Internship with USDA Albany Laboratory

Diana Jucha06/16/14 – 08/15/14Contra Costa College

Identifying mutations in wheat storage protein genes using SDS-PAGE Celiac disease is caused by

intolerance to gluten. Gluten proteins ( HMW-GS,

LMW-GS, and Gliadins) are the main determinants of dough quality.

Hypothesis: Deletion of gluten protein will cause change in quality of dough performance.

First Step: Induced mutagenesis in genes and identify deletions in gluten proteins.

Goal: Determine mutant lines of wheat that have low-gluten proteins while maintaining high dough quality.

MutagenesisBobWhite Protein Pattern

• Mutagenesis of wheat using fast-neutron radiation

• Protein pattern of glutenin

SDS-PAGESodium Dodecyl Sulfate PolyAcrylamide Gel Electrophoresis is what is used to screen for phenotypes of mutations of wheat storage protein genes by seed protein profiling.

Ground up seed

Ground seed in protein extraction buffer

Use SDS-PAGE to separate proteins

DJ 103

Gel picture

Results and Discussion

This research is beneficial to offer low-gluten options to those with gluten intolerances and general public to prevent growing gluten intolerance.

Screening total 59 lines of M2 wheat lines• 3 samples missing Bx7 and By9• 2 samples missing Ax2• 1 sample missing Ax2, Bx7, and By9

Acknowledgments Dr. Debbie Laudencia-Chingcuanco Cindy Chen Val Fan Dr. Katie Krolikowski Center for Science and Excellence

CitationsC van den Broeck, Hetty, Teun WJM van Herpen, Cees Schuit, Elma MJ

Salentijn, Liesbeth Dekking, Dirk Bosch, Rob J Hamer, Marinus JM Smulders, Ludovicus JWJ Gilissen, and Ingrid M van der Meer. "Removing celiac disease-related gluten proteins from bread wheat while retaining technological properties: a study with Chinese Spring deletion lines." : n. pag. Web. 1 Aug 2014.

Carroccioa, Antonio, Lidia Di Prima, Davide Noto, Francesca Fayer, Giuseppe Ambrosiano, Vincenzo Villanacci, Karen Lammers, Domenico Lafiandra, Enzo De Ambrogio, Gaetana Di Fede, Giuseppe Iacono, and Norberto Pogna. "Searching for wheat plants with low toxicity in celiac disease: Between direct toxicity and immunologic activation." Digestive and Liver Disease: n. pag. Web. 1 Aug. 2014. http://www.agricorner.com/wheat-procurement-campaign-2014-shahbaz-approves-3-5-million-metric-tons-target/http://videohive.net/item/wheat-background/5507066http://news.heartland.org/newspaper-article/2011/11/07/agribusiness-plants-seeds-even-bigger-crop-insurance-boondogglehttp://www.acs.org/content/acs/en/education/whatischemistry/landmarks/usda-flavor-chemistry.htmlhttp://logosandbrands.directory/usda-logo/http://www.american-school-search.com/safety/contra-costa-college