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A Novel ABA Insensitive mutant of Lotus japonicus with a Wilty Phenotype Displays Unaltered Nodulation Regulation

molecular Plant 2009年第3期2卷 487-499页

作者： bandana biswas pick kuen chan peter m.gressho.f ARC Centre of Excellence for Integrative Legume Research The University of Queensland St Lucia Brisbane QId 4072 Australia

An 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. （The name ＂Beyma＂ was taken from the Australian Aboriginal language, Wagiman, beyma, meaning ‘drying up＇.） The 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 insensitive phenotypes, 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 were similar to that of mG-20. Split root experiments on mG-20 showed that application of ABA on one side of the root inhibited nodulation locally but not systemically. We propose that ABA is not involved directly in systemic autoregulation of nodulation （AON）.

关键词： hormone biology water relations genetics development symbiosis legume.

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Classical Ethylene Insensitive mutants of the Arabidopsis EIN2 Orthologue Lack the Expected‘hypernodulation’ Response in Lotus japonicus

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Journal of Integrative Plant Biology 2013年第4期55卷 395-408页

作者： pick kuen chan bandana biswas peter m. gresshoff Australian Research Council (ARC) Centre of Excellence for Integrative Legume Research The University of Queensland ARC Centre of Excellence for Integrative Legume Research and School of Agriculture and food Sciences The University of Queensland

Three independent ethylene insensitive mutants were selected from an EmS- mutagenized population of Lotus japonicus mG-20 （miyakojima）. The mutants, called ＇Enigma＇, were mutated in the LjEIN2a gene from Lotus chromosome 1, sharing significant homology with Arabidopsis EIN2 （ethylene-insensitive2）. All three alleles showed classical ethylene insensitivity phenotypes （e.g., Triple Response）, but lacked the increased nodulation phenotype commonly associated with ethylene insensitivity. Indeed, all showed a marginal reduction in nodule number per plant, a phenotype that is enigmatic to sickle, an ethylene- insensitive EIN2 mutant in medicago truncatula. In contrast to wild type, but similar to an ETRI-1 ethylene ethylene-insensitive transgenic of L. japonicus, enigma mutants formed nodules in between the protoxylem poles, demonstrating the influence of ethylene on radial positioning. Suppression of nodule numbers by nitrate and colonisation by mycorrhizal fungi in the enigma-1 mutant were indistin- guishable from the wild-type mG-20. However, reflecting endogenous ethylene feedback, the enigma-1 mutant released more than twice the wild-type amount of ethylene, enigma-1 had a moderate reduction in growth, greater root mass （and lateral root formation）, delayed flowering and ripening, smaller pods and seeds. Expression analysis of ethylene-regulated genes, such as ETR1, NRL1 （neverripe-like 1）, and ElL3 in shoots and roots of enigma-1 and mG-20 illustrated that the ethylene-insensitive mutation strongly affected transcriptional responses in the root. These mutants open the possibility that EIN2 in L. japonicus, a determinate nodulating legume, acts in a more complex fashion possibly through the presence of a duplicated copy of LjEIN2.

关键词： ABA development ethylene legume nodulation symbiosis.

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