放线菌素D和环已亚胺对伊贝母胚性愈伤组织中胚性细胞团和球形胚发生及大分子代谢动态的影响  

The Influence of Actinonycin D and Cyloheximide on the Formation of the Masses of Embryogenic Cells and the Globular Embryoes and on Metabolic Kinesis of Macromolecule in Embryogenic Calli of F. pallidiflora Schrenk

作　　者：王仑山[1] 丁惠宾 林洁云 

机构地区：[1]兰州大学生物系,730000

出　　处：《Acta Genetica Sinica》1993年第2期159-166,共8页

基　　金：国家自然科学基金

摘　　要：伊贝母(F.pallidiflora Schrenk)胚性愈伤组织接种于NAA 1.0mg/L+6-BA2.0 mg/L的MS培养基上,在培养10天前可产生大量单细胞到多细胞胚性细胞团,培养10至15天,逐渐形成大量球形胚。利用这样一个实验体系,在培养0、1、2、3和4天后加入放线菌素D(AMD,20μg/ml)和环己亚胺(CHM,20μg/ml),继续培养至第6天,分析大分子代谢动态和观察胚性细胞团的形成情况;培养6和10天后加入同样浓度的AMD和CHM。继续培养至第15天,分析大分子代谢动态及观察球形胚形成情况。结果表明:(1)培养0、1、2、3和4天加入AMD的分别抑制胚性细胞团的100%、63%和45%,加入CHM的抑制100%、85%和75%,培养6和10天后加入CHM抑制球形胚的100%和75%;(2)DNA、RNA和蛋白质在胚性细胞团和球形胚形成时出现两个峰值,其中RNA变化剧烈,最早出现峰值。AMD和CHM分别抑制RNA和蛋白质的合成;(3)过氧化物酶同工酶带在胚性细胞团和球形胚形成过程中顺序表达,AMD和CHM分别在转录和转译水平上对其进行规律性抑制。根据以上结果,本文对伊贝母体细胞胚胎发生的机制进行了初步讨论。The embryogenic calli of F. pallidiflora Schrenk cultured on MS medium supplemented with MAA(1.0mg/L)+6-BA(2.0mg/L) produced a great number of the embryogenic cellular masses ranging from single cell to mudicells after 10 days in culture. If it was cultured for 10 to 15 days, a great number of the globular embryoes formed. Using this experimental system, AMD(20mg/ml) and CHM(20mg/ml) were added to the medium after cultured 0, 1, 2, 3, and 4 days respectively. When it went on culture for 6 days, the metabolic kinesis of ma-cromolecule was analysed and the formation of the masses of embryogenic cells was observed; another group of experiment is that after the subculture was cultured for 6 days and 10 days respectively, same concentration of AMD and CHM were added to the medium, when the culture continued for 15 days the metabolic kinesis of macromolecule was analysed the formation of the globular embryo was observed. The results are as follows: (1) Treatment with AMD at 0, 1, 2, 3 and 4 days inhibited 100%, 63% and 45% of the masses of embryogenic cells respectively. CHM inhibited 100%, 85% and 75% respectively. Treatment with CHM at 6 and 10 days inhibited 100% and 75% of the globular embryoes respectively. (2) DNA, RNA and protein appeared a two-peak curve during the formation of the masses of emhryogenic cells and globular embryoes respectively. The change of RNA is marked. The first peak curve of RNA appeared 3 days after the culture. AMD and CHM inhibited synthesis of RNA and protein respectively. (3) The peroxidase isoenzyme expressed in proper order in the course of the formation of the masses of embryogenic cells and the globular embryoes. Inhibition of AMD and CHM imposed on transcription and translation respectively.

关 键 词：伊贝母 放线菌素D 体细胞胚发生 

分 类 号：Q78[生物学—分子生物学]