Ti质粒T区tms基因的分离及其对高等植物分化的影响

根癌农杆菌Ti质粒的T区DNA带有致瘤基因,其基因1和基因2编码生长素吲哚乙酸生物合成途径中的两个酶。以pGV 354(pBR322质粒中插有Ti质粒C 58 T区DNA的HindⅢ15—HindⅢ22大片段)重组质粒出发,我们分离了基因1和基因2,并构建了带有卡那霉素抗性基因的重组质粒pBZ 692,通过基因载体pGV 3850,我们将基因1和基因2引入了高等植物。结果证明基因1和基因2能促使烟草、向日葵、土豆等转化组织分化长根,转化的根在MS_0培养基上能脱分化形成愈伤组织并自主生长,在转化的组织中有转化标记胭脂碱的存在。     

龙葵抗阿特拉津生物型叶绿体基因文库的建立

用对阿特拉津(Atrazine)除草剂抗性的龙葵生物型B_(12)株系作材料,制备叶绿体DNA。B_(12)株ctDNA(叶绿体DNA)经BamHI酶解,在0.7％琼脂糖凝胶电泳上呈现24条带,其中最大的片段为18.6kb,最小的片段为1kb。用pBR322作为载体,构建B_(12)株ctDNA BamHI片段文库。通过与探针的分子杂交,从中筛选出含有编码叶绿体32kd蛋白质的阿特拉津抗性基因的克隆pSB135和含有ATP合酶α亚单位基因的克隆pSB132。     

De novo synthesis of 3'-nucleotidase in germinating wheat embryo

The enzyme 3′-AMP nucleotidase was purified 2,500- to 5,000-fold from extracts of an acetone powder of wheat (Triticum aestivum) embryonic axes germinated for 40 hours. Sodium dodecyl sulfate acrylamide gel electrophoresis and chromatography on Biogel-P100 indicate that the enzyme is monomeric with a molecular weight of 39,000. Extracts of embryos germinated up to 6 hours have only 1% of the 40-hour level of enzyme activity. To see if the increase to 40 hours represents de novo synthesis, extracts were compared for their ability to react with a rabbit antibody prepared against the enzyme. In immunodiffusion tests, 40-hour extracts showed a strong precipitin line coincident with that of the purified enzyme, whereas no precipitation was observed with 1-hour extracts. When the enzyme present in 40-hour extracts was partially inactivated by EDTA, it still blocked the ability of the antibody to inhibit enzyme activity. Extracts of 1-hour embryos, in contrast, were not able to block the inhibitory activity of the antibody. Embryos allowed to take up ³⁵SO₄ between 40 and 46 hours of germination synthesized ³⁵S-labeled 3′-nucleotidase. In contrast, no radioactive protein synthesized by embryos during the first 6 hours of germination coincided on gel electrophoresis with the enzyme. These results indicate that the increase in 3′-nucleotidase activity is a consequence of de novo synthesis of the enzyme.     

大叶杨配囊及胚珠的形成和发育

本文应用细胞化学方法研究了大叶杨胚珠、胚囊的形成和发育过程中核酸、蛋白质及不溶性多糖的分布和消长。大孢子母细胞、大孢子四分体及功能大孢子中含较少不溶性多糖,但却含丰富的RNA和蛋白质。功能大孢子经分裂发育成八核的蓼型胚囊。四核胚囊开始积累细胞质多糖,成熟胚囊中除反足细胞外充满淀粉粒。反足细胞形成后不久即退化。助细胞具多糖性质的丝状器,受精前两个助细胞退化。卵细胞核对Feulgen反应呈负反应。二极核受精前由胚囊中部移向卵器,与卵器接触后融合形成次生核。发育早期的胚珠为厚珠心,双珠被。晚期,内珠被退化,故成熟胚珠为单珠被。四核胚囊时期,珠孔端珠心组织退化,胚囊伸向珠孔形成胚囊喙。合点端珠心组织含丰富的蛋白质和核酸,这一性质与绒毡层性质相似,可能涉及胚囊的营养运输。胚囊的营养来源于子房和胎座细胞内贮存的淀粉粒。     

黄山贝母和天目贝母的核型研究

本文首次报道了产于江西的黄山贝母(F.huangshanensis Y. K. Yanget C. J. Wu)和浙江的天目贝母(F. monantha Migo)的核型:前者为2n=24=2m 2sm 6st(4sc) 14t;后者为2n=24=2m 2sm 6st(4sc) 14t(2sc)。此外,与国内报道的其它贝母属植物的核型进行了比较和讨论。     

HBsAg重组DNA转化中国地鼠卵巢细胞二氢叶酸还原酶阴性突变株的细胞染色体变化及致瘤性

整合了含乙型肝炎病毒表面抗原(HBsAg)基因和dhfr基因的CHO-dhfr~-细胞,其染色体的畸变率和畸变类型都比亲代CHO-dhfr~-细胞高。但转化前后两系细胞的重要特性都未发生变异,即两者的染色体总数无差别,都是20条。两系细胞株接种裸鼠,均未发现有致瘤性。