Split-root systems applied to the study of the legume-rhizobial symbiosis: What have we learned?

Split-root system（SRS） approaches allow the differential treatment of separate and independent root systems, while sharing a common aerial part. As such, SRS is a useful tool for the discrimination of systemic（shoot origin）versus local（root/nodule origin） regulation mechanisms. This type of approach is particularly useful when studying the complex regulatory mechanisms governing the symbiosis established between legumes and Rhizobium bacteria. The current work provides an overview of the main insights gained from the application of SRS approaches to understand how nodule number（nodulation autoregulation） and nitrogen fixation are controlled both under non-stressful conditions and in response to a variety of stresses. Nodule number appears to be mainly controlled at the systemic level through a signal which is produced by nodule/root tissue, translocated to the shoot, and transmitted back to the root system, involving shoot Leu-rich repeat receptor-like kinases. In contrast, both local and systemic mechanisms have been shown to operate for the regulation of nitrogenase activity in nodules. Under drought and heavy metal stress, the regulation is mostly local,whereas the application of exogenous nitrogen seems to exert a regulation of nitrogen fixation both at the local and systemic levels.

A novel secreted protein, NISP1, is phosphorylated by soybean Nodulation Receptor Kinase to promote nodule symbiosis

Nodulation Receptor Kinase(NORK) functions as a co-receptor of Nod factor receptors to mediate rhizobial symbiosis in legumes, but its direct phosphorylation substrates that positively mediate root nodulation remain to be fully identified.Here, we identified a GmNORK-Interacting Small Protein(GmNISP1) that functions as a phosphorylation target of GmNORK to promote soybean nodulation. GmNORKα directly interacted with and phosphorylated GmNISP1. Transcription of GmNISP1 was strongly induced after rhizobial infection in soybean roots and nodules. GmNISP1 encodes a peptide containing 90 amino acids with a “DY” consensus motif at its N-terminus.GmNISP1 protein was detected to be present in the apoplastic space. Phosphorylation of GmNISP1 by GmNORKα could enhance its secretion into the apoplast. Pretreatment with either purified GmNISP1 or phosphorylation-mimic GmNISP1~(12D) on the roots could significantly increase nodule numbers compared with the treatment with phosphorylation-inactive GmNISP1~(12A).The data suggested a model that soybean GmNORK phosphorylates GmNISP1 to promote its secretion into the apoplast, which might function as a potential peptide hormone to promote root nodulation.

Reductive dechlorination of polychlorinated biphenyls is coupled to nitrogen fixation by a legume-rhizobium symbiosis

Chlorinated persistent organic pollutants, including polychlorinated biphenyls （PCBs）, represent a particularly serious environmental problem and human health risk worldwide. Leguminous plants and their symbiotic bacteria （rhizobia） are important components of the biogeochemical cycling of nitrogen in both agricultural and natural ecosystems. However, there have been relatively few detailed studies of the remediation of PCB-contaminated soils by legume-rhizobia symbionts. Here we report for the first time evidence of the reductive dechlorination of 2,4,4＇-trichlorobiphenyl （PCB 28） by an alfalfa-rhizobium nitrogen fixing symbiont. Alfalfa （Medicago sativa L.） inoculated with wild-type Sinorhizobiurn meliloti had significantly larger biomass and PCB 28 accumulation than alfalfa inoculated with the nitrogenase negative mutant rhizobium SmY. Dechlorination products of PCB 28, 2,4＇-dichlorobiphenyl （PCB 8）, and the emission of chloride ion （C1-） were also found to decrease significantly in the ineffective nodules infected by the mutant strain SmY. We therefore hypothesize that N2-fixation by the legume-rhizobium symbiont is coupled with the reductive dechlorination of PCBs within the nodules. The combination of these two processes is of great importance to the biogeochemical cycling and bioremediation of organochlorine pollutants in terrestrial ecosystems.