New staining method for fungal-infected plant tissues

Marques, João Paulo Rodrigues; Soares, Marli Kasue Misaki; Appezzato‐da‐Glória, Beatriz

doi:10.3906/bot-1204-9

Cited by 19 publications

(17 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Forty microsections were aligned in order on one glass slide, and five slides were made for each embedded block. Slides were stained by cotton blue and safranin O [ 26 ].…”

Section: Methodsmentioning

confidence: 99%

Observations on the Early Establishment of Foliar Endophytic Fungi in Leaf Discs and Living Leaves of a Model Woody Angiosperm, Populus trichocarpa (Salicaceae)

Huang

Zimmerman

Arnold

2018

JoF

View full text Add to dashboard Cite

Fungal endophytes are diverse and widespread symbionts that occur in the living tissues of all lineages of plants without causing evidence of disease. Culture-based and culture-free studies indicate that they often are abundant in the leaves of woody angiosperms, but only a few studies have visualized endophytic fungi in leaf tissues, and the process through which most endophytes colonize leaves has not been studied thoroughly. We inoculated leaf discs and the living leaves of a model woody angiosperm, Populus trichocarpa, which has endophytes that represent three distantly-related genera (Cladosporium, Penicillium, and Trichoderma). We used scanning electron microscopy and light microscopy to evaluate the timeline and processes by which they colonize leaf tissue. Under laboratory conditions with high humidity, conidia germinated on leaf discs to yield hyphae that grew epiphytically and incidentally entered stomata, but did not grow in a directed fashion toward stomatal openings. No cuticular penetration was observed. The endophytes readily colonized the interiors of leaf discs that were detached from living leaves, and could be visualized within discs with light microscopy. Although they were difficult to visualize within the interior of living leaves following in vivo inoculations, standard methods for isolating foliar endophytes confirmed their presence.

show abstract

“…Forty microsections were aligned in order on one glass slide, and five slides were made for each embedded block. Slides were stained by cotton blue and safranin O [ 26 ].…”

Section: Methodsmentioning

confidence: 99%

Observations on the Early Establishment of Foliar Endophytic Fungi in Leaf Discs and Living Leaves of a Model Woody Angiosperm, Populus trichocarpa (Salicaceae)

Huang

Zimmerman

Arnold

2018

JoF

View full text Add to dashboard Cite

show abstract

“…Lignified tissues are stained red by safranin O, whereas unlignified cellulose fibres are stained blue by astra blue (see also Metabolites and structural features). A double-staining method of safranin O with cotton blue has been used to identify fungal structures in plant tissues (Marques et al, 2013). With this staining method, fungal cell walls (blue) can be distinguished from the plant cell walls (orange).…”

Section: Individual Pathogen Stages Detected By Various Staining Methodsmentioning

confidence: 99%

Histological methods to detect the clubroot pathogen Plasmodiophora brassicae during its complex life cycle

Schuller

Ludwig‐Müller

2016

Plant Pathology

View full text Add to dashboard Cite

The clubroot pathogen Plasmodiophora brassicae is an obligate biotrophic protist that lives in close relationship with its host cell. The roots of the host plants are colonized and the plant growth is altered upon infection. While shoots can be stunted and show wilt symptoms after longer infection periods, the root system is converted to a tumorous root tissue, called 'clubroot', by alterations of the plant growth promoting hormones auxin, cytokinin and brassinosteroid. Because the life cycle occurs largely within the host cells, this leads to dramatic changes in host root morphology and anatomy. Thus, the identification of the respective protist structures in the host tissue by microscopy is challenging. Different staining methods as well as fluorescence and electron microscopy of thin sections can reveal specific life stages of P. brassicae and can yield additional information on the changes in the host tissues concerning, for example, cell wall properties. In addition, promoter-reporter fusions, immunostaining methods and in situ hybridization techniques can be used to gain additional information on the changes in the host roots.

show abstract

“…Lactophenol cotton blue-stained leaves demonstrated a network of mycelia in AB only, which was not observed in AB + SNP (Supplementary Figures S3A,B). Lactophenol cotton blue is a stain used to distinguish fungal mycelia from plant cells, which stains fungal hyphae in blue color, leaving plant cell wall unstained (Marques et al, 2013). Similarly, trypan blue is another stain used to distinguish between dead and live tissues (Fernández-Bautista et al, 2016).…”

Section: Evaluation Of Biosynthesized Silvermentioning

confidence: 99%

Omics-Based Mechanistic Insight Into the Role of Bioengineered Nanoparticles for Biotic Stress Amelioration by Modulating Plant Metabolic Pathways

Kumari

Pandey

Mishra

et al. 2020

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Bioengineered silver nanoparticles can emerge as a facile approach to combat plant pathogen, reducing the use of pesticides in an eco-friendly manner. The plants' response during tripartite interaction of plant, pathogen, and nanoparticles remains largely unknown. This study demonstrated the use of bioengineered silver nanoparticles in combating black spot disease caused by necrotrophic fungus Alternaria brassicicola in Arabidopsis thaliana via foliar spray. The particles reduced disease severity by 70-80% at 5 µg/ml without showing phytotoxicity. It elicited plant immunity by a significant reduction in reactive oxygen species (ROS), decreases in stress enzymes by 0.6-19.8fold, and emergence of autophagy. Comparative plant proteomics revealed 599 proteins expressed during the interaction, where 117 differential proteins were identified. Among different categories, proteins involved in bioenergy and metabolism were most abundant (44%), followed by proteins involved in plant defense (20%). Metabolic profiling by gas chromatography-mass spectroscopy yielded 39 metabolite derivatives in non-polar fraction and 25 in the polar fraction of plant extracts. It was observed that proteins involved in protein biogenesis and early plant defense were overexpressed to produce abundant antimicrobial metabolites and minimize ROS production. Bioengineered silver nanoparticles performed dual functions to combat pathogen attack by killing plant pathogen and eliciting immunity by altering plant defense proteome and metabolome.

show abstract

New staining method for fungal-infected plant tissues

Cited by 19 publications

References 24 publications

Observations on the Early Establishment of Foliar Endophytic Fungi in Leaf Discs and Living Leaves of a Model Woody Angiosperm, Populus trichocarpa (Salicaceae)

Observations on the Early Establishment of Foliar Endophytic Fungi in Leaf Discs and Living Leaves of a Model Woody Angiosperm, Populus trichocarpa (Salicaceae)

Histological methods to detect the clubroot pathogen Plasmodiophora brassicae during its complex life cycle

Omics-Based Mechanistic Insight Into the Role of Bioengineered Nanoparticles for Biotic Stress Amelioration by Modulating Plant Metabolic Pathways

Contact Info

Product

Resources

About