Upload
odetta
View
46
Download
1
Tags:
Embed Size (px)
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 Presentation
Citation preview
Analysis of Germination in Tomato Seeds
Tammy ChanDepartment of BiologyMentor: Dr. Hiro Nonogaki (Horticulture)
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
Endosperm
Testa
Embryo
Seed Morphology
Opposing Forces of Seed Germination
MechanicalResistance
EmbryoGrowth Potential
Mechanical Resistance
Testa
Endosperm
- - Man — Man — Man — Man — Man — Man — Man
Gal Gal—
—
Radicle
Endosperm Cap Composition
- - Man — Man — Man — Man — Man — Man — Man
Gal Gal—
—
β - Mannosidaseβ - Mannanase
α-Galactosidase
Sense Antisense
RNAi
Start Stop
DNARNA
Polymerase
RNAi
LeMAN2mRNA
antisense
Interference
Hypothesis: The endosperm cap of a tomato seed is weakened by the function of LeMAN2 mannanase during germination.
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
Characterization of RNAi plants
• Tissue Printing
To check mRNA:
To check enzyme activity:
•Gel Diffusion Assay
Paper towel
Positively-charged membrane
Half seed
Tissue Printing
Membrane
RNA
Tissue Printing
AP
Antibody
Detection Substrate
LeMAN2 RNA probe
DIG DIG DIG DIGDIG
Tissue Printing
WT Vector RNAi
WT
Vector
RNAi
Gel Diffusion Assay
Endosperm Cap
Radicle
Gel Diffusion Assay Results
Standard
Wildtype
Vector
RNAi
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
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