Plant & Animal Genomes XIII Conference

January 15-19, 2005
Town & Country Convention Center
San Diego, CA

TILLING In Soybean: A Reverse Genetic Tool For Functional Gene Analysis From Two Soybean Populations (Forrest And Williams82)

Khalid Meksem¹ , Faiza Tebbji¹ , Aziz Jamai¹ , Jennifer Cooper² , Robert Laport ² , Steve Henikoff ² , Abdelhafid Bendahmane³

¹  Plants and Microbes Genomics and Genetics lab; Department of Plant Soil and Agricultural Systems, Southern Illinois University at Carbondale, Room 176. Carbondale, IL 62901-4415. Fax: 618 453 7457; E-mail: meksemk@siu.edu; http://www.siu.edu/~meksem/

²  Seattle TILLING Project; Rm. 307 Hitchcock Hall Department of Biology; University of Washington 1521 NE Pacific; Seattle, WA 98195-001 http://tilling.fhcrc.org:9366/

³  INRA-URGV; CROPTIL Project; 2 Rue Gaston Crémieux CP 5708 91057 EVRY Cedex; FRANCE http://www.evry.inra.fr/

We demonstrated the feasibility of TILLING in soybean. Over the course of the past two years, the Meksem lab has played an integral role in the establishment of soybean EMS-TILLING libraries (from cv. Forrest and cv. Williams 82), which contain 5200 M2 families as of September 2004. TILLING data was generated from the analysis of the first 3000 DNAs extracted from soybean Forrest M2 plants, tested at 3 different locations, using 3 detection methods and pooling systems (NSF TILLING pilot project in collaboration with Dr. Henikoff, the European TILLING project CROPTIL in collaboration with Dr. Bendahmane, and Dr. Meksem’s lab).
Under the NSF TILLING pilot project, Dr. Meksem’s lab at Southern Illinois University in collaboration with Dr. Henikoff’s lab (Seattle, WA) selected 5 members of a soybean LRR-Kinase gene family as targets for TILLING to test the developed soybean EMS-mutagenized populations (including) the Rhg1 and Rhg4 genes. Using two subsets of 768 DNA samples from the soybean cv. Forrest M2 lines, we identified 2 to 4 point mutations per 1.5 Kbp of targeted DNA per 8-fold pool gel (representing 768 M2 plants).
Based on this data, we anticipate 2 to 4 mutants per 768 soybean M2 families, thereby screening a total of 6000 M2 families, may provide a series of 15 to 30 alleles within each 1.5Kb fragment of a target gene. A web page is dedicated to the project, www.soybeantilling.org