Development of Genetic Transformation, Tissue Culture, and Test Genes That Enable Accumulation of Polyamines Spermine in Corn and Tomato
Sustainable Agricultural Systems Laboratory
2013 Annual Report
1a.Objectives (from AD-416):
The first objective is to develop robust transformation system and tissue culture protocols for quickening flowering in corn and tomato. The second objective is to construct gene(s) fused to tissue/organ-specific promoters and test their transformation frequency. The third objective is to generate transgenic plants homozygous with the introduced gene(s), and analyze them at cellular and molecular levels.
1b.Approach (from AD-416):
The collaborator’s laboratory is adept in tissue culture technologies and transformation protocols. The collaborator will use gene constructs made in the ARS laboratory and develop transgenic corn and tomato plants. Because tissue-specific and other regulatable promoters, with and without fluorescent markers, will be employed for gene construction, their specificity of expression will be tested and validated at the whole plant level.
Developed and optimized protocols for one-dimensional and two-dimensional gel electrophoresis for separating proteins extracted from tomato fruit. We will use these protocols to perform proteomic analysis of fruits during ripening. The intent is to identify genes/proteins that are up-regulated or down-regulated in transgenic tomatoes expressing higher amounts of polyamines. Also, several box C/D snoRNA genes in tomato fruit were cloned and sequenced. The expression of these snoRNAs was found to decrease as the fruit ripened.