Arbuscular mycorrhiza (AM) is a widespread symbiosis formed by most land plants with fungi from Glomeromycotina subdivision. The main problem in study of AM fungi is the complication in identification, associated with high intra- and interspecific genetic polymorphism, as well as obligate status of AM fungi in relation to host plant. The methodology for AM fungi identification is constantly undergoing major changes. In the review the selection of optimal methods of molecular genetic identification for AM fungi is carried out. The sample preparation, selection of species-specific marker DNA fragments and primers design, amplification including nested PCR are considered. The prospects for cloning and next generation sequencing for AM fungi identification are analyzed and substantiated.
The review is aimed to analyze molecular mechanisms of carbohydrate transport during the formation of arbuscular mycorrhiza (AM), a widespread symbiosis of plants with Glomeromycotina subdivision fungi. Due to AM-symbiosis, plants receive microelements, mainly phosphorus, and fungi are supplied by products of carbon assimilation. The study of sugar transport mechanisms in plants as well as between plants and symbiont is methodologically difficult because of the obligatory status of AM fungi. The mechanisms of carbohydrate transport in leaf and root cells are concerned, particular interest is paid to transporters, specific to AM structures. Several resumptive schemes are designed. SWEET family of transporters (Sugars Will Eventually be Exported Transporters), including AM-specific uniporters are reviewed. We summarize results on expression of genes encoding transporter in cells of plants without AM, in AM-plant cells with arbuscules and AM-plant cells without arbuscules. The data on genes of MST proteins family (Monosaccharide Transporters) participating in direct transport of sugars from the soil to the foliar mycelium of AM fungi are considered.
Background. The work is aimed to solve actual problems in biology of arbuscular mycorrhiza (AM). Currently, a lot of mutants had been obtained in various plant model objects with defects in genes controlling AM development, however, the mechanisms controlling development of effective AM symbiosis are still unclear. Materials and methods. The authors conducted a mutagenesis in Medicago lupulina, a new convenient model plant for molecular-genetic studies. High mycotrophic M. lupulina line have early and high response to mycorrhization, high seed production, as well as signs of dwarfism under conditions without of AM and low level of phosphorus available for plants. This method allows visually to identify plant lines with defects in AM symbiosis. Results. 14 modes for mutagenesis by ethylmethanesulfonate were conducted. Usage of 3 mutagenesis modes allowed to obtain: productive M1 progeny with high part of viable seedlings (73.3%–86.0%); 1405 plants in M2 progeny. Conclusion. According to population analysis for mutant plants in M2 progeny (up to M9 generation) 15 plant lines were selected: one Myc– plant line unable to form AM symbiosis, 4 Pen– plant lines unable to form AM symbiosis, but characterized by appressoria formation; 3 Rmd– plant lines forming low-activity ineffective AM symbiosis; 3 Rmd– plant lines forming low-activity effective AM and 4 Rmd++ plant lines forming effective AM with high abundance of symbiotic structures (mycelium/arbuscules/vesicles) in the roots.
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