Isolation and Characterization of the Soybean Major Latex Homologue Msg, and Analysis of Its Pod -Specific Promoter
Stromvik, Martina Viktoria
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Permalink
https://hdl.handle.net/2142/87904
Description
Title
Isolation and Characterization of the Soybean Major Latex Homologue Msg, and Analysis of Its Pod -Specific Promoter
Author(s)
Stromvik, Martina Viktoria
Issue Date
2000
Doctoral Committee Chair(s)
Vodkin, Lila O.
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Plant Physiology
Language
eng
Abstract
The objective of this project was to identify and investigate a pod specific promoter from soybean for the potential use in disease resistance strategies. A random lambda cDNA clone, VS-107, isolated from a soybean pod library, showed elevated mRNA expression in soybean pods and it was used as a probe to retrieve the gene and promoter sequences from a soybean genomic lambda library. Computer analyses indicate that the gene, named Msg , belongs to the major latex protein family. Primers annealing at different positions 5' upstream of the gene, at the 3 'UTR and at the intron were used to obtain PCR fragments representing the flanking regions. The different regions were cloned into the binary vector pBI101 with a promoterless uidA (GUS) reporter gene. Fourteen deletion fragments of the 5' region were compared by particle bombardment of soybean pods and by transformation into Arabidopsis by Agrobacterium vacuum infiltration. The full length promoter is needed for maximal expression in floral and fruit tissues, guard cells and nodes. Surprisingly, the minimal promoter of 650 bp is only functional in Arabidopsis when the nosT is replaced by the Msg 3'UTR and it can be deleted without a change in expression pattern. A pathogen induction experiment on transgenic Arabidopsis plants did not show the Msg promoter to be induced after infiltration with Pseudomonas syringae. However, in soybean pods, the Msg transcript was localized by whole mount in situ hybridization to the mid pericarp layer, a continuous single cell layer believed to be involved in pathogen defense. The conclusion is that the Msg promoter is a strong soybean promoter that drives developmentally induced gene expression in tissues that are sites for pathogen defense.
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