Xylem Characterization Using Improved Pseudo-Schiff Propidium Iodide Staining of Whole Mount Samples and Confocal Laser-Scanning Microscopy

Coiro, Mario; Truernit, Elisabeth

doi:10.1007/978-1-4939-6722-3_11

Cited by 10 publications

(7 citation statements)

References 6 publications

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“…Sections of a leaf of Bowenia serrulata (W.Bull) Chamb. were stained and mounted according to Coiro and Truernit [45]. Cuticles were isolated by immersing leaf fragments in a 2:1 mixture of 30% hydrogen peroxide and 85% ethanol, warmed up to 60 °C until the leaf fragments turned transparent.…”

Section: Methodsmentioning

confidence: 99%

Eobowenia gen. nov. from the Early Cretaceous of Patagonia: indication for an early divergence of Bowenia?

Coiro

Pott

2017

BMC Evol Biol

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BackgroundEven if they are considered the quintessential “living fossils”, the fossil record of the extant genera of the Cycadales is quite poor, and only extends as far back as the Cenozoic. This lack of data represents a huge hindrance for the reconstruction of the recent history of this important group. Among extant genera, Bowenia (or cuticles resembling those of extant Bowenia) has been recorded in sediments from the Late Cretaceous and the Eocene of Australia, but its phylogenetic placement and the inference from molecular dating still imply a long ghost lineage for this genus.ResultsWe re-examine the fossil foliage Almargemia incrassata from the Lower Cretaceous Anfiteatro de Ticó Formation in Patagonia, Argentina, in the light of a comparative cuticular analysis of extant Zamiaceae. We identify important differences with the other member of the genus, viz. A. dentata, and bring to light some interesting characters shared exclusively between A. incrassata and extant Bowenia. We interpret our results to necessitate the erection of the new genus Eobowenia to accommodate the fossil leaf earlier assigned as Almargemia incrassata. We then perfom phylogenetic analyses, including the first combined morphological and molecular analysis of the Cycadales, that indicate that the newly erected genus could be related to extant Bowenia.Conclusion Eobowenia incrassata could represent an important clue for the understanding of evolution and biogeography of the extant genus Bowenia, as the presence of Eobowenia in Patagonia is yet another piece of the biogeographic puzzle that links southern South America with Australasia.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-017-0943-x) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Eobowenia gen. nov. from the Early Cretaceous of Patagonia: indication for an early divergence of Bowenia?

Coiro

Pott

2017

BMC Evol Biol

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“…Many previously published studies on traditional stains either targeted particular cellular components, such as lignin or suberin domains ( Brundrett et al 1988 , 1991 ; Lux et al 2005 , 2015 ; Ursache et al 2018 ), or the 3D cellular structure of tissues and organs with little topochemical detail ( Bougourd et al 2000 ; Kitin et al 2002 , 2003 ; Dubrovsky et al 2006 ; Coiro and Truernit 2017 ; Tofanelli et al 2019 ). Our goal is to present an integrative approach of both visualizing the tissue structure and the cellular chemistry while utilizing the advantages of fluorescence microscopy in terms of rapid and effective sample preparation and observation.…”

Section: Introductionmentioning

confidence: 99%

Direct fluorescence imaging of lignocellulosic and suberized cell walls in roots and stems

et al. 2020

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Background and Aims Investigating plant structure is fundamental in botanical science and provides crucial knowledge for the theories of plant evolution, ecophysiology, and for the biotechnological practices. Modern plant anatomy often targets the formation, localization and characterization of cellulosic, lignified, or suberized cell walls. While classical methods developed in the 60s are still popular, recent innovations in tissue preparation, fluorescence staining, and microscopy equipment offer advantages to the traditional practices for investigation of the complex lignocellulosic walls. Our goal is to enhance the productivity and quality of microscopy work by focusing on quick and cost-effective preparation of thick sections or plant specimen surfaces and efficient use of direct fluorescent stains. Methods We discuss popular histochemical microscopy techniques for visualization of cell walls, such as autofluorescence or staining with calcofluor, Congo red (CR), fluorol yellow (FY), safranin, and provide detailed descriptions of our own approaches and protocols. Key Results Autofluorescence of lignin in combination with CR and FY staining can clearly differentiate between lignified, suberized, and unlignified cell walls in root and stem tissues. Glycerol can serve as an effective clearing medium as well as the carrier of FY for staining of suberin and lipids allowing for observation of thick histological preparations. Three-dimensional imaging (3D) of all cell types together with chemical information by wide-field fluorescence or CLSM was achieved.

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“…The embedded specimens were sectioned on a Microm HM 355 rotary microtome using a conventional microtome knife D. Sections of mostly 2 µm were stained with ruthenium red and toluidine blue and mounted on microscope slides in Histomount. Non-embedded sections of leaflets were mounted in 80 % glycerol or Hoyer’s solution, prepared according to Coiro and Truernit (2017) . Sections of mature leaflets of D. edule and Bowenia spectabilis were subject to pseudo-Schiff propidium iodide (PS-PI) staining, a technique that allows confocal imaging of cell walls, as described in Coiro and Truernit (2017) , and then mounted in Hoyer’s solution.…”

Section: Methodsmentioning

confidence: 99%

“…Non-embedded sections of leaflets were mounted in 80 % glycerol or Hoyer’s solution, prepared according to Coiro and Truernit (2017) . Sections of mature leaflets of D. edule and Bowenia spectabilis were subject to pseudo-Schiff propidium iodide (PS-PI) staining, a technique that allows confocal imaging of cell walls, as described in Coiro and Truernit (2017) , and then mounted in Hoyer’s solution.…”

Section: Methodsmentioning

confidence: 99%

Stomatal development in the cycad family Zamiaceae

2021

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Background and Aims The gymnosperm order Cycadales is pivotal for our understanding of seed-plant phylogeny because of its phylogenetic placement close to the root node of extant spermatophytes and its combination of both derived and plesiomorphic character states. Although widely considered a ‘living fossil’ group, extant cycads display a high degree of morphological and anatomical variation. We investigate stomatal development in Zamiaceae to evaluate variation within the order and homologies between cycads and other seed plants. Methods Leaflets of seven species across five genera representing all major clades of Zamiaceae were examined at various stages of development using light microscopy and confocal microscopy. Key Results All genera examined have lateral subsidiary cells of perigenous origin that differ from other pavement cells in mature leaflets and could have a role in stomatal physiology. Early epidermal patterning in a “quartet” arrangement occurs in Ceratozamia, Zamia, and Stangeria. Distal encircling cells, which are sclerified at maturity, are present in all genera except Bowenia, which has relatively rapid elongation and differentiation of the pavement cells during leaflet development. Conclusions Stomatal structure and development in Zamiaceae highlights some traits that are plesiomorphic in seed plants, including the presence of perigenous encircling subsidiary cells, and shows a clear difference between the developmental trajectories of cycads and Bennettitales. Our study also shows an unexpected degree of variation among subclades in the family, potentially linked to differences in leaflet development and suggesting convergent evolution in cycads.

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Xylem Characterization Using Improved Pseudo-Schiff Propidium Iodide Staining of Whole Mount Samples and Confocal Laser-Scanning Microscopy

Cited by 10 publications

References 6 publications

Eobowenia gen. nov. from the Early Cretaceous of Patagonia: indication for an early divergence of Bowenia?

Eobowenia gen. nov. from the Early Cretaceous of Patagonia: indication for an early divergence of Bowenia?

Direct fluorescence imaging of lignocellulosic and suberized cell walls in roots and stems

Stomatal development in the cycad family Zamiaceae

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