The sterol biosynthesis inhibitor molecule fenhexamid impacts the vegetative compatibility of Glomus clarum.

The vegetative compatibility of the arbuscular mycorrhizal fungus (AMF) Glomus clarum MUCL 46238 was evaluated after continuous exposure to fenhexamid, a sterol biosynthesis inhibitor (SBI). Three lineages of this AMF were cultured in vitro for five generations in association with Ri T-DNA transformed carrot roots in the presence of 0, 5 or 10 mg l(-1) of fenhexamid. Whatever the AMF generation, fenhexamid at 5 and 10 mg l(-1) had no significant impact on the number of spores produced. However, vegetative compatibility tests (VCT) conducted with spores from the three lineages, in the presence of 10 mg l(-1) of fenhexamid, impacted the anastomosis process. At this concentration, the morphology of the germ tubes was modified. In addition, nitrotetrazolium-trypan blue staining revealed that 10 mg l(-l) of fenhexamid significantly reduced the probability of fusion between the germ tubes regardless of the culture conditions (i.e. absence or presence of fenhexamid) preceding the VCT. Our results demonstrated that spore production was not affected by fenhexamid, while anastomosis between germ tubes was decreased. This suggested that high concentrations, accumulation or repeated application of this SBI fungicide may impact the community structure of AMF in soil.

Impact of multispores in vitro subcultivation of Glomus sp. MUCL 43194 (DAOM 197198) on vegetative compatibility and genetic diversity detected by AFLP.

Vegetative compatibility and amplified fragment length polymorphism (AFLP) genotyping of in vitro multispores clonal lineages, issued from the same ancestor culture of the arbuscular mycorrhizal fungal strain MUCL 43194 and subcultured several generations in different locations, was assessed. Vegetative compatibility was studied by confronting the germ tubes of two spores from the same or different clonal lineages and stained with nitrotetrazolium blue-Trypan blue and diamidinophenylindole to detect hyphal fusions and nuclei, respectively. Further AFLP analysis of single spores was performed to assess the genetic profile and Dice similarity between clonal lineages. Germ tubes of spores distant by as many as 69 generations were capable of fusing. The anastomosis frequencies averaged 69% between spores from the same clonal lineage, 57% between spores from different clonal lineages, and 0% between spores belonging to different strains. The AFLP patterns showed similarities averaging 92% within clonal lineages and 86% between clonal lineages. Each spore presented unique genotype and some of them shared more markers with spores from different lineages than within the same lineage. We showed that MUCL 43194 maintained self-recognition for long periods of subcultures in vitro and that spores involved in compatibility tests had different genotypes. Our findings suggest that MUCL 43194 maintains genetic diversity by means of anastomoses.