Quantification of Mycorrhiza from Taiwania Using Real-Time PCR

Abstract

Scutellospora calospora and Glomus sp. were 2 dominant arbuscular mycorrhizal fungi (AMF) species extracted from rhizosphere soil of Taiwania (Taiwania cryptomerioides) tree stands near Feng-Gang Mountain nursery at Liukuei in Taiwan by wet sieving and sucrose centrifugation. To efficiently detect and quantify the population of the AMF, S. calospora and Glomus sp., associated with the roots of Taiwania, a real-time polymerase chain reaction (PCR) technique was used to amplify the 18S rRNA gene (ca. 649 bp) with specific primers, and it was detected using fluorescent SYBR(superscript ®) green I dye. Standard curves showed a linear relation (r^2=0.998 and r^2=0.999 each) between log values of the starting copy numbers of target sequences and real-time PCR threshold cycles. The real-time PCR quantification assay can also be used to monitor individual AMF communities in the greenhouse. The results demonstrate that the real-time PCR technique is a powerful universal tool for quantifying individual AMF species in modern mycorrhizal research.