Optimization of protoplast yields from the red algae Gracilaria dura (C. Agardh) J. Agardh and G. verrucosa (Huds.) Papenfuss

Gupta, Vishal; Kumar, Manoj; Kumari, Puja; Reddy, C. R. K.; Jha, Bhavanath

doi:10.1007/s10811-010-9579-6

Cited by 23 publications

(7 citation statements)

References 32 publications

(45 reference statements)

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“…We optimized the most efficient protoplasts generation with single enzymatic incubation and in shorter time periods in both microalgae species, in order to provide the fastest and most economically favorable way of generating microalgae cells with a partially removed cell wall, compared with previous studies on Dictyopteris prolifera , where a mixture of cellulose with other enzymes and longer time periods of treatment delivered the optimal cell wall digestion [ 34 ]. The shorter enzymatic period of treatment in our study is in agreement with multi-enzymatic treatments in Gracilaria and Ulva species [ 35 , 36 ]. Physiological and biochemical changes between the cell walls of various microalgae species rule the protoplast formation yields [ 37 ].…”

Section: Discussionsupporting

confidence: 88%

Magnetic Immobilization and Growth of Nannochloropsis oceanica and Scenedasmus almeriensis

Savvidou

Ferraro

Schinas

et al. 2021

Plants

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Microalgae are used in industrial and pharmaceutical applications. Their performance on biological applications may be improved by their immobilization. This study presents a way of cell immobilization using microalgae carrying magnetic properties. Nannochloropsis oceanica and Scenedasmus almeriensis cells were treated enzymatically (cellulase) and mechanically (glass beads), generating protoplasts as a means of incorporation of magnetic nanoparticles. Scanning electron microscopy images verified the successful cell wall destruction for both of the examined microalgae cells. Subsequently, protoplasts were transformed with magnetic nanoparticles by a continuous electroporation method and then cultured on a magnetic surface. Regeneration of transformed protoplasts was optimized using various organic carbon and amino acid supplements. Both protoplast preparation methods demonstrated similar efficiency. Casamino acids, as source of amino acids, were the most efficient compound for N. oceanica protoplasts regeneration in enzymatic and mechanical treatment, while for S. almeriensis protoplasts regeneration, fructose, as source of organic carbon, was the most effective. Protoplasts transformation efficiency values with magnetic nanoparticles after enzymatic or mechanical treatments for N. oceanica and S. almeriensis were 17.8% and 10.7%, and 18.6% and 15.7%, respectively. Finally, selected magnetic cells were immobilized and grown on a vertical magnetic surface exposed to light and without any supplement.

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Section: Discussionsupporting

confidence: 88%

Magnetic Immobilization and Growth of Nannochloropsis oceanica and Scenedasmus almeriensis

Savvidou

Ferraro

Schinas

et al. 2021

Plants

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“…Protoplast Isolation and Comet Assay. Protoplast isolation and purification were performed according to Gupta et al 23 Briefly, following IL treatment and washing, the algal fragments (300 mg FW) were chopped into small pieces of tissue (e1 mm thin) in natural seawater (NSW) in two replicates, each with 150 mg of tissue. The chopped tissues were then rinsed several times with NSW to remove debris.…”

Section: •àmentioning

confidence: 99%

Toxic Effects of Imidazolium Ionic Liquids on the Green Seaweed Ulva lactuca: Oxidative Stress and DNA Damage

Kumar

Trivedi²,

Reddy³

et al. 2011

Chem. Res. Toxicol.

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The green credentials of ionic liquids (ILs) are being increasingly questioned due to the growing evidence of their toxicity to aquatic ecosystems, although the mechanisms of toxicity are unknown. This study provides insights into the mechanism of toxicity and biological effects of 1-alkyl-3-methylimidazolium bromide [C(n)mim]Br (n = 4 to 16) on the marine macroalga Ulva lactuca. The cell viability of this alga during IL exposure was found to be negatively correlated to the chain length of the alkyl group. The IL ([C(12)mim]Br) exposure triggers the generation of reactive oxygen species (ROS viz. O(2)(•-), H(2)O(2), and OH(•)), damage of the membrane and DNA, and inhibition of antioxidant systems in the alga. The enhanced production of ROS and lipid peroxidation in the alga subjected to LC(50) concentration for 4 days was largely attributed to lipoxygenase (LOX) activity coupled with the induction of two LOX isoforms (~80 kDa and ~55 kDa). Pretreatment of the algal thallus with enzyme inhibitors such as diphenylene iodonium, sodium azide, cantharidin, and oxadiazoloquinoxalin-1-one, prior to [C(12)mim]Br exposure showed the regulation of ROS by the activation of membrane bound NADPH-oxidase and cytochrome oxidase. The IL exposure resulted in the accumulation of n-3 and n-6 fatty acids at 0.5 LC(50) concentration indicating the induction of desaturase enzymes. Furthermore, antioxidant enzyme activities such as superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR) were enhanced by 1.3-2.0-fold, while glutathione peroxidase (GSH-Px) diminished, together with a higher regeneration rate of reduced ascorbate and glutathione. The isoforms of antioxidant enzymes, namely, Mn-SOD (~85 kDa), APX (~125 and 45 kDa), and GR (~135 kDa) regulated differentially to IL exposure. The comet assay performed for the first time for seaweeds revealed the significant induction of DNA damage (>50-70% increase in % tail DNA over control) in alga exposed to ≥ LC(50) concentration.

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“…They have been extensively studied in many fields, such as proteomics, metabolomics, somatic hybridization, cybridization, and protoclonal variation studies (Reddy et al 2008a). In seaweed tissue culture, protoplast had been used to provide seed stock for cultivation purposes (Gupta et al 2011, Huddy et al 2013. Since plant cells are totipotent, many protoplasts can be isolated from a small fragment of seaweed thallus and cultured to produce seaweed seedlings (Huddy et al 2013).…”

Section: Protoplast Culturementioning

confidence: 99%

Recent advances in seaweed seedling production: a review of eucheumatoids and other valuable seaweeds

Jiksing¹,

Ongkudon²,

Thien³

et al. 2022

Algae

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Modern seaweed farming relies heavily on seedlings from natural beds or vegetative cuttings from previous harvests. However, this farming method has some disadvantages, such as physiological variation in the seed stock and decreased genetic variability, which reduces the growth rate, carrageenan yield, and gel strength of the seaweeds. A new method of seedling production that is sustainable, scalable, and produces a large number of high-quality plantlets is needed to support the seaweed farming industry. Recent use of tissue culture and micropropagation techniques in eucheumatoid seaweed production has yielded promising results in increasing seed supply and growing uniform seedlings in large numbers in a shorter time. Several seaweed species have been successfully cultured and regenerated into new plantlets in laboratories using direct regeneration, callus culture, and protoplast culture. The use of biostimulants and plant growth regulators in culture media increases the seedling quality even further. Seedlings produced by micropropagation grew faster and had better biochemical properties than conventionally cultivated seedlings. Before being transferred to a land-based grow-out system or ocean nets for farming, tissue-cultured seedlings were recommended to undergo an acclimatization process to increase their survival rate. Regular monitoring is needed to prevent disease and pest infestations and grazing by herbivorous fish and turtles during the farming process. The current review discusses recent techniques for producing eucheumatoid and other valuable seaweed farming materials, emphasizing the efficiency of micropropagation and the transition from laboratory culture to cultivation in land-based or open-sea grow-out systems to elucidate optimal conditions for sustainable seaweed production.

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Optimization of protoplast yields from the red algae Gracilaria dura (C. Agardh) J. Agardh and G. verrucosa (Huds.) Papenfuss

Cited by 23 publications

References 32 publications

Magnetic Immobilization and Growth of Nannochloropsis oceanica and Scenedasmus almeriensis

Magnetic Immobilization and Growth of Nannochloropsis oceanica and Scenedasmus almeriensis

Toxic Effects of Imidazolium Ionic Liquids on the Green Seaweed Ulva lactuca: Oxidative Stress and DNA Damage

Recent advances in seaweed seedling production: a review of eucheumatoids and other valuable seaweeds

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