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

Gaupels, F., Buhtz, A., Knauer, T., Deshmukh, S., Waller, F., van Bel, A. J. E., et al. (2008). Adaptation of aphid stylectomy for analyses of proteins and mRNAs in barley phloem sap. Journal of Experimental Botany, 59(12), 3297-3306. doi:10.1093/jxb/ern181.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-27A0-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-27A1-D
Genre: Journal Article

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 Creators:
Gaupels, F.1, Author
Buhtz, A.2, Author              
Knauer, T.1, Author
Deshmukh, S.1, Author
Waller, F.1, Author
van Bel, A. J. E.1, Author
Kogel, K. H.1, Author
Kehr, J.2, Author              
Affiliations:
1External Organizations, ou_persistent22              
2Micro- and Protein-Analysis, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society, ou_1753346              

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Free keywords: aphid barley cdna-aflp mrna phloem protein rhopalosiphum padi signalling stylectomy two-dimensional gel electrophoresis long-distance transport sieve-tube exudate ricinus-communis macromolecular trafficking in-vivo plants rice movement defense gene
 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 mu l 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 (wheat-induced resistance 1A)-like protein. Further, one-dimensional 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 phloem-mobile sucrose transporter. Notably, two-dimensional electrophoresis confirmed that there are > 250 phloem proteins awaiting identification in future studies.

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Language(s): eng - English
 Dates: 2008-07-182008
 Publication Status: Published in print
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 Identifiers: ISI: ISI:000259202900007
DOI: 10.1093/jxb/ern181
ISSN: 1460-2431 (Electronic) 0022-0957 (Linking)
URI: ://000259202900007 http://jxb.oxfordjournals.org/content/59/12/3297.full.pdf
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Title: Journal of Experimental Botany
Source Genre: Journal
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Pages: - Volume / Issue: 59 (12) Sequence Number: - Start / End Page: 3297 - 3306 Identifier: -