Abstract
Resistant germplasm resources are valuable for developing resistant varieties in agricultural production. However, recessive resistance genes are usually overlooked in hybrid breeding. Compared with dominant traits, however, they may confer resistance to different pathogenic races or pest biotypes with different mechanisms of action. The recessive rice bacterial blight resistance gene xa13, also involved in pollen development, has been cloned and its resistance mechanism has been recently characterized. This report describes the conversion of bacterial blight resistance mediated by the recessive xa13 gene into a dominant trait to facilitate its use in a breeding program. This was achieved by knockdown of the corresponding dominant allele Xa13 in transgenic rice using recently developed artificial microRNA technology. Tissue-specific promoters were used to exclude most of the expression of artificial microRNA in the anther to ensure that Xa13 functioned normally during pollen development. A battery of highly bacterial blight resistant transgenic plants with normal seed setting rates were acquired, indicating that highly specific gene silencing had been achieved. Our success with xa13 provides a paradigm that can be adapted to other recessive resistance genes.
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Acknowledgments
We thank Professor Shiping Wang for providing Xoo strain PXO99 and Professor Detlef Weigel for providing the pNW55 plasmid. This research was supported by National Special Key Project for Transgenic Breeding (2008ZX08010-002-1) and International Foundation for Science.
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Communicated by P. Ozias-Akins.
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Fig. S1 Schematic diagram of transfer DNA regions of the four final expression vectors. a Osrbcp-amiA; b Atrbcsp-amiA; c Osrbcp-amiB; d Atrbcp-amiB.
Fig. S2 Relative expression of Xa13 in the leaves of transgenic plants by qRT-PCR using Oligo(dT)18 and random hexamer primers. The relative quantity of Xa13 mRNA in the lines expressing amiA detected using random hexamer primers is higher than that using Oligo(dT)18 primer, but no similar phenomenon was observed in lines expressing amiB. Wt, Wild type.
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Li, C., Wei, J., Lin, Y. et al. Gene silencing using the recessive rice bacterial blight resistance gene xa13 as a new paradigm in plant breeding. Plant Cell Rep 31, 851–862 (2012). https://doi.org/10.1007/s00299-011-1206-8
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DOI: https://doi.org/10.1007/s00299-011-1206-8