|Klein, Robert - Bob|
Submitted to: Genome Research
Publication Type: Peer reviewed journal
Publication Acceptance Date: 4/19/2000
Publication Date: 6/1/2000
Citation: Klein, P.E., Klein, R.R., Cartinhour, S.W., Ulanch, P.E., Dong, J., Obert, J.A., Morishige, D.T., Schlueter, S.D., Childs, K.L., Ale, M. 2000. A high-throughput AFLP-based method for constructing integrated genetic and physical maps: Progress toward a sorghum genome map. Genome Research. 10:789-807. Interpretive Summary: Plant genetics is a field of scientific research whose main purpose is to totally understand at the most complex and minute detail (the genetic level) just how plants live and function. With such understanding, better plant varieties can be developed to greatly improve the production of human foodstuffs and animal feedstuffs worldwide. We have conducted research on sorghum (a type of grass whose seed or grain is used both as a human food and animal feed), using very sophisticated and complex techniques of molecular biology. Results from our work will be very useful in aiding other scientists (plant breeders, etc.) to develop better plant varieties. Our work makes it easier to determine how molecular components of plants called genes are arranged within the plant. Knowing this arrangement of genes, referred to as a physical map, will be very helpful in manipulating sorghum to produce more productive and healthy varieties. The techniques we have developed will also be useful in working with important agricultural plants other than sorghum.
Technical Abstract: The construction of an integrated genetic and physical map of the sorghum genome is a primary goal of our sorghum genome project. To help accomplish this task, we developed a new high-throughput method for building contigs and locating clones on the sorghum genetic map. This involved pooling 24,576 sorghum clones in six different matrices to create 184 pools of DNA. .DNA fragments from each pool were amplified using amplified fragment lengt polymorphism technology with each set amplifying 28 single copy DNA markers that were useful for identifying overlapping clones. Analysis of a sorghum mapping population using the same genetic markers located 200 of the contigs on the sorghum genetic map. Restriction endonuclease fingerprinting of the entire collection of sorghum clones was applied to test and extend the contigs constructed using this genetic marker-based methodology. Analysis of the fingerprint data allowed for the identification of 3,366 contigs, each containing an average of 5 clones. Thus, the combination of fingerprinting and genetic marker-based contig assembly and mapping provides a reliable, high-throughput method for building an integrated genetic and physical map of the sorghum genome.