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

Article


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. https://doi.org/10.1007/s00299-006-0191-9
Article Title

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

ERA Journal ID2645
Article CategoryArticle
AuthorsKhanna, Harjeet K. (Author) and Daggard, Grant E. (Author)
Journal TitlePlant Cell Reports
Journal Citation25 (12), pp. 1336-1346
Number of Pages11
Year2006
Place of PublicationGermany
ISSN0721-7714
1432-203X
Digital Object Identifier (DOI)https://doi.org/10.1007/s00299-006-0191-9
Web Address (URL)https://link.springer.com/article/10.1007/s00299-006-0191-9
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.

Keywordsantifreeze protein, codon optimisation, frost tolerance, signal peptide, transgenic, wheat
Contains Sensitive ContentDoes not contain sensitive content
ANZSRC Field of Research 2020300105. Genetically modified field crops and pasture
300406. Crop and pasture improvement (incl. selection and breeding)
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Byline AffiliationsQueensland University of Technology
Centre for Systems Biology
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