Institute of Molecular Plant Sciences, Clusius Laboratory, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands
Induction of the formation of root nodule primordia in legume roots by symbiotic rhizobia is probably preceded by a change in plant hormone physiology. We used a Vicia sativa (vetch) split root system to study the effect of inoculation with rhizobia or purified Nod factors (lipochitin oligosaccharides, LCOs) on polar auxin transport in roots. Addition of R. leguminosarum bv. viciae, the infective symbiote of vetch, to roots of its host plant reduced polar auxin transport capacity of these roots within 24 h, in contrast to addition of non-nodulating, Sym plasmid-cured rhizobia. Addition of purified vetch-specific LCOs (NodRlv-IV/V[18:4,Ac]) caused a transient reduction in as little as 4 h after application, while after 16 h a second, stronger, and prolonged inhibition was observed that lasted at least 48 h. This reduction of auxin transport capacity was in the same order of magnitude as inhibition by N-(1-naphthyl)phthalamic acid (NPA). Purified LCOs (NodRm-IV[16:2,Ac,S]) from Sinorhizobium meliloti, the symbiote of alfalfa, and chitopentaose were inactive, which indicates a specific effect of LCOs produced by R. leguminosarum bv. viciae. Auxin transport inhibition was restricted to the apical nodulation-susceptible part of the roots, whereas the upper parts of the roots showed no difference in auxin transport after treatment. The effect could be observed with as low as 10-9 M NodRlv-IV/V[18:4,Ac] LCOs. Reduction of auxin transport by LCOs could not be inhibited by nitrate. Since inhibition of auxin transport capacity preceded the first root cortical cell divisions that result in root primordium formation, our results suggest a direct relationship between LCOs, polar auxin transport, and root nodule initiation, consistent with the hypothesis of U. Mathesius, H. R. M. Schlaman, H. P. Spaink, C. Sautter, B. G. Rolfe, and M. A. Djordjevic (Plant J. 14:23--34, 1998). However, nonmitogenic NodRlv-IV/V[18:1,Ac] showed a similar effect, which suggests that mitogenicity results from additional effects, in concert with auxin transport inhibition.
