Date of Award

2012

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Clore, Amy

Keywords

Maize, Endosperm, In Situ Hybridization

Area of Concentration

Biology

Abstract

This thesis research was a part of a larger research effort funded by the National Science Foundation to study the regulation of early endosperm development in Zea mays L. As part of the larger study, a group of inbred lines generated around the world are being studied to look for variations in development as compared to the standard reference line, B73. A second major goal of the grant is to identify key regulator genes and their patterns of localization. Preliminary visualization of sections of the line CML322 seemed to indicate precocious kernel (including of the endosperm) development. Digital imaging and morphometry were undertaken for this thesis to systematically test this hypothesis. We confirmed that the CML322 endosperm does grow precociously under greenhouse conditions in Florida. The CML322 line was then used to test the efficacy of a new generation of hyper-labeled oligonucleotide probes for in situ hybridization. Specifically, a probe complementary to the transcript encoding the 27 kilodalton (kDa) gamma zein storage protein was employed as a test case for determining the conditions needed to assess the expression patterns of genes currently being identified by the larger project. Results showed that these probes are useful for probing transcripts located in maize tissue and may be used in the future to assess the expression of target genes such as marker genes and transcription factors within maize tissue. Unfortunately, this particular type of oligonucleotide probe has now become unavailable; however, if/when it does become available again (as the vendor says it will), the optimal conditions for this tissue have been determined and can be modified as needed for each probe. Even if other types of probes, such as complimentary RNA probes, need to be used, some of the conditions identified here will be useful.

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