Arbuscular mycorrhiza in mini‐mycorrhizotrons: first contact of Medicago truncatula roots with Glomus intraradices induces chalcone synthase

Summary • The association of plants with arbuscular mycorrhizal (AM) fungi is widespread in nature, but little is known about molecular aspects of this symbiosis. Particularly during the early stages of the AM symbiosis, it is difficult to monitor growth of the two partners, to dissect gene expression patterns and to correlate them with plant, fungal or symbiosis development. • A new system, the ‘mini-mycorrhizotron’, was established to cultivate seedlings of Medicago truncatula in mycorrhizal symbiosis with Glomus intraradices under gnotobiotic conditions. This system allows natural growth of the symbiotic partners and permits the continuous noninvasive observation of the development of plant and fungus under a microscope. • The mini-mycorrhizotron was used to determine the stage of induction of a mycorrhiza-related gene detected by differential display-reverse transcription-PCR, namely a novel chalcone synthase (Mt-chs1). The gene is induced in roots at the stage of the first fungal contact. • The mini-mycorrhizotron allowed identification and cloning of a symbiosis-related gene, and the correlation between its expression and the developmental stage of the symbiosis was established. This provides a useful tool for molecular and developmental studies of the early stages of AM symbioses. Mt-chs1 nucleotide sequence can be found at the Genbank data base (accession no. AJ277211). The following sequences were also used: Glycine maxchs (X52097), chs1 (X54644); chs2 (X65635), chs3 (X53958), chs5 (L07647), chs6 (L03352), chs7 (M98871); Pisum sativumchs (X80007), chs1 (D10661), chs2 (X63334), chs3 (D88261), chs4 (D88260), chs5 (D88262), chs7 (D88263); Medicago sativachs1 (L02901), chsI (X68106), chs2 (L02902), chs4–1 (U01018), chs8 (L02904), chs9 (L02905), 12–1 (U01021); Medicago truncatulachs1, this work (AJ277211); Trifolium subterraneumchs1 (M91193), chs2 (M91194), chs3 (L24515), chs4 (L24516), chs5 (L24517), chs6 (M91195); Phaseolus vulgarischs (X06411) and Vigna unguiculatachs (X74821).