analysis of germination in tomato seeds
DESCRIPTION
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. - PowerPoint PPT PresentationTRANSCRIPT
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Analysis of Germination in Tomato Seeds
Tammy ChanDepartment of BiologyMentor: Dr. Hiro Nonogaki (Horticulture)
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Importance of tomato seeds
• Agriculture and food source
•Antioxidant
• Carries genetic information
http://www.bereagardens.org/images/tomato_field.jpgModified from: http://groupenadatoulon.lautre.net/IMG/jpg/Tomato-gene.jpghttp://www.umassvegetable.org/images/soils_crops_pest_mgt/crop/tomato1.jpg
•Tomatina
Modified from: http://upload.wikimedia.org/wikipedia/commons/8/88/Bright_red_tomato_and_cross_section02.jpg
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Endosperm
Testa
Embryo
Seed Morphology
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Opposing Forces of Seed Germination
MechanicalResistance
EmbryoGrowth Potential
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Mechanical Resistance
Testa
Endosperm
- - Man — Man — Man — Man — Man — Man — Man
Gal Gal—
—
Radicle
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Endosperm Cap Composition
- - Man — Man — Man — Man — Man — Man — Man
Gal Gal—
—
β - Mannosidaseβ - Mannanase
α-Galactosidase
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Sense Antisense
RNAi
Start Stop
DNARNA
Polymerase
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RNAi
LeMAN2mRNA
antisense
Interference
Hypothesis: The endosperm cap of a tomato seed is weakened by the function of LeMAN2 mannanase during germination.
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Seed Germination Curve
Seed Germination of Tomato Seeds
0
20
40
60
80
100
120
0 20 40 60 80 100 120
Time (hours)
Seed Germination (%)
Series1
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Characterization of RNAi plants
• Tissue Printing
To check mRNA:
To check enzyme activity:
•Gel Diffusion Assay
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Paper towel
Positively-charged membrane
Half seed
Tissue Printing
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Membrane
RNA
Tissue Printing
AP
Antibody
Detection Substrate
LeMAN2 RNA probe
DIG DIG DIG DIGDIG
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Tissue Printing
WT Vector RNAi
WT
Vector
RNAi
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Gel Diffusion Assay
Endosperm Cap
Radicle
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Gel Diffusion Assay Results
Standard
Wildtype
Vector
RNAi
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Chen and Bradford (2000) Plant Physiol 124: 1265-1274
Imbibition Time (h)
Pu
nctu
re F
orc
e (
N)
Endosperm cap
Radicle Tip
Texture analyzer
Puncture Force Analysis
WT
RNAi
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Acknowledgments
Howard Hughes Medical Institute
Dr. Hiro Nonogaki
Dr. Kevin Ahern
Seed Biology Lab
H2 O
Dr. Po-Pu (Eric) LiuDr. Masa AsahinaJessica KristofJing SunJennifer Coppersmith
Jen Hiro JingMe
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