Arbuscular mycorrhiza (AM) represents a symbiotic mutualistic association between most land plants and Glomeromycota fungi. AM fungi develops specialized intraradical and highly branched structures, called arbuscules, where bidirectional exchange of nutrients between plant and fungi partners occurs, improving plant growth and fitness. Transcriptional reprogramming and hormonal regulation are necessary for the formation of the arbuscules. SlDLK2, a member of the third clade from the DWARF14 family of α, β-hydrolases closely related to the strigolactone receptor D14, is a negative regulator of arbuscule branching in tomato, but the underlying mechanisms are unknown. We explored the possible role of SlDLK2 on the regulation of hormonal balance. RNA-seq analysis was performed on roots from composite tomato plants overexpressing SlDLK2 and in control plants transformed with the empty vector. Analysis of transcriptomic data predicted that significantly repressed genes were enriched for genes related to hormone biosynthesis pathways, with a special relevance of carotenoid/apocarotenoid biosynthesis genes. Stable transgenic SlDLK2 overexpressing (OE) tomato lines were obtained, and hormone contents were analyzed in their roots and leaves. Interesting significant hormonal changes were found in roots of SlDLK2 OE lines with respect to the control lines, with a strong decrease on jasmonic acid and ABA. In addition, SlDLK2 OE roots showed a slight reduction in auxin contents and in one of the major strigolactones in tomato, solanacol. Overall, our results suggest that the negative regulation of AM symbiosis by SlDLK2 is associated with the repression of genes involved in the biosynthesis of AM-promoting hormones.

• Klíčová slova
• DLK2, arbuscular mycorrhiza, plant hormones, tomato, transcriptomics,
• Publikační typ
• časopisecké články MeSH

The REQUIRED FOR ARBUSCULAR MYCORRHIZATION1 (RAM1) transcription factor from the GRAS family is well known for its role as a master regulator of the arbuscular mycorrhizal (AM) symbiosis in dicotyledonous and monocotyledonous species, being essential in transcriptional reprogramming for the development and functionality of the arbuscules. In tomato, SlGRAS27 is the putative orthologue of RAM1 (here named SlRAM1), but has not yet been characterized. A reduced colonization of the root and impaired arbuscule formation were observed in SlRAM1-silenced plants, confirming the functional conservation of the RAM1 orthologue in tomato. However, unexpectedly, SlRAM1-overexpressing (UBIL:SlRAM1) plants also showed decreased mycorrhizal colonization. Analysis of non-mycorrhizal UBIL:SlRAM1 roots revealed an overall regulation of AM-related genes and a reduction of strigolactone biosynthesis. Moreover, external application of the strigolactone analogue GR244DO almost completely reversed the negative effects of SlRAM1 overexpression on the frequency of mycorrhization. However, it only partially recovered the pattern of arbuscule distribution observed in control plants. Our results strongly suggest that SlRAM1 has a dual regulatory role during mycorrhization and, in addition to its recognized action as a positive regulator of arbuscule development, it is also involved in different mechanisms for the negative regulation of mycorrhization, including the repression of strigolactone biosynthesis.

• Klíčová slova
• Arbuscular mycorrhiza, GRAS, RAM1, strigolactones, tomato, transcriptional regulation,
• MeSH
• kořeny rostlin mikrobiologie   metabolismus   genetika   MeSH
• mykorhiza * fyziologie   MeSH
• regulace genové exprese u rostlin MeSH
• rostlinné proteiny * metabolismus   genetika   MeSH
• Solanum lycopersicum * mikrobiologie   genetika   metabolismus   MeSH
• symbióza MeSH
• transkripční faktory * metabolismus   genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• rostlinné proteiny * MeSH
• transkripční faktory * MeSH

The Oryza sativa (rice) carotenoid cleavage dioxygenase OsZAS was described to produce zaxinone, a plant growth-promoting apocarotenoid. A zas mutant line showed reduced arbuscular mycorrhizal (AM) colonization, but the mechanisms underlying this behavior are unknown. Here, we investigated how OsZAS and exogenous zaxinone treatment regulate mycorrhization. Micromolar exogenous supply of zaxinone rescued root growth but not the mycorrhizal defects of the zas mutant, and even reduced mycorrhization in wild-type and zas genotypes. The zas line did not display the increase in the level of strigolactones (SLs) that was observed in wild-type plants at 7 days post-inoculation with AM fungus. Moreover, exogenous treatment with the synthetic SL analog GR24 rescued the zas mutant mycorrhizal phenotype, indicating that the lower AM colonization rate of zas is caused by a deficiency in SLs at the early stages of the interaction, and indicating that during this phase OsZAS activity is required to induce SL production, possibly mediated by the Dwarf14-Like (D14L) signaling pathway. OsZAS is expressed in arbuscule-containing cells, and OsPT11prom::OsZAS transgenic lines, where OsZAS expression is driven by the OsPT11 promoter active in arbusculated cells, exhibit increased mycorrhization compared with the wild type. Overall, our results show that the genetic manipulation of OsZAS activity in planta leads to a different effect on AM symbiosis from that of exogenous zaxinone treatment, and demonstrate that OsZAS influences the extent of AM colonization, acting as a component of a regulatory network that involves SLs.

• Klíčová slova
• Oryza sativa, OsPT11, GR24, apocarotenoids, arbuscular mycorrhizal symbiosis, in situ hybridization, strigolactones, zaxinone, zaxinone synthase,
• MeSH
• dioxygenasy * metabolismus   MeSH
• karotenoidy metabolismus   MeSH
• kořeny rostlin metabolismus   MeSH
• mykorhiza * metabolismus   MeSH
• rýže (rod) * metabolismus   MeSH
• symbióza fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dioxygenasy * MeSH
• karotenoidy MeSH