investigation of the barren stalk1 gene lineage in graminoid poales daniel woods

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Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

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Page 1: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Investigation of the BARREN STALK1 gene lineage in graminoid Poales

Daniel Woods

Page 2: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

rice corn

Page 3: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Outline

1) Introduction into field of Evolutionary Developmental Genetics and Grasses

2)What’s BARREN STALK1

3) Hypothesis/aims

4) Preliminary Data

Page 4: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

rice

corn

Graminoid Poales

?

phylogenetics

morphologicaldevelopment

molecular evolution

functional analyses

gene expression

Page 5: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods
Page 6: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods
Page 7: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Chasmanthium (CEN) inflorescence development

Page 8: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods
Page 9: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

barren stalk1 (ba1) phenotypeWild-type tassel ba1-ref WT meristem ba1-mum1

Gallavotti, A., et al. (2004). Nature 432(7017): 630-5

Gallavotti, A., et al. (2004). Nature 432(7017): 630-5

Page 10: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Rice (OsLAX)Maize (ZmBA1)

BA1/LAX expression via in situ hybridization

Gallavotti, A., et al. (2004), Nature 432(7017): 630-5 Komatsu, K., et al. (2003). Proc Natl Acad Sci USA 100(20): 11765-70

Page 11: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Hypotheses:

1) BA1 duplication event occurred at base of grass family

2) BA1 is expressed in branching primordia throughout inflorescence development

Page 12: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Specific aims to test hypothesis

1) Isolate BA1 bHLH genes from diverse array of grasses and near grass relatives

2) Generate phylogenies to pinpoint duplication event

3) Document inflorescence development using Scanning Electron Microscopy (SEM)

4) Gene Expression Analysis via in situ hybridization

Page 13: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

BEP PACCMAD

Page 14: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Gene Isolation and Preparation

Extractions Primer Design

Polymerase Chain Reaction(Semi-nested)

Gel Extraction

Ligation

Clone

PCR screen

Alkaline lysis

Sequence(Sanger)

Page 15: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods

Generate Phylogenies

Maximum Parsimony

Maximum Likelihood

Bayesian

Page 16: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods
Page 17: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods
Page 18: Investigation of the BARREN STALK1 gene lineage in graminoid Poales Daniel Woods