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Document Type

Thesis - University Access Only

Award Date

2013

Degree Name

Master of Science (MS)

Department

Biology and Microbiology

First Advisor

Jai S. Rohila

Abstract

Wheat is a commercially important crop of the USA and consumed worldwide. North Dakota, Montana and South Dakota are major wheat producing states of the USA. Climatic changes cause fluctuations in its yield from year to year. These north central regions of the USA are no exceptions to the global trend of climatic variability. In order to sustain the increasing demand for wheat and to avert the risk for famine in the near future, FAO has estimated to double the crop production by 2040. In the 20th century conventional plant breeding has played a crucial role in bringing up green revolution, but in 21st century the use of in vitro tools of biotechnology has taken modern plant breeding programs to the pinnacle of success. Transgenics have successful stories in many commercial crops such as rice, maize, barley, sorghum, soybean, and canola. Despite all this progress genetic engineering is still at its infancy in wheat. Compared to conventional techniques of selective breeding, the genetic engineering approach is faster in transferring the desired trait into the wheat genome. Our current study focusses on exploiting genetic engineering to integrate and express a foreign gene in the wheat genome. Biolistics and Agrobacterium-mediated genetic transformation (AMGT) are the two most promising tools to achieve these goals. AMGT has some advantages over the biolistics (such as a low copy number), thus AMGT is a method of choice for us. The purpose of this study was to develop a robust protocol for in vitro manipulations, AMGT and plantlet regeneration of wheat suitable for South Dakota (SD) breeders. During the study 20 different wheat genotypes along with four different A. tumefaciens strains were evaluated. pCAMBIA-1305.02 carrying GUSPlus as a reporter gene and hygromycin as plant selection gene was used for wheat transformation. Further protocol was optimized using three cultivars of spring and winter wheat each. Stable integration and expression of GUS gene in T0 plants was confirmed by molecular analysis. The highest transformation efficiency of embryogenic calli derived from immature embryos was recorded to be 0.74±0.02%.

Library of Congress Subject Headings

Wheat--Genetics
Agrobacterium tumefaciens
Wheat--Genetic engineering
Plant tissue culture

Description

Includes bibliographical references

Format

application/pdf

Number of Pages

208

Publisher

South Dakota State University

Rights

In Copyright - Educational Use Permitted
http://rightsstatements.org/vocab/InC-EDU/1.0/

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