importance of microrna for seedling development in arabidopsis thaliana jennifer coppersmith...
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![Page 1: Importance of microRNA for seedling development in Arabidopsis thaliana Jennifer Coppersmith Department of Biochemistry and Biophysics Mentor: Dr. Hiro](https://reader036.vdocuments.us/reader036/viewer/2022062320/56649d615503460f94a4319b/html5/thumbnails/1.jpg)
Importance of microRNA for seedling
development in Arabidopsis thaliana
Jennifer CoppersmithDepartment of Biochemistry and BiophysicsMentor: Dr. Hiro Nonogaki (Horticulture)
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Significance of Seed Development
pgrc3.agr.ca/images/seeds.gif
Agriculture and Food
Why Arabidopsis?
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Arabidopsis Seed Structure
Embryo
Endosperm
Testa
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Overview of Experiment
T DNA insert
SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE13 (mSPL13)
SPL13
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microRNA
• 21-24nt in length
• Gene regulation at the mRNA level
www.qiagen.com/about/PressReleases/images/9.jpg
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RISCRISC
miRNA
microRNA
mRNA
miRNA
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RISC
miRNA
microRNA mutation
mRNA
Mutation
Hypothesis: SPL13 needs to be down regulated by microRNA156 to maintain normal developmental program for germination or seedling establishment.
GUGCUCUCUCUCUUCUGUCA CACGAGUGAGAGAAGACAGU
GUGCUCUCUCUCUUCUGUCA CACGAGUGAGAGAAGACAGU
A A A A
SPL13
miR156
mSPL13
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WT Cont mSPL13
rRNA
SPL13
Overaccumulation of SPL13
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Mutant Phenotype
Cot
Cot
4 leaf seedling (normal) 2 leaf seedling (delayed)
Cot
Cot
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2-leaf seedlings4-leaf seedlings
Developmental Delay
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Temporal Seedling Arrest
WTmSPL13
CONTROL
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0 d
Shoot Apical Meristem (SAM)
2 d
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Storage Protein and Meristem
Meristem
Embryo
Cot
Hypocotyl
Radical
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WT
1 d 3 d2 d 4 d
mSPL13
mSPL13WT
3 d
mSPL13WT
2 d
mSPL13WT
4 dMeristem Development
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mSPL13WT
8 d
Visible Difference
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Analysis
WT mSPL13
SPL13
Based on phenotype, down regulation of SPL13 via microRNA is essential for proper meristem
development.
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Determine Potential Targets of SPL13
miRNA156
SPL13 SPL13
Promoter Gene X
X
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AcknowledgementsHiro Nonogaki
Kevin Ahern
Howard Hughes Medical Institute
Lab Members:
Masa Asahina
Tammy Chan
Po-Pu Liu
Jessica Kristof
Piotr Pupel
Jing Sun
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SQUA
SQUAMOSA ( or APETALA1)
10 bp conserved stretch
19-mer core seq
- 419 - 393
- 455 - 429
AP1
T T C G A C G T C C G T A C A A C A A G T C T T T A T
C C A G T G G T C C G T A C A A T G T T A C T T A C C
SBP1 SBP2SPL13
?
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Normal Embryogenesis in mSPL13
Coppersmith JL, Hewitt JR, Nonogaki H, unpublished results