Abstract
Maize (Zea mays spp. mays) is a staple crop for more than 900 million people. The seeds or kernels provide a rich source of calories because ~70 % of the weight is carbohydrate, mostly in the form of starch. The content and composition of starch are complex traits controlled by many genes, offering multiple potential targets for intervention. We used a multigene engineering approach combining the overexpression of Bt2, Sh2, Sh1 and GbssIIa (to enhance the activity of sucrose synthase, AGPase and granule-bound starch synthase) with the suppression of SbeI and SbeIIb by RNA interference (to reduce the activity of starch branching enzyme). Maize plants expressing all six genes plus the selectable marker showed a 2.8–7.7 % increase in the endosperm starch content and a 37.8–43.7 % increase in the proportion of amylose, which was significant compared to untransformed control plants. We also observed improvements in other agronomic traits, such as a 20.1–34.7 % increase in 100-grain weight, a 13.9–19.0 % increase in ear weight, and larger kernels with a better appearance, presumably reflecting the modified starch structure within the kernels. Our results confirm that multigene engineering applied to the starch biosynthesis pathway can not only modulate the quality and quantity of starch but can also improve starch-dependent agronomic traits.
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Acknowledgments
This work was supported by the National Transgenic Maize Project #2009ZX08003-024B and Nature Science Foundation of Jilin Province #20080567 (to JP), and the National Nature Science Foundation #31170259 (to XQ), and #31070269 (to CZ). We thank Dr. Robert Henry from Southern Cross University Australia, for kindly providing the endosperm-specific promoters, and to Prof. Paul Christou of Universitat de Lleida for his constructive comments which improved the manuscript.
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Lili Jiang and Xiaoming Yu contributed equally to this work.
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11248_2013_9717_MOESM1_ESM.tiff
Fig. S1. Expression and silencing cassettes used for maize transformation. The expression of Sh1, Bt2, Sh2 and GbssIIa was driven by different monocot endosperm-specific promoters: barley B1-hordein, rice glutelin-1, barley ISA and wheat high molecular weight glutenin, respectively. The two RNAi constructs were driven by a barley D-hordein promoter. The bar gene was controlled by the maize ubiquitin promoter (Ubi). (TIFF 1148 kb)
11248_2013_9717_MOESM2_ESM.tif
Fig. S2. PCR analysis of genomic DNA from T1 plants transformed with expression cassettes containing Sh1 (a), Bt2 (b), Sh2 (c), GbssIIa (d), SbeI (e) and SbeIIb (f). (TIFF 1575 kb)
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Jiang, L., Yu, X., Qi, X. et al. Multigene engineering of starch biosynthesis in maize endosperm increases the total starch content and the proportion of amylose. Transgenic Res 22, 1133–1142 (2013). https://doi.org/10.1007/s11248-013-9717-4
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DOI: https://doi.org/10.1007/s11248-013-9717-4