锯叶棕体细胞胚胎发生及植株再生体系的建立

Somatic Embryogenesis and Establishment of Plant Regenerative System of Saw Palmetto

从锯叶棕成熟种子中成功实现体细胞胚胎发生和植株再生。成熟种子放在MS培养基上培养,加入0.15%活性炭,452μmol/L 2,4-D和14.7μmol/L N6-2i P,所有成熟合子胚培养5周后长出体胚簇,12周后,外植体转移到2,4-D浓度减到90.4μmol/L的MS培养基中进行体胚增殖,之后体胚转移到含有0.9,9μmol/L TDZ或没有生长调节剂的基本培养基中转化成小植株。9μmol/L TDZ对植株再生是无效的,然而,在0.9μmol/L TDZ中作次培养有12%胚胎能长成完整植株,没有生长调节剂中35%胚胎长出嫩枝,这些嫩枝转移到含有22.2μmol/L NAA的培养基中全部都会生根。picloram和2,4-D对锯叶棕幼苗愈伤的诱导效果最佳,6-BA次之,对子叶来说,picloram和2,4-D对愈伤的诱导起不到任何作用,而对茎尖的诱导效果最好。

Somatic embryogenesis and plant regeneration is achieved through saw palmetto（Serenoa repens）. Embryos are cultured on Skoog medium with 0.15%（w/V） activated charcoal, 452 μmol/L 2,4-dichlorophenoxyacetic acid（2,4-D） and 14.7μmol/L N6-（2-isopentenyl） adenine（2i P）. After culture initiation for 5 weeks, clusters of somatic embryos is developed. After 12 weeks, explants are transferred to the same medium with the amount of 2,4-D reduced to 90.4 μmol/L which resulted in somatic embryo proliferation. Somatic embryos are then transferred to the basal medium containing 0.9, 9 μmol/L thidiazuron（TDZ） or no growth regulator for conversion into plantlets. The 9 μmol/L TDZ treatment is ineffective for plant regeneration. However, 12% of the embryos subcultured on 0.9 μmol/L TDZ are able to produce complete plantlets. Shoot production was obtained from 35% of the embryos subcultured in the absence of growth regulators. Shoot production is obtained from 35% of the embryos subcultured in the absence of growth regulators. Rooting（100%） is achieved after these shoots are transferred into medium containing 22.2μmol/L α-naphthaleneacetic acid（NAA）. Picloram and 2,4-D is better for cal us induction of saw palm than 6-BA. Picloram and2,4-D treatment are ineffective for calus induction of cotyledon, but best for stem tip.