Imaging of native early embryogenic tissue
of Scots pine (<i>Pinus sylvestris</i> L.) by ESEM

Hřib, J.; Vooková, B.; Neděla, Vilém

doi:10.1515/biol-2015-0028

Cited by 5 publications

(3 citation statements)

References 22 publications

(26 reference statements)

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“…Our paper underlines the wide usability and advantages of the ESEM which can be used not only as a tool for the imaging of conventionally treated or highly susceptible fresh biological samples 28,29 and polymers 30 , but also for fast, effective and inexpensive in-situ preparation allowing repetitive observation and elemental analysis of plant samples using our new ELTM. The in-situ preparation process can be controlled and directly modified according to sample specificity.…”

Section: Discussionmentioning

confidence: 89%

In-situ preparation of plant samples in ESEM for energy dispersive x-ray microanalysis and repetitive observation in SEM and ESEM

Tihlaříková

Neděla

Đorđević

2019

Sci Rep

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The Extended Low Temperature Method (ELTM) for the in-situ preparation of plant samples in an environmental scanning electron microscope enables carrying out repetitive topographical and material analysis at a higher resolution in the vacuum conditions of a scanning electron microscope or in the low gas pressure conditions of an environmental scanning electron microscope. The method does not require any chemical intervention and is thus suitable for imaging delicate structures rarely observable with common treatment methods. The method enables both sample stabilization as close to their native state as possible, as well as the transfer of the same sample from a low vacuum to an atmospheric condition for sample storage or later study. It is impossible for wet samples in the environmental scanning electron microscope. Our studies illustrate the high applicability of the ELTM for different types of plant tissue, from imaging of plant waxes at higher resolution, the morphological study of highly susceptible early somatic embryos to the elemental microanalysis of root cells. The method established here provides a very fast, universal and inexpensive solution for plant sample treatment usable in a commercial environmental scanning electron microscope equipped with a cooling Peltier stage.

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Section: Discussionmentioning

confidence: 89%

In-situ preparation of plant samples in ESEM for energy dispersive x-ray microanalysis and repetitive observation in SEM and ESEM

Tihlaříková

Neděla

Đorđević

2019

Sci Rep

Self Cite

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“…On the other hand, the shear force generated by stirring can destroy the aggregation of ET, and promote either the formation or lysis of ET; excessive stirring can cause damage to cell development [26]. Therefore, the appropriate orbiting speed is important for the normal proliferation of ET in suspension culture [27]. In Korean pine suspension culture, the orbiting speed significantly affects the proliferation of ET, and the amount of proliferation gradually decreases as the orbiting speed increases [14,22].…”

Section: Discussionmentioning

confidence: 99%

Suspension Culture and Somatic Embryogenesis of Korean Pine

Peng¹,

Gao²,

Wang³

et al. 2022

Phyton

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Korean pine is an important afforestation tree species in Northeast China, which has a high ecological and economic value. Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as explants has been successful, it cannot be applied to automation and large-scale production. Therefore, we urgently need a method that can increase the output of somatic embryos (SEs) to meet the needs of large-scale production. We used Korean pine 1-1 and 1-100 cell lines as research materials to evaluate the effects of inoculum-density, culture time, orbiting speed, vessel volume, plant growth regulator (PGR) concentration, and carbon source on the proliferation of embryogenic tissue (ET). The somatic embryogenesis ability of ET cultured in different liquid suspension media was also evaluated. We found that during liquid suspension culture of Korean pine ET, the sedimented cell volume (SCV), fresh weight (FW) and dry weight (DW) were affected by inoculumdensity, culture time, orbiting speed, 2,4-D concentration, 6-BA concentration and carbon source type. Fourty mg ⋅ mL −1 ET were transferred to a 200 mL Erlenmeyer flask containing 20 mL liquid medium, and cultured at 100 rpm/min for 14 days to obtain the maximum proliferation. In addition, we also found that SCV, FW and DW were higher when PGRs were reduced in the liquid suspension medium. The substitution of maltose for sucrose resulted in slow growth of cultures and limited SE yield (13 SEs g −1 FW). Although culture proliferation was high at 50 rpm, SE yield was inhibited by 48% compared with 100 rpm (50 rpm = 33 SEs g −1 FW; 100 rpm/min = 70 SEs g −1 FW). Cultivation in low-concentration PGR(1.15 μM ⋅ L −1 2,4-D, 0.25 μM ⋅ L −1 6-BA) and sucrose liquid medium at 100 rpm/min (80 SEs g −1 FW) could not only promote culture proliferation but also increase SE yield. The determination of the suspension culture scheme of Korean pine ET provides a reference for further expansion to bioreactor culture in the future and lays a foundation for the automation and scale of somatic embryogenesis of Korean pine.

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“…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].…”

Section: Introductionmentioning

confidence: 99%

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

Korinek

Mikulka

Hřib

et al. 2017

Measurement Science Review

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The paper describes the visualization of the cells (ESEs) and mucilage (ECMSN) in an embryogenic tissue via magnetic resonance imaging (MRI) relaxometry measurement combined with the subsequent multi-parametric segmentation. The computed relaxometry maps T1 and T2 show a thin layer (transition layer) between the culture medium and the embryogenic tissue. The ESEs, mucilage, and transition layer differ in their relaxation times T1 and T2; thus, these times can be used to characterize the individual parts within the embryogenic tissue. The observed mean values of the relaxation times T1 and T2 of the ESEs, mucilage, and transition layer are as follows: 1469 ± 324 and 53 ± 10 ms, 1784 ± 124 and 74 ± 8 ms, 929 ± 164 and 32 ± 4.7 ms, respectively. The multi-parametric segmentation exploiting the T1 and T2 relaxation times as a classifier shows the distribution of the ESEs and mucilage within the embryogenic tissue. The discussed T1 and T2 indicators can be utilized to characterize both the growth-related changes in an embryogenic tissue and the effect of biotic/abiotic stresses, thus potentially becoming a distinctive indicator of the state of any examined embryogenic tissue.

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Imaging of native early embryogenic tissue of Scots pine (Pinus sylvestris L.) by ESEM

Cited by 5 publications

References 22 publications

In-situ preparation of plant samples in ESEM for energy dispersive x-ray microanalysis and repetitive observation in SEM and ESEM

In-situ preparation of plant samples in ESEM for energy dispersive x-ray microanalysis and repetitive observation in SEM and ESEM

Suspension Culture and Somatic Embryogenesis of Korean Pine

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

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