Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani|
Shu, Canwei; Sun, Si; Chen, Jieling; Chen, Jianyi & Zhou, Erxun
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), guanidiniumthiocyanate (GTC) andmodified Trizol, were compared in this study.
Results: The electrophoresis results showed that it failed to extract total RNA from the sclerotia usingmodified 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–PVPmethods 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.
Polyvinylpyrrolidone; Rhizoctonia solani; RT-PCR; Sclerotia; Sodium dodecyl sulfate; Total RNA extraction