1. “CSI” Plant Style: From the Laboratory to the Wheat Field

2. CAP Grant Discover new, valuable, genes and QTLs Discover new, valuable, genes and QTLs Deploy them into adapted varieties around the country Deploy them into adapted varieties around the country Use marker-assisted selection strategies to enhance gene deployment Use marker-assisted selection strategies to enhance gene deployment

3. CAP Grant 16 institutions around the country coordinate breeding efforts 16 institutions around the country coordinate breeding efforts 4 year grant started in 2006 4 year grant started in 2006 Over 20 cultivars/germplasm already released Over 20 cultivars/germplasm already released (http://maswheat.ucdavis.edu) (http://maswheat.ucdavis.edu)

4. Impact of Wheat in Washington In 2006 Excess of 2.2 million acres planted Excess of 2.2 million acres planted 81% winter wheat, 19% spring wheat 81% winter wheat, 19% spring wheat Over 140 million bushels harvested Over 140 million bushels harvested Net return of $615 million Net return of $615 million

5. From Problem to Lab to Wheat Field

6. What Traditional Problems Do We Face

7. New Characteristics Needed Drought tolerance Drought tolerance Disease and Insect resistance/tolerance Disease and Insect resistance/tolerance Herbicide resistance/tolerance Herbicide resistance/tolerance High yield High yield Excellent end-use quality (High GPC) Excellent end-use quality (High GPC)

8. Definitions DNA—Deoxyribo nucleic acid that carries the genetic information in a cell DNA—Deoxyribo nucleic acid that carries the genetic information in a cell Gene—a sequence of DNA that occupies a specific location and determines a particular characteristic Gene—a sequence of DNA that occupies a specific location and determines a particular characteristic

9. Definitions Allele-one member of a pair of genes that occupies a specific location on DNA Allele-one member of a pair of genes that occupies a specific location on DNA A= a= A= a= = homozygous dominant = homozygous dominant = homozygous recessive = homozygous recessive = heterozygous = heterozygous

10. Punett Square

11. Crossing Plants are identified which have the trait and which need the trait Plants are identified which have the trait and which need the trait Cross is made to introgress trait Cross is made to introgress trait Survivor, no game X Rock-star yielder, dies Survivor, no game X Rock-star yielder, dies 10 Seeds (all ) 10 Seeds (all ) 25 ( ) 50 ( ) 25 ( ) 25 ( ) 50 ( ) 25 ( ) Self pollination

33. Crossing Cross is made to introgress trait Cross is made to introgress trait Survivor, no game X Rock-star yielder, dies Survivor, no game X Rock-star yielder, dies 10 Seeds (all ) 10 Seeds (all ) 25 ( ) 50 ( ) 25 ( ) 25 ( ) 50 ( ) 25 ( ) Self pollination Survivor, Rock-star yielder?

34. How Do We Determine Which Seeds Carry The Trait We Are Interested In?

36. So, which of you plants are resistant to fungal diseases?

37. I am!

38. Definitions Molecular Markers—DNA segments that can be used as flags to track genes Molecular Markers—DNA segments that can be used as flags to track genes Linkage—occurs when particular genes are inherited jointly Linkage—occurs when particular genes are inherited jointly Linked Markers—markers which are closely associated with a gene Linked Markers—markers which are closely associated with a gene

39. DNA Gene Marker We know the DNA sequence of the marker Sometimes we do not know the sequence of the gene we are looking for We can use the marker to indirectly select for the gene of interest

40. Tightly Linked Marker Gene Marker Gene Marker Non-linked Marker 95% confident 50% confident

41. Marker-Assisted Selection Identify marker which is linked to gene Identify marker which is linked to gene We can extract DNA from the plants and look for the presence of the marker We can extract DNA from the plants and look for the presence of the marker This will tell us which have the trait and which do not This will tell us which have the trait and which do not

43. Picture of harvesting tissue Picture of harvesting tissue

44. 12 34

45. + 95% Ethanol

47. PCR Explanation Polymerase Chain Reaction Polymerase Chain Reaction Used to amplify DNA via enzymatic replication Used to amplify DNA via enzymatic replication Allows a small amount of DNA to be used for analysis Allows a small amount of DNA to be used for analysis PCR reaction PCR reaction PCR reaction PCR reaction http://www.danquinnart.com/wheatcap/index.htm http://www.danquinnart.com/wheatcap/index.htm

49. Gel Electrophoresis Allows separation of PCR product Allows separation of PCR product DNA inserted into agarose gel DNA inserted into agarose gel DNA fragments travel with current DNA fragments travel with current Smaller fragments will travel faster than large fragments Smaller fragments will travel faster than large fragments

57. Gel Analysis

62. So, which of you plants are resistant to fungal diseases?

63. Evaluation After MAS has identified which plants carry the trait, further evaluation is done in the field to make sure the plants are productive After MAS has identified which plants carry the trait, further evaluation is done in the field to make sure the plants are productive Time to release a variety is shortened Time to release a variety is shortened Why would this be important? Why would this be important?

64. Zak + 1 gene

65. Washington Farmers Tara 2002 Shepherds Grain WSU Food Services Crossing Trait Selection From Laboratory to Wheat Field

66. Acknowledgements