Open AccessGene Silencing by Expression of Hairpin RNA in Lotus japonicus Roots and Root Nodules
Author(s) -
Hirotaka Kumagai,
Hiroshi Kouchi
Publication year - 2003
Publication title -
molecular plant-microbe interactions
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.565
H-Index - 153
eISSN - 1943-7706
pISSN - 0894-0282
DOI - 10.1094/mpmi.2003.16.8.663
Subject(s) - small hairpin rna , agrobacterium , lotus japonicus , gene silencing , biology , rna silencing , lotus corniculatus , root nodule , gene , rna interference , gene expression , microbiology and biotechnology , rna , transformation (genetics) , coding region , lotus , genetics , botany , symbiosis , mutant , bacteria
We investigated the efficacy of self-complementary hairpin RNA (hpRNA) expression to induce RNA silencing in the roots and nodules of model legume Lotus japonicus, using hairy root transformation mediated by Agrobacterium rhizogenes. Transgenic lines that express β-glucuronidase (GUS) by constitutive or nodule-specific promoters were supertransformed by infection of A. rhizogenes harboring constructs for the expression of hpRNAs with sequences complementary to the GUS coding region. GUS activity in more than 60% of the hairy roots was decreased or silenced almost completely. Silencing of the GUS gene was also observed in symbiotic nodules formed on hairy roots in both early and late stages of nodule organogenesis. These results indicate that transient RNA silencing by hairy root transformation provides a powerful tool for loss-of-function analyses of genes that function in roots and root nodules.