速生湿地松良种胚性愈伤组织诱导与增殖

Induction and proliferation of embryogenic callus on improved varieties of fast-growing Pinus elliottii

关键词:  湿地松, 未成熟合子胚, 体细胞胚胎发生, 胚性愈伤组织

Key words:  Pinus elliottii, immature zygotic embryo, somatic embryogenesis, embryonic callus

摘要:

为使速生湿地松良种快速大规模繁殖,对其胚性愈伤组织进行诱导和增殖优化研究。该文以1代湿地松种子园中10个速生湿地松优良无性系(基因型)的未成熟合子胚为外植体,系统研究基因型、合子胚发育阶段、基本培养基、植物生长调节剂种类和浓度等不同因子对胚性愈伤组织诱导效率的影响,探讨胚性愈伤组织的增殖条件。结果表明:基因型、合子胚发育阶段、基本培养基及植物生长调节剂种类和浓度等均对胚性愈伤组织诱导有不同程度的影响。10个基因型均诱导出了胚性愈伤组织,其中以2号基因型诱导率最高,达到25.37%; 合子胚发育阶段中,以处于多胚阶段的球果诱导率最高; 四种基本培养基中,以在DCR基本培养基上诱导效果最佳; 植物生长调节剂浓度及组合中以2,4-D 2.0 mg·L^-1 + KT 2.5 mg·L^-1诱导率最高,达到27.78%。湿地松胚性愈伤组织增殖的最佳培养基为在DCR基本培养基上添加0.5 mg·L^-1 2,4-D、1.0 mg·L^-1 KT、500 mg·L^-1 CH和300 mg·L^-1 L-谷氨酰胺。综上研究结果,为湿地松良种进一步开展成熟胚诱导及植株再生奠定了基础。

Abstract:

In order to make the rapid and large-scale propagation of fast-growing varieties of Pinus elliottii, the induction and proliferation of embryogenic callus were studied. The immature zygotic embryos(with endosperm)of ten P. elliottii families were used to systematically study the effects of different factors on the induction efficiency of embryogenic callus, including genotype, the developmental stage of zygotic embryos, basic medium, types and concentrations of plant growth regulator(PGR). At the same time, the proliferation conditions of embryogenic callus were discussed. The results were as follows: The genotype, the developmental stage of zygotic embryos, basic medium and types and concentrations of plant growth regulator(PGR)all had different degrees of influence on the induction of embryogenic callus. Embryogenic callus were induced in all ten genotypes, of which the induction rate of Genotype 2 was the highest, reaching 25.37%; In the stage of zygotic embryo development, the induction rate of cones at the multiple embryo stage was the highest; Among the four basic media, the induction effect on DCR basic medium was the best; The induction rate of 2,4-D 2.0 mg·L^-1+KT 2.5 mg·L^-1 was the highest, reached 27.78%. The proliferation of embryogenic callus for P. elliottii was achieved on DCR medium, supplemented with 2,4-D(0.5 mg·L^-1), KT(1.0 mg·L^-1), CH(500 mg·L^-1)and Gln(300 mg·L^-1). This study laid a foundation for further developing mature embryo induction and plant regeneration of fast-growing varieties of P. elliottii.