Anther culture and haploid plant regeneration in purple coneflower (Echinacea purpurea L.)

Abstract

Anthers containing microspores at the uninucleate stage were excised from capitula of field grown plants of purple coneflower, Echinacea purpurea, and cultured on medium conducive to callus growth. In callus induction cultures, N6 basal medium was more effective than Murashige and Skoog (MS), and a combination of benzyladenine (BA) at 2.22 lM with naphthaleneacetic acid (NAA) at 0.054 lM was more effective than 2,4-dichlorophenoxyacetic acid (2,4-D) alone at 4.52, 9.05 and 13.57 lM. Callus induction rate as high as 85.8% was achieved, and no statistically significant differences were found among cultures with various densities of 20, 40, 60, 80 and 100 anthers per bottle each filled with 40-ml medium. Shoots were regenerated from calluses on MS medium containing 2.22 lM BA and various concentrations of NAA, with the highest regeneration rate of 95.24% obtained when 0.27 lM NAA was applied. Although a large portion of the regenerated shoots had prominent symptom of vitrification, some normal shoots could be easily rooted on MS medium containing 0.054 lM NAA. Thirty regenerated plants were randomly selected and 19 of them were confirmed to be haploid by observation of chromosome number of root-tip cells.

Estimation of the Cloning Potential in Six Selected Genotypes of Purple Coneflower (Echinacea PurpureaL.)

“…The methods adopted in the present experiments for the preparation of medium and for maintenance of the cultures were basically as those described before (15,22).…”

New basal media for half-anther culture of Anthurium andreanum Linden ex André cv. Tropical

“…2 (Hoagland and Arnon 1950), N 6 , Schenk and Hildebrand (Schenk and Hildebrandt 1972), Woody Plant Medium (Lloyd and McCown 1981), and Quoirin and Lepoivre (Quoirin andLepoivre 1977) (Teixeira da Silva et al 2005). These studies mostly indicate that basal medium is a key factor in developing plant anther culture protocols and maximize the response of anther explants cultured for organogenic events including callus and embryo induction, shoot and root regeneration, and embryo germination (Zhao et al 2006;Koleva-Gudeva et al 2007;Gorecka et al 2009;Sutan et al 2010). …”

“…MS was the most effective for callus induction in Lycopersicon esculentum Mill. (Shtereva et al 1998), liquid PG-96 for timothy (Guo et al 1999), N 6 for Echinacea purpurea (Zhao et al 2006), Linsmeir and Skoog (LS, Linsmaier and Skoog 1965) and NN for pepper (Koleva-Gudeva et al 2007), and PGR-free B5 medium for carrot (Gorecka et al 2009). These studies indicate that anther culture is still dependent on the plant genotype and selection of basal medium.…”

Haploid and Double Hapoids in Ornamentals – A Review

“…All resulting plants were diploid or tetraploid and it was determined that the plants originated from the anther wall (Fu et al, 2008). Zhao et al, (2006) developed 19 haploid plant of Echinacea purpurea by anther culture. They found that, in callus induction cultures, N6 basal medium was more effective than MS media, and a combination of BA at 2.22 µM with NAA at 0.054 µM was more effective than 2,4-D alone.…”