大麦胚性悬浮细胞系的研究

Studies on Embryogenic Cell Suspension Cultures of Barley

本文报道了2个均质分散的大麦胚性悬浮细胞系的建立程序及其生物学特性,并用干涉相差显微术和荧光显微术检测了悬浮细胞的质量和活力。悬浮细胞在改良CC培养基中培养2星期,其干重可以增加10倍,其干重加倍时间为3天,水解酪蛋白和L-脯氨酸能促进它的生长,接种细胞密度能显著影响生长、缩短继代周期,促进分裂。2个细胞系均能通过体细胞胚胎发生途径再生植株,但IM细胞系的植株再生能力远高于RG细胞系。随着继代次数的增加,2个细胞系均出现一定频率的染色体数目和结构的变异,染色体的变异程度与悬浮细胞的植株再生能力有一定的关系。建立的2个细胞系为大麦遗传操作和新技术育种提供了良好的实验材料。

Two fast-growing, well-dispersed and homogeneous cell suspension lines were established from embryogenic calli of two different barley varieties.The quality and viability of suspension cells obtained were carefully observed using interference phase contrast microscope and fluorescence microscope. Cell suspension cultures, maintained in modified CC medium, showed a 10—fold increase in dry weight after two weeks with a doubling time of about 3 days. Addition of L-proline and casein—hydrolysate in the medium had positive effect on the growth of cell cultures. Subculture interval could affect mitotic index. Both cell lines were able to regenerate plants by somatic embryogonesis. but cell line IM showed much higher regeneration capacity than cell line RG.Comparatively high efficiencies of variations in chromosome number and structure were found in both cell lines.but cell line RG showed much higher chromosomal abnormalities than IM.The correlation between karyotype and morphogenic capacity was noticed.