Taxonomic identification of arbuscular mycorrhizal (AM) fungal spores extracted directly from the field is sometimes difficult because spores are often degraded or parasitized by other organisms. Single-spore inoculation of a suitable host plant allows for establishing monosporic cultures of AM fungi. This study aimed to propagate AM fungal spores isolated from maize soil using single spores for morphological characterization. First, trap cultures were established to trigger the sporulation of AM fungal species. Second, trap cultures were established with individual morphotypes by picking up only one spore under a dissecting microscope and transferring it to a small triangle of sterilized filter paper, which was then carefully inoculated below a root from germinated sorghum seeds in each pot and covered with a sterile substrate. All pots were placed in sunbags and maintained in a plant growth room for 120 days. Spores obtained from single spore trap cultures from each treatment, maize after oats (MO), maize after maize (MM), maize after peas (MP), and maize after soybean (MS), were extracted using the sieving method. Healthy spores were selected for morphological analysis. Direct PCR was conducted by crushing spores in RNAlater and applying three sets of primer pairs: ITS1 × ITS4, NS31 × AML2, and SSUmcf and LSUmBr. Nucleotide sequences obtained from Sanger sequencing were aligned on MEGA X. The phylogenetic tree showed that the closest neighbors of the propagated AM fungal species belonged to the genera Claroideoglomus, Funneliformis, Gigaspora, Paraglomus, and Rhizophagus. The morphological characteristics were compared to the descriptive features of described species posted on the INVAM website, and they included Acaulospora cavernata, Diversispora spurca, Funneliformis geosporus, Funneliformis mosseae, Gigaspora clarus, Gigaspora margarita, Glomus macrosporum, Paraglomus occultum, and Rhizophagus intraradices. These findings can provide a great contribution to crop productivity and sustainable management of the agricultural ecosystem. Also, the isolate analyzed could be grouped into efficient promoters of growth and mycorrhization of maize independent of their geographical location.

The use of arbuscular mycorrhizal fungi (AMF) as biofertilizer in agriculture is a sustainable approach to fertilization. The first step in the production of AMF biofertilizer is inoculation of mycotrophic plants with a composite of soil and native plant roots, containing potentially viable AMF spores from natural habitats, to a trap culture. A single host plant or a consortium of host plants can be used to propagate AMF spores. However, the difference in the comparative efficiency of mono- and co-cultivated host plants used for the production of AMF spores and the maintenance of original AMF community composition has not been well elucidated. Here, we prepared trap culture with nutrient-poor soil from coastal sand dune vegetation collected during the dry season when the AMF spore density and relative abundance of Glomeromycota ITS2 sequences were significantly higher (p = <0.05) than in the wet season. The AMF communities in the soil were mainly composed of Glomus spp. Maize (Zea mays L.) and/or Sorghum (Sorghum bicolor (L.). Moench) were grown in trap cultures in the greenhouse. Our results demonstrated that co-cultivation of the host plants increased the production of AMF spores but, compared to mono-cultivation of host plants, did not better sustain the native AMF community compositions in the coastal sand dune soil. We propose that the co-cultivation of host plants in a trap culture broadens AMF-host plant compatibilities and thus sustains the symbiotic association of the natively diverse AMF. Therefore, the results of this study suggest that further research is needed to confirm whether the co-culturing of more than one host plant is as efficient a strategy as using a monoculture of a single host plant.

OBJECTIVE: Arbuscular mycorrhizal fungi (AMF) are often regarded as non-specific symbionts, but some AMF communities show host preference in various ecosystems including vineyards. Grapevine plants are very responsive to AMF colonization. Although these fungi have potentially significant applications for sustainable agricultural ecosystems, there is a gap in knowledge regarding AMF-grapevine interactions worldwide and especially in New Zealand. This study focused on identifying AMF taxa colonizing grapevines in New Zealand vineyards and investigated the effect of grapevine rootstocks on AMF community diversity and composition.
RESULTS: Denaturing gradient gel electrophoresis (DGGE) and trap cultures were used to characterize the AMF communities. Grapevine roots from three vineyards and nine rootstocks were analysed by DGGE and used in trap cultures for AMF recovery. Trap cultures allowed the recovery of six AMF spore morphotypes that belonged to Ambispora sp., Claroideoglomus sp., Funneliformis sp. and Glomus sp. Bands excised, reamplified and sequenced from the DGGE were assigned to Glomus sp., Rhizophagus sp. and Claroideoglomus sp. The AMF community analyses demonstrated that rootstock significantly (P < 0·05) influenced the AMF community composition in all sites.
CONCLUSIONS: The study showed that for a comprehensive identification of AMF, both results from trap culture and molecular work were needed and that the rootstock cultivar was the main driver of the arbuscular mycorrhizal community colonizing the roots.
CONCLUSIONS: This study provides a firm foundation for future research exploring the beneficial use of AMF in enhancing grapevine production and sustainability.