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Analysis of the intercellular fluid proteome of sugarcane roots inoculed with Glomus clarum.

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Author(s):
Simão Lindoso de Souza
Total Authors: 1
Document type: Master's Dissertation
Press: Piracicaba. , ilustrações, tabelas.
Institution: Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC)
Defense date:
Examining board members:
Marcio Rodrigues Lambais; Claudia de Mattos Bellato; Elke Jurandy Bran Nogueira Cardoso
Advisor: Marcio Rodrigues Lambais
Field of knowledge: Agronomical Sciences - Agronomy
Indexed in: Banco de Dados Bibliográficos da USP-DEDALUS; Biblioteca Digital de Teses e Dissertações - USP
Location: Universidade de São Paulo. Biblioteca Central da Escola Superior de Agricultura Luiz de Queiroz; ESALQBC/t633.61 79765; S729a; Universidade de São Paulo. Biblioteca do Centro de Energia Nuclear na Agricultura; CENA /(043) 1368; S729a
Abstract

The mechanisms controlling arbuscular mycorrhizae (AM) development at different soil P concentrations are not understood. It has been proposed that proteins secreted in the root apoplast may have important roles in AM regulation. The analyses of the intercellular fluid (IF) proteome from sugarcane roots inoculated with arbuscular mycorrhizal fungi, grown at low or high P conditions, compared to the IF proteome from not-inoculated roots may contribute to the understanding of the mechanisms controlling AM development, and was the aim of this work. Sugarcane seedlings were inoculated with Glomus clarum, Glomus etunicatum or Gigaspora rosea, and grown in sterilized substrate containing 20 or 200 mg P kg -1 . Eight weeks after inoculation, the plants were harvested and the following parameters evaluated: shoot dry weight, nutrients in the shoots and intraradical fungal growth. The results showed that intraradical colonization by G. clarum and G. etunicatum at high P conditions was significantly lower than at low P. Equal amounts of proteins were used to compare the proteome from the IF of not-inoculated and G. clarum inoculated roots, at low and high P conditions, using 2D-PAGE. The proteome analyses revealed the predominance of acidic proteins in the IF of sugarcane. A total of 49 proteins were detected in the IF of sugarcane at low P soil concentration. From those, 8.2% were induced and 10.2% suppressed (³ 50%) in the IF of roots inoculated with G. clarum, as compared to not-inoculated controls. Thirteen proteins were detected only in the IF of mycorrhizal roots, and represent putative mycorrhizins or extracellular fungal proteins induced at low P conditions. A total of 56 proteins were detected in the IF of sugarcane at high P conditions. From those, 8.9% were induced and 16.1% suppressed (³ 50%) in the IF of roots inoculated with G. clarum, as compared to not-inoculated controls. Twelve proteins were detected only in the IF of mycorrhizal roots, and represent putative mycorrhizins or extracellular fungal proteins, induced at high P conditions. Comparing the proteomes from the IF of non-inoculated sugarcane roots at low and high P conditions, 16 proteins were detected only at low P conditions, whereas 24 proteins were detected only at high P conditions. Comparing the proteomes from the IF of sugarcane roots inoculated with G. clarum at low and high P conditions, 12 proteins were detected only at low P conditions, whereas 5 proteins were detected only at high P conditions. Those proteins may be, direct or indirectly, involved in the development and/or efficiency of AM. Further characterization of those proteins by mass spectrometry and/or N-terminal sequencing would contribute to the determinations of their possible functions in AM. (AU)