Abstract
Regeneration into haploid plantlets had been obtained in spring onion using flower and ovary culture. Flowers and ovaries were cultured into media using two protocols (A and B) and the ability to produce callus or somatic embryogenesis were invistigated. Flowers around 3.0-5.0 mm long were collected, whole flower or ovary which were excised from flowers using dissecting microscope were cultured into BDS medium supplemented 2.0 mg L⁻¹ 2, 4-D and 2.0 mg L⁻¹ myo-inositol (protocol A) or into BDS media supplemented 2.0 mg L⁻¹ BAP fortified with 100 g L⁻¹ 2,4-0 and 2.0 mg L⁻¹ sucrose, 200 mg L⁻¹ BAP for 14 days, then sub-cultured by regeneration medium (BDS) supplemented with 1.0 mg L⁻¹ NAA and 2.0 mg L⁻¹ 2iP (protocol B). Embryos were emerged from ovary wall after around 4-5 months, high frequency of embryogenesis induction was produced from ovaries that were using protocol A. While the less percentage observed from flower culture using protocol B.
Highlights
Spring onion (Allium fistulosum L.) is a biennial monocotyledonous plant is one of the most important vegetable crops around the world in terms of crop value.Spring onion is grown widely in Cameron Highlands in the Tringkap and Kea Farm areas (Ramasamy, 1991)
Plant material and sterilization: Whole umbel of spring onion (Allium fistulosum L.) was taken from donor plants when the flowers were at 3-4 days before anthesis (Fig. 1), individual flower 3-5 mm with pedicel were removed and sterilized with 70% alcohol for 45 sec surface-sterilized with 5% sodium hypochlorite (NaClO) with a few drops of tween 20 for 15 min and rinsed four times with sterile distilled water
Culture media: Two protocols were used in this study, first protocol (A) described by Jakse and Bohanes (2003) was followed in this study, whole flower buds or ovaries after excised from flower were cultured in Petri dish 100×15 mm were transferred to 7 cm plastic pots containing peat moss and covered with plastic cap having small hole in top for gaseous exchange for one weeks and we started remove the cap for one hour per days and increase uncovered time daily two, three and four hours for two weeks, the cap was removed
Summary
Spring onion (Allium fistulosum L.) is a biennial monocotyledonous plant is one of the most important vegetable crops around the world in terms of crop value. Spring onion is grown widely in Cameron Highlands in the Tringkap and Kea Farm areas (Ramasamy, 1991). The demand for specific varieties accommodate to local environment is too high since onion is a photoperiodically sensible plant and forms bulbs only after special environmental conditions among genotypes. According to the inbreeding method, two types of onion varieties are cultivated: hybrid varieties and open pollinated (Jakse and Bohanec, 2003). Onion ranking coming second after tomato in the list of vegetables cultured worldwide, It is estimated that over 90 million tons on 4.7 million ha (FAO., 2013). The production of F1 hybrids is considered one of the main goals in onion breeding program. The main restriction of this field is the length of the time needed to produce inbred
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