Title

Functional Characterization Of A Newly Identified Nicotiana Tabacum Sulfate Transporter Gene NtaSULTR2 Involved In Sulfate Transport And Distribution

Author(s)

Adam Herbert

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Botany

Presentation Type

Mentored

Mentor

Mentor: Hong Luo, Department of Genetics and Biochemistry, Clemson University

Abstract

Sulfur, one of six macronutrients required by plants, is the rudimental component of cysteine, methionine and sulfolipid. It also serves as the active center of many enzymes. In soil, inorganic sulfate (SO42-) is the most common sulfur source for plants. Over the past twenty years, sulfate transporters involved in sulfate uptake and distribution have been well studied in plants. Fourteen sulfate transporter genes, classified into five groups, were identified in Arabidopsis thaliana. To date, twelve sulfate transporter genes have been identified in rice, Oryza sativa. However, only one sulfate transporter gene, NtaSULTR2 has been identified and cloned in tobacco, Nicotiana tabacum. NtaSULTR2 is regulated by a sulfate responding micro-RNA, with repression of NtaSULTR2 expression in transgenic tobacco leading to impaired sulfate homeostasis. The purpose of this study is to functionally characterize the gene NtaSULTR2. A micro-RNA resistant NtaSULTR2 cDNA, rNtaSULTR2 was created using a site-directed mutagenesis method. rNtaSULTR2 was ligated into the plasmid pHL721 in order to overexpress the cDNA. rNtaSULTR2 was also ligated into the plasmid pHL722 for RNA-interference, repressing NtaSULTR2. These two vectors were then transformed into Agrobacterium tumefaciens LBA4404 by electroporation. They will be transformed into tobacco using the leaf disc transformation method. This experiment could help solve issues with soil having decreasing sulfur levels by allowing the tobacco to have a higher uptake of sulfur.

Start Date

4-11-2015 11:00 AM

End Date

4-11-2015 11:15 AM

COinS
 
Apr 11th, 11:00 AM Apr 11th, 11:15 AM

Functional Characterization Of A Newly Identified Nicotiana Tabacum Sulfate Transporter Gene NtaSULTR2 Involved In Sulfate Transport And Distribution

Sulfur, one of six macronutrients required by plants, is the rudimental component of cysteine, methionine and sulfolipid. It also serves as the active center of many enzymes. In soil, inorganic sulfate (SO42-) is the most common sulfur source for plants. Over the past twenty years, sulfate transporters involved in sulfate uptake and distribution have been well studied in plants. Fourteen sulfate transporter genes, classified into five groups, were identified in Arabidopsis thaliana. To date, twelve sulfate transporter genes have been identified in rice, Oryza sativa. However, only one sulfate transporter gene, NtaSULTR2 has been identified and cloned in tobacco, Nicotiana tabacum. NtaSULTR2 is regulated by a sulfate responding micro-RNA, with repression of NtaSULTR2 expression in transgenic tobacco leading to impaired sulfate homeostasis. The purpose of this study is to functionally characterize the gene NtaSULTR2. A micro-RNA resistant NtaSULTR2 cDNA, rNtaSULTR2 was created using a site-directed mutagenesis method. rNtaSULTR2 was ligated into the plasmid pHL721 in order to overexpress the cDNA. rNtaSULTR2 was also ligated into the plasmid pHL722 for RNA-interference, repressing NtaSULTR2. These two vectors were then transformed into Agrobacterium tumefaciens LBA4404 by electroporation. They will be transformed into tobacco using the leaf disc transformation method. This experiment could help solve issues with soil having decreasing sulfur levels by allowing the tobacco to have a higher uptake of sulfur.