白皮松成熟合子胚的胚性愈伤组织诱导研究

Research on Induction of Embryogenic Callus from Mature Zygotic Embryo of Pinus bungeanaas

[目的]建立白皮松体细胞胚胎发生快速繁殖体系,诱导出胚性愈伤组织。[方法]以白皮松成熟合子胚为外植体,以DCR作为基本培养基,研究不同浓度激素对外植体发育和胚性愈伤组织得率的影响。同时在不同激素浓度下对培养4周的外植体外形以及胚性愈伤组织进行了观察与分析。[结果]培养基中加入2,4-D可以使得外植体生长减慢,并出现愈伤组织,6-BA则使外植体的子叶变长,膨大并玻璃化。当2,4-D浓度为8mg/L、6-BA浓度为2mg/L时,胚性愈伤组织的诱导效果较好,诱导率平均为40.625%,该愈伤组织为白色半透明状,细胞小而细胞质浓,说明高浓度2,4-D和低浓度6-BA有利于白皮松成熟胚的胚性愈伤组织的获得。[结论]合适的激素种类和浓度是诱导白皮松成熟胚产生胚性愈伤组织的关键因素。

[Objective] The study aimed to establish the rapid propagation system of Pinus bugeana through somatic embryogenesis so as to induce the embryogenic callus.[Method] With the mature zygotic embryo of P.bungeanaas as the explants and DCR as the basic medium,the effects of different hormone concn.on the development of the explants and the induction rate of embryogenic callus were studied and the observation and analysis were made on the shape of explant cultured for 4 weeks and the embryogenic callus under the different hormone concn.meanwhile.[Result] The observation on the explants showed adding 2,4-D in DCR medium could make the explants grow slower and produce the callus,while 6-BA could make the cotyledon of the explant become longer,expanding and vitrification.8 mg/L 2,4-D and 2 mg/L 6-BA could get better induction of embryogenic callus,with the induction rate of 40.625%.The embryogenic callus was white-translucent appearance and the cell was small with dense cytoplasm,showing that high concn.of 2,4-D and low concn.of 6-BA were beneficial to embryogenic callus induction.[Conclusion] The suitable hormone kinds and concn.were the key factors on inducing the embryogenic callus from the mature embryo of P.bungeanaas.