Androgenesis in the Bioenergy Plant Miscanthus sinensis: From Calli Induction to Plant Regeneration

Miscanthus spp. is a valuable energy grass with a high biomass production, making it a natural source for first‐ and second‐generation fuels. Therefore, the aim of this paper was to investigate a methodology for the haploid production of diploid Miscanthus sinensis Andersson by anther culture. In three experiments the effect of genotype, induction medium, culture beginning date, and cold pretreatment were investigated. The experiments resulted in effective callus induction and plant regeneration. The genotype, culture medium, and culture induction date had significant influence on the number of induced calli whereas cold spikelet pretreatment reduced the culture effectiveness rates. A delay of culture initiation provided a higher callus yield together with enhanced triploid participation in a group of regenerates. Overall, in the culture of 27,300 anthers, 1887 calli were induced, and regeneration gave 2064 plants. In the group of 1523 examined plants, there were 186 haploids and 472 diploids. The comparative genetic analyses of the parental genotypes and selected anther‐derived plants were conducted using the simple sequence repeats (SSRs). For all of the analyzed androgenic plants, SSR allele segregation allowed for the confirmation of homozygosity. The alleles represented balanced (e.g., 1:3) as well as unbalanced (e.g., 5:0) segregations. Plants with calli from the same origins carried the same SSR alleles.