EJPAU 2008. Urbańska A. MICROSCOPIC VISUALIZATION AND ELECTROPHORETIC SEPARATION OF APHID PEROXIDASE

Electronic Journal of Polish Agricultural Universities (EJPAU) founded by all Polish Agriculture Universities presents original papers and review articles relevant to all aspects of agricultural sciences. It is target for persons working both in science and industry,regulatory agencies or teaching in agricultural sector. Covered by IFIS Publishing (Food Science and Technology Abstracts), ELSEVIER Science - Food Science and Technology Program, CAS USA (Chemical Abstracts), CABI Publishing UK and ALPSP (Association of Learned and Professional Society Publisher - full membership). Presented in the Master List of Thomson ISI.

2008
Volume 11
Issue 4

Topic:

Biology

ELECTRONIC
JOURNAL OF
POLISH
AGRICULTURAL
UNIVERSITIES

Urbańska A. 2008. MICROSCOPIC VISUALIZATION AND ELECTROPHORETIC SEPARATION OF APHID PEROXIDASE, EJPAU 11(4), #31.
Available Online: http://www.ejpau.media.pl/volume11/issue4/abs-31.html

MICROSCOPIC VISUALIZATION AND ELECTROPHORETIC SEPARATION OF APHID PEROXIDASE

Anna Urbańska
Department of Biochemistry and Molecular Biology, University of Natural Sciences and Humanities in Siedlce, Poland

ABSTRACT

Localization of the enzyme, peroxidase (POD)
in the gelling saliva (the solid stylet sheath) and in the midgut of aphid, Sitobion
avenae F. and Rhopalosiphum padi L. is recognized in this paper, by
the sensitive staining procedure with 3,3', 5,5' – tetramethylbenzidine and H₂O₂,
(TMBZ-H₂O₂). POD activity is not identified in soluble
fraction of salivary proteins diffused around of the stylet sheath and released
in the watery saliva, and that is extremely easly visualized with the Coomassie
Brillant Blue R-250 reagent both in the agarose gel and on the sucrose syrup
layer punctured by aphids. Moreover, localization, quantification and molecular
weight of aphid POD isoenzymes after their separation by sodium dodecyl sulfate
– polyacrylamide gel electrophoresis (SDS–PAGE) and staining by TMBZ–H₂O₂/Coomassie
Blue are shown. Generally, for salivary glands, midgut and whole aphid organism
two POD bands/isoenzymes are established. Real significance of aphid POD for
formation of stylet sheath, elimimination of H₂O₂ generated
in aphid tissues as result of red-ox metabolism and detoxication/toxication of
phenolic allelochemicals is specified. TMBZ – H₂O₂ appears
as excellent/suitable technique for the microscopical visulization of POD reaction
in various aphid tissues and agarose or polyacrylamide gels.

Key words: peroxidase (POD), POD activity, protein, aphid, salivary secretions, stylet sheath, watery saliva, midgut, isoenzyme, TMBZ-H₂O₂ technique, SDS–PAGE, Coomassie Blue stain.

Anna Urbańska
Department of Biochemistry and Molecular Biology,
University of Natural Sciences and Humanities in Siedlce, Poland
B. Prusa 12, 08-110 Siedlce, Poland
phone: +48 25 643 12 22,
fax: +48 25 644 59 59
email: annau@uph.edu.pl

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