Adaptation of aphid stylectomy for analyses of proteins and mRNAs in barley phloem sap

Gaupels, Frank and Buhtz, Anja and Knauer, Torsten and Deshmukh, Sachin and Waller, Frank and Van Bel, Aart J. E. and Kogel, Karl-Heinz and Kehr, Julia (2008). Adaptation of aphid stylectomy for analyses of proteins and mRNAs in barley phloem sap. "Journal of Experimental Botany", v. 59 (n. 12); pp. 3297-3306. ISSN 0022-0957. https://doi.org/10.1093/jxb/ern181.

Description

Title: Adaptation of aphid stylectomy for analyses of proteins and mRNAs in barley phloem sap
Author/s:
  • Gaupels, Frank
  • Buhtz, Anja
  • Knauer, Torsten
  • Deshmukh, Sachin
  • Waller, Frank
  • Van Bel, Aart J. E.
  • Kogel, Karl-Heinz
  • Kehr, Julia
Item Type: Article
Título de Revista/Publicación: Journal of Experimental Botany
Date: September 2008
ISSN: 0022-0957
Volume: 59
Subjects:
Freetext Keywords: Aphid, barley, cDNA-AFLP, mRNA, phloem, protein, Rhopalosiphum padi, signalling, stylectomy, twodimensional gel electrophoresis.
Faculty: Otros Centros UPM
Department: Biotecnologia [hasta 2014]
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

Sieve tubes are transport conduits not only for photoassimilates but also for macromolecules and other compounds that are involved in sieve tube maintenance and systemic signalling. In order to gain sufficient amounts of pure phloem exudates from barley plants for analyses of the protein and mRNA composition, a previously described stylectomy set-up was optimized. Aphids were placed in sealed cages, which, immediately after microcauterization of the stylets, were flooded with water-saturated silicon oil. The exuding phloem sap was collected with a capillary connected to a pump. Using up to 30 plants and 600 aphids (Rhopalosiphum padi) in parallel, an average of 10 ml of phloem sap could be obtained within 6 h of sampling. In first analyses of the macromolecular content, eight so far unknown phloem mRNAs were identified by cDNA-amplified fragment length polymorphism. Transcripts in barley phloem exudates are related to metabolism, signalling, and pathogen defence, for example coding for a protein kinase and a pathogen- and insect-responsive WIR1A (wheatinduced resistance 1A)-like protein. Further, onedimensional gel electrophoresis and subsequent partial sequencing by mass spectrometry led to the identification of seven major proteins with putative functions in stress responses and transport of mRNAs, proteins, and sugars. Two of the discovered proteins probably represent isoforms of a new phloemmobile sucrose transporter. Notably, two-dimensional electrophoresis confirmed that there are >250 phloem proteins awaiting identification in future studies.

More information

Item ID: 3027
DC Identifier: http://oa.upm.es/3027/
OAI Identifier: oai:oa.upm.es:3027
DOI: 10.1093/jxb/ern181
Official URL: http://jxb.oxfordjournals.org/cgi/content/full/59/12/3297
Deposited by: Memoria Investigacion
Deposited on: 12 May 2010 10:59
Last Modified: 20 Apr 2016 12:36
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