A combinatory approach for analysis of protein sets in barley sieve-tube samples using EDTA-facilitated exudation and aphid stylectomy

Gaupels, Frank; Knauer, Torsten; Bel, Aart J. E. van

doi:10.1016/j.jplph.2007.07.023

Cited by 40 publications

(30 citation statements)

References 56 publications

(63 reference statements)

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“…However, separate sampling of EFP and fascicular phloem exudates of pumpkin using microdissection revealed large differences in the sugar and protein composition between both phloem systems (Zhang et al, 2010). In this report, this finding was confirmed by employing aphid stylectomy for the collection of pure phloem sap (Gaupels et al, 2008a(Gaupels et al, , 2008c. Pumpkin stylet exudates were compared with exudates from cut petioles and stems displaying different SDS-PAGE band patterns between the two sample types (Fig.…”

Section: Resultssupporting

confidence: 51%

“…Pumpkin phloem latex was either snap frozen in liquid N 2 for metabolite analyses or mixed with the same volume of phloem buffer if not otherwise stated (50 mM Tris-HCl, pH 7.8, 0.1% b-mercaptoethanol;McEuen and Hill, 1982). Aphid stylectomy was used for phloem sampling according to Gaupels et al (2008aGaupels et al ( , 2008c with the aphid Macrosiphum euphorbiae. The time until sample gelation was measured after the addition of 3 volumes of alkaline buffer (25 mM HEPES/NaOH, pH 7.8) to the phloem latex (80 mL).…”

Section: Plant Material Sampling and Sample Gelation Assaymentioning

confidence: 99%

“…SDS-PAGE, Coomassie blue staining, and silver staining were performed according to Gaupels et al (2008bGaupels et al ( , 2008c. ImageJ was applied for the quantification of SDS-PAGE bands.…”

Section: Sds-page and Western-blot Analysesmentioning

confidence: 99%

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Deciphering Systemic Wound Responses of the Pumpkin Extrafascicular Phloem by Metabolomics and Stable Isotope-Coded Protein Labeling

et al. 2012

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In cucurbits, phloem latex exudes from cut sieve tubes of the extrafascicular phloem (EFP), serving in defense against herbivores. We analyzed inducible defense mechanisms in the EFP of pumpkin (Cucurbita maxima) after leaf damage. As an early systemic response, wounding elicited transient accumulation of jasmonates and a decrease in exudation probably due to partial sieve tube occlusion by callose. The energy status of the EFP was enhanced as indicated by increased levels of ATP, phosphate, and intermediates of the citric acid cycle. Gas chromatography coupled to mass spectrometry also revealed that sucrose transport, gluconeogenesis/glycolysis, and amino acid metabolism were up-regulated after wounding. Combining ProteoMiner technology for the enrichment of low-abundance proteins with stable isotope-coded protein labeling, we identified 51 wound-regulated phloem proteins. Two Sucrose-Nonfermenting1-related protein kinases and a 32-kD 14-3-3 protein are candidate central regulators of stress metabolism in the EFP. Other proteins, such as the Silverleaf Whitefly-Induced Protein1, Mitogen Activated Protein Kinase6, and Heat Shock Protein81, have known defensive functions. Isotope-coded protein labeling and western-blot analyses indicated that Cyclophilin18 is a reliable marker for stress responses of the EFP. As a hint toward the induction of redox signaling, we have observed delayed oxidation-triggered polymerization of the major Phloem Protein1 (PP1) and PP2, which correlated with a decline in carbonylation of PP2. In sum, wounding triggered transient sieve tube occlusion, enhanced energy metabolism, and accumulation of defense-related proteins in the pumpkin EFP. The systemic wound response was mediated by jasmonate and redox signaling.

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Section: Resultssupporting

confidence: 51%

Section: Plant Material Sampling and Sample Gelation Assaymentioning

confidence: 99%

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Deciphering Systemic Wound Responses of the Pumpkin Extrafascicular Phloem by Metabolomics and Stable Isotope-Coded Protein Labeling

et al. 2012

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“…This allows for the collection of undiluted phloem sap, so that one can measure directly the concentrations of the amino acids and other phloem components. However, the volume collected from each stylet is typically 10 to 150 nL [61,[63][64][65], and evaporation from these very small samples can alter the component concentrations [59,62,66].…”

Section: Introductionmentioning

confidence: 99%

A speciation model of essential trace metal ions in phloem

Harris

Sammons

Grabiak

2012

Journal of Inorganic Biochemistry

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“…Ten exudate droplets were dissolved in 600l Millipore water and were stored at -20°C. Exudate droplets of phloem sap collected from R. padi on H. vulgare were sampled as described previously (Gaupels et al, 2008) after six hours of stylet exudation. Droplets were also dissolved to 600l with Millipore water and stored at -20°C.…”

Section: Introductionmentioning

confidence: 99%

Plant cues for aphid navigation in vascular tissues

Hewer

Will

Bel

2010

Journal of Experimental Biology

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SUMMARY The ability of aphids to detect and find sieve tubes suggests that aphids receive cues for sieve-tube recognition by taking samples. Specific natural conditions such as pH value, sugar species and concentration, viscosity, and oxygen pressure may enable sieve-tube detection. We tested the preference of Megoura viciae and Myzus persicae for potential plant-borne orientation parameters in artificial choice-chamber systems. Both species preferred sucrose (in comparison with fructose, glucose, raffinose or sorbitol) at concentrations of 15–22.5% (over a tested range of 0–22.5%) and at approximately pH 7 (over a tested range of pH 5–8). This preference matches the composition of the sieve-tube sap of their host plants. Likewise, Rhopalosiphum padi (normally found on barley plants with sucrose in the phloem sap) and Macrosiphum euphorbiae (normally found on pumpkin plants with raffinose-family oligosaccharides in the phloem sap) showed a significant preference for sucrose. In the absence of sucrose, however, M. euphorbiae strongly preferred raffinose. No clear preference for any carbohydrate was observed for Macrosiphum rosae and Aphis pomi (both normally found on plants with various amounts of sorbitol in the phloem sap). Electrical penetration graph (EPG) measurements of M. persicae feeding on artificial diets confirmed that sieve tubes are recognized solely by a combination of carbohydrate abundance and a neutral to slightly alkaline pH.

show abstract

A combinatory approach for analysis of protein sets in barley sieve-tube samples using EDTA-facilitated exudation and aphid stylectomy

Cited by 40 publications

References 56 publications

Deciphering Systemic Wound Responses of the Pumpkin Extrafascicular Phloem by Metabolomics and Stable Isotope-Coded Protein Labeling

Deciphering Systemic Wound Responses of the Pumpkin Extrafascicular Phloem by Metabolomics and Stable Isotope-Coded Protein Labeling

A speciation model of essential trace metal ions in phloem

Plant cues for aphid navigation in vascular tissues

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