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Document Type

Thesis - University Access Only

Award Date

2015

Degree Name

Master of Science (MS)

Department

Biology and Microbiology

First Advisor

Jai S. Rohila

Abstract

Senescence in plants is a genetically programmed sequence of biochemical and physiological changes, but little is known about the key players for it in the perennial biomass based biofuel grasses. Leaf proteome of early- and late- senescence germplasm of PCG and SG was analyzed around the senescence period via 2D-DIGE technology combined with Mass Spectrometry. Upon analyzing the manually filtered differentially expressed proteins of after to before senescence, 9 unique proteins were assigned to PCG, and 18 proteins to SG. Before the senescence a constitutive overexpression of 14-3-3 like protein, and GST was observed in ‘late senescing’ germplasm of PCG. Antagonistically, before the onset of senescence, the RAB protein, and SAM synthase were found up regulated in ‘early senescing’ germplasm of PCG compared to ‘late senescing’ germplasm. Before the onset of senescence, three key enzymes of Calvin cycle: Transketolase, Sedoheptulose-1,7-bisphosphatase, and Fructose-bisphosphate aldolase were found to be down-regulated in ‘early senescing’ PCG compared to its ‘late senescing’ germplasm. Overall for after to before senescence in both PCG and SG, ten common proteins were found significantly differentially expressed that included 5 up regulated and 5 down-regulated proteins. xiv A robust tissue culture system was also developed for PCG for its use in future biotechnology projects. Callus of PCG was induced by culturing sterilized mature seeds on the callus induction medium (D3.2), which contained Murashige and Skoog basal medium with 3% sucrose. Additionally, 3 mgL-1 2,4-D, 1 mgL-1 IAA, 7.8 µM CuSO4, 30 µM Silver thiosulphate (STS), and 6 g L-1 Agar were also added in the medium. For regeneration, Murashige and Skoog based medium (D-Reg) which contained Murashige and Skoog basal and sucrose along with 1 mgL-1 BAP, 0.5 mgL-1 GA, 0.2 mgL-1, NAA, 7.8 µM CuSO4, 30 µM STS, and 6 gL-1 Agar was formulated. Copper and STS were found to have beneficial effects on the callus induction and regeneration processes of PCG. After callus induction on D3.2 medium and 3 successive subcultures, calli were transferred onto the D-Reg medium for shoot regeneration. For rooting, half strength Murashige and Skoog with 3% sucrose and 10 µgL-1 NAA was found optimal.

Library of Congress Subject Headings

Spartina--Aging
Switchgrass--Aging
Proteomics
Plant tissue culture
Energy crops
Biomass energy.

Description

Includes bibliographical references ( pages 21-31)

Format

application/pdf

Number of Pages

114

Publisher

South Dakota State University

Rights

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

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