太子参试管微块根形成及其应用初探

太子参组培苗单茎节段插入含45g·L^-1蔗糖的MS培养基中,于60μmol·m^-2·s^-1光照强度下培养120d后,其节部位长出不定根膨大形成与太子参体内块根相似、重量约为块根1/20的微块根,平均每株可形成1.54个微块根。微块根移植入大田后,其生长特性和物候期与大田块根相似,都有低温萌芽、高温休眠的特性;开花结果、不定根膨大、块根成熟等也相似。同时,组培中的微块根还具有易培养、耐贮存、体积小、易移栽和繁殖系数高等优点,在生产中具有很大的应用潜力。     

基因工程综合实训教学改革与创新

为培养学生的岗位职业能力,实现与岗位零对接,基因工程综合实训以工作任务为线索确定教学内容,以产品生产流程为载体设计教学过程,以职业能力为依据改革教学方法,以发展性评价为手段,强化技能形成。实践证明,"基因工程综合实训"教学改革,有利于培养学生的动手能力和实践创新能力,有利于促进以职业能力培养为核心的教育。     

蓝猪耳离体受精初试

用体内-体外方法分离了蓝猪耳精细胞。用酶解和解剖方法分离了其成熟卵细胞。分离的精、卵细胞用电融合介导尝试了体外诱导融合。在合适的渗透压（6％甘露醇）和合适的氯化钙（0．04％CaCl2·2H2O）溶液中,用交流电场为30～35V10-25s使精、卵细胞排队;用直流电场400～600V45-50μs65脉冲穿孔条件可诱导30％的精、卵细胞融合和70％以上的卵细胞之间的融合。尝试了人工合子的单细胞培养但未获成功。诱导蓝猪耳精、卵细胞融合的条件与玉米和水稻不同。     

金线莲外植体筛选及愈伤组织诱导研究

以金线莲茎段、叶片、茎片、不定芽为试材,分别在添加6-BA、ZT、NAA、KT 5个不同处理的MS及1/2MS培养基上培养,诱导愈伤组织。结果表明,以茎段、茎片、不定芽为外植体,在MS + 6-BA 2.0 mg/L + NAA0.5 mg/L、MS + NAA 2.0 mg/L + KT 0.1 mg/L和MS + 6-BA 2.0 mg/L + NAA 0.5 mg/L + ZT 0.2 mg/L培养基中培养,均能成功诱导愈伤组织。不定芽为诱导愈伤组织最佳外植体,最佳培养基为MS + 6-BA 2.0 mg/L + NAA 0.5 mg/L + KT 0.2 mg/L。     

Molecular Analysis of Atypical Family 18 Chitinase from Fujian Oyster Crassostrea angulata and Its Physiological Role in the Digestive System

Chitinolytic enzymes have an important physiological significance in immune and digestive systems in plants and animals, but chitinase has not been identified as having a role in the digestive system in molluscan. In our study, a novel chitinase homologue, named Ca-Chit, has been cloned and characterized as the oyster Crassostrea angulate. The 3998bp full-length cDNA of Ca-Chit consisted of 23bp 5-UTR, 3288 ORF and 688bp 3-UTR. The deduced amino acids sequence shares homologue with the chitinase of family 18. The molecular weight of the protein was predicted to be 119.389 kDa, with a pI of 6.74. The Ca-Chit protein was a modular enzyme composed of a glycosyl hydrolase family 18 domain, threonine-rich region profile and a putative membrane anchor domain. Gene expression profiles monitored by quantitative RT-PCR in different adult tissues showed that the mRNA of Ca-Chit expressed markedly higher visceral mass than any other tissues. The results of the whole mount in-situ hybridization displayed that Ca-Chit starts to express the visceral mass of D-veliger larvae and then the digestive gland forms a crystalline structure during larval development. Furthermore, the adult oysters challenged by starvation indicated that the Ca-Chit expression would be regulated by feed. All the observations made suggest that Ca-Chit plays an important role in the digestive system of the oyster, Crassostrea angulate.     

烟草小孢子母细胞减数分裂过程中微管分布变化

应用间接免疫荧光标记技术和激光共聚焦扫描显微镜成像技术观察了烟草小孢子母细胞减数分裂过程中微管的分布变化。在减数分裂前期,小孢子母细胞中的微管较短,随机分散在细胞质中。在减数分裂中期,细胞质中微管形成纺锤体,控制染色体的分布。进入减数分裂I后期,部分纺锤体微管将两组染色体拉向两级。在减数分裂Ⅱ中期,细胞中的微管又形成两个纺锤体。在减数分裂Ⅱ后期,纺锤体微管解聚为微管蛋白分散在细胞质中。胞质分裂发生在四个细胞核形成之后,通过细胞核之间的质膜向内缢缩分隔四个细胞核,产生四个小孢子。     

九龙江河口区nirS型反硝化细菌多样性及系统发育学分析

【目的】结合16S rRNA基因克隆文库和nirS基因克隆文库的分析,揭示九龙江河口区nirS型反硝化细菌多样性。【方法】选取九龙江河口区一富营养化采样点,分别采集水样及沉积物样品,进行理化因子的测定并提取细菌总DNA。以水样DNA构建16S rRNA基因克隆文库,以沉积物DNA构建nirS基因克隆文库,分析微生物群落结构的多样性并构建系统发育树。【结果】从16S rRNA基因克隆文库中获得86条有效序列,按97%的序列相似性划分为53个OTU,分别属于Proteobacteria门、Planctomycetes门、Bacteroidetes门、Actinobacteria门、Firmicutes门和Chloroflexi门。其中属于Proteobacteria门OTU的克隆子占克隆数的62.9%,是最优势的类群,分属于Alphaproteobacteria、Betaproteobacteria、Gammaproteobacteria和Deltaproteobacteria纲等。从nirS基因克隆文库中获得190条有效序列,翻译为氨基酸序列后,按82%的序列相似性划分为60个OTU,并定位到属的水平。其中Proteobacteria门是最优势的类群,占文库克隆子总数的71.6%,包括Alphaproteobacteria纲(5.8%)、Betaproteobacteria纲(49.0%)和Gammaproteobacteria纲(16.9%)。nirS基因克隆文库中丰度最高的OTU与GenBank中的一株可培养反硝化菌Thauera sp. R-26906具有100%的序列相似性。【结论】九龙江河口区的微生物以及亚硝酸盐还原酶基因(nirS)具有丰富的多样性。大部分NirS序列在GenBank中的最相似序列来源于河口、海湾等相似的环境。     

2′-epi-2′-O-Acetylthevetin B extracted from seeds of Cerbera manghas L. induces cell cycle arrest and apoptosis in human hepatocellular carcinoma HepG2 cells

2′-epi-2′-O-Acetylthevetin B (GHSC-74) is a cardiac glycoside isolated from the seeds of Cerbera manghas L. We have demonstrated that GHSC-74 reduced the viability of HepG2 cells in a time- and dose-dependent manner. The present study was designed to explore cellular mechanisms whereby GHSC-74 led to cell cycle arrest and apoptosis in HepG2 cells. Cell cycle flow cytometry demonstrated that HepG2 cells treated with GHSC-74 (4 μM) resulted in S and G2 phase arrest in a time-dependent manner, as confirmed by mitotic index analysis. G2 phase arrest was accompanied with down-regulation of CDC2 and Cyclin B1 protein. Furthermore, GHSC-74-induced apoptotic killing, as demonstrated by DNA fragmentation, DAPI staining, and flow cytometric detection of sub-G1 DNA content in HepG2 cells. GHSC-74 treatment resulted in a significant increase in reactive oxygen species, activation of caspase-9, dissipation of mitochondrial membrane potential, and translocation of apoptosis-inducing factor (AIF) from the mitochondrion to the nucleus in HepG2 cells. Nevertheless, after GHSC-74 exposure, no significant Fas and FasL up-regulation was observed in HepG2 cells by flow cytometry. In addition, treatment with antioxidant N-acetyl-l-cysteine (NAC) and broad-spectrum caspase inhibitor z-VAD-fmk partially prevented apoptosis but did not abrogate GHSC-74-induced nuclear translocation of AIF. In conclusion, we have demonstrated that GHSC-74 inhibited growth of HepG2 cells by inducing S and G2 phase arrest of the cell cycle and by triggering apoptosis via mitochondrial disruption including both caspase-dependent and -independent pathways, and ROS generation.     

Defects in cytoskeletal microtubule deployment of microsporocytes contribute to fertility loss in genic male-sterile Chinese cabbage

The microtubular cytoskeleton of male-sterile Chinese cabbage was examined to characterize cytoplasmically based defects during microsporogenesis of fertile and sterile microsporocytes. At the onset of meiosis, microtubules (MTs) in fertile microsporocytes were short and anisotropically oriented in the microsporocyte cytoplasm. As the microsporocytes entered metaphase I, the MTs constructed a bisymmetrical spindle characterized by conspicuous kinetochore fibers closely associated with chromosomes in the medial plane. During anaphase I, interzonal MTs become conspicuous between the two sets of chromosomes and the polar regions become more distant as spindle MTs are depleted, essentially disappearing at telophase I. Radially distributed MTs increased and the microsporocyte entered meiosis II, producing two spindles at angles to one another within the wall of the microsporocyte. Indicative of the completion of anaphase II is the formation of a field of aligned MTs between two non-sister nuclei, after which the cytoplasm produced centripetal furrows, meeting in the center of the cell and dividing it into four microspores at the completion of cytokinesis. In sterile microsporocytes, however, an abnormal arrangement of MTs occurred at the conclusion of anaphase II. Although two spindles formed, the angle and the boundary between the spindles were not maintained. At the onset of telophase II, the two spindles migrated to a central region and laterally fused in irregular orientations in which the decondensing chromatin of the non-sister nuclei may form separate or merged nuclei, followed by irregular cytokinesis. The result of meiosis was 41.8 % two binuclear products, and 58.2 % one diploid and one binuclear sterile products.     

福建沿海岛屿水鸟考察报告

野外共考察了福建沿海岛屿57个,记录到水鸟16科94种,其中18种在岛屿上有见繁殖,统计到水鸟总量近17000只.文中对一些全球受威胁物种,如黑嘴端凤头燕鸥Sterna bernsteini、黑脸琵鹭Platalea minor、黄嘴白鹭Egretta eulophotes、黑嘴鸥Larus saundersi等种类在福建沿海的状况做了进一步论述.