The importance of AAE in enhancing
The importance of AAE3 in enhancing the survival of plants and yeast when confronted with certain environmental stresses has been documented in recent reports. In response to biotic stress such as oxalate-secreting micro-organisms, AAE3 was found to reduce the inhibitory growth effects of the secreted oxalate on yeast . In plants, AAE3 was determined to reduce the susceptibility of plants to oxalate-secreting phytopathogens [1,3,7]. In addition, two studies utilizing different plant species reported the importance of AAE3 in regulating aluminum tolerance [, , ]. An interesting aspect of these two latter studies is that in one report, a reduction in AAE3 resulted in a decrease in aluminum sensitivity , while in the other report, an increase in AAE3 resulted in a decrease in aluminum sensitivity . This apparent discrepancy will need to be clarified with additional studies before any firm conclusion can be drawn.
Thus far, the importance of AAE3 in plant growth has been confined to a single report in Arabidopsis . Characterization of an Ataae3 T-DNA mutant showed that lack of AAE3 AZ20 kinase resulted in reduction in vegetative growth, seed mucilage production, seed germination, and an increase in calcium oxalate accumulation in Arabidopsis. Whether such phenotypes are specific to Arabidopsis or whether a reduction in AAE3 expression would result in similar phenotypes in other plants remains unknown.
To expand our understanding of the impact of AAE3 in plant growth and development, we report here the characterization of Mtaae3 RNAi knock-down and Mtaae3 Tnt1 knock-out mutants. Comparisons of these determined phenotypes to the Ataae3 mutants provide additional insights into the responsiveness of each developmental phenotype to changes in the level of AAE3 gene expression. Like the Ataae3 T-DNA knock-out mutant, the Mtaae3 RNAi knock-down mutant showed an increase in calcium oxalate accumulation but lacked other Ataae3 mutant phenotypes. This apparent discrepancy was clarified through the phenotypic characterization of a Mtaae3 Tnt1 knock-out mutant, which displayed phenotypes similar to Ataae3 T-DNA knock-out mutant. These findings show that the level of AAE3 gene expression is important in determining the severity of the exhibited phenotypes.
Materials and methods
Results and discussion
Acknowledgements The contents of this publication do not necessarily reflect the views or policies of the U.S. Department of Agriculture, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. This work was supported by the U.S. Department of Agriculture, Agricultural Research Service, under Cooperative agreement number 58-3092-5-001. Development of M. truncatula Tnt1 insertion lines and reverse genetics screenings were supported by National Science Foundation, USA (DBI 0703285 & IOS-1127155) and The Noble Research Institute.
Enzymes Enzymes are highly specific biocatalysts involved in the chemical reactions taking place inside the living cell. They are generally protein molecules but, often contain a non-protein component known as a cofactor which is necessary for the catalytic activity. Enzymes are widely used in industries, clinical diagnosis and scientific research . Enzyme specificity is due to stereochemical and chemical affinity towards substrate which is achieved by three different point of interaction between substrate and enzyme. Substrate binds to the enzyme at a specific site to form an enzyme-substrate complex. The region of the enzyme containing the substrate binding site and catalytic site is known as the active site. The stability of the enzyme-substrate complex determines whether a substrate will be converted into product by a particular enzyme or not . Enzymes that contain a single polypeptide chain are known as monomeric enzymes and those having two or more chains are known as oligomeric enzymes. Allosteric enzymes are mainly oligomeric and they are regulated through feedback mechanism. These enzymes contain different catalytic and regulatory sites. Different polypeptide subunits combine to form an oligomeric enzyme. Multienzyme complexes are required to catalyze sequence of reactions without releasing intermediate products .