Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani

Background Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) is one of the most important pathogens of rice (Oryza sativa L.) that causes severe yield losses in all rice-growing regions. Sclerotia, formed from the aggregation of hyphae, are important structures in the life cycles of R. solani...

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Autores principales:	Shu,Canwei, Sun,Si, Chen,Jieling, Chen,Jianyi, Zhou,Erxun
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2014
Materias:	Polyvinylpyrrolidone Rhizoctonia solani RT-PCR Sclerotia Sodium dodecyl sulfate Total RNA extraction
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100009
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spelling	oai:scielo:S0717-345820140001000092014-04-07Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solaniShu,CanweiSun,SiChen,JielingChen,JianyiZhou,Erxun Polyvinylpyrrolidone Rhizoctonia solani RT-PCR Sclerotia Sodium dodecyl sulfate Total RNA extraction Background Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) is one of the most important pathogens of rice (Oryza sativa L.) that causes severe yield losses in all rice-growing regions. Sclerotia, formed from the aggregation of hyphae, are important structures in the life cycles of R. solani and contain a large quantity of polysaccharides, lipids, proteins and pigments. In order to extract high-quality total RNA from the sclerotia of R. solani, five methods, including E.Z.N.A.™ Fungal RNA Kit, sodium dodecyl sulfate (SDS)-sodium borate, SDS-polyvinylpyrrolidone (PVP), guanidinium thiocyanate (GTC) and modified Trizol, were compared in this study. Results The electrophoresis results showed that it failed to extract total RNA from the sclerotia using modified Trizol method, whereas it could extract total RNA from the sclerotia using other four methods. Further experiments confirmed that the total RNA extracted using SDS-sodium borate, SDS-PVP and E.Z.N.A.™ Fungal RNA Kit methods could be used for RT-PCR of the specific amplification of GAPDH gene fragments, and that extracted using GTC method did not fulfill the requirement for above-mentioned RT-PCR experiment. Conclusion It is concluded that SDS-sodium borate and SDS-PVP methods were the better ones for the extraction of high-quality total RNA that could be used for future gene cloning and expression studies, whereas E.Z.N.A.™ Fungal RNA Kit was not taken into consideration when deal with a large quantity of samples because it is expensive and relatively low yield.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.17 n.1 20142014-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100009en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Polyvinylpyrrolidone Rhizoctonia solani RT-PCR Sclerotia Sodium dodecyl sulfate Total RNA extraction
spellingShingle	Polyvinylpyrrolidone Rhizoctonia solani RT-PCR Sclerotia Sodium dodecyl sulfate Total RNA extraction Shu,Canwei Sun,Si Chen,Jieling Chen,Jianyi Zhou,Erxun Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani
description	Background Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) is one of the most important pathogens of rice (Oryza sativa L.) that causes severe yield losses in all rice-growing regions. Sclerotia, formed from the aggregation of hyphae, are important structures in the life cycles of R. solani and contain a large quantity of polysaccharides, lipids, proteins and pigments. In order to extract high-quality total RNA from the sclerotia of R. solani, five methods, including E.Z.N.A.™ Fungal RNA Kit, sodium dodecyl sulfate (SDS)-sodium borate, SDS-polyvinylpyrrolidone (PVP), guanidinium thiocyanate (GTC) and modified Trizol, were compared in this study. Results The electrophoresis results showed that it failed to extract total RNA from the sclerotia using modified Trizol method, whereas it could extract total RNA from the sclerotia using other four methods. Further experiments confirmed that the total RNA extracted using SDS-sodium borate, SDS-PVP and E.Z.N.A.™ Fungal RNA Kit methods could be used for RT-PCR of the specific amplification of GAPDH gene fragments, and that extracted using GTC method did not fulfill the requirement for above-mentioned RT-PCR experiment. Conclusion It is concluded that SDS-sodium borate and SDS-PVP methods were the better ones for the extraction of high-quality total RNA that could be used for future gene cloning and expression studies, whereas E.Z.N.A.™ Fungal RNA Kit was not taken into consideration when deal with a large quantity of samples because it is expensive and relatively low yield.
author	Shu,Canwei Sun,Si Chen,Jieling Chen,Jianyi Zhou,Erxun
author_facet	Shu,Canwei Sun,Si Chen,Jieling Chen,Jianyi Zhou,Erxun
author_sort	Shu,Canwei
title	Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani
title_short	Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani
title_full	Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani
title_fullStr	Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani
title_full_unstemmed	Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani
title_sort	comparison of different methods for total rna extraction from sclerotia of rhizoctonia solani
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2014
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100009
work_keys_str_mv	AT shucanwei comparisonofdifferentmethodsfortotalrnaextractionfromsclerotiaofrhizoctoniasolani AT sunsi comparisonofdifferentmethodsfortotalrnaextractionfromsclerotiaofrhizoctoniasolani AT chenjieling comparisonofdifferentmethodsfortotalrnaextractionfromsclerotiaofrhizoctoniasolani AT chenjianyi comparisonofdifferentmethodsfortotalrnaextractionfromsclerotiaofrhizoctoniasolani AT zhouerxun comparisonofdifferentmethodsfortotalrnaextractionfromsclerotiaofrhizoctoniasolani
_version_	1718441888293322752

Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani

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