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New techniques for studying competition by Rhizobia and for assessing nitrogen fixation in the field
Authors:Kate J Wilson  Mark B Peoples  Richard A Jefferson
Institution:(1) CAMBIA (Center for the Application of Molecular Biology to International Agriculture), GPO Box 3200, 2601 Canberra, ACT, Australia;(2) CSIRO Division of Plant Industry, GPO Box 1600, 2601 Canberra, ACT, Australia
Abstract:One of the key factors limiting the proper assessment and use of rhizobial strains in the field is the lack of suitable methodology to screen the success of individual isolates in competing for nodule occupancy with different cultivars of legumes and in different soil and agronomic conditions. The use of marker genes enables individual rhizobial strains to be identified by a simple colour assay, thus enabling a dramatic increase in throughput of strain screening. One such marker system for rhizobial ecology, the GUS system, is already in use to facilitate rapid screening of rhizobial isolates. Other markers, which will allow the competitive behaviour of several strains to be studied at once, are under development.Likewise, breeding of the host legume for a high efficiency of nitrogen fixation is hampered by the difficulty in assessing this property. The method which currently gives the highest throughput of analysis, and has been successfully used in soybean breeding programs, is the ureide technique. However, it remains somewhat laborious for use in routine breeding programs. In this paper we discuss the potential use of reporter genes to provide information on the relative levels of ureides and other nitrogenous compounds in plants growing in the field. This would greatly increase the rate at which this trait could be scored, and would thus enable routine assays for increased symbiotic nitrogen fixation for breeding or management purposes in legume crops such as soybean (Glycine max) and common bean (Phaseolus vulgaris).
Keywords:bioindicators  legumes  microbial ecology  molecular tools  N2 fixation  Rhizobium
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