Identification of new potential regulators of the Medicago truncatula-Sinorhizobium meliloti symbiosis using a large-scale suppression subtractive hybridization approach.

摘要: We set up a large-scale suppression subtractive hybridization (SSH) approach to identify Medicago truncatula genes differentially expressed at different stages of the symbiotic interaction with Sinorhizobium meliloti, particular interest for regulatory genes. constructed 7 SSH libraries covering successive from Nod factor signal transduction S. meliloti infection, nodule organogenesis, and functioning. Over 26,000 clones were screened by two rounds macroarray hybridizations. In all, 3,340 clones, corresponding whose expression was potentially affected, selected, sequenced, ordered into 2,107 tentative gene clusters, including 767 MtS clusters new M. total, 52 encoding potential proteins, transcription factors (TFs) other elements cascades, identified. The pattern some them analyzed quantitative reverse-transcription polymerase chain reaction in wild-type – mutants blocked before or after infection. Three genes, coding TFs bHLH WRKY families C2H2 zinc-finger protein, respectively, found be upregulated, following inoculation, infection-defective mutant lin, whereas also root-hair-curling hcl. role these early steps is discussed.