Molecular and biological studies in Medicago truncatula plant mutant line by Tnt1

Abstract

In this research thesis, we characterized a novel Medicago truncatula mutant, assigned it as black nodule mutant (blkn). This mutant is a Tnt1-retrotransposon mutant, Tnt1 is Nicotiana tabacum retrotransposon which is replicated via an RNA copy and integrated in Medicago truncatula genome. blkn is obtained from Noble Foundation Tnt1 -mutant collection. This line was produced in vitro through A. tumefactions transformation. blkn mutant is achieving a null function root nodules phenotype and exhibited about double contents of phenolic compound than the wild type, the mutated nodules displayed abnormality in infection and nitrogen fixation zones. The first characterization step for our mutant was using ABI 310 Genetic Analyzer to compare the polymorphism percentage between blkn and wild type. The Genetic Analyzer was used with a novel modification as we used the Tnt1 borders-specific oligonucleotide primers instead of using the universal oligonucleotide primers along with florescence labeled