| Literature DB >> 33873725 |
Noam Alkan1,2, Vijay Gadkar1,2, Joel Coburn1,3, Oded Yarden4, Yoram Kapulnik1.
Abstract
• A rapid method to quantify the colonization of the arbuscular mycorrhizal fungus (AMF) Glomus intraradices in planta using quantitative real-time polymerase chain reaction (qRT-PCR) technique. • Specific PCR primers for the fungus (28S rDNA sequence) and host root tissue (chitinase and chalcone synthase gene) were developed and their respective specificity determined. • The plant specific primers for Lycopersicon esculentum, Medicago truncatula amplified linearly over a concentration range of: 6.4 pg to 20 ng. The G. intraradices-specific primer amplified as low as 1 pg of its target DNA, which allowed us to detect a single spore of the fungus. High degrees of correlation were obtained when threshold cycle (Ct) was plotted against vesicular, hyphal and total colonization using microscopically quantified host roots. • This is the first report of the application of the qRT-PCR technique for quantification of AMF colonization in planta. The success of its application should open up the possibility of its wider application in AM research.Entities:
Keywords: Glomus intraradices; Medicago truncatula; arbuscular mycorrhiza; in vitro culture; quantitative real-time PCR (qRT-PCR); tomato
Year: 2004 PMID: 33873725 DOI: 10.1046/j.1469-8137.2004.00975.x
Source DB: PubMed Journal: New Phytol ISSN: 0028-646X Impact factor: 10.151