2008 Annual Report
1a.Objectives (from AD-416)
Objective 1: Identify a core set of molecular markers tailored for systematic characterization of the genetic diversity within and among Gossypium germplasm accessions that will be maintained under the sister project 6202-21000-032-00D.
Objective 2: Maintain and enhance CottonDB as a user-friendly tool for the cotton research community.
Sub-objective 2.A: Maintain and enhance CottonDB, including development of user friendly public interfaces.
Sub-objective 2.B: Develop bioinformatic software and tools to assist both users and curators of CottonDB.
Objective 3: Collaborate with other public sector researchers to construct and integrate physical and genetic maps of G. hirsutum.
Sub-objective 3.A: Develop cotton genetic maps that contain PCR-based DNA markers.
Sub-objective 3.B: Develop cotton physical maps that contain large-insert BAC clones.
Sub-objective 3.C: Integrate cotton genetic and physical maps with EST unigene information.
Objective 4: Identify key genes and genomic regions of cotton for use in developing cotton germplasm resources that exhibit desirable/improved agronomic and fiber traits.
Sub-objective 4.A: Apply genomic and bioinformatic tools to identify and characterize QTLs or alleles from cotton genetic resources, maintained under the sister project 6202-21000-032-00D, that govern key agronomic or fiber traits.
Sub-objective 4.B: Apply the preceding information to identify superior parents for developing breeding populations with novel sources of variability for traits of interest.
Sub-objective 4.C: Recombine and select the preceding breeding populations to accumulate desirable QTLs and alleles in enhanced cotton breeding lines.
1b.Approach (from AD-416)
To develop a portable core set of markers for cotton (Objective 1), new SSR and SNP markers will be developed from cotton BAC libraries and other genomic DNA templates. From the markers created, a core set of 208 markers will be carefully selected from the saturated genome map of tetraploid cotton (TM-1 x 3-79) with 8 markers from each of 26 chromosomes. Each of these core markers will have a high polymorphism information content (PIC) value to be determined on a standardized core germplasm panel consisting of 12 diverse Gossypium genotypes. These markers will be evenly distributed on the cotton genome, with every chromosome arm having 4 core markers at approximately 15-cM intervals. Data from marker development will be stored and made available in the CottonDB database. CottonDB, a tool for the research community, will be enhanced through continued migration of its information content to a relational structure, improved display pages, and direct record-to-record links between internet databases to integrate information into a larger virtual database (Objective 2). To enrich the delivered content and streamline users' searches for specific information, work will integrate related data from multiple databases. Solutions developed by other genome databases will be adapted and implemented to this project's databases where appropriate. To construct and integrate physical and genetic maps, genetic mapping of TM-1 BAC-derived and other markers will be conducted using the TM-1 x 3-79 RI population. Diagnostic DNA markers will be identified that are capable of detecting polymorphism in intraspecific populations, and these markers will be used to genotype the entire TM-1 x 3-79 RI mapping population. A score matrix will be generated from the genotyping experiments and merged with the existing mapping database to perform linkage analysis via MapMaker and/or JOINMAP software programs. Recombination frequencies will be converted into map distances (cM). Approximately 500 SSR and 500 SNP markers will be added to the existing genetic map that contains 1,200 SSR markers to obtain an average resolution of 1-2 cM per marker. Integration of cotton genetic and physical maps will be achieved by anchoring framework genetic markers to TM-1 BAC contigs, and locating BAC-derived markers to the TM-1 x 3-79 RI map (Objective 3). Comparisons of genetic and physical map tools (CMap and IntegratedMap) will allow for consolidation of all structural and physical genomic information. In order to utilize the growing numbers of QTLs reported in cotton, work will validate those QTL by aligning genomic locations and comparing genetic effects (Objective 4). Information for QTLs of interest will be related among comparable studies in cotton and will be obtained from a variety of sources, including published accounts and database records. Once specific chromosomal regions containing genes that make a significant contribution to the expression of a complex phenotype of interest are identified, fine-mapping of the most promising genomic regions will be used to identify polymorphisms in coding and/or regulatory regions.
This is a new project that replaced 6202-21000-025-00D and which is continuing and expanding upon the work of the precursor project. During FY 2008, significant progress was made in developing genomic sequence resources for Upland cotton. Quality DNA sequences were obtained of nearly 200 minimum tiling path (MTP) clones, selected from a pair of homoeologous chromosomes (12 and 26) of Upland cotton. Sequence assembly and annotation are currently being analyzed to reveal the genomic relation, gene content, and function. Progress continues on migrating CottonDB to a new database architecture; new sets of cotton genomic and genetic data including DNA markers, maps, and DNA sequences have been added to the database. Work is in the early stages on a new project to characterize the genetic variability within improved world-wide cotton cultivars contained in the Project's Cotton Germplasm Collection. (NP 301, Components 1, 2, Problem Statements 1.B, 2.B)
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