Ca2+在茉莉酸甲酯诱导拟南芥气孔关闭中的信号转导作用

以拟南芥叶片下表皮为材料 ,分别用表皮生物分析法和激光扫描共聚焦显微镜成像技术 ,研究茉莉酸甲酯 (JA Me)促进气孔关闭过程中胞质Ca2 浓度的变化及其与气孔关闭的关系。结果表明 ,10 - 7到 10 - 3mol L的JA Me处理能促进拟南芥叶片的气孔关闭 ,其中 ,10 - 5mol L是最适浓度。用 10 - 5mol L的JA Me处理5min ,胞质Ca2 浓度从静息态的 10 5nmol L增加到 332 0nmol L ;质膜Ca2 通道阻断剂LaCl3和Ca2 螯合剂EGTA均能明显地降低JA Me对气孔关闭的促进作用。由此推测 ,胞质Ca2 可能是JA Me促进气孔关闭的重要信号转导因子     

木兰科植物种子采集与贮运技术

木兰科植物树型优美、花大芳香,用途广泛,深受人们喜爱。近年全国掀起了种植木兰科植物的热潮。但由于大家对木兰科植物种子的生物学特性缺乏全面了解,许多技术措施不当,造成大量烂种,损失不小。本人根据多年的工作经验,就种子采集处理、贮藏和运输中的几个技术问题,谈谈个人见解,供同行参考。     

cDNA-AFLP技术在植物表达分析上的应用

阐述了一种mRNA指纹图谱技术,即cDNA_AFLP技术的原理和程序; 该技术具有重现性高、准确可靠、效率高等特点,可广泛应用于植物遗传标记分析、基因表达特性研究以及分离植物基因等,具有良好的发展前景。     

黑杨萎蔫叶片挥发性物质的成分分析

采用水蒸气蒸馏法和气体吸附法提取黑杨萎蔫叶片的挥发性气味物质,用二氯甲烷做萃取剂,气相色谱_质谱联用技术分离和鉴定萃取到的气味物质的化学组分,结果表明,水蒸气蒸馏法共得到19个峰,鉴定出结构的有16个峰,未鉴明的3个峰;主要组分为4号峰的(Z)_3_己烯醇,相对含量为28.71%,其次为14号峰的邻羟基苯甲醛,相对含量为10.35%。气体吸附法共分离到20个峰,鉴定出结构的有17个峰;主要组分分别为6、7、8号峰的(E)_2_己烯醛、(Z)_3_己烯醇、(E)_2_己烯醇,相对含量分别为20.78%、29.12%、29.22%。两种方法所得到的相同组分为9种,相似性为46.15%。     

抑制差减杂交技术在细菌基因差异表达研究中的应用

同种细菌基因组差异及基因差异表达的鉴定与研究具有重要的分子生物学意义。某一菌株所具有的而另一菌株所缺乏的基因可能决定菌株重要的遗传特征。抑制差减杂交技术是近年来新建立起来的一种快速鉴别差异表达基因的新方法,本文就该方法的原理及在原核细菌中的应用作一综述。     

绵羊胞内单精子注射技术

In this study, the possibility of sheep transgenesis by intracytoplasmic sperm injection (ICSI) was assessed. In experiment 1, activation of ovine oocytes matured in vitro in preparation for ICSI has been investigated with 3.42 mmol/L Ca2+ treatment, ionomycin alone and ionomycin followed by 6-dimethylaminopurine (DMAP) after 3-h delay (group 1, 2 and 3, respectively). After activation, the oocytes were then cultured in SOFaaBSA medium. Cleavage rates were significantly (P<0.05) different among three groups (18.4%, 91.8% and 71.7%, respectively). In additional culture, no parthenotes in group 1, whereas 11% and 17.4% in group 2 and 3 developed to the blastocyst stage. Therefore we used the third activation method in the following ICSI tests. In experiment 2, development of ovine oocytes after ICSI was investigated. Thawed semen from two rams was separated by Percoll centrifugation and was used for ICSI or in vitro fertilization (IVF) trails. A total of 71.8% of oocytes reached the 2-cell stage following living sperm injection, which was significantly (p>0.05) different from those following IVF (41.4%) and sham-ICSI (30.2%). After seven days' culture, no sham-injected oocytes developed into the blastocyst stage, although 7% in ICSI and 16.1% in IVF-oocytes developed into the blastocyst stage, but there was no significant difference in ICSI and IVF groups (p>0.05). In the further study, the possibility of sheep transgenesis by ICSI was assessed. After coinjection of ovine oocytes matured in vitro with dead sperm cold to -20 degrees C and exogenous DNA encoding green fluorescent protein (GFP), seventy-three percent of coinjected oocytes developed to 2-cell stage (33/45) and two of them were transgene-expressing embryos. Among ten embryos at the 16-cell stage, all embryonic cells in one transgenic embryo still expressed GFP. Four coinjected blastocysts were thawed and transferred to the uterine of the two progesterone-synchronized recipient ewe. No pregnancies were detected on the 60th day. These results suggested sheep transgenic embryos could be produced by ICSI and further studies should be performed.