Background: The dried sclerotium of medicinal fungus
Polyporus umbellatus
(Pers.) Fries has many
pharmacological functions such as diuretic and anticancer activity, in which high-content
polysaccharides may play an important role. However, RNA isolation is difficult in filamentous fungi and
lacking in
P. umbellatus.
Results: Five methods for RNA extraction from five strains collected from four
provinces were assessed for their ability to recover a high-quality RNA applicable for sequence-related
amplification polymorphism (SRAP) PCR and GDP-D-mannose pyrophosphorylase (GMP) gene
expression profiles. Both A260/A280 and A260/A230 ratios of the best Trizol Plus + RNAiso-mate for
Plant Tissue method are around 2 with a yield of 1122.00 ± 0.21 ng μl
-1. The Trizol method also
showed good quality with the yield 469.60 ng μl
-1. The SRAP PCR amplified clear and polymorphic
bands in all five cDNA samples transcribed from RNA by using primer Me4-Em4. GMP gene fragment
(1251 bp) was successfully amplified by RT-PCR, suggesting the integrity of isolated RNA.
Conclusion:All these results showed that the total RNA isolated by this protocol is of sufficient quality
for subsequent molecular applications.
