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柑橘愈伤组织是细胞融合、遗传转化等生物技术研究的良好材料。本实验室通过近20年的工作,先后诱导和保存了近80份不同种类和品种的柑橘愈伤组织。这些愈伤组织在MT基本培养基上继代保存。在长期保存过程中,部分材料仍然保持较好的胚胎发生能力。为研究愈伤组织在继代过程中的染色体变异趋势以及这种变异对胚胎发生能力的影响,本文选择有代表性的35份材料作为试材,在4年中连续3次使用流式细胞仪,对细胞DNA含量进行测定。结果发现,待测的35种愈伤组织均存在部分细胞DNA含量加倍的现象,而且,随着时间的推移,除佩奇橘柚、沙漠蒂甜橙、鲁斯脐橙、印度酸橘等4种愈伤组织的DNA含量加倍的细胞有减少的趋势外,其他愈伤组织的DNA含量加倍的细胞均有增加的趋势。数据分析表明,在待测的35种愈伤组织中,有71.4%的基因型的愈伤组织细胞出现了多倍体细胞增加的趋势。对这35种柑橘愈伤组织进行体细胞胚诱导,发现只有9种愈伤组织具有不同程度的胚胎发生能力,其他26种愈伤组织不能再生。相关性分析表明,柑橘愈伤组织细胞DNA含量增加的细胞比例与体细胞胚胎发生能力之间的相关性较小,相关系数为r=-0.10(P<0.01),未达到显著水平。另外,柑橘胚性愈伤组织继代时间与体细胞胚胎发生能力之间的相关性也不显著。
Citrus callus is a good material for biotechnology research such as cell fusion and genetic transformation. Nearly 20 years of work in our laboratory has induced and preserved nearly 80 different types and varieties of citrus calli. These callus were subcultured on MT minimal medium. In the long-term preservation process, some materials still maintain good embryogenic capacity. In order to study the chromosome variation of callus in the process of subculture and the effect of this variation on embryogenesis ability, 35 representative materials were chosen as the test material in this study. Flow cytometry , The cell DNA content was determined. The results showed that some of the 35 kinds of callus tested had the phenomenon of doubling the DNA content of some cells. Moreover, with the passage of time, petiole, orange peel, Cells with doubled DNA content in callus had a tendency to decrease, and those with doubled DNA content in other calluses tended to increase. Data analysis showed that among the 35 callus tested, 71.4% of the genotype callus cells tended to have polyploid cells increased. Somatic embryos were induced in these 35 kinds of citrus callus, and found that only 9 kinds of callus had different degrees of embryogenic ability, and the other 26 kinds of callus could not regenerate. Correlation analysis showed that the correlation between the proportion of cells with increased DNA content in callus and the ability of somatic embryogenesis was relatively small with a correlation coefficient of r = -0.10 (P <0.01), which did not reach the significant level. In addition, there was no significant correlation between citrus embryogenic callus subculture time and somatic embryogenesis ability.