高产人参寡糖素培养细胞变异克隆系的筛选

SCREENIG OF VARIANT CLONE LINE WITH HIGH YIELDING OLIGOSACCHARIN FROM SUPENSION CULTURE CELL OF PANAX GINSENG

用２ｍｍｏｌ／Ｌ的ＭＮＮＧ处理经过滤的人参悬浮培养细胞１小时后，细胞存活率下降显著，细胞克隆植板率只是对照组的１０．１２％。经细胞平板克隆共获得克隆系１５１株，其中很多克隆系在转移培养中生长缓慢，甚至不生长而死亡。经分析可供测定的克隆系生长和寡糖素含量的差异，对１１株寡糖素含量较高克隆系经连续１０代继代培养观察，选出一株稳定高产人参寡糖素优良克隆系ＰＧＭＢ－３７，其平均生长速率是０．５５８ｇＤＷＬ￣（－１）ｄ￣（－１）为亲本的１．５倍，平均寡糖素含量是１４．６７％ＤＷ，平均寡糖素产率是２Ａ５６ｇ／Ｌ，分别比亲本高７０％和１５６％，并且它的过氧化物酶同工酶谱特征与亲本之间也呈稳定性差异。

The suspension cells of Panax ginseng which had been filtrated,were treated with mutagenic compound,N-methyl-N-nitro-N-nitrosoguanidine at 2 mmol/ L,for 1 hour.Cell viability was remarkablely decreased and the formation of cell clone was only 10.12% of control. Cells treated were plated on a nutrient agar medium. More than 150 clone lines were obtained af-ter plating culture of 60 days, of which many clone lines grew slow or stopped growing and died during transplantation culture. On the light of the stability of 11 clone lines with high yield of oligosaccharin obtained on their differences in eell growth and oligosaccharin content among all clone lines picked out petri dishes, a stable clone line PGMB-37 with high yield of oligosaccharin had been selected through 10 generations of successive subculture. The mean growth rate of clone line PGMB-37 was 0.558 gDWL￣(-1)d￣(-1),and was about 1.5-fold to its parent,and its mean oligosaccharin content and yield were 1 4.67 % DW and 2.456 g / L,which were near 70 % and 156 % higher than those of its parent,respectively.The patterns of isoperoxidases of clone line PGMB-37 were different from those of its parent,and these differences were also stable during successive subculture.