Quantification of roots in soil, plant-fungal symbioses and in mixed plant populations with real-time PCR

Abstract

Plant-soil and underground plant-plant interactions have direct implications for the conservation of biodiversity, plant productivity and understanding of local ecosystem responses to global environmental change. Study of associated organisms in the soil, symbiosis and in mixed plant populations, is limited by difficulties in the species quantification and sensitive detection. We have designed and tested new PCR primers and hydrolysis probes for TaqMan assays that target the nuclear ribosomal gene plant DNA. Species-specific probes and universal primers were developed for the monocots and dicots plant species and evaluated for specificity and sensitivity in TaqMan assays on DNA extracted from soil and in mixed plant populations. The assays were sensitive and able to quantify rear targets in mixed plant populations. This approach appears to offer a quick and sensitive way to study roots in soil, symbiosis and in mixed plant populations composition and could provide insights into root function and responses not readily provided by other methods. These features identify the new primer sets developed in this study as useful complements to other universal primers for the study of plant diversity and community composition.