High frequency production of haploid embryos in asparagus anther culture

Summary

A method for obtaining a high frequency of haploid asparagus embryos through anther culture was developed. Flowers collected from plants in the field in July, August and September 1990, for the genotype G203, were stored at 5°C for 24 h. Anthers were placed on Murashige and Skoog medium (MS) containing 500 mg l ⁻¹ casein hydrolysate, 800 mg l⁻¹ glutamine, 2 mg l ⁻¹ NAA, 1 mg l ⁻¹ BA and 5 % sucrose at 32 °C in the dark for three to four weeks to induce calli. Calli were then grown at 25 °C with a 16 h photoperiod for three to four weeks. Developing embryos and calli were transferred to embryo maturation medium, MS containing 6% sucrose, 0.1 mg l ⁻¹ NAA, 0.1 mg l ⁻¹ kinetin and 0.65 mg l ⁻¹ ancymidol, for four weeks. More than 50% of the recovered mature embryos germinated on MS containing l mg l ⁻¹ GA₃. Anthers with microspores at the late-uninucleate stage had the highest frequency of total and embryogenic calli formation, 40% and 15%, respectively. Each embryogenic callus usually produced 10–15 embryos. Aproximately 75 plants per 100 anthers cultured were recovered: 76% haploid, 22% diploid and 2% triploid. High temperature was critical for the induction of embryogenic callus.