Vir gene inducers in Dunaliella salina ; an insight in to the Agrobacterium -mediated genetic transformation of microalgae

Simon, Daris Pazhukunnel; Narayanan, Anila; Gouda, K. G. Mallikarjun; Ravi, Saranya

doi:10.1016/j.algal.2015.06.007

Cited by 9 publications

(5 citation statements)

References 28 publications

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“…But, D-glucose and AS alone do not cause any significant increase in the transformation rate. Several studies have reported that microalgae are able to secrete several small molecules, including phenolics into the culture medium during their growth [ 32 – 34 ]. It is likely that some of these molecules are capable of inducing the Agrobacterium Vir genes, explaining the transformation as well as Vir gene induction.…”

Section: Discussionmentioning

confidence: 99%

Synergistic Action of D-Glucose and Acetosyringone on Agrobacterium Strains for Efficient Dunaliella Transformation

Srinivasan

Gothandam

2016

PLoS ONE

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An effective transformation protocol for Dunaliella, a β-carotene producer, was developed using the synergistic mechanism of D-glucose and Acetosyringone on three different Agrobacterium strains (EHA105, GV3101 and LBA4404). In the present study, we investigated the pre-induction of Agrobacterium strains harboring pMDC45 binary vector in TAP media at varying concentrations of D-glucose (5 mM, 10 mM, and 15mM) and 100 μM of Acetosyringone for co-cultivation. Induction of Agrobacterium strains with 10 mM D-glucose and 100 μM Acetosyringone showed higher rates of efficiency compared to other treatments. The presence of GFP and HPT transgenes as a measure of transformation efficiency from the transgenic lines were determined using fluorescent microscopy, PCR, and southern blot analyzes. Highest transformation rate was obtained with the Agrobacterium strain LBA4404 (181 ± 3.78 cfu per 106 cells) followed by GV3101 (128 ± 5.29 cfu per 106 cells) and EHA105 (61 ± 5.03 cfu per 106 cells). However, the Agrobacterium strain GV3101 exhibited more efficient single copy transgene (HPT) transfer into the genome of D. salina than LBA4404. Therefore, future studies dealing with genetic modifications in D. salina can utilize GV3101 as an optimal Agrobacterium strain for gene transfer.

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Section: Discussionmentioning

confidence: 99%

Synergistic Action of D-Glucose and Acetosyringone on Agrobacterium Strains for Efficient Dunaliella Transformation

Srinivasan

Gothandam

2016

PLoS ONE

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“…Bacterium, Agrobacterium tumefaciens is used in this method to transfer DNA segment into the target cells by natural infection process. The T-DNA sequence (part of Ti plasmid) of the bacterium, which allows insertion of the desired DNA construct is utilized for the transformation ( Pazhukunnel et al, 2015 ; Banerjee et al, 2016 ) This method has been utilized in transformation of microalgal species like Chlamydomonas reinhardtii , Haematococcus lacustris , Chlorella sp, Dunaliella bardawil , Dunaliella salina , Symbiodinium sp, Nannochloropsis sp. and Parachlorella kessleri .…”

Section: Algal Transformation Methodsmentioning

confidence: 99%

Genetic Engineering of Microalgae for Secondary Metabolite Production: Recent Developments, Challenges, and Future Prospects

Sreenikethanam

Raj

Banu

et al. 2022

Front. Bioeng. Biotechnol.

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Microalgae are highly diverse photosynthetic organisms with higher growth rate and simple nutritional requirements. They are evolved with an efficiency to adapt to a wide range of environmental conditions, resulting in a variety of genetic diversity. Algae accounts for nearly half of global photosynthesis, which makes them a crucial player for CO2 sequestration. In addition, they have metabolic capacities to produce novel secondary metabolites of pharmaceutical, nutraceutical and industrial applications. Studies have explored the inherent metabolic capacities of microalgae with altered growth conditions for the production of primary and secondary metabolites. However, the production of the targeted metabolites at higher rates is not guaranteed just with the inherent genetic potentials. The strain improvement using genetic engineering is possible hope to overcome the conventional methods of culture condition improvements for metabolite synthesis. Although the advanced gene editing tools are available, the gene manipulation of microalgae remains relatively unexplored. Among the performed gene manipulations studies, most of them focus on primary metabolites with limited focus on secondary metabolite production. The targeted genes can be overexpressed to enhance the production of the desired metabolite or redesigning them using the synthetic biology. A mutant (KOR1) rich in carotenoid and lipid content was developed in a recent study employing mutational breeding in microalgae (Kato, Commun. Biol, 2021, 4, 450). There are lot of challenges in genetic engineering associated with large algal diversity but the numerous applications of secondary metabolites make this field of research very vital for the biotech industries. This review, summarise all the genetic engineering studies and their significance with respect to secondary metabolite production from microalgae. Further, current genetic engineering strategies, their limitations and future strategies are also discussed.

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“…A binary vector consisting of additional virulence genes from a Ti plasmid to enhance the transformation frequency is refered to as a super-binary vector (Komori and Komari 2011). Vir gene inducers refer to factors that facilitate the expression of vir genes which include, amongst others, acetosyringone, vanillin caffeic acid (Simon et al 2015). A ternary system makes use of an accessory plasmid that is a virulence helper plasmid to help carry additional virulence gene cluster.…”

Section: Agrobacterium-mediated Transformationmentioning

confidence: 99%

“…Acetosyringone is a prominent phenolic compound that contributes greatly to the effective transformation of Agrobacterium. Synergistic actions of various other compounds responsible for transformation may include vanillin, vanillic acid, caffeic acid, gallic acid, and coumarin as in the case of the microalga Dunaliella salina (Simon et al 2015). Additionally, a down-regulation of o-methyltransferases has been reported to reduce the susceptibility of plants to agro-infection by lowering the virulence of Agrobacterium (Maury et al 2010).…”

Section: Agrobacterium-mediated Transformationmentioning

confidence: 99%

A discussion on cotton transformation during the last decade (2010–2021); an update on present trends and future prospects

et al. 2021

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The introduction of genetically modified (GM) cotton in 1996 in the US and its worldwide spread later rejuvenated cotton production in many parts of the world. The evolution is continued since then and currently, the 3rd and fourth generation of same GM cotton is grown in many parts of the world. The GM cotton introduced in 1996 was simple Bt cotton that expressed a single Cry1Ac gene, the later generation carried multiple Cry genes along with the genes controlling herbicide tolerance. Current day GM cotton does not only give stable resistance against lepidopteran insects but also facilitates the farmers to spray broad-spectrum herbicides without harming the crop. The evolution of GM cotton is continued both on the basic and applied side and interventions have been introduced during the last decade. Earlier the cotton transformation was limited to Cocker strains which are getting possible in many other varieties, too. It is successful with both gene gun, and Agrobacterium and inplanta transformation has made it a routine activity. Apart from overexpression studies for various purposes including biotic, abiotic, and quality traits, RNAi and genome editing are explored vigorously. Through this review, we have tried to explore and discuss various interventions for improving transformation protocols, the applications of cotton transformation, and future strategies being developed to get maximum benefits from this technology during the last decade.

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Vir gene inducers in Dunaliella salina ; an insight in to the Agrobacterium -mediated genetic transformation of microalgae

Cited by 9 publications

References 28 publications

Synergistic Action of D-Glucose and Acetosyringone on Agrobacterium Strains for Efficient Dunaliella Transformation

Synergistic Action of D-Glucose and Acetosyringone on Agrobacterium Strains for Efficient Dunaliella Transformation

Genetic Engineering of Microalgae for Secondary Metabolite Production: Recent Developments, Challenges, and Future Prospects

A discussion on cotton transformation during the last decade (2010–2021); an update on present trends and future prospects

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