【作者】 高娃；

【导师】 白玉娥； 叶冬梅；

【作者基本信息】 内蒙古农业大学 ， 森林培育， 2008， 硕士

【摘要】 论文以沙地云杉成熟合子胚为材料,研究了沙地云杉体细胞胚胎发生过程中的生理生化变化,着重探讨了多胺在体胚发生中的作用机理。1.首先对沙地云杉体细胞胚发生系统的优化培养条件进行了研究表明:诱导初始愈伤组织的最佳培养基为;SH +2,4-D(0.1㎎/L)+6-BA(0.1㎎/L);分化胚性愈伤组织的最佳培养基为; LS+ABA(5.0mg/L)+PEG4000(50.0g/L)加入50.0g/LPEG4000可改善胚性愈伤组织的质量,诱导出的胚性愈伤组织黄绿色、色泽亮丽、松散、颗粒状、大小均匀一致。胚性愈伤组织诱导周期为60d,第50d左右需继代扩繁。2.沙地云杉胚性愈伤组织体细胞胚胎发生过程中,SOD酶活性较高,而POD活性较低。附加外源Put促进体细胞胚发生过程中POD酶活性上升,SOD酶活性降低。附加多胺抑制剂时鱼雷胚时期的SOD酶活性较高,随着胚的成熟SOD酶活性逐渐下降,而POD酶活性一直保持上升状态。3.体细胞胚胎发生初期(多细胞原胚时期)及成熟阶段(球形胚、心形胚、鱼雷胚及子叶胚时期)内源PA含量不同;附加30mg/L的外源Put可以提高内源PA的含量,促进球形胚的发生。特别是对内源Put和Spm的影响较大。附加外源Put后球形胚时期内源PA含量明显增加,但体细胞胚成熟阶段内源PA含量下降。维持较高水平的Put/Spd和Put/Spm是细胞柄团前期发育所需要的,但不利于体细胞胚成熟。4.CHA和MGBG 2种抑制剂处理的结果基本相似。单独附加加抑制剂(CHA或MGBG)时,只对内源Spd有抑制作用,增加内源Spm和Put含量;多胺抑制剂和外源Put同时处理时,明显降低内源Spm和Put含量。因而判断了抑制剂(CHA或MGBG)和Put同时处理时才对内源Spm和Put含量起到抑制作用。单独附加抑制剂(CHA或MGBG)时,是通过抑制内源Spd而降低总多胺含量。更多还原

【Abstract】 The physiological and biochemical changes were studied during somatic embryogenesis in Picea mongolica (W.D.Xu) mature zygote embryo, and roles of polyamines on somatic embryogenesis and the mechanism were investigated.1. The conditions for somatic embryogenesis in Asparagus were optimized. The most suitable medium were: SH+2,4-D(0.1㎎/L)+6-BA (0.1㎎/L)for primary calluses induction and LS+ABA(5.0 mg/L)+PEG4000(50.0g/L) for embryogenic calluses induction. Whitish-yellow, bright, fresh, incompact and coincident particle embryogenic calli were obtained on medium supplemented with 50.0 g/LPEG4000 putrescine. Embryogenic calli can be induced within 60 days, and calli need be transfered to fresh medium within 50 days for subculture.2. Our test result showed that: The SOD activity is high, but the POD activity is low in process of embryogenic calli somatic embryogenesis in Picea mongolica(W.D.Xu).The POD enzyme activity is rising,the SOD activity is reducing when add exogenous hormone Put in the process of somatic embryogensis.when add the inhibitor the Torpedo-shaped embryo times SOD activity is high,the SOD activeness is low with mature of the embryo.The POD activeness maintains at the rise when the process of similarly inhibitor.3. It is different that the endogene PA content of the somatic embryogenesis and the mature each stage.(Globular embryo,Torpedo-shaped embryo,Heart-shaped embryoid and Cotyledon embryo) 30mg/L exogenous hormone Put may enhance endogene PA’s content.The endogene PA’s content increases obviously in the sphere embryogenesis times when add exogenous hormone Put.The somatic embryogensis mature period endogene PA’s content drops. It is need that maintenance high level’s Put/Spd and Put/Spm are clavate pre-embryo growth,does not favor to the somatic cell embryo to be mature.4. 2 inhibitor (CHA and MGBG) process analysis result basic similar. When independent increase inhibitor (CHA or MGBG), only has the inhibitory action to endogene Spd, increases endogene Spm and the Put content; When with the Put simultaneous working, reduces endogene Spm and the Put content obviously. Thus the judgment inhibitor (CHA or MGBG) and the Put simultaneous working is only then plays the inhibitory action to endogene Spm and the Put content. The independent inhibitor (CHA or MGBG) process, through suppresses endogene Spd to reduce the total multi-amine content.更多还原