Characterization of the Embryogenic Tissue of the Norway Spruce Including a Transition Layer between the Tissue and the Culture Medium by Magnetic Resonance Imaging

Magnetic resonance imaging (MRI) is a non-invasive method widely applied in the study of molecules. The MRI approach is frequently employed not only in medicine but also in biological, biochemical, and chemical research. Most of the papers describing the use of MRI to investigate plant physiology have emphasized certain aspects of anatomy or functional morphology [1]. In plant biology, MRI supports several major activities, namely, the research of the water and mineral compounds transported within a plant [2], [3], the determination of plant metabolites [4], [5], the investigation of cellular processes [6], and the examination of the growth and development of plants [7]. MRI is also instrumental towards monitoring water changes in early somatic embryos (ESEs). Šupálková et al. [8] examine the influence of the ESEs of spruce treated with cadmium and/or lead ions for twelve days using multi-instrumental analytical procedures; these authors employ image analysis to estimate the growth, a fluorimetric sensor to detect the viability of enzyme-treated ESEs, and the MRI technique to facilitate non-destructive measurement of the volume of the ESEs.

 The interaction between the cells plays a fundamental role in the growth and development of multicellular organisms. In this context, let us note that
the structural integrity of plants was described by, for example, Šebánek et al. [9]; it may also be mentioned that the extracellular matrix (ECM) issue pertains to Dostál’s [10] topic of the structural integrity of plant organism [11].

At the early stage of their development, the ESEs are covered with a special extracellular cell wall layer referred to as the extracellular matrix surface network (ECMSN), [12]. Neděla et al. [13], [14], [15] utilized an environmental scanning electron microscope (ESEM) to describe the ECM and/or ECMSN in conifers (spruce, pine, and fir) at the native stage. Arabinogalactan-proteins (AGPs) are progressively accumulated within the ECMSN, a well known marker covering the embryogenic cells during the embryo development. Specific AGPs are essential in somatic embryogenesis and exhibit the capability of directing the development of the cells [16]. It is possible that AGPs may be present as adhesives in the middle lamella to cement the cell-to-cell contact and they may also be involved in the adhesion of the callus cell clumps [17]. Two homogeneous AGPs were purified from the Norway spruce callus cells via ion-exchange and gel-permeation chromatography followed by enzymatic treatment [18]. The function of AGPs was recently summarized in a review by Seifert and Roberts [19].

Our article further extends a related, previously published paper [20], where the authors conducted 4 experiments: 1) the designing of methods to assess the water amount in relevant somatic embryos; 2) a comparison of the SNR in images acquired at different magnetic flux density values of the basic magnetic field; 3) monitoring the tissue growth via various techniques; and 4) the segmentation of two subjectively distinguishable regions in the tissue, with each of these exhibiting a specific T2 relaxation time. Our research report develops the work carried out by Mikulka et al., especially as regards the image processing methodology. In the given context, the aim of the present article is to utilize MRI relaxometry in order to visualize the relaxation times of the early embryogenic tissue (callus) of the Norway spruce.

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