Somatic hybridization between the zinc accumulator Thlaspi caerulescens and Brassica napus

Abstract 

Somatic hybrids between the zinc hyperaccumulator Thlaspi caerulescens and Brassica napus were produced by electrofusion of protoplasts isolated from each species. Optimization of electrofusion parameters yielded interspecies heteroplasmic fusion rates of up to 13%. Hybrids were selected by screening the growing calli for Zn tolerance. In addition, a second novel selection technique was developed based on the observation that a high proportion of hybrid microcalli grown in liquid media did not adhere to the wall of the culture vessel, while microcalli derived from parental cells did. Seventeen from a total of 64 regenerated plants were conclusively verified as hybrids by AFLP DNA analysis. The hybrid plants were grown in soil for up to 4 months, and at least five flowered. Several of these hybrids survived when grown on high-zinc media.These hybridsaccumulated levels of zinc and cadmium that would have been toxic for B. napus. The data indicate that transfer of the trait for metal hyperaccumulation in plants is possible through somatic hybridization.