Selection of transformation-efficient barley genotypes based on TFA (transformation amenability) haplotype and higher resolution mapping of the TFA loci
The genetic substitution of transformation amenability alleles from ‘Golden Promise’ can facilitate the development of transformation-efficient lines from recalcitrant barley cultivars.
Barley (Hordeum vulgare) cv. ‘Golden Promise’ is one of the most useful and well-studied cultivars for genetic manipulation. In a previous report, we identified several transformation amenability (TFA) loci responsible for Agrobacterium-mediated transformation using the F2 generation of immature embryos, derived from ‘Haruna Nijo’ × ‘Golden Promise,’ as explants. In this report, we describe higher density mapping of these TFA regions with additional SNP markers using the same transgenic plants. To demonstrate the robustness of transformability alleles at the TFA loci, we genotyped 202 doubled haploid progeny from the cross ‘Golden Promise’ × ‘Full Pint.’ Based on SNP genotype, we selected lines having ‘Golden Promise’ alleles at TFA loci and used them for transformation. Of the successfully transformed lines, DH120366 came the closest to achieving a level of transformation efficiency comparable to ‘Golden Promise.’ The results validate that the genetic substitution of TFA alleles from ‘Golden Promise’ can facilitate the development of transformation-efficient lines from recalcitrant barley cultivars.
KeywordsAgrobacterium tumefaciens Doubled haploid Hordeum vulgare (barley) Single nucleotide polymorphism Transformation
We thank Tanya Filichkin and Laura Helgerson for their parts in developing the Oregon Promise population, Yuka Motoi (IPSR, Okayama Univ.) for technical assistance, and Dr. Yukihiro Ito (Tohoku Univ., Japan) for providing the pBUH3 vector. This work was supported by the Spanish Ministry of Science and Innovation (Project AGL2015-69435-C3-2-R) to L.C., the Gatsby Foundation and BBSRC Institutional Strategic Programme (BB/J004553/1) to M.M., and JSPS KAKENHI, Grant Numbers 24880025 and 16K18634 to H.H.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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