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

Permanent Link: http://ncf.sobek.ufl.edu/NCFE004876/00001

Material Information

Title: REVERSE GENETICS AND REAL-TIME PCR TO IDENTIFY GENES THAT REGULATE EARLY MAIZE KERNEL DEVELOPMENT
Physical Description: Book
Language: English
Creator: True, Jillian
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2013
Publication Date: 2013

Subjects

Subjects / Keywords: Endosperm
Embryo
Genetics
Development
Developmental
Biology
Maize
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

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.
Statement of Responsibility: by Jillian True
Thesis: Thesis (B.A.) -- New College of Florida, 2013
General Note: Online version not currently available.
Electronic Access: RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE
Bibliography: Includes bibliographical references.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Local: Faculty Sponsor: Clore, Amy

Record Information

Source Institution: New College of Florida
Holding Location: New College of Florida
Rights Management: Applicable rights reserved.
Classification: local - S.T. 2013 T86
System ID: NCFE004876:00001

Permanent Link: http://ncf.sobek.ufl.edu/NCFE004876/00001

Material Information

Title: REVERSE GENETICS AND REAL-TIME PCR TO IDENTIFY GENES THAT REGULATE EARLY MAIZE KERNEL DEVELOPMENT
Physical Description: Book
Language: English
Creator: True, Jillian
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2013
Publication Date: 2013

Subjects

Subjects / Keywords: Endosperm
Embryo
Genetics
Development
Developmental
Biology
Maize
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

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.
Statement of Responsibility: by Jillian True
Thesis: Thesis (B.A.) -- New College of Florida, 2013
General Note: Online version not currently available.
Electronic Access: RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE
Bibliography: Includes bibliographical references.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Local: Faculty Sponsor: Clore, Amy

Record Information

Source Institution: New College of Florida
Holding Location: New College of Florida
Rights Management: Applicable rights reserved.
Classification: local - S.T. 2013 T86
System ID: NCFE004876:00001

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