Open Access Highly Accessed Research article

Concerted suppression of all starch branching enzyme genes in barley produces amylose-only starch granules

Massimiliano Carciofi1, Andreas Blennow2*, Susanne L Jensen23, Shahnoor S Shaik2, Anette Henriksen4, Alain Buléon5, Preben B Holm1 and Kim H Hebelstrup1*

Author Affiliations

1 Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark

2 Department of Plant Biology and Biotechnology, VKR Research Centre for Pro-Active Plants, Faculty of Life Sciences, University of Copenhagen, Frederiksberg, Denmark

3 KMC, Herningvej 60, Brande, 7330, Denmark

4 The Protein Chemistry Group, Carlsberg Laboratory, Copenhagen, Denmark

5 UR1268 Biopolymeres Interactions Assemblages, INRA, Nantes, F-44300, France

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BMC Plant Biology 2012, 12:223  doi:10.1186/1471-2229-12-223

Published: 21 November 2012

Additional files

Additional file 1:

SBE target sequences. Target sequence of SBEI, SBEIIa and SBEIIb for the chimeric SBE RNAi construct.

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Additional file 2:

Transgene expression analysis. Relative expression levels of selection marker gene (Hpt) and transgenic hairpin construct analysed by RT qPCR in leaves of control and transgenic T1 lines (three technical replicates each). SE is indicated.

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Additional file 3:

Differential scanning calorimetry table. Differential scanning calorimetry (DSC) in aqueous suspension of starch extracted from control and from SBE RNAi4.1 lines. ΔH: Change in enthalpy due to starch thermal dissolution in water.

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Additional file 4:

PCR of SBE RNAi 4.9 T1genomic DNA. A PCR was performed using the primers Hairpin Fw and Rev (Figure 1a) to detect presence of the transgene hairpin in the genomic DNA of SBE RNAi 4.9 T1 grains. Primers for GAPDH was used as a positive control. The experiment was conducted in triplicate showing the same result.

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Additional file 5:

β-Glucan content. β-Glucan content in SBE RNAi4.1, SBE RNAi4.9 and control T2grains. β-glucan content is reported in percent dry weight (average values of two biological and six technical replicates).

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Additional file 6:

Polarization microscopy. Bright field microscopy image (left) and polarized microscopy image (right) of control starch granules (a, b) and amylose-only granules from line SBE RNAi4.1 (c, d).

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Additional file 7:

Yield. (a) Average number of spikes per plant calculated in all plants from a semi-field trial (sample population: 50 plants each line). (b) Average number of grains per spike in 25 plants per line from a semi-field trial. (c) Average single grain mass calculated for 25 plants per line from a semi-field trial. (d) Average yield per plant in milligram of grain produced calculated in a sample of 25 plants per line from semi-field trial. (e) Correlation coefficients (r) and significance level as P values for the three yield components: average yield per plant, average number of spikes per plant and average grain mass.

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Additional file 8:

Germination frequency. Germination frequency in soil of barley grains from SBE RNAi 4.1 and control lines.

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Additional file 9:

Primers. Primers for RT qPCR.

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