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题名: 第一部分:金花茶胚状体中脱落酸和玉米素核苷的高压液相色谱测定及ABA对胚状体发生的影响。第二部分:金花茶体细胞胚发育的超微结构的观察
作者: 彭艳华
学位类别: 硕士
答辩日期: 1988
授予单位: 中国科学院昆明植物研究所
授予地点: 中国科学院昆明植物研究所
导师: 庄承纪
中文摘要: 用HPLC法测定了金花茶(Camellia Chrysantha(Hu)Tuyama)胚状体形成的继代培养的两类材料(I:具有正常产生次级胚能力;II:产生次级胚能力极差)的ABA和ZR含量。并以ABA为外源激素进行胚状体发生的培养。HPLC柱材料为Zorbax ODSS.ABA以冰醋酸--乙醇--水溶液(1:78:22)作流动相,保留时间4.33;ZR以加几滴硫酸钠(0.05M)的乙脯-甲醇-水溶液(10:70:30)作流动相,保留时间为5.15.HPLC之前的提、初纯化的回收率:ABA为64.8%;ZR为57.3%。测得结果,第I类材料玉米素核苷为2.69ng/g.fWt,ABA为69.55ng/g.fWt;第二类材料ZR为2.44ng/g.fWt,ABA为10.72ng/g.fWt。在含ABA(0.1μM、1μM、10μM、50μM)的培养基上,胚状体形成频率、数目、鲜重增加均优于对照。而且随ABA浓度升高胚状体形成频率和数目增高,但鲜重增加降低。而后材料转到不含激素的培养基上胚状体生长发育良好,并且不同浓度的ABA处理之间鲜重几乎没有差别。观察了金花茶(Camellia Chrysantha(Hu)Tuyama)胚状体发育各阶段的超微结构。前胚期及球形期细胞原生质丰富,线粒体、高尔基体、内质网数目多,液泡少而小且散生。随发育时期的推时,细胞器数目减少,而液泡化程度增高。心形期细胞壁内出现许多形状不一、大小不等的小泡结构,细胞壁电子密度不均匀。鱼雷期细胞壁内小泡结构较均等,而且胞间连丝极为丰富,此阶段开始出现淀粉粒。子叶期淀 粉粒相当普遍,一般成堆靠近细胞壁或胞核附近,叶绿体中也可见较多的淀粉粒、小油滴,虽然有基质片层结构,但无基粒片层结构。讨论了结构变化与细胞分化、胚状体发育的关系。
英文摘要: A EPLG method was used to aquantify ABA and ZR in two kind materials of somatic embryogenesis in Camellia Chrysantha (HU) Tuyama. Among the materials, one can normally produce the secondary embryos, the other can do it litte. Furthermore, the culture of the somatic embryogenesis was undertaken in the medium with ABA (0.1μm; 1μm; 10μm; 50μm.). ABA and ZR were determinated by HPLC on coium packed with Zorbax ODS. Of those, the former was completed under the mobile phase, a mixed solution of glacial acid--menthancl--water (1:78:22); flow rate, 0.6ml/min; wave length 254nm; detector, spd-1. The latter, mobile phase, a with a few drops Na2SO4 (0.05m) mixed solution of acetonitrile--enthanol--water (10:70:30) flow rate, 0.9ml/min; wave length 270nm, detector, SPD-1. External standard method was used. Recovery rates (External standard method) of ABA and ZR before HPLC was resp--ectively 64.8% and 57.3%. The determinated results as follows: the first material, the contend of ZR is 2.69ng/g.fwt, and ABA 69.55ng/g.fwt; the second material, ZR is 2.4ng/g.fwt, and ABA10.72ng/g.fwt. The frequency and unmber of somatic embryogenesis in the medium with ABA were higher than those in the contrast, so was the fresh weight. Noreover, the concertration of ABA increased in posstitive proportion to the frequency and unmber, but inverse dependence on the fresh weight. After 35 days, the materials transfered to the medium without hormones the somatic embryiods developed well. No difference was found among treated by a series of concentra-tion of ABA. The present experiment probably indicated that in somatic embryogenesis ZR is not indispensable but ABA is promotive.
语种: 中文
内容类型: 学位论文
URI标识: http://ir.kib.ac.cn/handle/151853/834
Appears in Collections:昆明植物所硕博研究生毕业学位论文_学位论文

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