Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants

Abstract

BackgroundInsertional mutagenesis represents one of the most effective ways to acquire information about a plant gene’s function. However, it is hindered by the autosomal genome being diploid and therefore, most mutations being recessive. The problem is addressed by inducing the transposition during anther culture so that selected mutations can be transmitted and then regenerated to a homozygous state.ResultsTo this end, we treated transgenic rice floral tissues containing the inducible transposon with an inducer, salicylic acid. Excision events were detected in regenerated calli and subsequent plantlets. DNA blot and PCR assay were used to determine the homogeneity of knockout mutants. About 5% of the mutants containing transposition events were homozygous. Furthermore, the inducible transposon was active during calli regeneration.ConclusionsThis strategy could be applicable to improve transposition efficiency in microspore development stages to create stable di-haploid mutants in plants.Electronic supplementary materialThe online version of this article (doi:10.1186/1999-3110-55-27) contains supplementary material, which is available to authorized users.

Embryo Rescue in Plant Breeding

“…COKC was introduced into rice plants and activated by salicylic acid (SA) to trigger transposition events. The assumption was that transposition could be induced during anther culture, allowing selected mutations to be transmitted and recovered in the homozygous state in DH regenerated plants [381]. Yang and Charng [381] used five independent transgenic lines, each containing a single copy of COKC, as the plants from which anthers were taken for callus production and regeneration of DH plants.…”

“…The assumption was that transposition could be induced during anther culture, allowing selected mutations to be transmitted and recovered in the homozygous state in DH regenerated plants [381]. Yang and Charng [381] used five independent transgenic lines, each containing a single copy of COKC, as the plants from which anthers were taken for callus production and regeneration of DH plants. The authors observed that inducible transposon was active during callus regeneration, resulting in around 5% of the mutants possessing transposon events in the homozygous condition.…”