The effect of EDTA and EDDS on lead uptake and localization in hydroponically grown Pisum sativum L.

Pisum sativum plants were treated for 3 days with an aqueous solution of 100 lM Pb(NO3)2 or with a mixture of lead nitrate and ethylenediaminetetraacetic acid (EDTA) or [S,S]-ethylenediaminedisuccinic acid (EDDS) at equimolar concentrations. Lead decline from the incubation media and its accumulation and localization at the morphological and ultrastructural levels as well as plant growth parameters (root growth, root and shoot dry weight) were estimated after 1 and 3 days of treatment. The tested chelators, especially EDTA, significantly diminished Pb uptake by plants as compared to the lead nitrate-treated material. Simultaneously, EDTA significantly enhanced Pb translocation from roots to shoots. In the presence of both chelates, plant growth parameters remained considerably higher than in the case of uncomplexed Pb. Considerable differences between the tested chelators were visible in Pb localization both at the morphological and ultrastructural level. In Pb?EDTA-treated roots, lead was mainly located in the apical parts, while in Pb?EDDS-exposed material Pb was evenly distributed along the whole root length. Transmission electron microscopy and EDS analysis revealed that in meristematic cells of the roots incubated in Pb?EDTA, large electron-dense lead deposits were located in vacuoles and small granules were rarely noticed in cell walls or cytoplasm, while after Pb?EDDS treatment metal deposits were restricted to the border between plasmalemma and cell wall. Such results imply different ways of transport of those complexed Pb forms.The X-ray microanalysis was performed in the Laboratory of Electron Microscopy, Nencki Institute of Experimental Biology, Warsaw, Poland at the equipment installed within the project sponsored by the EU Structural Funds: Centre of Advanced Technology BIM—Equipment purchase for the Laboratory of Biological and Medical Imaging. Conflict of interest T

Is callose a barrier for lead ions entering Lemna minor L. root cells?

Plants have developed a range of strategies for resisting environmental stresses. One of the most common is the synthesis and deposition of callose, which functions as a barrier against stress factor penetration. The aim of our study was to examine whether callose forms an efficient barrier against Pb penetration in the roots of Lemna minor L. exposed to this metal. The obtained results showed that Pb induced callose synthesis in L. minor roots, but it was not deposited regularly in all tissues and cells. Callose occurred mainly in the protoderm and in the centre of the root tip (procambial central cylinder). Moreover, continuous callose bands, which could form an efficient barrier for Pb penetration, were formed only in the newly formed and anticlinal cell walls (CWs); while in other CWs, callose formed only small clusters or incomplete bands. Such an arrangement of callose within root CWs inefficiently protected the protoplast from Pb penetration. As a result, Pb was commonly present inside the root cells. In the light of the results, the barrier role of callose against metal ion penetration appears to be less obvious than previously believed. It was indicated that induction of callose synthesis is not enough for a successful blockade of the stress factor penetration. Furthermore, it would appear that the pattern of callose distribution has an important role in this defence strategy

Distinct expression, localization and function of two Rab7 proteins encoded by paralogous genes in a free-living model eukaryote

Rab7 GTPases are involved in membrane trafficking in the late endosomal/lysosomal pathway. In Paramecium octaurelia Rab7a and Rab7b are encoded by paralogous genes. Antipeptide antibodies generated against divergent C-termini recognize Rab7a of 22.5 kDa and Rab7b of 25 kDa, respectively. In 2D gel electrophoresis two immunoreactive spots were identified for Rab7b at pI about 6.34 and about 6.18 and only one spot for Rab7a of pI about 6.34 suggesting post-translational modification of Rab7b. Mass spectrometry revealed eight identical phosphorylated residues in the both proteins. ProQ Emerald staining and ConA overlay of immunoprecipitated Rab7b indicated its putative glycosylation that was further supported by a faster electrophoretic mobility of this protein upon deglycosylation. Such a post-translational modification and substitution of Ala140 in Rab7a for Ser140 in Rab7b may result in distinct targeting to the oral apparatus where Rab7b associates with the microtubular structures as revealed by STED confocal and electron microscopy. Rab7a was mapped to phagosomal compartment. Absolute qReal-Time PCR analysis revealed that expression of Rab7a was 2.6-fold higher than that of Rab7b. Upon latex internalization it was further 2-fold increased for Rab7a and only slightly for Rab7b. Post-transcriptional gene silencing of rab7a suppressed phagosome formation by 70 % and impaired their acidification. Ultrastructural analysis with double immunogold labeling revealed that this effect was due to the lack of V-ATPase recruitment to phagolysosomes. No significant phenotype changes were noticed in cells upon rab7b silencing. In conclusion, Rab7b acquired a new function, whereas Rab7a can be assigned to the phagolysosomal pathway

Diminishing benefits of urban living for children and adolescents’ growth and development

Optimal growth and development in childhood and adolescence is crucial for lifelong health and well-being1–6. Here we used data from 2,325 population-based studies, with measurements of height and weight from 71 million participants, to report the height and body-mass index (BMI) of children and adolescents aged 5–19 years on the basis of rural and urban place of residence in 200 countries and territories from 1990 to 2020. In 1990, children and adolescents residing in cities were taller than their rural counterparts in all but a few high-income countries. By 2020, the urban height advantage became smaller in most countries, and in many high-income western countries it reversed into a small urban-based disadvantage. The exception was for boys in most countries in sub-Saharan Africa and in some countries in Oceania, south Asia and the region of central Asia, Middle East and north Africa. In these countries, successive cohorts of boys from rural places either did not gain height or possibly became shorter, and hence fell further behind their urban peers. The difference between the age-standardized mean BMI of children in urban and rural areas was <1.1 kg m–2 in the vast majority of countries. Within this small range, BMI increased slightly more in cities than in rural areas, except in south Asia, sub-Saharan Africa and some countries in central and eastern Europe. Our results show that in much of the world, the growth and developmental advantages of living in cities have diminished in the twenty-first century, whereas in much of sub-Saharan Africa they have amplified