplast isolation and regeneration protocols are well established for several ulvophycean taxa ( Ulva , Monostroma , Chaetomorpha and Bryopsis ); 219,221–223 while a detailed model system for high‐throughput macroalgal nursery, based on protoplast‐derived propagation technologies for large‐scale aquaculture of Ulva species has been recently proposed 224 …”

Section: Future Perspectivesmentioning

confidence: 99%

“…219,220 Green macroalgae may be in the forefront of protoplast mediated propagation technologies. Protoplast isolation and regeneration protocols are well established for several ulvophycean taxa (Ulva, Monostroma, Chaetomorpha and Bryopsis); 219,[221][222][223] while a detailed model system for high-throughput macroalgal nursery, based on protoplast-derived propagation technologies for largescale aquaculture of Ulva species has been recently proposed. 224 Macroalgal aquaculture is expected to gain traction in emerging markets, particularly in Europe where large-scale consortium projects/networks have been joining efforts to boost macroalgae aquaculture (eg GENIALG; PHYCOMORPH), also aiding in the development and publication of guidelines towards macroalgae aquaculture sustainability.…”

Section: Future Per S Pec Tive Smentioning

confidence: 99%

The underexplored potential of green macroalgae in aquaculture

Moreira

Cruz

Marques

et al. 2021

Reviews in Aquaculture

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Green macroalgae (Chlorophyta) currently represent a residual fraction (<1%) of global seaweed biomass production landings. In turn, red (Rhodophyta) and brown (Ochrophyta) macroalgae dominate the remaining percentage of aquaculture production, exceeding 32 million tonnes per annum. However, the industry relies on a relatively low number of species, in which as few as seven macroalgal genera collectively represent the bulk of global production metrics. At present, innovation and increased sustainability of the industry calls for diversification of macroalgal species/strains in aquaculture to counteract potential adverse effects ensuing from genetic impoverishment, decreased resilience to disease and climate change. Despite the dominance of red and brown seaweed regarding production figures, aquaculture of green macroalgae has witnessed an increasing trend in productivity and diversification over the last decades, particularly in Asia, where green seaweed taxa often occupy specific market niches in the food sector. Furthermore, growing interest in green seaweeds in aquaculture has been highlighted for different applications in emerging western markets (eg IMTA, biorefineries, food delicacies), owing to a unique diversity of cytomorphologies, ecophysiological traits, propagation capacities and bioactive compounds featured by this group of macroalgae. Cultivation technologies are relatively well developed, but sustainability assessments are scarce and required to unlock the potential of green seaweeds. Although it is likely that green macroalgae will remain occupying specialised market niches, in which high‐value products are favoured, we argue that aquaculture of chlorophytan taxa presents itself as a compelling option under the current quest for commercial diversification of products and expansion of the sector.

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“…We isolated the protoplasts from laboratory culture samples using a simple mixture of commercial enzymes. The reason for choosing cultured thalli was that they offer an advantage over field samples in terms of applicability, as the former meets the need for protoplast transformation regarding genetic background [24]. So far, protoplast isolation in S. lomentaria and other Dictyopteris species has been performed on field materials [23,25,26].…”

Section: Introductionmentioning

confidence: 99%

Protoplast isolation from Dictyopteris pacifica and Scytosiphon lomentaria, using a simple commercial enzyme preparation

Avila-Peltroche

Won

Cho

2021

Journal of Genetic Engineering and Biotechnology

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Background Protoplasts (i.e., naked plant cells) can be used for in vitro manipulations and genetic improvement in cultivars with economic value. During the last decade, protoplast research in economic brown algae has been scarce, and it is usually hampered by the use of non-commercial enzymes or crude extracts for isolating protoplasts. Dictyopteris pacifica is part of a brown algal genus well known by its wide chemical diversity and biological properties. Scytosiphon lomentaria is an edible brown seaweed with antioxidant, antitumor, and antiviral properties. So far, there are no protoplast isolation protocols using commercial enzymes for these two economic brown algae. In this study, we obtained protoplasts from cultured samples of D. pacifica and S. lomentaria using commercially available enzymes. Additionally, we investigated the effects of Driselase inclusion and Ca-chelation pre-treatment on protoplast yields in order to optimize the conditions for protoplast preparations. Results Protoplasts were isolated from Dictyopteris pacifica and Scytosiphon lomentaria using the commercially available Cellulase Onozuka RS (1%) and Alginate lyase (4 U mL−1), and short incubation time (4 h). Driselase did not show significant effects on protoplast production in both species. Ca-chelation pre-treatment only increased the number of protoplasts in D. pacifica. Under optimal conditions, the protoplast yields from D. pacifica and S. lomentaria were 4.83 ± 2.08 and 74.64 ± 32.49 × 106 protoplasts g−1 fresh weight, respectively. The values obtained for S. lomentaria were 2–3 orders of magnitude higher than previously reported. Conclusions Our results show that high protoplast yields can be obtained from D. pacifica and S. lomentaria using a simple mixture of commercial enzymes (Cellulase RS and Alginate lyase) and short incubation time (4 h). This work also represents the first report of protoplast isolation in D. pacifica. The method proposed here can help to expand protoplast technology in more brown algal species.

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High efficiency of protoplast preparation for artificially cultured Ulva prolifera (Ulvophyceae, Chlorophyta)

Cited by 10 publications

References 20 publications

Comparative Genomics of Pseudomonas sp. Strain SI-3 Associated With Macroalga Ulva prolifera, the Causative Species for Green Tide in the Yellow Sea

Comparative Genomics of Pseudomonas sp. Strain SI-3 Associated With Macroalga Ulva prolifera, the Causative Species for Green Tide in the Yellow Sea

The underexplored potential of green macroalgae in aquaculture

Protoplast isolation from Dictyopteris pacifica and Scytosiphon lomentaria, using a simple commercial enzyme preparation

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