Importance of controls for the demonstration of carbohydrates in electron microscopy with the silver methenamine or the thiocarbohydrazide‐silver proteinate methods

Courtoy, R.; Simar, Léopold

doi:10.1111/j.1365-2818.1974.tb03929.x

Cited by 95 publications

(34 citation statements)

References 13 publications

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“…Roots were harvested weekly and prepared for transmission electron microscopy (TEM) and scanning electron microscopy (SEM) as described previously (Duddridge & Read, 1982). Some sections were stained for periodate-sensitive carbohydrates using Thiery's procedure (Thiery, 1967) and the controls used by Courtoy & Simar (1974). Specimens were examined for TEM using an AEI 801 and for SEM using a Cambridge Stereoscan 150.…”

Section: Methodsmentioning

confidence: 99%

The Development and Ultrastructure of Ectomycorrhizas

Duddridge

1986

New Phytologist

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SUMMARYThe effect of the presence of an exogenous carbon source in the synthesis medium on compatible and incompatible interactions between Suillus grevillei (Klotzsch) Sing, and a number of ectomycorrhizal hosts has been studied at the ultrastructural level. Comparisons are made with the results of the previous study in which carbohydrate was omitted. Observations suggest that tbe availability of an external source of carbon to the fungus upsets the balance of the symbiosis in favour of the fungal partner, even in compatible interactions, causing changes in the host cell wall and at the bost-fungus interface. The rate and degree of colonization of the root surface is also influenced by the presence of carbohydrate in the synthesis medium. These results suggest that any in vitro studies undertaken on compatibility and specificity in ectomycorrhizal associations should omit carbohydrate from the medium or use an aseptically synthesized donor host as the source of fungal inoculum.

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Section: Methodsmentioning

confidence: 99%

The Development and Ultrastructure of Ectomycorrhizas

Duddridge

1986

New Phytologist

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“…The stained sections were kept in darkness until they were examined. To exclude the possibility of unspecific staining, several control treatments (TCH-SP, PA-SP, and SP) were performed by omitting one or two steps of the standard PA-TCH-SP method as described (Courtoy and Simar, 1974).…”

Section: Temmentioning

confidence: 99%

Autophagy Contributes to Leaf Starch Degradation

et al. 2013

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Transitory starch, a major photosynthetic product in the leaves of land plants, accumulates in chloroplasts during the day and is hydrolyzed to maltose and Glc at night to support respiration and metabolism. Previous studies in Arabidopsis thaliana indicated that the degradation of transitory starch only occurs in the chloroplasts. Here, we report that autophagy, a nonplastidial process, participates in leaf starch degradation. Excessive starch accumulation was observed in Nicotiana benthamiana seedlings treated with an autophagy inhibitor and in autophagy-related (ATG) gene-silenced N. benthamiana and in Arabidopsis atg mutants. Autophagic activity in the leaves responded to the dynamic starch contents during the night. Microscopy showed that a type of small starch granule-like structure (SSGL) was localized outside the chloroplast and was sequestered by autophagic bodies. Moreover, an increased number of SSGLs was observed during starch depletion, and disruption of autophagy reduced the number of vacuole-localized SSGLs. These data suggest that autophagy contributes to transitory starch degradation by sequestering SSGLs to the vacuole for their subsequent breakdown.

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“…Ultrathin sections collected on gold grids were stained for polysaccharides (PATAg test) according to the method of Thiery (1967); and for cystine-containing proteins (SWIFT test) according to the method of Swift (1968). The controls for the PATAg test employed non-oxidized thiocarbohydrazide-silver proteinate treated sections, as recommended by Courtoy & Simar (1974). The controls for SWIFT test were made by reduction with benzyl thiol, and alkylation with iodoacetate followed by treatment with silver-methenamine reagent.…”

Section: Tissue Processingmentioning

confidence: 99%

Infectivity of pine and eucalypt isolates of Pisolithus tinctorius (Pers.) Coker & Couch on roots of Eucalyptus urophylla S. T. Blake in vitro

Lei¹,

Lapeyrie

Malajczuk

et al. 1990

New Phytologist

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SUMMARYAn ultrastructural comparison of early stages of mycorrhiza formation on Eucalyptus urophylla was carried out using two isolates of P. tinctorius, one cultured from a sporocarp collected under pine and the other cultured from a sporocarp collected under eucalypt. Major differences at the fungus-root interface were observed 2 and 4 d following inoculation. Eucalypt root cells in contact with the isolate from pine developed a thickened cell wall, whereas the isolate from eucalypt was associated with an accumulation of extracellular fibrils. The fibrils stained positively for carbohydrates and protein. They were absent with the isolate from pine. Four days after inoculation with the isolate from eucalypt, acid phosphatase activity was detected along the fungal plasmalemma at the interface with host cells. This enzyme activity was not detected with the isolate from pine. Vacuolar acid phosphatase activity was detected in both fungal isolates when in contact with Eucalyptus roots. It is suggested that host recognition of ectomycorrhizal symbionts can be manifested within hours of inoculation with the fungal partner.

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Importance of controls for the demonstration of carbohydrates in electron microscopy with the silver methenamine or the thiocarbohydrazide‐silver proteinate methods

Cited by 95 publications

References 13 publications

The Development and Ultrastructure of Ectomycorrhizas

The Development and Ultrastructure of Ectomycorrhizas

Autophagy Contributes to Leaf Starch Degradation

Infectivity of pine and eucalypt isolates of Pisolithus tinctorius (Pers.) Coker & Couch on roots of Eucalyptus urophylla S. T. Blake in vitro

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