CONSTRUCTION AND APPLICATIONS OF A MYCORRHIZAL ARBUSCULAR SPECIFIC cDNA LIBRARY.

To exploit the potential benefits of mycorrhizas, we need to investigate the processes that occur in these symbiotic interactions, particularly in the arbuscular compartment where nutrients are exchanged between the plant and the fungus. Progress in this area is restricted due to the intricacy and complexity of this plant-fungus interface and many techniques that have been employed successfully in other plants and animal systems cannot be used. An effective approach to study processes in arbuscules is to examine transcript composition and dynamics. We applied laser capture microdissection (LCM) to isolate approximately 3000 arbuscules from Glomus intraradices colonised Me- dicago truncatula roots. Total RNA was extracted from microdissected arbuscules and subjected to T7 RNA polymerase-based linear amplification. Amplified RNA was then usedfor construction of a cDNA library. The presence and level of enrichment of mycorrhiza-specific transcripts was determined by quantitative Real-time and conventional PCR. To improve enrichment a cDNA library subtraction was performed. Complementation of yeast mutants deficient in the uptake of.potassium, phosphate, sulphate, amino acids, ammonium and of a Mn²âºsensitive strain, demonstrates the functionality of our cDNA library.

The influence of protein kinase KIN10 gene expression on root phenotype of Arabidopsis thaliana root system in condition of energy stress.

It was shown phenotypic changes in the root system of seedlings Arabidopsis thaliana in transgenic lines with overexpression and suppressed gene expression of serine-threonine protein kinase KIN10 in conditions of energy shortage and under normal conditions. The normal growth and development of KIN10 overexpressing plants with in energy deficiency conditions were detected. The significant inhibition of the plant development under normal conditions for these plant lines was obsereved. The levels of KIN10 gene expression under normal conditions in different organs A. thaliana, particularly in the roots, stems, leaves and flowers were analyzed. The highest level expression of the gene was fixed in the leaves.