1a.Objectives (from AD-416)
The objectives of this cooperative research project are to enhance our knowledge regarding grass development including, development in response to biotic and abiotic stress through the use of genetic and genomic tools in sorghum and other cereal grasses.
1b.Approach (from AD-416)
In order to identify candidate genes, sample materials will be collected from different sorghum tissues and treatments and used for the generation and sequencing of expression libraries; and QPCR analysis.
During FY 2011, research focused on extension of past development and analysis of qPCR methods in sorghum to monitor expression of both the mature miRNA and the corresponding precursor miRNA, from which it is derived. In addition, the group began to examine total RNA expression in sorghum using next generation sequencing technology (specifically Illumina’ DGE-tagging protocol). Information about miRNA expression related to different stress treatments and the function of their gene targets will be invaluable for understanding the role of these non-coding RNAs in post-transcriptional gene regulation and for our ability to manipulate stress tolerance in plants.
Using Sorghum as a model, 8-day old hydroponically grown sorghum seedlings are subjected to various stress conditions. Most of our work this past year has focused on abscisic acid (ABA) stress. The ABA treatment is used as a proxy for abiotic stress such as drought. In this treatment, 8-day old sorghum seedlings are subjected to 125µM ABA for 24hr and then both roots and shoots are harvested for RNA analysis. In the last year, the work has focused on analyses and submission of a publication summarizing the results of the work. In addition, the project has contributed to library and sequencing support for RNA seq and whole genome bisulfite libraries that will be used to support structural and functional annotations for sorghum.
Research conducted under this agreement was monitored through monthly conference calls and in person meetings between the ADODR and the Cooperator.