Filter strip as a method of choice for apoplastic fluid extraction from maize roots

Maksimović, Jelena Dragišić; Živanović, Branka D.; Maksimović, Vuk; Mojović, Miloš; Nikolić, Miroslav; Vučinić, Željko

doi:10.1016/j.plantsci.2014.03.009

Cited by 14 publications

(17 citation statements)

References 60 publications

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“…The activity of MDH detected in AF extracted from un-inoculated and inoculated roots was up to 1.5% compared to the total soluble protein extract from roots (Supplementary Table 1 ). MDH levels below 2% are considered suitable for plant apoplast studies (Dannel et al, 1995 ; Dragišić Maksimović et al, 2008 , 2014 ; Yang et al, 2015 ).…”

Section: Resultsmentioning

confidence: 99%

The Apoplastic Secretome of Trichoderma virens During Interaction With Maize Roots Shows an Inhibition of Plant Defence and Scavenging Oxidative Stress Secreted Proteins

et al. 2018

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In Nature, almost every plant is colonized by fungi. Trichoderma virens is a biocontrol fungus which has the capacity to behave as an opportunistic plant endophyte. Even though many plants are colonized by this symbiont, the exact mechanisms by which Trichoderma masks its entrance into its plant host remain unknown, but likely involve the secretion of different families of proteins into the apoplast that may play crucial roles in the suppression of plant immune responses. In this study, we investigated T. virens colonization of maize roots under hydroponic conditions, evidencing inter- and intracellular colonization by the fungus and modifications in root morphology and coloration. Moreover, we show that upon host penetration, T. virens secretes into the apoplast an arsenal of proteins to facilitate inter- and intracellular colonization of maize root tissues. Using a gel-free shotgun proteomics approach, 95 and 43 secretory proteins were identified from maize and T. virens, respectively. A reduction in the maize secretome (36%) was induced by T. virens, including two major groups, glycosyl hydrolases and peroxidases. Furthermore, T. virens secreted proteins were mainly involved in cell wall hydrolysis, scavenging of reactive oxygen species and secondary metabolism, as well as putative effector-like proteins. Levels of peroxidase activity were reduced in the inoculated roots, suggesting a strategy used by T. virens to manipulate host immune responses. The results provide an insight into the crosstalk in the apoplast which is essential to maintain the T. virens-plant interaction.

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Section: Resultsmentioning

confidence: 99%

The Apoplastic Secretome of Trichoderma virens During Interaction With Maize Roots Shows an Inhibition of Plant Defence and Scavenging Oxidative Stress Secreted Proteins

et al. 2018

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“…The findings that zeamines biosynthesis is sensitive to nutrient conditions under in vitro conditions may also be relevant to the in planta pathogen-plant interactions, given that zeamines are the key factors associated with the pathogenesis of D. zeae . Studies in some monocotyledon plants showed that sugars, amino acids and organic acids are the main organic compounds [28] . However, the root fluid composition in rice is still unknown.…”

Section: Discussionmentioning

confidence: 99%

Production of Novel Antibiotics Zeamines through Optimizing Dickeya zeae Fermentation Conditions

et al. 2014

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Dickeya zeae strain EC1 was recently shown to produce a new type of phytotoxins designated as zeamine and zeamine II, which are potent wide-spectrum antibiotics against Gram-positive and Gram-negative bacterial pathogens, suggesting their promising potential as clinical medicines. In this study, the optimized medium composition and culture conditions for biosynthesis of novel antibiotics zeamines have been established by using response surface methodology, largely increasing the yield of zeamines from original about 7.35 µg·mL−1 in minimal medium to about 150 µg·mL−1 in LS5 medium. The study identified the major factors contributing to zeamines production, which include nitrate, sucrose, asparaginate, mineral elements Mg2+ and K+, and optimized amount of phosphate. In addition, the results showed that overexpression of zmsK in D. zeae strain EC1 could further increase zeamines yield to about 180 µg·mL−1 in LS5 medium. The findings from this study could facilitate further characterization and utilization of these two novel antibiotics, and also provide useful clues for understanding the regulatory mechanisms that govern D. zeae virulence.

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“…Here, GC-MS analysis detected zero or trace levels of G-6-P in AWF extractions. By contrast, in cut maize root sections, G-6-P was detected after AWF extraction at all centrifugation speeds and increased in concentration at higher speeds, indicating cytoplasmic contamination 10 .…”

Section: Representative Resultsmentioning

confidence: 68%

The Infiltration-centrifugation Technique for Extraction of Apoplastic Fluid from Plant Leaves Using <em>Phaseolus vulgaris</em> as an Example

O’Leary

Rico

McCraw

et al. 2014

JoVE

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The apoplast is a distinct extracellular compartment in plant tissues that lies outside the plasma membrane and includes the cell wall. The apoplastic compartment of plant leaves is the site of several important biological processes, including cell wall formation, cellular nutrient and water uptake and export, plant-endophyte interactions and defence responses to pathogens. The infiltration-centrifugation method is well established as a robust technique for the analysis of the soluble apoplast composition of various plant species. The fluid obtained by this method is commonly known as apoplast washing fluid (AWF). The following protocol describes an optimized vacuum infiltration and centrifugation method for AWF extraction from Phaseolus vulgaris (French bean) cv. Tendergreen leaves. The limitations of this method and the optimization of the protocol for other plant species are discussed. Recovered AWF can be used in a wide range of downstream experiments that seek to characterize the composition of the apoplast and how it varies in response to plant species and genotype, plant development and environmental conditions, or to determine how microorganisms grow in apoplast fluid and respond to changes in its composition.

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Filter strip as a method of choice for apoplastic fluid extraction from maize roots

Cited by 14 publications

References 60 publications

The Apoplastic Secretome of Trichoderma virens During Interaction With Maize Roots Shows an Inhibition of Plant Defence and Scavenging Oxidative Stress Secreted Proteins

The Apoplastic Secretome of Trichoderma virens During Interaction With Maize Roots Shows an Inhibition of Plant Defence and Scavenging Oxidative Stress Secreted Proteins

Production of Novel Antibiotics Zeamines through Optimizing Dickeya zeae Fermentation Conditions

The Infiltration-centrifugation Technique for Extraction of Apoplastic Fluid from Plant Leaves Using <em>Phaseolus vulgaris</em> as an Example

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