应用滋养和包埋技术进行籼稻原生质体培养和植株再生

Protoplast Culture and Plant Regeneration of Indica Rice by Nurse Culture and Encapsulation Technique

关键词:  籼稻  原生质体培养  植板率  植株再生

Key words:  indica rice  protoplast culture  plating efficiency  plant regeneration

摘要:

从几个籼稻品种不同发育阶段的幼穗分离出密度平均为2.2×10⁵个/mL~4.6×10⁵个/mL的原生质体。将这些原生质体包埋在藻酸钠颗粒里并分别在粳稻愈伤组织滋养下培养和没有愈伤组织滋养。在这两种培养条件下原生质体稳定,分裂频率分别达到48.4%和38.8%,群体存活率达到5.3%和4.7%,愈伤组织绿苗分化率达到71.6%和62.3%。这两种方法优于常用的琼脂糖包埋法和琼脂糖颗粒法。

Abstract:

Protoplasts with densities of 2.2×10⁵/mL to 4.6×10⁵/mL on average were isolated from different developing stages of young panicles of indica rice.The technique in which the isolated protoplasts encapsulated in sodium alginate beads and nursed on calli of japonica rice showed most efficiency. Another method of protoplasts encapsulated in sodium alginate beads without nurse feeder was also successful. Protoplasts were stable and their division and colony establishment in sodium alginate beads with nurse feeder were shown up to 48.4% and 5.3%,respectively, and without nurse feeder up to 38.8% and 4.7%, respectively.The both methods were superior to agarose entranpment and agarose bead methods with the latter being a standard method. The two successfully improved methods would be useful for gene transfer and other biotechnological studies in indica rice.