Plant material and culture conditions have been described (7). Briefly, a diploid tissue culture line, WOO1C, of wild carrot, Daucus carota L., was maintained in Murashige and Skoog medium supplemented with 0.1 mg of 2,4-dichlorophenoxy... Plant material and culture
conditions have been described (7). Briefly, a diploid tissue
culture line, WOO1C, of wild carrot, Daucus carota L., was
maintained in Murashige and Skoog medium supplemented
with 0.1 mg of 2,4-dichlorophenoxyacetic acid (2,4-D). To regenerate
plants from the culture, callus tissue was transferred
to the same medium devoid of 2,4-D (embryogenic medium).
All experiments were performed with cultures grown in liquid
medium. For convenience, cultures grown in 2,4-D-containing
medium are referred to as "callus cultures", and those grown
in medium without 2,4-D as "embryo cultures."
The growth of callus cultures was measured by the sidearmturbidity
method (7). The number of cells at any time point of
growth was estimated from turbidity in a Klett-Summerson
calorimeter and was expressed in arbitrary units. For example,
Klett 100 corresponds to ="2 X 106 cells per ml. The relationship
is linear up to Klett 150 in our Klett-Summerson colorimeter.
Suspension cultures ofWOOLC cell line are normally maintained
at cell densities between 106 and 107 cells per ml in 0.1
mg of 2,4-D per liter (high-density culture) in shake flasks.
During a quantitative study on embryogenesis, we found that
maximal embryo production can be achieved by first incubating
the culture at high density in medium without 2,4-D for one
generation time and then diluting it to 2-3 x 104 cells per ml
(low-density culture). To compare cultures of comparable density,
callus cultures and embryo cultures were subjected to the
same procedure. To initiate low-density embryo or callus cultures,
an 8-day-old high-density culture grown at the logarithmic
phase was washed three times with fresh callus or embryogenic
medium and resuspended at 8 x 105 cells per ml for one
generation (3 days) in its corresponding medium. It was then
diluted to 2 x 104 cells per ml; 20 ml of the culture was incubated
in a plastic Petri dish (9 cm in diameter) at 240C. The
morphogenetic events of the cultures were examined and photographed
under a dissecting microscope.
请不要用google的那个,我看不懂~~
植物材料与文化
条件已被描述( 7 ) 。简单来说,二倍体组织
文化线, woo1c ,野生萝卜,胡萝卜,是
保持murashige和skoog培养基
用0.1毫克的2,4 -二氯苯氧乙酸( 2,4 -四) 。再生
植物从文化,愈伤组织被移送
到同一个中等空洞2,4三维(胚中等) 。
所有实验均与文化成长在液体
中等。为方便起见,文化的成长,在2,4三维含
中等称为"愈伤组织培养" ,而那些成长
在培养基2,4三维"胚胎文化" 。
生长愈伤组织培养,是衡量该sidearmturbidity
法( 7 ) 。细胞数量在任何时间点
增长估计从浊度在klett -萨默森
热量表,并有人表示,在任意单位。举例来说,
klett 100对应= " 2 × 106细胞每毫升。关系
是直线上升,以klett 150我们klett -萨默森色。
悬浮培养ofwoolc细胞系通常保持
在细胞密度之间的106和107细胞每毫升0.1
毫克的2,4三维每公升(高密度文化) ,在摇瓶。
在定量研究胚胎发生后,我们发现
Z大的胚胎生产,可以取得diyi孵化
文化,在高密度的媒体2,4三维1
一代人的时间,然后再稀释至2-3 x 104个细胞中每毫升
(低密度文化) 。比较文化比较的密度,
愈伤组织培养及胚胎文化遭到了
同样的程序。发起低密度胚或愈伤组织培养,
为期8天的岁高密度培养的速度增长,对数
diyi阶段被冲至3倍,与新鲜愈伤组织或胚
中型及悬浮于8 × 105个细胞每毫升1
一代人( 3天)在其相应的媒介。当时
稀释至2 × 104个细胞,每毫升, 20毫升的文化孵育
在一个塑料皮氏培养皿( 9公分) ,在240c 。该
形态发生的事件,其文化的人进行检查和拍照
根据解剖显微镜