Addition of Colchicine to Wheat Anther Culture Media to Increase Doubled Haploid Plant Production

Abstract

Chromosome doubling is critical for obtaining doubled-haploid plants from wheat (Triticum aestivum L.) anther culture. The most common doubling method applies colchicine to the plant. However, colchicine is phytotoxic and can induce a high frequency of plant death. In this experiment, anthers from two wheat genotypes ('Pavon 76' and 'Centurk') were placed on nine embryoid initiation media having three sugar sources (maltose, sucrose, and maltose + glucose) with three colchicine concentrations (0.0, 0.1, and 0.2 g • 1'). Wheat starch was used as a gelling agent. After three days, the anthers were washed and moved to fresh media without colchicine. Increasing the colchicine concentration decreased the number of embryoids produced from 77,4 embryoids/100 anthers to 29.9 embryoids/100 anthers, but did not significantly affect the frequency of plant regeneration (0.49 green plants/embryoid to 0.40 green plants/embryoid), and increased the frequency of doubled-haploid plants (19.0 doubledhaploid plants/100 green plants to 72.3 doubledhaploid plants/100 green plants). Considering the total number of doubled-haploid plants produced, low levels of colchicine added to the initiation media were very effective.

Enhanced induction of microspore embryogenesis after n-butanol treatment in wheat (Triticum aestivum L.) anther culture

“…Toxicity, along with genotypic effects, are the main drawbacks when applying these substances (Navarro-Alvarez et al 1994;Zamani et al 2003). In the present work, toxicity was not detected at the low concentrations (0.1-0.2%) of n-butanol applied, as previously reported by Dhonukshe et al 2003 andGardiner et al 2003.…”

Plant Regeneration from Anther Culture of Panax ginseng

“…In our experiment, high callus induction rate obtained when we used 3% of sucrose as a carbon source. High concentration (6-12%) of maltose improved embryoid induction and shoot regeneration from anthers of wheat, barley and potato (Finnie et al, 1989;Navarro-Alvarez et al, 1994;Batty and Dunwell, 1989). However, ginseng anthers could not show high level of callus induction in different concentration of maltose compared with sucrose.…”

“…Maltose has recently been found to be superior to sucrose for embryoid induction and plant regeneration in anther/microspore culture of barley and wheat (Finnie et al, 1989;Navarro-Alvarez et al, 1994). The type and concentration of sugar on callus induction and shoot regeneration in anther culture of ginseng has not previously been investigated.…”

Recent advances and application of doubled haploids in wheat breeding

“…Colchicine has been added directly to the anther culture induction medium at concentrations of about 0.2 g/l (500 _M). Anthers were transferred to colchicine free medium after 72 h, resulting in a frequency of fertile plants up to about 70% (Barnabas et al, 1991;Navarro-Alvarez et al, 1994). Alternatively, Ouyang et al (1994) cultured pollen calli on colchicine containing medium during regeneration and reported an average increase in the frequency of fertile plants from 17% in the control to 54% with the best in vitro treatment.…”

“…Spontaneous rates of chromosome doubling among plants derived from microspores of wheat are relatively low. In many experiments, only 15 to 20% of the plants obtained are capable of seed set by selfing without a treatment for chromosome doubling (Hansen et al, 1988;Navarro-Alvarez et al, 1994;Ouyang et al, 1994). To regain fertility, the number of wheat chromosomes need to be doubled following haploid embryo rescue and seedling formation.…”