A proteomics approach towards understanding blast fungus infection of rice grown under different levels of nitrogen fertilization

Abstract

Proteins extracted from leaf blades of rice plants infected with blast fungus, Magnaporthe grisea, were separated by two-dimensional polyacrylamide gel electrophoresis. The separated proteins were electroblotted onto a polyvinylidene difluoride membrane, and 63 proteins were analyzed by a gas-phase protein sequencer. The N-terminal amino acid sequences of 33 out of 63 proteins were determined in this manner. N-terminal regions of the remaining proteins could not be sequenced. The internal amino acid sequences of 12 proteins were determined by sequence analysis of peptides obtained by the Cleveland peptide mapping method. The amino acid sequences were compared with those of known plant and animal protein sequences to understand the nature of these proteins. As expected, leaf blades revealed predominantly the presence of photosynthetic proteins. Using this experimental approach named as proteome analysis, the functional proteins during blast fungus infection of rice with different levels of nitrogen nutrient were analyzed. Twelve proteins which appeared to change with different levels of nitrogen nutrient were identified. It was revealed that the level of ribulose-1,5-bisphosphate carboxylase/oxygenase was increased by top-dressing with nitrogen nutrient. Additionally, the pathogenesis related protein were observed following blast fungus infection using immunoblot analysis. It was conjectured that these proteins might be involved in incompatible interaction in rice plants following blast fungus infection. The information obtained on the amino acid sequences and antibodies interaction is expected to be helpful in predicting the function of these proteins.

Expression and Function of Proteins during Development of the Basal Region in Rice Seedlings

“…Studies of environmental stimuli have focused on searching for marker proteins in response to stimuli, such as treatment with the plant hormones jasmonic acid (23), brassinolide (24), or gibberellin (19,25); growth under salt (26); drought (27); ozone stress (28); and infection with blast fungus (29,30) or Rice yellow mottle virus (31).…”

A Proteomics Approach to Investigating Promotive Effects of Brassinolide on Lamina Inclination and Root Growth in Rice Seedlings.

“…[13][14][15][16][17] The techniques of proteome analysis have been employed to study alterations in rice cellular protein expression in response to various stimuli or as a result of differentiation and development. 18,19) The study of partial amino acid sequence analysis of plant proteins will contribute greatly to the field of molecular biology for the identification of proteins through homology search. The information thus obtained from amino acid sequencing of these proteins will be helpful in predicting the function of the proteins and for molecular cloning in future experiments.…”

Métodos e estratégias em proteômica e suas aplicações na área vegetal

“…Estudos na área de estresses abióticos, como seca, temperatura (SULE et al, 2004) e salinidade (DANI et al, 2005), e estresses bióticos, como doenças (KONISHI et al, 2001) e pragas, podem empregar a análise de proteínas diferencialmente expressas, com o intuito principal de fornecer base à descoberta de novos marcadores moleculares. Muitas dessas proteínas podem revelar funções consistentes com a resposta ao estresse.…”