Involvement of glutathione in β-cyclodextrin-hemin complex-induced lateral root formation in tomato seedlings

β-cyclodextrin-hemin complex (β-CDH) was shown to induce lateral root (LR) formation in tomato. However, the molecular mechanism is still elusive. In this report, the role of reduced glutathione (GSH) in the induction of lateral root triggered by β-CDH was investigated. Similar to the responses of β-CDH, exogenously applied with 0.1 mΜ GSH not only increased endogenous GSH content determined by spectrophotography and the monochlorobimane (MCB)-dependent fluorescent analysis, but also induced, thereafter, LR formation. Meanwhile, both β-CDH- and GSH-induced lateral root primordia (LRP) exhibited a similar accelerated anatomic structure. Above inducible responses were blocked significantly when the L-buthionine-(S,R)-sulfoximine (BSO), a potent and specific inhibitor of the enzyme catalyzing the first step of GSH biosynthesis, was separately applied. Upon β-CDH treatment, the changes of endogenous GSH content determined by spectrophotography and fluorescent analysis were consistent with the transcripts of two GSH synthetic genes, GSH1 and GSH2 encoding γ-glutamyl cysteine synthetase and glutathione synthetase, respectively. Exogenously applied with β-CDH could rescue N-1-naphthylphthalamic acid (NPA; IAA depletion)-triggered inhibition of LR formation. Further molecular evidence revealed that both β-CDH and GSH modulated gene expression of cell cycle regulatory genes (CYCA2;1, CYCA3;1, CYCD3;1, and CDKA1) and auxin signaling genes (ARF7 and RSI-1), six marker genes responsible for LR formation. By contrast, above changes were sensitive to the co-treatment with BSO. All together, these results suggest a role for GSH in the regulation of tomato LR development triggered by β-CDH.