The use of fluorescence microscopy to monitor root development in micropropagated explants

Nicholas, J. R.; Gates, Phil; Grierson, Don

doi:10.1080/14620316.1986.11515721

Cited by 11 publications

(9 citation statements)

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“…Polysaccharides were stained with periodic acid shift reagent-PAS [ 79 ] counterstained with 0.02% Toluidine Blue for general structure, and proteins with 0.25% Naphtol Blue Black in 1% acetic acid [ 80 ]. Also 0.07% calcofluor white for cellulose [ 48 ] and other polysaccharides [ 52 ], 0.01% auramine in 0.05M phosphate buffer for cutin and lipids [ 81 ], and 0.01% acridine orange in 0.03% phosphate buffer, pH7.4 [ 82 ] were used to observe the stylar morphology.…”

Section: Methodsmentioning

confidence: 99%

Glycoprotein composition along the pistil of Malus x domestica and the modulation of pollen tube growth

2014

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BackgroundThe characteristics of pollen tube growth are not constant, but display distinct patterns of growth within the different tissues of the pistil. In the stigma, the growth rate is slow and autotrophic, whereas in the style, it is rapid and heterotrophic. Very little is known about the interactions between these distinct maternal tissues and the traversing pollen tube and the role of this interaction on the observed metabolism. In this work we characterise pollen tube growth in the apple flower and look for differences in glycoprotein epitope localization between two different maternal tissues, the stigma and the style.ResultsWhile immunocytochemically-detected arabinogalactan proteins were present at high levels in the stigma, they were not detected in the transmitting tissue of the style, where extensins were abundant. Whereas extensins remained at high levels in unpollinated pistils, they were no longer present in the style following pollen tube passage. Similarily, while abundant in unpollinated styles, insoluble polysaccharides such as β-glucans, were depleted in pollinated pistils.ConclusionsThe switch from autotropic to heterotrophic pollen tube growth correlates spatially with a change of glycoprotein epitopes between the stigma and the style. The depletion of extensins and polysaccharides following pollen tube passage in the style suggest a possible contribution to the acceleration of heterotrophic pollen tube growth, which would imply an active contribution of female tissues on prezygotic male–female crosstalk.

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Section: Methodsmentioning

confidence: 99%

Glycoprotein composition along the pistil of Malus x domestica and the modulation of pollen tube growth

2014

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“…To observe pollen tubes and callose, squashed preparations and paraffin-embedded material sectioned at 10 µm were stained with 0·1 % aniline blue in 0·1 n PO 4 K 3 ( Currier, 1957 ; Linskens and Esser, 1957 ). Sections were also stained with 0·01 % acridine orange in 0·03 m phosphate buffer to observe DNA and RNA ( Nicholas et al , 1986 ; Dudley et al , 1987 ), with 0·01 % auramine O in 0·05 m phosphate buffer to observe cutine and suberine ( Heslop-Harrison, 1977 ) and with 0·07 % calcofluor in water for cellulose ( Hughes and McCully, 1975 ). For general histological examination, paraffin-embedded material was stained with a mixed staining in the following order: 0·1 % aniline blue in 0·1 n PO 4 K 3 , 0·01 % acridine orange in water, 0·01 % auramine O in water and 0·01 % calcofluor in water, for 10 min each at 40 °C to accelerate staining and drying.…”

Section: Methodsmentioning

confidence: 99%

The progamic phase of an early-divergent angiosperm, Annona cherimola (Annonaceae)

2009

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Background and AimsRecent studies of reproductive biology in ancient angiosperm lineages are beginning to shed light on the early evolution of flowering plants, but comparative studies are restricted by fragmented and meagre species representation in these angiosperm clades. In the present study, the progamic phase, from pollination to fertilization, is characterized in Annona cherimola, which is a member of the Annonaceae, the largest extant family among early-divergent angiosperms. Beside interest due to its phylogenetic position, this species is also an ancient crop with a clear niche for expansion in subtropical climates.MethodsThe kinetics of the reproductive process was established following controlled pollinations and sequential fixation. Gynoecium anatomy, pollen tube pathway, embryo sac and early post-fertilization events were characterized histochemically.Key ResultsA plesiomorphic gynoecium with a semi-open carpel shows a continuous secretory papillar surface along the carpel margins, which run from the stigma down to the obturator in the ovary. The pollen grains germinate in the stigma and compete in the stigma-style interface to reach the narrow secretory area that lines the margins of the semi-open stylar canal and is able to host just one to three pollen tubes. The embryo sac has eight nuclei and is well provisioned with large starch grains that are used during early cellular endosperm development.ConclusionsA plesiomorphic simple gynoecium hosts a simple pollen–pistil interaction, based on a support–control system of pollen tube growth. Support is provided through basipetal secretory activity in the cells that line the pollen tube pathway. Spatial constraints, favouring pollen tube competition, are mediated by a dramatic reduction in the secretory surface available for pollen tube growth at the stigma–style interface. This extramural pollen tube competition contrasts with the intrastylar competition predominant in more recently derived lineages of angiosperms.

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“…For this purpose, five flowers for each day of pollination were fixed 24 h after pollination in 2.5% (v/v) glutaraldehyde in 30 mmol/L phosphate buffer (Sabatini, Bench, and Barrnett, 1963) at pH 6.8, dehydrated in an ethanol series, and embedded in Historesin (Reichert-Jung, Heidelberg, Germany). Sections, 2 m thick, were stained with 0.07% calcofluor white for cellulose (Hughes and McCully, 1975), 0.01% auramine in 0.05 M phosphate buffer (pH 7.8) for cutin (Heslop-Harrison, 1977), and acridine orange (0.01%) in 0.03% phosphate buffer (Nicholas, Gates, and Grierson, 1986) for a general overview of cytoplasm and intercellular matrix. For a general histological examination of the tissue, these stains were also used in combination.…”

Section: Plant Materials and Pollination Procedures-pear Trees (Pyrus mentioning

confidence: 99%

Asynchronous development of stigmatic receptivity in the pear (Pyrus communis; Rosaceae) flower

Sanzol

Rallo

Herrero

2003

American J of Botany

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While stigma anatomy is well documented for a good number of species, little information is available on the acquisition and cessation of stigmatic receptivity. The aim of this work is to characterize the development of stigma receptivity, from anthesis to stigma degeneration, in the pentacarpellar pear (Pyrus communis) flower. Stigma development and stigmatic receptivity were monitored over two consecutive years, as the capacity of the stigmas to offer support for pollen germination and pollen tube growth. In an experiment where hand pollinations were delayed for specified times after anthesis, three different stigmatic developmental stages could be observed: (1) immature stigmas, which allow pollen adhesion but not hydration; (2) receptive stigmas, which allow proper pollen hydration and germination; and (3) degenerated stigmas, in which pollen hydrates and germinates properly, but pollen tube growth is impaired soon after germination. This developmental characterization showed that stigmas in different developmental stages coexist within a flower and that the acquisition and cessation of stigmatic receptivity by each carpel occur in a sequential manner. In this way, while the duration of stigmatic receptivity for each carpel is rather short, the flower has an expanded receptive period. This asynchronous period of receptivity for the different stigmas of a single flower is discussed as a strategy that could serve to maximize pollination resources under unreliable pollination conditions.

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The use of fluorescence microscopy to monitor root development in micropropagated explants

Cited by 11 publications

References 0 publications

Glycoprotein composition along the pistil of Malus x domestica and the modulation of pollen tube growth

Glycoprotein composition along the pistil of Malus x domestica and the modulation of pollen tube growth

The progamic phase of an early-divergent angiosperm, Annona cherimola (Annonaceae)

Asynchronous development of stigmatic receptivity in the pear (Pyrus communis; Rosaceae) flower

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