Dataset Information

Cellular Fractionation and Nanoscopic X-Ray Fluorescence Imaging Analyses Reveal Changes of Zinc Distribution in Leaf Cells of Iron-Deficient Plants.

ABSTRACT: Multilevel interactions among nutrients occur in the soil-plant system. Among them, Fe and Zn homeostasis in plants are of great relevance because of their importance for plant and human nutrition. However, the mechanisms underlying the interplay between Fe and Zn in plants are still poorly understood. In order to elucidate how Zn interacts with Fe homeostasis, it is crucial to assess Zn distribution either in the plant tissues or within the cells. In this study, we investigated the subcellular Zn distribution in Fe-deficient leaf cells of cucumber plants by using two different approaches: cellular fractionation coupled with inductively coupled plasma mass spectrometry (ICP/MS) and nanoscopic synchrotron X-ray fluorescence imaging. Fe-deficient leaves showed a strong accumulation of Zn as well as a strong alteration of the organelles' ultrastructure at the cellular level. The cellular fractionation-ICP/MS approach revealed that Zn accumulates in both chloroplasts and mitochondria of Fe deficient leaves. Nano-XRF imaging revealed Zn accumulation in chloroplast and mitochondrial compartments, with a higher concentration in chloroplasts. Such results show that (i) both approaches are suitable to investigate Zn distribution at the subcellular level and (ii) cellular Fe and Zn interactions take place mainly in the organelles, especially in the chloroplasts.

SUBMITTER: Vigani G

PROVIDER: S-EPMC6085429 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Cellular Fractionation and Nanoscopic X-Ray Fluorescence Imaging Analyses Reveal Changes of Zinc Distribution in Leaf Cells of Iron-Deficient Plants.

Vigani Gianpiero G Bohic Sylvain S Faoro Franco F Vekemans Bart B Vincze Lazlo L Terzano Roberto R

Frontiers in plant science 20180803

Multilevel interactions among nutrients occur in the soil-plant system. Among them, Fe and Zn homeostasis in plants are of great relevance because of their importance for plant and human nutrition. However, the mechanisms underlying the interplay between Fe and Zn in plants are still poorly understood. In order to elucidate how Zn interacts with Fe homeostasis, it is crucial to assess Zn distribution either in the plant tissues or within the cells. In this study, we investigated the subcellular ...[more]

PMID: 30123229

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Project description:BackgroundMany metals are essential for plants and humans. Knowledge of metal distribution in plant tissues in vivo contributes to the understanding of physiological mechanisms of metal uptake, accumulation and sequestration. For those studies, X-rays are a non-destructive tool, especially suited to study metals in plants.ResultsWe present microfluorescence imaging of trace elements in living plants using a customized benchtop X-ray fluorescence machine. The system was optimized by additional detector shielding to minimize stray counts, and by a custom-made measuring chamber to ensure sample integrity. Protocols of data recording and analysis were optimised to minimise artefacts. We show that Zn distribution maps of whole leaves in high resolution are easily attainable in the hyperaccumulator Noccaea caerulescens. The sensitivity of the method was further shown by analysis of micro- (Cu, Ni, Fe, Zn) and macronutrients (Ca, K) in non-hyperaccumulating crop plants (soybean roots and pepper leaves), which could be obtained in high resolution for scan areas of several millimetres. This allows to study trace metal distribution in shoots and roots with a wide overview of the object, and thus avoids making conclusions based on singular features of tiny spots. The custom-made measuring chamber with continuous humidity and air supply coupled to devices for imaging chlorophyll fluorescence kinetic measurements enabled direct correlation of element distribution with photosynthesis. Leaf samples remained vital even after 20 h of X-ray measurements. Subtle changes in some of photosynthetic parameters in response to the X-ray radiation are discussed.ConclusionsWe show that using an optimized benchtop machine, with protocols for measurement and quantification tailored for plant analyses, trace metal distribution can be investigated in a reliable manner in intact, living plant leaves and roots. Zinc distribution maps showed higher accumulation in the tips and the veins of young leaves compared to the mesophyll tissue, while in the older leaves the distribution was more homogeneous.

Project description:Background and aimsAllometric scaling between stomata and xylem for terrestrial woody plants is a widely observed pattern that may be constrained by water transport. Floating-leaved plants, a particular life form of aquatic plants, have leaves in direct contact with both air and water and a poorly developed xylem that may not be limited by water supply as for terrestrial plants. However, whether such an allometric scaling relationship still exists in floating-leaved plants has not been explored.MethodsWe analysed 31 floating-leaved species/varieties with a range in leaf area covering six orders of magnitude. For all 31 floating-leaved plants, we studied the allometric relationships between leaf area and petiole transverse area, and between total stomatal area and petiole vascular area.Key resultsThe slopes of both relationships were similar to the slope of the allometric relationship (1.23) between total stomatal area and xylem area of 53 terrestrial plants. However, for ten of them with xylem that can be clearly defined, the strong positive relationship between total stomatal area and petiole xylem area had a significantly smaller slope than that of terrestrial plants (0.64 vs. 1.23). Furthermore, after considering phylogeny, the scaling relationships between total stomatal area and petiole traits in floating-leaved plants remained significant.ConclusionsWe speculated that for floating-leaved plants, the hyperallometric relationship (slope >1) between the construction of leaf/stoma and petiole was promoted by the high demand for photosynthesis and thus more leaves/stomata. While the hypoallometric relationship (slope <1) between stomatal and xylem area was related more to hydraulic processes, the selection pressure on stomata was lower than xylem of floating-leaved plants. Allometric relationships among the hydraulic traits on water transport of aquatic plants are the result of natural selection to achieve maximum carbon gain, which is similar to terrestrial plants.

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Cellular Fractionation and Nanoscopic X-Ray Fluorescence Imaging Analyses Reveal Changes of Zinc Distribution in Leaf Cells of Iron-Deficient Plants.

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Cellular Fractionation and Nanoscopic X-Ray Fluorescence Imaging Analyses Reveal Changes of Zinc Distribution in Leaf Cells of Iron-Deficient Plants.

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