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• W86976467 abstract "Legumes are unique in their ability to form a symbiotic relationship with soil bacteria called rhizobia which reside in root nodules and fix atmospheric nitrogen. This is beneficial to the environment as the need for nitrogen fertiliser is eliminated or at least reduced. However, biological, like industrial nitrogen fixation is energetically expensive and the plant has developed several mechanisms, both local and systemic (autoregulation of nodulation), to control the number of nodules. Both processes, the formation of the nodules (and nitrogen fixation) as well as the control of nodule number, have been extensively studied. Plant hormones are involved in all known processes of plant development including nodulation. Roles of auxins and cytokinins in particular are well documented. Involvement of the plant hormone abscisic acid (ABA) in nodulation is less known. ABA plays a critical role in physiological processes such as germination, seed dormancy, growth, lateral root formation, desiccation and environmental stress responses. ABA may also be also involved in various stages of nodulation such as growth of nodules, autoregulation of nodule number as well as desiccation of nodules. This project was undertaken to further understand the role of ABA in nodulation by working on the model legume Lotus japonicus and its bacterial partner Mesorhizobium loti. The study was done using three approaches; (i) isolation and characterisation of an ABA insensitive mutant, (ii) creation of a transgenic line with ABA insensitivity conferring gene abi1 from Arabidopsis, and (iii) physiological studies, including the effect of external application of ABA. The ABA insensitive mutant Beyma was isolated in Lotus japonicus MG-20 from an EMS mutagenesis population using root growth inhibition to applied ABA as the screening criterion. This stable mutant that segregates as a dominant Mendelian mutation, is insensitive to ABA induced inhibition of germination, vegetative growth, stomatal opening as well as nodulation. Tissue ABA levels were normal, suggesting a ‘sensitivity’ rather than ‘biosynthesis’ mutation. It is slow growing (50-70 % of wild type MG-20) and has a near-constitutive wilty phenotype associated with its inability to regulate stomatal opening. Whilst showing a wide range of ABA insensitivephenotypes, Beyma did not show alteration of nodule number control, as in the absence of added ABA, the number and patterning (but not size) of nodules formed in the mutant was similar to MG-20. Transgenic lines containing the abi1, ABA insensitive mutant gene from Arabidopsis was created in Lotus japonicus Gifu B-129. The presence of the transgene was confirmed by PCR. The lines generated showed ABA insensitive phenotypes, such as defective stomatal opening leading to wiltyness and frequent drying of branches and leaves. However, like Beyma the transgenic lines did not show alteration in nodule number control but did show an alteration in nodule size. Through physiological studies it was found that for germination, ABA sensing differed in different ecotypes of Lotus japonicus. Gifu was more sensitive to ABA than MG-20 and this reduced sensitivity in MG-20 was clearly due to a single dominant locus as confirmed by the segregation ratio in an F1 population. Variations in germination sensitivity to ABA were also observed between various mutants of both Gifu and MG-20. There was lesser effect of ABA in shoot growth in MG-20 and its mutants, though Gifu and its mutants were somewhat more sensitive, again emphasising the difference in sensitivity between the two ecotypes. This same trend was also observed with root growth. In Lotus japonicus, stimulation of lateral root growth was only observed in MG-20 and MG-20 mutants, and not in Gifu and its mutants. A change in the orientation of growth was observed in both Gifu and MG-20 possibly due to gravitropic response to ABA. Gifu was again seen to be less sensitive when a nodulation study was done in closed jars, leading to the possibility that sensing of ABA for germination maybe through a different pathway to nodulation. Split root experiments demonstrated that application of ABA to one side of the root, inhibited nodulation locally but not systemically. Stomata of such plants were completely closed, yet the root portion not exposed to the inhibitory level of ABA nodulated normally. This result along with the fact that both the ABA insensitive mutant and the Atabi1 transgenics do not show nodule number defects lead to the proposal that ABA is not involved directly in systemic autoregulation of nodulation (AON)." @default.
• W86976467 created "2016-06-24" @default.
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• W86976467 date "2008-06-01" @default.
• W86976467 modified "2023-09-27" @default.
• W86976467 title "Genetic and Physiological Characterisation of ABA Insensitivity in the Model Legume Lotus japonicus" @default.
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