Co-Authors:
Alkan, N., Dept. of Agronomy and Nat. Resources, Institute of Field and Garden Crops, Volcani Center, PO Box 6, Bet Dagan 50 250, Israel
Gadkar, V., Dept. of Agronomy and Nat. Resources, Institute of Field and Garden Crops, Volcani Center, PO Box 6, Bet Dagan 50 250, Israel
Coburn, J., Dept. of Agronomy and Nat. Resources, Institute of Field and Garden Crops, Volcani Center, PO Box 6, Bet Dagan 50 250, Israel, Department of Biology, University of Concordia, 1455 de Maisonneuve Boulevard, West Montreal, Que. H3G 1M8, Canada
Yarden, O., Dept. Plant Pathol. and Microbiol., Fac. Agric., Food Environ. Qual. S., Hebrew University of Jerusalem, Rehovot 76 100, Israel
Kapulnik, Y., Dept. of Agronomy and Nat. Resources, Institute of Field and Garden Crops, Volcani Center, PO Box 6, Bet Dagan 50 250, Israel
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.
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