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Improvement of wheat (Triticum aestivum L.) anther culture methods for doubled haploid plant production
Abstract
The production of doubled haploids by anther culture is usually accomplished by three steps: the culture of anthers to produce embryos, plant regeneration and chromosome doubling. In these experiments, different sugars were used in initiation and regeneration media with wheat starch as gelling agent. For embryo initiation, anthers from three wheat genotypes were cultured in Liang's 85D12 initiation media with seven sugars at 0.26 M/L and 2,4 D (2 mg/L), NAA (1 mg/L), glutamine (254 mg/L), and wheat starch (5% W/V). Maltose was the best sugar (105 embryos/100 anthers), followed by glucose (47 embryos/100 anthers) and maltose + glucose (37 embryos/100 anthers). These sugars were superior to the standard sugar sucrose (24 embryos/100 anthers). Maltose was superior to sucrose for each genotype. Galactose and mannose were ineffective in embryo initiation. For regeneration the group of embryos produced by each sugar was divided in two groups; the first group was transferred to a regeneration medium with the same sugar used in initiation (SI), and the second one to a sucrose regeneration media. The regeneration media included Liang's 85D12 salts, sugars at 0.06 M/L, and wheat starch at 7% W/V. Highly significant differences were observed among genotypes, SI-sugars and SR-sugars. Pavon 76 was the best for plant regeneration (36.7%), followed by Centurk (30.8%), and Arapahoe (1.7%). Embryos produced by SI-maltose regenerated higher frequency of plants. SR-maltose and SR-sucrose with SI-maltose produced higher number of plants than any other sugar combination. The number of albinos and the frequency of spontaneous doubling also were affected by genotypes and sugars. Pavon 76 produced more albinos (8.4%) and doubled haploids (23%) than Centurk (3.4% and 13% respectively). SR-glucose produced the highest frequency of albinos (14.5%) and SR-Maltose + Glucose induced the highest frequency of spontaneous doubling (27.5%). For the third experiment two wheat genotypes, three colchicine concentrations (0.0, 0.1, and 0.2 g/L), and three sugars (maltose, sucrose, and maltose + glucose) were used. Increasing colchicine concentration decreased the number of embryos produced, but not the frequency of plant regeneration. Considering the total number of doubled haploid plants produced, low levels of colchicine added to the media were very effective in chromosome doubling. Differences were found between genotypes and among sugars. Pavon 76 was superior to Centurk and maltose was superior to sucrose and maltose + glucose.
Subject Area
Plant propagation|Agronomy|Botany
Recommended Citation
Navarro-Alvarez, Willy, "Improvement of wheat (Triticum aestivum L.) anther culture methods for doubled haploid plant production" (1992). ETD collection for University of Nebraska-Lincoln. AAI9308190.
https://digitalcommons.unl.edu/dissertations/AAI9308190