摘要
对苜蓿（品种农宝）的下胚轴外植体进行离体培养建立了再生体系。结果表明．子叶和下胚轴切段在附加0．2～1.0mg／LIAA和2．0mg／L6-BA或2．0mg／LKT的MS培养基上培养．均能诱导出愈伤组织．但子叶所诱导的愈伤组织不具有分化能力。下胚轴切段培养5～7d即可诱导出愈伤组织．并分化出绿色芽点．在原培养基上进而可分化出大量的丛生芽。在含0．2mg／L IAA和2．0mg／L6-BA的MS培养基上．其分化率最高可达到42．6％。这些芽在含0．2mg／LIAA和0．5mg／LKT的MS培养基上长成2cm左右的幼苗时。将其切下转至1／2MSo培养基上，培养10d即可诱导生根．得到再生植株。随机选取实生苗和下胚轴愈伤组织再生苗进行RAPD分析．结果表明，所用的50个随机引物中有5个扩增出差异性条带，甚至再生植株之间也有1条引物扩增的条带有差异．这说明经组织培养所得到的再生植株与实生苗相比。在DNA水平上发生了一定程度变异。并且这种变异也存在于再生植株之间。
A regeneration system was established from hypoeotyl explants of Medicago sativa eultivar Nongbao. The calli were easily induced from the hypocotyl and cotyledon segments cultured on MS medium containing 0.2 -1.0 mg/L IAA, 2.0 mg/L 6-BA or 2.0 mg/L KT. The calli induced from cotyledon segments had no differentiation eapability; while many green buds emerged from hypoeotyl segments after culturing for 5-7 d and developed into a number of clusters of young shoots. The highest differentiation frequency of buds was up to 42.6% on the MS medium containing 0.2 mg/L IAA and 2.0 mg/L 6-BA. When the shoots had elongated to about 2 cm on an MS medium with 0.2 mg/L IAA and 0.5 mg/L KT, they were cut and transferred 1/2 MSo medium. The roots were produced 10 d later. Five of fifty random primers amplified DNA polymorphism bands in RAPD analysis of seed-derived seedlings and of in vitro regenerated plantlets taken at random. One primer even amplified the difference between the regenerated plants. This suggested that there was some variation in DNA in plants of M. sativa regenerated in vitro.
出处
《草业学报》
CSCD
2006年第5期115-121,共7页
Acta Prataculturae Sinica
基金
陕西省自然科学研究计划项目(2004C110)
陕西省教育厅专项科研基金(JH04230)资助
关键词
紫花苜蓿
子叶
下胚轴
再生植株
RAPD
Medicago sativa
cotyledon
hypocotyl
plant regeneration
RAPD
