Comparative Analysis of Three Different Total Nucleic Acid Extraction Protocols for the Diagnosis of Geminiviruses in Squash (<i>Cucurbita moschata</i>)

Hernández, E.; Mora-Umaña, Floribeth; Albertazzi, Federico J.; Karkashian, James P.; Ramírez, Pedro José Rueda

doi:10.1111/j.1439-0434.2011.01850.x

Cited by 3 publications

(6 citation statements)

References 22 publications

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“…Los ácidos nucleicos totales de las muestras de tomate y arvenses se extrajeron según el protocolo descrito por Dellaporta et al (1983), con las modificaciones detalladas por Hernández et al (2012). La presencia de begomovirus fue evaluada inicialmente utilizando la técnica de hibridación molecular por dot blot según lo detallan Hernández et al (2012).…”

Section: Detección De Begomovirusunclassified

“…La presencia de begomovirus fue evaluada inicialmente utilizando la técnica de hibridación molecular por dot blot según lo detallan Hernández et al (2012).…”

Section: Detección De Begomovirusunclassified

“…A partir de los productos de RCA se realizó una reacción en cadena de la polimerasa (PCR) de punto final con dos diferentes pares de iniciadores: el primer par PAR1c715 y PAL1v1978 permitieron amplificar una región de aproximadamente 1,4 kb de la parte superior del componente A (Top-A) del genoma del virus (Rojas et al, 1993), y los iniciadores PAV494 y PAC1048 que amplificaron la región más conservada del gen que codifica la proteína de cubierta (gen cp) (Wyatt y Brown, 1996). Las reacciones de PCR se realizaron según las condiciones descritas por Hernández et al (2012).…”

Section: Detección De Begomovirusunclassified

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Detección de crinivirus y begomovirus en plántulas de tomate y arvenses asociadas a semilleros

et al. 2017

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The aim of this work was to detect plant infections caused by Tomato chlorosis virus (ToCV) and begomovirus in tomato plantlets, and in growing weeds around nursery greenhouses. During one year, starting in April 2008, 168 leaf tissue samples were collected, 90 tomato plantlets and 78 weeds from three different nurseries in Cartago province, Costa Rica. Reverse transcription and real time polymerase chain reaction (qRT-PCR) was used to determine that 18,9% of tomato plantlets and 7,7% of weeds were infected with ToCV virus. Begomoviruses were detected using Dot Blot hybridization and non-radioactive probe. Next, hybridization results were confirmed using Rolling Circle Amplification (RCA) followed by PCR, using universal primers. None tomato plantlet resulted positive when tested, but there were six weeds infected; in fact, Phytolacca icosandra and Brassica sp. were both coinfected with ToCV virus. These results suggest that ToCV infected tomato plantlets when commercialized, could serve as way of virus introduction to other country regions. Finally, weeds growing around greenhouses have shown to be potential viral sources of ToCV and begomovirus.

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Section: Detección De Begomovirusunclassified

“…La presencia de begomovirus fue evaluada inicialmente utilizando la técnica de hibridación molecular por dot blot según lo detallan Hernández et al (2012).…”

Section: Detección De Begomovirusunclassified

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Detección de crinivirus y begomovirus en plántulas de tomate y arvenses asociadas a semilleros

et al. 2017

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“…The plant tissue samples were lyophilised and stored at À70 C (10-15 mg). DNA extractions were carried out by the method of Dellaporta et al (1983) modified by Hernández et al (2012).…”

Section: Introductionmentioning

confidence: 99%

Begomovirus diversity in tomato crops in Costa Rica

Valverde‐Méndez,

Hernández,

Matamoros

et al. 2023

Annals of Applied Biology

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Begomoviruses (Geminiviridae family) are characterized by their high recombination rate and a wide range of hosts, making their control difficult. In Costa Rica, various species of bipartite begomoviruses have been reported, which are Pepper golden mosaic virus (PepGMV), Tomato yellow mottle virus (ToYMoV), Tomato leaf curl Sinaloa virus (ToLCSiV) and the monopartite begomovirus Tomato yellow leaf curl virus (TYLCV). Since the TYLCV first report in Costa Rica, neither additional knowledge has been produced on how this begomovirus has spread in the country's territory nor on the distribution of the other bipartite species. A total of 429 tomato samples collected during the years 2015–2016 were used to study these aspects. Each sample was georeferenced and analysed with various techniques such as nucleic acid hybridization, polymerase chain reaction (PCR) and sequencing for the begomoviruses previously reported in Costa Rica. It was found that the presence/absence of the different species can vary, depending on the province. TYLCV is present in the six provinces analysed in this work, with a proportion from 3.7 to 86.6 per cent. Alajuela, Cartago, and Heredia are the provinces most affected by tomato‐infecting begomoviruses. Fourteen different haplotypes of TYLCV were detected, but all were identified as TYLCV‐IL. The distribution of TYLCV was related to the presence of the whitefly Bemisia tabaci MED, especially in the country's main tomato production areas. This information allows the phytosanitary surveillance services to develop strategies for the integrated management of the disease and to contribute data to the genetic improvement programmes of the crop.

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“…Improving nucleic acid extraction essentially involves enhancing the quantity of extracted nucleic acid while maintaining its purity. This indicates that continuous optimization and improvement of nucleic acid extraction technology is of great significance [ 3 , 16 , 17 ]. As the biochemical characters of nucleic acids are similar [ 3 ], DNA extraction was selected as the model in this study.…”

Section: Introductionmentioning

confidence: 99%

Improved DNA extraction on bamboo paper and cotton is tightly correlated with their crystallinity and hygroscopicity

Lei

2022

PLoS ONE

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DNA extraction, a vital pre-requisite for most biological studies, continues to be studied extensively. According to some studies, DNA shows a certain degree of absorbability on filter paper made of plant fiber-based adsorbent material. However, the principle underlying such specific adsorption as well as plant species associated with plant fiber-based adsorbents and optimized extraction conditions have not yet been studied. This study demonstrates the tight correlation between crystallinity and hygroscopicity in plant fiber-based adsorbents used for DNA extraction and proposes the concept of DNA adsorption on plant fiber-based adsorbents, for the first time. We also explored optimal extracting and eluting conditions and developed a novel plant fiber-based DNA extraction method that was quadruple times more powerful than current approaches. Starting with the screening of various types of earthed plant fiber-based adsorbents, we went on to mine new plant fiber-based adsorbents, bamboo paper and degreased cotton, and succeeded in increasing their efficiency of DNA extraction to 4.2 times than that of current approaches. We found a very strong correlation between the crystallinity and hygroscopicity of plant fiber-based adsorbents which showed efficiency for DNA extraction, and thus propose a principle that potentially governs such specific adsorption processes, in the hope that this information may guide related multidisciplinary research studies in the future. Nanodrop, electrophoresis and PCR were selected to demonstrate the quantity, quality, integrity and utility of the extracted DNA. Furthermore, crystallinity, hygroscopicity, pore size distribution and composition of plant fiber-based adsorbents were studied to explore their correlation in an attempt to understand the principle underlying this particular type of adsorption. The findings of this study may be further extended to the extraction of other types of nucleic acids with similar biochemical properties.

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Comparative Analysis of Three Different Total Nucleic Acid Extraction Protocols for the Diagnosis of Geminiviruses in Squash (Cucurbita moschata)

Cited by 3 publications

References 22 publications

Detección de crinivirus y begomovirus en plántulas de tomate y arvenses asociadas a semilleros

Detección de crinivirus y begomovirus en plántulas de tomate y arvenses asociadas a semilleros

Begomovirus diversity in tomato crops in Costa Rica

Improved DNA extraction on bamboo paper and cotton is tightly correlated with their crystallinity and hygroscopicity

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