| Description: | PURPOSE. The production and use of trinitrotulene (TNT) since the early part of this century has resulted in the widespread contamination of military bases, manufacturing facilities and wastewater in these areas. We aim to engineer transgenic plants to monitor environmental contamination by TNT. This will be accomplished by identifying TNT responsive promoters and fusing these promoters with GFP. When TNT is present in the soil, the plant will produce GFP, which can be remotely sensed.RATIONALE. TNT is a xenobiotic nitoaromatic compound for which very few biological systems have evolved specialized pathways. Recently however, plants were shown to be capable of degrading TNT. The sequential reduction of the nitro groups is thought to be mediated by non specific enzymes that reduce low potential electron carriers such as glutathion reductase, cytochrome P-450, thioredoxin reductase. To date their inducibility by TNT has not been investigated, and no other TNT inducible plant gene has been reported. We propose to identify genes responsive to TNT using microarrays.EXPERIMENTAL DESIGN. To identify TNT doses affecting growth and development of Arabidopsis ecotype Col1, seeds were plated under sterile conditions on Arabidopsis MS media. After incubation at 4 degrees for 4 days, the plates were transferred to 25 degrees with a photoperiod of 16 hours. Nine days later plantlets were transferred to Magenta boxes containing 50 ml Arabidopsis MS media with TNT concentrations of 0, 1, 5, 10 and 20 uM. Toxicity was assessed as percentage of root length, rosette length and diameter relative to control 10 days after transfert to Magenta boxes. We observed that a low concentration of 1 uM TNT stimulated root and rosette growth. Higher concentrations inhibited growth, with a concentration of 10 uM reducing growth by 50%. We therefore plan to investigate the expression profile at concentrations of 1 and 10 uM TNT. With 2 technical replicates for each dose, the experiment will require the use of 4 slides.PERSPECTIVE. At 1 and 10 uM TNT we expect to find genes that are specific to the pathways affected by TNT as well as genes involved in a general stress response. A substractive comparison with existing databases will be conducted. We are also planning to investigate other nitro-substituted explosives at UNCG when we will be equipped with a scanner. Ultimately we will be able to narrow down the candidate promoters to be fused to GFP and assessed in plants. |
NASCArrays Information at The BAR
Welcome to NASCArrays information at the BAR. This page hosts meta-information from the NASCArrays service (2002-2013). This information was parsed
from text files available on the NASCArrays site. NASCArrays data is on iPlant server. To download experiment data from iPlant, please click on the
experiment number. To download the CEL files, please click on the ftp link.