Legumes form symbioses with arbuscular mycorrhiza (AM) fungi and nitrogen fixing root nodule bacteria. Intracellular root infection by either endosymbiont is controlled by the activation of the calcium and calmodulin-dependent kinase (CCaMK), a central regulatory component of the plant's common symbiosis signaling network. We performed a microscopy screen for Lotus japonicus mutants defective in AM development and isolated a mutant, nena, that aborted fungal infection in the rhizodermis. NENA encodes a WD40 repeat protein related to the nucleoporins Sec13 and Seh1. Localization of NENA to the nuclear rim and yeast two-hybrid experiments indicated a role for NENA in a conserved subcomplex of the nuclear pore scaffold. Although nena mutants were able to form pink nodules in symbiosis with Mesorhizobium loti, root hair infection was not observed. Moreover, Nod factor induction of the symbiotic genes NIN, SbtM4, and SbtS, as well as perinuclear calcium spiking, were impaired. Detailed phenotypic analyses of nena mutants revealed a rhizobial infection mode that overcame the lack of rhizodermal responsiveness and carried the hallmarks of crack entry, including a requirement for ethylene. CCaMK-dependent processes were only abolished in the rhizodermis but not in the cortex of nena mutants. These data support the concept of tissue-specific components for the activation of CCaMK.
Due to the physico-chemical similarities of caesium (Cs+) to potassium (K+) on the one hand and strontium (Sr2+) to calcium (Ca2+) on the other hand, both elements can easily be taken up by plants and thus enter the food chain. This could be detrimental when radionuclides such as 137Cs and 90Sr are involved. In this study, both genetic and physiological aspects of Cs+ and Sr2+ accumulation in Arabidopsis thaliana were investigated using 86 Arabidopsis accessions and a segregating F2 population of the low Cs+ accumulating Sq-1 (Ascot, UK) crossed with the high uptaking Sorbo (Khurmatov, Tajikistan). Hydroponically grown plants were exposed to subtoxic levels of Cs+ and Sr2+ using radioactive isotopes as tracers. In the natural accessions shoot concentration of Cs+ as well as Sr2+ varied about 2-fold, whereas its heritability ranged for both ions between 0.60 and 0.73. Shoot accumulation of Cs+ and Sr2+ could be compromised by increasing concentrations of their essential analogues K+ and Ca2+, respectively, causing a reduction of up to 80%. In the case of the segregating F2/F3 population Sq-1×Sorbo, this study identified several QTL for the trait Cs+ and Sr2+ accumulation, with main QTL on chromosomes 1 and 5. According to the correlation and discrimination surveys combined with QTL-analysis Cs+ and Sr2+ uptake seemed to be mediated mostly via non-selective cation channels. A polymorphism, affecting amino acids close to the K+-pore of one candidate, CYCLIC-NUCLEOTIDE-GATED CHANNEL 1 (CNGC1), was identified in Sorbo and associated with high Cs+ concentrating accessions.
The non-essential cation caesium (Cs þ ) is assimilated by all organisms. Thus, anthropogenically released radiocaesium is of concern to agriculture. Cs þ accumulates owing to its chemical similarity to the potassium ion (K þ ). The apparent lack of a Cs þ -specific uptake mechanism has obstructed attempts to manipulate Cs þ accumulation without causing pleiotropic effects. Here we show that the SNARE protein Sec22p/SEC22 specifically impacts Cs þ accumulation in yeast and in plants. Loss of Saccharomyces cerevisiae Sec22p does not affect K þ homeostasis, yet halves Cs þ concentration compared with the wild type. Mathematical modelling of the uptake time course predicts a compromised vacuolar Cs þ deposition in sec22D. Biochemical fractionation confirms this and indicates a new feature of Sec22p in enhancing non-selective cation deposition. A developmentally controlled loss-of-function mutant of the orthologous Arabidopsis thaliana SEC22 phenocopies the reduced Cs þ uptake without affecting plant growth. This finding provides a new strategy to reduce radiocaesium entry into the food chain.
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