Isolation and expression analysis of genes induced by water deficit stress in loblolly pine (Pinus taeda L.) /

Water deficit stress (WDS) is one of the most important

Bibliographic Details
Main Author: Padmanabhan, Veeraragavan
Format: Thesis Book
Language:English
Published: [Place of publication not identified] : [publisher not identified] ; 1996.
Subjects:
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Description
Summary:Water deficit stress (WDS) is one of the most important
factors affecting survival and growth of loblolly pine (Pinus
taeda). A study was undertaken in order to isolate and
characterize WDS-responsive genes for genetic engineering of
loblolly pine trees for marginal and semi-arid regions.
During this study period, four WDS-responsive cDNAs (1p3,
1p8, 1p15, 1p20) were isolated and characterized. These
clones show distinct patterns of gene expression under WDS
conditions. Sequence analysis revealed that 1p3 is
homologous to a stress related gene. The putative LP8
protein is a calcium-binding protein. The 1p15 is proposed
to code for an H2A component of histones. The predicted LP20
protein is similar to Wilm's tumor suppresser protein.
Southern blot and RT-PCR analysis revealed that 1p3 belongs
to a small multigene family. The genomic clone of 1p3 (1p3-
1) was isolated, characterized and sequenced. The sequence
comparison showed that 1p-3-1 is a related member of 1p3 gene
family. RT-PCR using common primers led to the
identification of two additional members of the gene family.
All the four members of the gene family show distinct
patterns of expression under WDS situations. The
transcription start site for the 1p3-1 genomic clone was
mapped. The genomic clone had 1026 bases of promoter region
with 591 bases in the 31 untranslated region. It has a
putative intron, spanning a region of 275-452 bp. The
promoter region of the 1p3-1 genomic clone was
translationally fused to a uidA reporter gene. Transient
expression studies using the P. ellioti cell suspension
cultures revealed that 1p3(P) was induced in response to WDS.
Exo-deletion analysis of 1p3(P) suggested the presence of a
negative regulatory element between the -1026 and -745
region. The activity of 1p3(P) was observed in various
tissues in transgenic tobacco in the T0 generation. This
study is another step towards understanding the WDS-response
in loblolly pine in achieving our long term goal of
genetically engineered tree species for increased WDS
tolerance.
Item Description:Vita.
"Major Subject: Plant Physiology".
Physical Description:xiv, 144 leaves : illustrations ; 28 cm.
Issued also on microfiche from University Microfilms Inc.
Bibliography:Includes bibliographical references: pages 118-143.