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以栓皮栎实生苗叶片为外植体诱导体细胞胚胎发生,调查碳水化合物、渗透剂和植物生长调节剂对体胚增殖、成熟和萌发的影响,建立体胚增殖、成熟和萌发的培养基方案。叶片外植体在附加1.0mg·L-1NAA和0.5mg·L-1BA的起始培养基上诱导形成前胚性团块。这些胚性团块在增殖培养基上培养6周,在附加1mg·L-1BA、0.25mg·L-1NAA和3%蔗糖的MS培养基上增殖效果最好。将单个体胚转接到成熟培养基上进行培养,蔗糖浓度对栓皮栎体胚成熟以及后续的萌发有显著影响。成熟培养基中附加5%的蔗糖,体胚成熟率和萌发率分别达到63.5%和33.8%。虽然在成熟培养基中附加ABA有利于体胚成熟,但对体胚的进一步萌发没有促进作用。为了提高萌发率,成熟体胚在附加植物生长调节剂的萌发培养基上进行培养,以及进行预冷处理。成熟体胚4°C冷处理没有促进胚根和上胚轴的发育萌发。在附加0.5mg·L-1BA和0.25mg·L-1IBA的1/2MS萌发培养基上,体胚萌发率达到65.9%,再生植株转化率达到9.4%。
Somatic embryogenesis was induced by leaves of Quercus variabilis seedlings. The effects of carbohydrates, osmotic agents and plant growth regulators on the proliferation, maturation and germination of somatic embryos were investigated. The medium for the proliferation, maturation and germination of somatic embryos was established . Leaf explants induced the formation of pre-embryogenic lumps on the initial medium supplemented with 1.0 mg · L-1 NAA and 0.5 mg · L-1 BA. These embryogenic lumps were cultured on proliferation medium for 6 weeks and proliferated best on MS medium supplemented with 1 mg · L -1 BA, 0.25 mg · L -1 NAA and 3% sucrose. Single embryos were transferred to mature medium for culture, sucrose concentration on Quercus variabilis somatic embryo maturation and subsequent germination have a significant impact. In mature medium supplemented with 5% sucrose, the rate of somatic embryo maturation and germination reached 63.5% and 33.8% respectively. Although ABA supplementation in mature media favored somatic embryo maturation, it did not promote further somatic embryogenesis. In order to increase the germination rate, mature somatic embryos are cultured on the germination medium supplemented with plant growth regulators and subjected to a precooling treatment. Cold treatment of mature somatic embryos at 4 ° C did not promote the germination of radicle and hypocotyls. On 1 / 2MS germination medium supplemented with 0.5mg · L-1BA and 0.25mg · L-1IBA, germination rate reached 65.9% and regeneration rate reached 9.4%.