以未成熟合子胚为外植体,建立黑松体细胞胚胎发生和植株再生体系。研究基本培养基、2,4-D和6-BA对胚性愈伤组织诱导的影响,探讨ABA和PEG对体细胞胚成熟及植株再生的影响。结果表明: 在添加4 mg·L-12,4-D +2 mg·L-16-BA的DCR培养基上胚性愈伤组织诱导率为3.33%; 在添加30 mg·L-1 ABA +25 g·L-1 PEG的培养基DCR上,每克愈伤组织体细胞胚数量达72个,体细胞胚的萌发率为63.8%、植株转化率为43.5%,高质量的黑松体胚能提高体胚萌发率和植株转化率,再生植株1个月移栽成活率为50%。该研究可为抗病黑松的快速大规模繁殖提供一条新途径。
A somatic embryogenesis system from immature zygotic embryos of Pinus thunbergii was established. The effects of basic medium, 2,4-D and 6-BA on embryogenic callus induction were studied, and the effects of ABA and PEG on somatic embryo maturation and plant regeneration were investigated. The results showed that cultured on DCR medium supplemented with 4 mg·L-12,4-D and 2 mg·L-16-BA, the embryogenic callus induction rate was 3.33%; and on DCR medium containing 30 mg·L-1 ABA and 25 g·L-1 PEG, the number of somatic embryo per gram embryogenic callus was up to 72, and the somatic embryo germination rate was 63.8% and plantlet conversion rate from the germinated somatic embryos was 43.5%. High quality somatic embryos could improve the germination rate and plantlet conversion rate. Up to 50% of the transplanted plantlets were successfully survived one month in soil. This study would provide a new way for rapid and large-scale propagation of wilt-resistant P. thunbergii.
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