Monitoring of Crosstalk Between Jasmonate and Auxin in the Framework of Plant Stress Responses of Roots.

Over the last few years, it became more and more evident that plant hormone action is to great parts determined through their sophisticated crosstalk, rather than by their isolated activities. Thus, the parallel analysis of interconnected phytohormones in only very small amounts of tissue developed to an important issue in the field of plant sciences. In the following, a highly sensitive and accurate method is described for the quantitative analysis of the plant hormones jasmonic acid and indole-3-acetic acid in the model plant Arabidopsis thaliana. The described methodology is, however, not limited to the analysis of Arabidopsis samples but can also be applied to other plant species. The presented method is optimized for the working up of as little as 20-50 mg of plant tissue. Thus, it is well suited for the analysis of plant hormone contents in plant tissue of only little biomass, such as roots. The presented protocol facilitates the implementation of the method into other laboratories that have access to appropriate laboratory equipment and comparable state-of-the-art gas chromatography-mass spectrometry (GC-MS) technology.

Highly Sensitive Salicylic Acid Quantification in Milligram Amounts of Plant Tissue.

Technical advances in mass spectrometry constantly raise the bar for analyzing trace amounts of plant hormones in only very small amounts of tissue. Here, a highly sensitive and accurate method is described for the quantitative analysis of the plant hormone salicylic acid not only in the model plant Arabidopsis thaliana but also in other plant species. The presented method is optimized for the working up of as little as 20 to 50 mg of plant tissue. The discussed protocol and the utilized laboratory equipment facilitate the implementation of the method into other laboratories that possess access to adequate state-of-the-art gas chromatography-mass spectrometry (GC-MS) equipment.

Characterization of Four Bifunctional Plant IAM/PAM-Amidohydrolases Capable of Contributing to Auxin Biosynthesis.

Amidases [EC 3.5.1.4] capable of converting indole-3-acetamide (IAM) into the major plant growth hormone indole-3-acetic acid (IAA) are assumed to be involved in auxin de novo biosynthesis. With the emerging amount of genomics data, it was possible to identify over forty proteins with substantial homology to the already characterized amidases from Arabidopsis and tobacco. The observed high conservation of amidase-like proteins throughout the plant kingdom may suggest an important role of theses enzymes in plant development. Here, we report cloning and functional analysis of four, thus far, uncharacterized plant amidases from Oryza sativa, Sorghum bicolor, Medicago truncatula, and Populus trichocarpa. Intriguingly, we were able to demonstrate that the examined amidases are also capable of converting phenyl-2-acetamide (PAM) into phenyl-2-acetic acid (PAA), an auxin endogenous to several plant species including Arabidopsis. Furthermore, we compared the subcellular localization of the enzymes to that of Arabidopsis AMI1, providing further evidence for similar enzymatic functions. Our results point to the presence of a presumably conserved pathway of auxin biosynthesis via IAM, as amidases, both of monocot, and dicot origins, were analyzed.