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Starch branching enzyme IIb in wheat is expressed at low levels in the endosperm compared to other cereals and encoded at a non-syntenic locus

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Abstract

Studies of maize starch branching enzyme mutants suggest that the amylose extender high amylose starch phenotype is a consequence of the lack of expression of the predominant starch branching enzyme II isoform expressed in the endosperm, SBEIIb. However, in wheat, the ratio of SBEIIb and SBEIIa expression are inversely related to the expression levels observed in maize and rice. Analysis of RNA at 15 days post anthesis suggests that there are about 4-fold more RNA for SBE IIa than for SBE IIb. The genes for SBE IIa and SBE IIb from wheat are distinguished in the size of the first three exons, allowing isoform-specific antibodies to be produced. These antibodies were used to demonstrate that in the soluble fraction, the amount of SBE IIa protein is two to three fold higher than SBIIb, whereas in the starch granule, there is two to three fold more SBE IIb protein amount than SBE IIa. In a further difference to maize and rice, the genes for SBE IIa and SBE IIb are both located on the long arm of chromosome 2 in wheat, in a position not expected from rice–maize–wheat synteny.

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Abbreviations

CS:

Chinese spring

dNTP:

Deoxynucleotide triphosphate

DPA:

Days post anthesis

GST:

Glutathione S-transferase

NT:

Nullisomic tetrasomic

ORF:

Open reading frame

PBS:

Phosphate buffered saline

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Acknowledgements

The authors acknowledge Sandra Hey, Rothamsted Experimental Station, Harpenden, Herts, UK AL5 2JQ for kindly providing the CS cDNA expression library and Biogemma UK for the financial support for conducting of this work. The authors also acknowledge the valuable input of Tanya Phongkham, CSIRO Plant Industry in some aspects of the work.

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Correspondence to Sadequr Rahman.

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Regina, A., Kosar-Hashemi, B., Li, Z. et al. Starch branching enzyme IIb in wheat is expressed at low levels in the endosperm compared to other cereals and encoded at a non-syntenic locus. Planta 222, 899–909 (2005). https://doi.org/10.1007/s00425-005-0032-z

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