Abstract
Genetic mutagenesis is a very efficient tool in studying genes function. Because of great benefits of legumes as human food and animal feed worldwide, we used a model plant Medicago truncatula for identification gene function related to nitrogen fixation process. Our mutant is a Medicago mutant line contains a tobacco Tnt1 retro-transposon mobile element with the two Long Terminal Repeats (LTR) inserted within the genome. Our mutant is predicted to contain a mutation in gene/s belonging to symbiotic interaction between legume and rhizobia. A novel technique was used based on using fluorescent oligonucleotide primers against oligonucleotide primers for Tnt1-LTRs of our mutant. This novel protocol was very successful in detection the polymorphism between our mutant line and the wild variant R108 using Biosystems 310 Genetic Analyzer. Electropherograms of the mutant line and wild type gave a total of 561 well- resolved AFLP peaks, 357of which were polymorphic peaks and 204 were monomorphic peaks. This novel technique enables the calculation percentage of polymorphism between the mutant line and the wild type. Additionally primers combinations amplified more bands from others to detect polymorphism between the plants
Highlights
Legumes played vital roles in improving agriculture and considered as onethird of the world’s major crop production
Plant materials The mutant line of Medicago truncatula was provided from Noble Foundation Tnt1 mutant collection that transformed from M. truncatula R108-1 ecotype [23]
Pre-selective amplification was achieved using preselective oligonucleotide primer pair and the product was run on 1.5% agarose gel
Summary
Legumes played vital roles in improving agriculture and considered as onethird of the world’s major crop production. They represent about 27% of the world’s crop production. Animals depend on forage legume plants as major source of food like Medicago stavia and Trifolium spp [1]. This group of plants has unique character as they have ability to make symbiotic relationship with soil bacteria called rhizobia. Rhizobia are beneficial soil bacteria that belong to gram negative group [2] In this relation legumes can fix nitrogen in specific organ called root nodule [1,3]. Nitrogen fixation process occurs naturally during growing this family of plants and so we can decrease our usage of industrial nitrogen fertilizers and related ecosystem pollution [1]
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