Targeted expression of redesigned and codon optimised synthetic gene leads to recyrstallisation inhibition and reduced electrolyte leakage in spring wheat at sub-zero temperatures

Khanna, Harjeet K. and Daggard, Grant E. (2006) Targeted expression of redesigned and codon optimised synthetic gene leads to recyrstallisation inhibition and reduced electrolyte leakage in spring wheat at sub-zero temperatures. Plant Cell Reports, 25 (12). pp. 1336-1346. ISSN 0721-7714

Abstract

[Abstract]: Antifreeze proteins (AFPs) adsorb to ice crystals
and inhibit their growth, leading to non-colligative freezing
point depression. Crops like spring wheat, that are highly
susceptible to frost damage, can potentially be made frost
tolerant by expressing AFPs in the cytoplasm and apoplast
where ice recrystallisation leads to cellular damage. The
protein sequence for HPLC-6 α-helical antifreeze protein
fromwinter flounderwas rationally redesigned after removing
the prosequences in the native protein. Wheat nuclear
gene preferred amino acid codons were used to synthesize
a recombinant antifreeze gene, rAFPI. Antifreeze protein
was targeted to the apoplast using a Murine leader peptide
sequence from the mAb24 light chain or retained in
the endoplasmic reticulum using C-terminus KDEL sequence.
The coding sequences were placed downstream of
the rice Actin promoter and Actin-1 intron and upstream
of the nopaline synthase terminator in the plant expression
vectors. Transgenic wheat lines were generated through
micro projectile bombardment of immature embryos of
spring wheat cultivar Seri 82. Levels of antifreeze protein
in the transgenic lines without any targeting peptide were
low (0.06–0.07%). The apoplast-targeted protein reached
a level of 1.61% of total soluble protein, 90% of which
was present in the apoplast. ER-retained protein accumulated
in the cells at levels up to 0.65% of total soluble
proteins. Transgenic wheat line T-8 with apoplast-targeted antifreeze protein exhibited the highest levels of antifreeze
activity and provided significant freezing protection even
at temperatures as low as −7◦C.


Statistics for USQ ePrint 3541
Statistics for this ePrint Item
Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Awaiting Author's version. Deposited in accordance with the copyright policy of the publisher.
Depositing User: epEditor USQ
Faculty / Department / School: Historic - Faculty of Sciences - No Department
Date Deposited: 16 Nov 2007 05:30
Last Modified: 02 Jul 2013 22:54
Uncontrolled Keywords: antifreeze protein, codon optimisation, frost tolerance, signal peptide, transgenic, wheat
Fields of Research : 10 Technology > 1001 Agricultural Biotechnology > 100105 Genetically Modified Field Crops and Pasture
07 Agricultural and Veterinary Sciences > 0703 Crop and Pasture Production > 070305 Crop and Pasture Improvement (Selection and Breeding)
Identification Number or DOI: doi: 10.1007/s00299-006-0191-9
URI: http://eprints.usq.edu.au/id/eprint/3541

Actions (login required)

View Item Archive Repository Staff Only