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Tomato Seed Development and Germination

Tomato Seed Development and Germination. Theresa Nguyen Department of Biology Mentor: Dr. Hiro Nonogaki. Why Seeds?. Agriculture Food source rich in antioxidants Carries genetic information http://union.osu.edu/mgarden/images/tomato.jpg http://birdsandbeesonline.com/tomato.gif. Embryo.

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Tomato Seed Development and Germination

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  1. Tomato Seed Development and Germination Theresa Nguyen Department of Biology Mentor: Dr. Hiro Nonogaki

  2. Why Seeds? • Agriculture • Food source rich in antioxidants • Carries genetic information http://union.osu.edu/mgarden/images/tomato.jpg http://birdsandbeesonline.com/tomato.gif

  3. Embryo Endosperm Testa Tomato Seed Morphology

  4. Seed Germination Embryo Growth Potential Mechanical Resistance Endosperm Cap

  5. Gal Gal — — - - Man — Man — Man — Man — Man — Man — Man Seed Germination Radicle Endosperm Cap

  6. Gal Gal — — - - Man — Man — Man — Man — Man — Man — Man Endosperm Cap Degradation α-Galactosidase β - Mannanase β - Mannosidase β – Mannanase enzyme is encoded by LeMAN2 gene.

  7. TISSUE PRINTING LeMAN2 Gene LeMAN2 is highly expressed in the endosperm cap

  8. AS2 C2C2-GATA1 0.5 0.5 0.6 0.6 0.45 0.45 0.4 0.4 0.5 0.5 0.35 0.35 0.4 0.4 0.3 0.3 Relative mRNA levels Relative mRNA levels 0.25 0.3 0.3 0.2 0.2 0.2 0.2 0.15 0.15 0.1 0.1 0.1 0.05 0 0 0 0 12h 24h 36h 48h 12h 24h 36h 48h C2C2-GATA2 MBF 0.08 0.05 0.07 0.045 0.04 0.06 0.035 0.05 Relative mRNA levels 0.03 0.04 0.025 Relative mRNA levels 0.02 0.03 0.015 0.02 0.01 0.01 0.005 0 0 12h 24h 48h 36h 12h 24h 36h 48h Imbibition Time (h) Imbibition Time (h) Stage Specific Expression Asymmetric Leaves 2 Imbibition Time (h) Imbibition Time (h) Multi-protein Bridging Factor

  9. 0.05 0.05 0.04 0.04 0.03 0.03 Relative mRNA levels Relative mRNA levels 0.02 0.02 0.01 0.01 0 0 12 24 36 Imbibition Time (h) Imbibition Time (h) Stage Specific Expression 0.12 LeMAN2 C2C2-GATA2 0.1 0.08 Relative mRNA levels 0.06 0.04 0.02 0 1224 36 Imbibition Time (h)

  10. Endosperm cap (EC) Lateral endosperm (LE) Cotyledon (C) Root (R) Tissue Specificity

  11. AS2 0.4 0.25 C2C2-GATA1 0.3 0.2 0.15 Relative mRNA levels 0.2 Relative mRNA levels 0.1 0.1 0.03 MBF 0.05 0 EC LE C R 0.04 0 C2C2-GATA2 Seed Tissues EC LE C R 0.02 Relative mRNA levels Seed Tissues 0.03 Relative mRNA levels 0.01 0.02 0.01 0 EC LE C R Seed Tissues 0 EC LE C R Seed Tissues Tissue Specific Expression Asymmetric Leaves 2 Multi-protein Bridging Factor

  12. 0.04 LeMAN2 0.03 Relative mRNA levels 0.02 0.01 0 EC LE C R 0.04 C2C2-GATA2 Seed Tissues 0.03 Relative mRNA levels 0.02 0.01 0 EC LE C R Seed Tissues Tissue Specific Expression LeMAN2 Notice the localization of gene expression in the endosperm cap.

  13. Hypothetical Regulation • GATA2 gene is upstream of LeMAN2 genein tomato

  14. Amino Acid Sequence Alignment GATA2 (tomato) AGP1 (tobacco) http://www.plantcare.com/oldSite/httpdocs/images/namedImages/Tomatoes.jpg http://upload.wikimedia.org/wikipedia/commons/a/ae/Nicotiana_Tobacco_Plants_1909px.jpg

  15. AGACCAA Myb AGP1 AGP1 Pathway in Tobacco

  16. Myb AGACCAA Myb GATA2 Promoter LeMAN2 Prediction • GATA2 is the regulatory protein that binds indirectly to promoter region of LeMAN2

  17. 86.793kD MBP GATA2 Protein Expression • GATA2 = 35.993 kD • MBP = 50.8 kD pMal vector

  18. – + Molecular Marker 209 kD 124 80 49.1 34.8 28.9 20.6 7.1 Results • Fusion protein MBP-GATA2 is clearly induced with Isopropyl β-D-1-thiogalactopyranoside (IPTG) MBP-GATA2

  19. Microarray Analysis • Gene Chip® • Massive approach versus gene-by-gene • Tomato chip contains ≈10,000 genes http://www.colorado.edu/news/reports/renewable-energy/images/genechip.jpg

  20. Gene Functional Classification • There was a total of 166 endosperm cap specific genes

  21. Acknowledgments • Dr. Kevin Ahern • Howard Hughes Medical Institute • Dr. Hiro Nonogaki • Seed Biology Lab • Wioletta Pluskota, Ph.D • Masa Asahina, Ph.D • George W. Bassel II, Ph.D • Arun Kumar • Tammy Chan • Jing Sun • Jen Coppersmith • Natalya Goloviznina Thank you!

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