Stimulation of nodulation in Medicago truncatula by low concentrations of ammonium: quantitative reverse transcription PCR analysis of selected genes.
Although mineral nitrogen generally has negative effects on nodulation in legume-rhizobia symbioses, low concentrations of ammonium stimulate nodulation in some legumes. In this study, the effects of ammonium and nitrate on growth, nodulation and expression of 2 nitrogen transport and 12 putative nodulation-related genes of the model symbiosis of Medicago truncatula - Sinorhizobium meliloti are investigated. After 3 weeks of hydroponic growth, whole-plant nodulation was enhanced in all the ammonium treatments and up to three-fold in the 0.5 mM treatment compared with the zero-nitrogen control. Specific nodulation (nodules g(-1) root dry weight) was greatly stimulated in the 0.1 and 0.5 mM NH4+ treatments, to a lower extent in the 0.1 mM NO3- treatment, and inhibited in all other treatments. Expression of the 14 selected genes was observed at 0, 6, 12 and 24 h after exposure to rhizobia and nitrogen. Expression of nitrogen transporter genes increased significantly, but responses of the three genes putatively associated with symbiosis signaling/nodule initiation were mixed. There were infrequent responses of genes coding for an ABA-activated protein kinase or a gibberellin-regulated protein, but an ethylene-responsive element-binding factor showed increased expression in various treatments and sampling times. Three auxin-responsive genes and three cytokinin-responsive genes showed varied responses to ammonium and nitrate. This study indicates that low concentrations of ammonium stimulate nodulation in M. truncatula, but the data were inconclusive in verifying the hypothesis that a relatively high ratio of cytokinin to auxin in roots may be an underlying mechanism in this stimulation of nodulation.
Fei
Houman
H
Department of Biology, Saint Mary's University, Halifax, Nova Scotia, Canada.
Vessey
J Kevin
JK
D003583:Cytokinins / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D007210:Indoleacetic Acids / Q000378:metabolism; D046913:Medicago truncatula / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D009566:Nitrates / Q000494:pharmacology; D009584:Nitrogen / Q000378:metabolism; D010937:Plant Growth Regulators / Q000378:metabolism; D055170:Plant Root Nodulation / Q000187:drug effects*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*; D018749:RNA, Plant / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053204:Root Nodules, Plant / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D016962:Sinorhizobium meliloti / Q000502:physiology; D013559:Symbiosis
D003583:Cytokinins; D007210:Indoleacetic Acids; D009566:Nitrates; D010937:Plant Growth Regulators; D000644:Quaternary Ammonium Compounds; D018749:RNA, Plant; D009584:Nitrogen