Project Number: 5442-21000-030-04
Project Type: Reimbursable

Start Date: Jul 01, 2003
End Date: Sep 30, 2006

Objective:
Complete genomic targeting and high-resolution mapping of the Tsn1 gene. Develop a BAC contig spanning the Tsn1 locus. Identify candidate genes for Tsn1 and isolate the Tsn1 gene sequence. Verify the cloning of Tsn1 by complementation experiments and comparisons with Tsn1 mutants.

Approach:
AFLP-bulked segregant analysis will be used to identify DNA fragments near the Tsn1 locus. Such fragments will be converted to RFLP markers and used for high-resolution mapping in populations consisting of approximately 2,500 gametes. Markers closely flanking the gene will be used to screen a Langdon durum BAC library. Positive BAC clones will be fingerprinted and contiged to identify those clones belonging to chromosome 5B that may span the Tsn1 locus. Selected BAC's will then be sequenced and surveyed for putative low-copy segments, which can be used for mapping and chromosome walking. Once a delineated segment containing Tsn1 is identified, it will be surveyed for open reading frames (ORF's). The ORF's will be compared to the sequences of the corresponding ORF's in Tsn1 mutant genotypes to identify the Tsn1 gene. In addition, candidate ORF's will also be transformed into 'Bobwhite' Tsn1 mutant genotype for complementation. Transgenic plants will be screened with Ptr ToxA to determine the functional Tsn1 gene.

Project Team

Faris, Justin

Project Annual Reports

FY 2003

Related National Programs

Plant, Microbial & Insect Genetic Res., Genomics, & Genetic Improv. I (301)   Plant Diseases (303)