Identification of highly transformable wheat genotypes for mass production of fertile transgenic plants

Abstract

The efficiency of wheat biolistic transformation systems strongly depends on the bombardment parameters, the condition of the donor plant, and the plant genotype chosen for the transformation process. This paper analyzes the transformation efficiency of the 129 wheat sister lines generically called 'Bobwhite', originally obtained from the cross 'Aurora'//'Kalyan'/'Bluebird 3'/'Woodpecker'. A number of factors influencing the transformation were examined, such as the ability to produce embryogenic callus, regeneration in selection medium, and overall transformation performance. Of the 129 genotypes evaluated, eight demonstrated transformation efficiencies above 60% (60 independent transgenic events per 100 immature embryos bombarded). Among the eight genotypes identified, we studied agronomic characteristics such as earliness to identify the most adaptable line(s) for different lab conditions. 'Bobwhite' SH 98 26 was identified as a super-transformable wheat line.

Tissue culture efficiency of wheat species with different genomic formulas

“…Likewise, the culture efficiency of immature embryo tissues of the other wheat species with AABB genome (T. dicoccum 'Runo', T. turgidum and T. polonicum) as well as of hexaploid T. spelta (AABBDD genome) was good (about 10-12 green plant per initial explants). This performance is usually regarded as sufficient for inclusion in genetic engineering programs for polyploid wheat cultivars (Pellegrineschi et al 2002, He et al 2010. The results of our study contradict findings of Yang at al.…”

“…Diploid wheat species were T. monococcum L. (einkorn, AA) and T. sinskajae A. To induce plant regeneration from immature embryo culture we used a conventional two-step protocol that included 4-week cultivation of explants in the dark on media containing 2,4-D (2,4-dichlorophenoxyacetic acid) following plant differentiation in the light on media without growth regulators (Fennel et al 1996, Machii et al 1998, Pellegrineschi et al 2002, Chauhan et al 2007, Miroshnichenko et al 2013. Immature embryos were isolated from caryopses of greenhousegrown wheat plants 11-15 days after anthesis.…”

“…al 1998, Barro et al 1999, Bohorova et al 2001, He and Lazzeri 2001, Pellegrineschi et al 2002, Eudes et al 2003, Vendruscolo et al 2008, Miroshnichenko et al 2013) and for reliable and efficient genetic engineering (Mamrutha et al 2014, Jones 2015. However, the cultivation of donor plants to ensure a regular supply of immature embryos is labor-intensive and requires a lot of time and space.…”

Map-based isolation of the leaf rust disease resistance gene Lr10 from the hexaploid wheat ( Triticum aestivum L.) genome

“…pUbiT10rga2-1A, which contains a 3-kb T10rga2-1A full-length cDNA with 65 bp upstream and 203 bp downstream of the coding sequence was generated by using a similar strategy. A total of 350 immature embryos of the Bobwhite accession SH 98 56 (18) were cotransformed with pUBiT10rga1, pUbiT10rga2-1A, and a plasmid containing the selectable phosphomannose isomerase marker (19) by using the PDS-1000͞He biolistic particle delivery System (Bio-Rad). Regeneration and selection of the transformed plants were performed as described (18,19).…”

“…A total of 350 immature embryos of the Bobwhite accession SH 98 56 (18) were cotransformed with pUBiT10rga1, pUbiT10rga2-1A, and a plasmid containing the selectable phosphomannose isomerase marker (19) by using the PDS-1000͞He biolistic particle delivery System (Bio-Rad). Regeneration and selection of the transformed plants were performed as described (18,19). Eight independent T0 transgenic lines containing either the two transgenes or only one of the transgenes were obtained.…”