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Analysis of Germination in Tomato Seeds

Analysis of Germination in Tomato Seeds. Tammy Chan Department of Biology Mentor: Dr. Hiro Nonogaki (Horticulture). Importance of tomato seeds. Agriculture and food source. Carries genetic information. Antioxidant. Tomatina.

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Analysis of Germination in Tomato Seeds

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  1. Analysis of Germination in Tomato Seeds Tammy Chan Department of Biology Mentor: Dr. Hiro Nonogaki (Horticulture)

  2. Importance of tomato seeds • Agriculture and food source • Carries genetic information • Antioxidant • Tomatina http://www.umassvegetable.org/images/soils_crops_pest_mgt/crop/tomato1.jpg Modified from: http://groupenadatoulon.lautre.net/IMG/jpg/Tomato-gene.jpg Modified from: http://upload.wikimedia.org/wikipedia/commons/8/88/Bright_red_tomato_and_cross_section02.jpg http://www.bereagardens.org/images/tomato_field.jpg http://www.telegraph.co.uk/news/graphics/2006/12/29/yearpix/060830tomatina.jpg

  3. Seed Morphology Endosperm Testa Embryo

  4. Opposing Forces of Seed Germination Embryo Growth Potential Mechanical Resistance

  5. Gal Gal — — - - Man — Man — Man — Man — Man — Man — Man Mechanical Resistance Testa Radicle Endosperm

  6. Gal Gal — — - - Man — Man — Man — Man — Man — Man — Man Endosperm Cap Composition α-Galactosidase β - Mannanase β - Mannosidase

  7. RNA Polymerase RNAi Start Stop Antisense Sense DNA

  8. antisense RNAi LeMAN2 mRNA Interference Hypothesis: The endosperm cap of a tomato seed is weakened by the function of LeMAN2 mannanase during germination.

  9. Seed Germination Curve

  10. Characterization of RNAi plants To check mRNA: • Tissue Printing To check enzyme activity: • Gel Diffusion Assay

  11. Tissue Printing Positively-charged membrane Half seed Paper towel

  12. AP Antibody DIG DIG DIG DIG DIG LeMAN2 RNA probe Detection Substrate Tissue Printing RNA Membrane

  13. Tissue Printing WT Vector RNAi WT Vector RNAi

  14. Gel Diffusion Assay Endosperm Cap Radicle

  15. Gel Diffusion Assay Results Standard Wildtype Vector RNAi

  16. Puncture Force Analysis Endosperm cap RNAi Radicle Tip Puncture Force (N) WT Imbibition Time (h) Texture analyzer Chen and Bradford (2000) Plant Physiol 124: 1265-1274

  17. H2O Acknowledgments Dr. Kevin Ahern Howard Hughes Medical Institute Dr. Hiro Nonogaki Seed Biology Lab Dr. Po-Pu (Eric) Liu Dr. Masa Asahina Jessica Kristof Jing Sun Jennifer Coppersmith Me Jing Jen Hiro

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