Theses and Dissertations

Author

Ramesh Katam

Issuing Body

Mississippi State University

Advisor

Matta, Frank B.

Committee Member

Spiers, James M.

Committee Member

Braswell, John H.

Committee Member

Silva, Juan L.

Committee Member

Sheikh, Mehboob B.

Other Advisors or Committee Members

Stringer, Stephen J.

Date of Degree

1-1-2008

Document Type

Dissertation - Open Access

Major

Horticulture

Degree Name

Doctor of Philosophy

College

College of Agriculture and Life Sciences

Department

Department of Plant and Soil Sciences

Abstract

Water stress affects vine productivity, disease tolerance, and enological characteristics of grape. Florida Hybrid Bunch grape are developed through hybridization of local grape spp with Vitis vinifera. These cultivars are mostly grown in southeast region of United States. Water deficit conditions resulted due to failure of rains in the region has developed concern among Florida grape growers to increase water use efficiency of grape. The goal of this research is to identify genes and proteins differentially expressed in response to water stress and to correlate these changes with enological characteristics. Investigating transcripts and proteins will allow us to correlate them and confirm the involvement of specific genes responding to stress. Florida hybrid bunch ‘Suwannee’ grape plants were maintained under green house conditions. Water stress was induced by withholding irrigation. The leaf samples were collected from both irrigated and stressed plants at 5, 10, 15 and 20 day interval. We generated over 200 Subtractive Hybridization PCR products from control and water stressed leaf tissues. Cloning, sequencing and transcript analysis revealed that, 54 genes related to drought and defense regulated pathways out of 125 characterized transcripts. Proteins were extracted from leaf tissue with trichloroacetic acid /acetone and separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The proteins were sequenced in LC/Mass Spectrophotometer. The most important differentially expressed genes include sucrose synthase, actin, isoprene synthase, ABF3, SNF1 related protein kinase, WRKY type transcription factors, AP2, ASR2, glyoxalase I and, cytochrome b which play significant role in cell permeability, transportation, photosynthesis and, maintenance in osmotic stress. We have found that ribulose bisphosphate carboxylase and phosphoribulokinase, which play major role in photosynthesis, were suppressed in response to water stress in Florida hybrid bunch. The results suggested that water stress affects expression of cDNAs associated with defense and drought regulated functions. Such profiling studies will be used to explicate specific pathways disconcerted by water deficit treatments, and in the identification of varietal differences.

URI

https://hdl.handle.net/11668/17100

Comments

subtractive hybridization||proteomics||genomics||grape||DDRT||2D electrophoresis

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