抗松针褐斑病湿地松体细胞的悬浮培养

Suspension Culture of Somatic Cells of Pinus elliottii Against Brown Spot Needle Blight of Pine

为了获得大量的抗松针褐斑病胚性愈伤组织试验材料,完善抗性湿地松体胚发生技术,以抗松针褐斑病湿地松胚性愈伤组织为材料,探究抗性湿地松体细胞悬浮培养的较适条件,建立抗性湿地松体细胞的悬浮培养体系。研究结果显示:悬浮培养的胚性细胞较分散,利于胚性细胞充分获取营养,可产成熟体胚153个/g,而固体培养的胚性细胞产胚仅为56个/g。抗性湿地松悬浮细胞的较佳培养周期为7-9 d,较适宜的基本培养基为1/4 DCR,麦芽糖添加量为15 g/L,NAA添加量为0.5 mg/L;较佳的继代添加比为1/2;同时研究还发现不同基因型的抗性湿地松悬浮细胞在相同的培养条件下,细胞活力有一定差异,其中无性系13#-9活力最佳。

In order to obtain a large number of embryogenic calluses of Pinus elliottii being resistant to brown spot needle blight of pine and to improve the technique of somatic embryogenesis from the resistant embryogenic callus,the suitable conditions of somatic cell suspension culture of P.elliottii were studied,and the suspension culture system of somatic cells of P.elliottii resistant to brown spot needle blight of pine was established.The results showed that suspension culture of embryogenic cells was more dispersed,which was conducive for embryonic cells to have full nutrition,and 153 mature somatic embryos per gram was produced,the embryogenic cells produced in solid culture per gram were only 56.The optimal culture period of suspension cells of resistant P.elliottii was 7-9 d,the suitable basic medium was 1/4 DCR,the dosage of maltose was 15 g/L,NAA was 0.5 mg/L,and the optimal subculture addition ratio was 1/2.Meanwhile under the same culture conditions,the viabilities of suspension cells of resistant P.elliottii in different genotypes varied,and that of clone 13#-9 was the best in this experiment.