Direct colony PCR for rapid identification of varied microalgae from freshwater environment

Radha, Sudhakar; Fathima, Anwar Aliya; Sellamuthu, Iyappan; Ramya, Mohandass

doi:10.1007/s10811-012-9895-0

Cited by 40 publications

(20 citation statements)

References 25 publications

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“…Molecular identification of microalgae. Colony PCR protocol was followed for all the species as per previous study (Radha et al, 2013). The internal transcribed spacer (ITS-2) region of each isolate was amplified using universal primer (Hall et al, 2010).…”

Section: Experimental Materials and Methodsmentioning

confidence: 99%

Agrobacterium-Mediated Transformation of Three Freshwater Microalgal Strains

Sanitha¹,

Radha²,

Fatima³

et al. 2014

Pol J Microbiol

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Microalgal transformation has gained interest in recent years. Agrobacterium-mediated transformation remains as the most efficient method for the development of transgenic plants and microalgae due to its wide host range, inexpensive procedure and transfer of large segments of DNA. In the present study, three different microalgal species were isolated from freshwater environment and identified based on the morphological characteristics and ITS-2 region amplification. Agrobacterium-mediated transformation was successful for the isolates Chlorella sp., Ankistrodesmus sp and Scenedesmus bajacalifornicus. Gene integration and expression was confirmed by PCR amplification of hptII and GUS histochemical assay. A. tumifaciens contamination was checked by amplification of npt II gene (kanamycin resistant) which lies outside the T-border. Based on GUS assay, transformation efficiencies were found to be 12.25% for Chlorella sp. 2.96% for Scenedesmus bajacalifornicus and 3.5% for Ankistrodesmus sp.

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Section: Experimental Materials and Methodsmentioning

confidence: 99%

Agrobacterium-Mediated Transformation of Three Freshwater Microalgal Strains

Sanitha¹,

Radha²,

Fatima³

et al. 2014

Pol J Microbiol

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“…La identificación molecular de microalgas utilizando secuencias de ADN podría proporcionar los medios para identificarlas de forma consistente y rápida, independientemente de la etapa de la vida y fenotipo (Hadi et al, 2016). Recientemente se han desarrollado protocolos eficaces para la identificación de cepas de microalgas utilizando varios marcadores moleculares, entre los que destacan los genes de la subunidad mayor de la ribulosa-1,5-bisfosfato carboxilasa/oxigenasa (rbcL), el espaciador interno transcrito 2 (ITS-2) y el gen del ARNr 18S (Radha et al, 2013). En el presente estudio, se revisó la clasificación de algunas cepas de microalgas de la colección del Centro de Investigación en Biotecnología del Instituto Tecnológico de Costa Rica (CIB-ITCR) utilizando datos de la secuencia de ADN codificante para la subunidad pequeña del ARN ribosomal (SSU rDNA) y se reportó las primeras anotaciones moleculares sobre la diversidad de microalgas de Costa Rica.…”

Section: Genéticaunclassified

Identificación de una colección de microalgas aisladas de Costa Rica mediante secuenciación de ADNr 18S

et al. 2018

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RESUMENLas microalgas componen un diverso grupo polifilético de microorganismos fotosintéticos. Debido a su potencial biotecnológico, los estudios para aislar e identificar nuevas cepas han incrementado, por lo que es necesario el desarrollo de nuevas técnicas para su correcta identificación y clasificación. Utilizando herramientas de biología molecular, en este estudio se analizó el gen del ADNr 18S de 12 cepas microalgales aisladas de diferentes regiones de Costa Rica, resultando seis pertenecientes a la clase Trebouxiophyceae, tres a Chlorophyceae, dos a Prymnesiophyceae y una a Cyanidiophyceae. Este estudio reporta por primera vez la identificación molecular de cepas microalgales aisladas de Costa Rica, resaltando la diversidad de estos microorganismos en el país.Palabras clave: biotecnología microalgal, filogenia, identificación molecular. ABSTRACTMicroalgae integrate a diverse polyphyletic group of photosynthetic microorganisms. Due to its biotechnological potential, studies to isolate and identify new strains have increased, hence, it is necessary to develop new techniques for their correct identification and classification. Using molecular biology tools, this study analyzed the 18S rDNA gene from 12 microalgal strains isolated from different regions of Costa Rica, resulting in six strains belonging to the class Trebouxiophyceae, three to Chlorophyceae, two to Prymnesiophyceae and one to Cyanidiophyceae. This study reports for the first time the molecular identification of microalgal strains isolated from Costa Rica, highlighting the diversity of these microorganisms in the country.

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“…While cellular morphology may be used as a preliminary guide for identification, molecular techniques are also required. Amplification of SSU rDNA (18S), revealed that RGd-1 had the closest sequence similarity (96%) to Lemnicola hungarica with 99% sequence coverage [20,21]. Full length ITS amplification was additionally performed to obtain greater resolution for strain identification, the results of which revealed 94% sequence similarity with its closest match, Sellaphora pupula.…”

Section: Diatom Isolation and Molecular Characterizationmentioning

confidence: 99%

Combining multiple nutrient stresses and bicarbonate addition to promote lipid accumulation in the diatom RGd-1

Moll

Gardner²,

Eustance

et al. 2014

Algal Research

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Algal biofuels represent a renewable, potentially viable, solution to mitigate transportation fuel demands. A novel diatom strain, RGd-1, isolated from Yellowstone National Park, produces high concentrations of lipids that can be converted to biodiesel. To increase the cell concentration and determine optimal conditions for growth, RGd-1 was grown without added Si, in the presence of four Si concentrations within the soluble range (0.5-2 mM), and one above the soluble range (2.5 mM). Medium Si concentrations and intracellular triacylglycerol (TAG) content were monitored daily by inductively coupled plasma mass spectrometry and Nile Red fluorescence, respectively (end-point TAG values were measured using gas chromatography). Si depletion with or without combined nitrate (NO 3 −) limitation was shown to induce TAG accumulation. Additionally, the effects of sodium bicarbonate (NaHCO 3) supplementation were examined on cultures grown using two NO 3 − concentrations (2.94 and 1 mM NO 3 −), which also resulted in increased TAG accumulation. It was determined that utilizing a combination of two independent physiological stresses and HCO 3 − supplementation resulted in the highest total and per cell TAG accumulation. 1 Triacylglycerol (TAG), monoacylglycerol (MAG), diacylglycerol (DAG), fatty acid methyl esters (FAME), dissolved inorganic carbon (DIC), Yellowstone National Park (YNP), Field Emission-Scanning Electron Microscopy (FE-SEM), inductively coupled plasma-mass spectrometry (ICP-MS), ion chromatography (IC), photosynthetically active radiation (PAR), dry cell weight (DCW), ash free dry weight (AFDW), biofuel potential (BP), gas chromatography-mass spectrometry (GC-MS), gas chromatography-flame ionization detection (GC-FID), relative fluorescence units (rfu), internally transcribed spacer (ITS). 2 The dissolved forms of silica are monomeric and dimeric silicic acid (Si(OH)4), which diatoms incorporate into their cell walls in the solid form of silica (SiO2). In an attempt to simplify this matter, the dissolved and solid forms of silicon will herein be referred to as Si and silica, respectively.

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Direct colony PCR for rapid identification of varied microalgae from freshwater environment

Cited by 40 publications

References 25 publications

Agrobacterium-Mediated Transformation of Three Freshwater Microalgal Strains

Agrobacterium-Mediated Transformation of Three Freshwater Microalgal Strains

Identificación de una colección de microalgas aisladas de Costa Rica mediante secuenciación de ADNr 18S

Combining multiple nutrient stresses and bicarbonate addition to promote lipid accumulation in the diatom RGd-1

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