概要: Receptor-like kinases (RLKs) are key cell signalling components. The rice ARBUSCULAR RECEPTOR-LIKE KINASE 1 (OsARK1) functions during a stage post-arbuscule development in the arbuscular mycorrhizal (AM) association and belongs to an undefined subfamily of RLKs. Our phylogenetic analysis revealed that ARK1 has an ancient paralogue, ARK2. Single ark2 and ark1ark2 double mutants in rice showed a non-redundant AM symbiotic function for OsARK2. Global transcriptomics identified a set of genes co-regulated by the two RLKs suggesting OsARK1 and OsARK2 to orchestrate symbiosis in a common pathway. ARK lineage proteins harbour a newly identified SPARK domain in their extracellular regions which underwent parallel losses in ARK1 and ARK2 in monocots. This protein domain has ancient origins in the green lineage and defines additional overlooked groups of putative cell surface receptors
项目整体设计: Four different genotypes (ark1-2,ark2,ark1ark2,wild type) of rice were subjected to mycorrhizal fungi R. irregularis. Ten biological replicates were generated for each genotype. To study the effects of mutants ark1 and ark2, the transcriptomes were analyzed against the wild type controls.
Plants were watered every second day, the first two weeks with reverse osmosis water (R-O H2O) followed by low Pi fertilization every other watering day with half-strength Hoagland solution containing 25 μM KH2PO4.
处理方案:
The AM fungal model species R. irregularis (DAOM197198) was employed for all inoculation assays. Spores extracted from Agrobacterium rhizogenes transformed carrot hairy root cultures (Bécard and Fortin, 1988) were used. Aliquoted inoculum was applied onto the substrate of half-filled cones (12 cm depth, 2.5 cm diameter), after which sand substrate was replenished and germinated seedlings transferred to the containers.
提取方案:
RNA was extracted using the TRIzol method
建库方案:
1 μg RNA per sample was used for library preparation generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, Hitchin, United Kingdom) following manufacturer’s instructions. Library quality was monitored on the Agilent Bioanalyzer 2100 system
