Molecular diversity of arbuscular mycorrhizal fungi colonising Hyacinthoides non-scripta (bluebell) in a seminatural woodland

Arbuscular mycorrhizal (AM) fungi form symbiotic associations with plant roots. Around 150 species have been described and it is becoming clear that many of these species have different functional properties. The species diversity of AM fungi actively growing in roots is therefore an important component of ecosystem diversity. However, it is difficult to identify AM fungi below the genus level from morphology in planta, as they possess few informative characters. We present here a molecular method for identifying infrageneric sequence types that estimate the taxonomic diversity of AM fungi present in actively growing roots. Bluebell roots were sampled from beneath two different canopy types, oak and sycamore, and DNA sequences were amplified from roots by the polymerase chain reaction with fungal-specific primers for part of the small subunit ribosomal RNA gene. Restriction fragment length polymorphism among 141 clones was assessed and 62 clones were sequenced. When aligned, discrete sequence groups emerged that cluster into the three families of AM fungi: Acaulosporaceae, Gigasporaceae and Glomaceae. The sequence variation is consistent with rRNA secondary structure. The same sequence types were found at both sampling times. Frequencies of Scutellospora increased in December, and Acaulospora increased in abundance in July. Sites with a sycamore canopy show a reduced abundance of Acaulospora, and those with oak showed a reduced abundance of Glomus. These distribution patterns are consistent with previous morphological studies carried out in this woodland. The molecular method provides an alternative method of estimating the distribution and abundance of AM fungi, and has the potential to provide greater resolution at the infrageneric level.