REVERSE GENETICS AND REAL-TIME PCR TO IDENTIFY GENES THAT REGULATE EARLY MAIZE KERNEL DEVELOPMENT

Jillian True

Date of Award

2013

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Clore, Amy

Keywords

Endosperm, Embryo, Genetics, Development, Developmental, Biology, Maize

Area of Concentration

Biology, Chemistry

Abstract

The maize kernel consists mostly of endosperm, a tissue rich in starch and protein that provides nutrition to the germinating seedling and those who eat corn, as well as an embryo and outer covering called the pericarp. A reverse genetics approach was adopted to study how kernel development is regulated. Previously, the transcriptomes (or comprehensive sets of all RNA molecules produced reflecting gene expression levels) of kernels from 0-12 days after pollination (DAP) were determined using the technique of deep sequencing (RNA-Seq). An in silico analysis was conducted on the results revealing that genes encoding putative transcription factors had potentially interesting expression patterns. The expression patterns of these genes were confirmed in the present study by real-time reverse-transcriptase polymerase chain reaction (real-time RT-PCR) of kernel cDNAs collected at 0-12 DAP, consistent with roles of these genes in development. Uniform Mu seed stocks harboring mutator transposable element insertions in these genes of interest were planted, and each line was genotyped using gene-specific and Mu-insertion-specific primers. The harvested ears were examined for potential phenotypes including kernel color, area, volume, and density. There was one putative phenotype observed out of five harvested lines. One quarter of the kernels on a heterozygous ear were an unusual peachy-tan color and had altered embryo morphology. As a follow-up to my portion of this larger study, further investigations of this possible phenotype were conducted by the Yadegari laboratory at the University of Arizona revealing a likely function in embryo development despite higher expression levels of the identified gene in the endosperm versus the embryo in wild-type kernels. Potential explanations for this apparent discrepancy will be discussed.

Rights

The author has not granted New College of Florida the nonexclusive right to archive, make accessible, and distribute for educational purposes this work in whole or in part in all forms of media, now or hereafter known. The copyright of this work remains with the author.

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