彩色马蹄莲品种‘Parfait’多倍体诱导及其生物学特征变化

以彩色马蹄莲品种‘Parfait’（Zantedeschiahybrid‘Parfait’）离体丛生芽块为实验材料,对其多倍体诱导过程中秋水仙素和二甲基亚砜（DMSO）浓度以及浸泡时间进行分析,并比较了多倍体与二倍体植株在叶形指数、气孔特征、叶绿素含量和染色体数的差异,最终通过回归分析确定最佳诱导条件。结果显示：随秋水仙素质量体积分数的提高及浸泡时间的缩短,各处理组的丛生芽存活率逐渐增加且均低于对照,而多倍体诱导率逐渐降低且均显著高于对照。综合考虑丛生芽存活率和多倍体诱导率等因素,根据回归分析确定‘Parfait’多倍体诱导的最佳条件为：丛生芽块在含质量体积分数0.20%秋水仙素和体积分数0.10%DMSO的MS液体培养基中浸泡24h,多倍体诱导率可达50.02%。比较分析结果表明：多倍体植株的叶片长度、厚度和长宽比分别为二倍体植株的1.23、1.19和2.93倍,保卫细胞的长度和宽度以及每气孔叶绿体数分别为二倍体植株的1.90、1.96和2.03倍,叶绿素a和总叶绿素含量分别为二倍体植株的1.28和1.17倍;但多倍体植株的叶宽和气孔密度均较小,分别仅为二倍体植株的42.08%和61.55%。除叶绿素b含量外,多倍体植株的其他生物学特性均与二倍体植株差异显著。染色体计数结果显示：获得的多倍体大多为四倍体,染色体数为2n=64,同时还得到了一些嵌合体和六倍体。研究结果表明：彩色马蹄莲品种‘Parfait’多倍体植株的多数生物学特性优于二倍体植株,且其对环境的适应性更强。     

S100β as an early biomarker of excitotoxic damage in spinal cord organotypic cultures

S100β is a cytoplasmic calcium‐binding protein mainly expressed by glia and considered to be a useful biomarker for brain or spinal cord injury. Indeed, clinical studies suggest that the S100β concentration in serum or cerebrospinal fluid may predict lesion outcome and prognosis. The relation of S100β levels to damage severity and its timecourse remains, however, unclear. This study used a validated in vitro model of spinal cord injury induced by kainate‐mediated excitotoxicity to investigate these issues. After 22 days in vitro, rat organotypic spinal cord slices were subjected to one transient application (1 h) of 1 or 100 μM kainate followed by washout. While the lower kainate concentration did not evoke neuronal loss or S100β increase, the larger concentration elicited 40% neuronal death, no change in glial number and a delayed, significant rise in extracellular S100β that peaked at 24 h. This increase was associated with a stronger expression of the S100β protein as indicated by western blotting and immunohistochemistry. Application of the microtubule disrupting agent colchicine did not change the rise in S100β induced by kainate, an effect blocked by the glutamate receptor antagonists CNQX and APV. Our data suggest that excitotoxicity was followed by release of S100β perhaps from a readily releasable pool through a mechanism independent of microtubule assembly. The raised extracellular level of S100β appeared to reflect glial reactivity to the kainate‐evoked lesion in accordance with the view that this protein may be involved in tissue protection and repair after acute injury.

     

秋水仙素对洋葱根尖细胞的诱变效应

通过三种浓度的秋水仙素对洋葱根尖进行控时处理,结果发现,该药剂能使洋葱根尖细胞有丝分裂指数下降,促进洋葱根尖膨大,导致染色体加倍,并产生畸变细胞。     

Induction of tetraploidy in Buddleia globosa

The aim was to produce a tetraploid form of Buddleia globosa to facilitate introgression of yellow flower colour into B. davidii, which is naturally tetraploid. Protocols were established for the micropropagation of B. globosa and tetraploid plants were obtained by application in vitro of colchicine to pre-cultured excised nodal sections. Three concentrations of colchicine were applied (0.01%, 0.05% and 0.1% w/v) for 1, 2 or 3 days. At 0.01% tetraploids were produced only after 2 days of application. All other treatments produced at least one tetraploid. The colchicine technique was extremely effective: of 29 lines tested, 19 were tetraploid and 5 were mixoploid. The vegetative characteristics of these tetraploids are described and the flowering characteristics of the three that flowered. This revised version was published online in August 2006 with corrections to the Cover Date.     

黄芩组织培养同源四倍体的诱导

应用组织培养技术对黄芩进行多倍体诱导,结果表明：组织培养条件下,在培养基中添加一定浓度和秋水仙素,或者把带有绿色芽点的黄芩愈伤组织经０．２％秋水仙素溶液浸泡一定时间后再进行培养,均可诱发黄芩多倍体的产生,但以后效果较好,诱导率可达４０．０％,通过试管苗根尖染色体显微观察,鉴定出５０多个黄芩同源四倍体,为今后优良品种的选育打下基础。     

Microtubule‐dependent changes in morphology and localization of chloride transport proteins in gill mitochondria‐rich cells of the tilapia,Oreochromis mossambicus

The tilapia (Oreochromis mossambicus) is a euryhaline fish exhibiting adaptive changes in cell size, phenotype, and ionoregulatory functions upon salinity challenge. Na⁺/Cl^? cotransporter (NCC) and Na⁺/K⁺/2Cl^? cotransporter (NKCC) are localized in the apical and basolateral membranes of mitochondria‐rich (MR) cells of the gills. These cells are responsible for chloride absorption (NCC) and secretion (NKCC), respectively, thus, the switch of gill NCC and NKCC expression is a crucial regulatory mechanism for salinity adaptation in tilapia. However, little is known about the interaction of cytoskeleton and these adaptive changes. In this study, we examined the time‐course of changes in the localization of NKCC/NCC in the gills of tilapia transferred from fresh water (FW) to brackish water (20‰) and from seawater (SW; 35‰) to FW. The results showed that basolateral NKCC disappeared and NCC was expressed in the apical membrane of MR cells. To further clarify the process of these adaptive changes, colchicine, a specific inhibitor of microtubule‐dependent cellular regulating processes was used. SW‐acclimated tilapia were transferred to SW, FW, and FW with colchicine (colchicine‐FW) for 96 h. Compared with the FW‐treatment group, in the MR cells of colchicine‐FW‐treatment group, (1) the average size was significantly larger, (2) only wavy‐convex‐subtype apical surfaces were found, and (3) the basolateral (cytoplasmic) NKCC signals were still exhibited. Taken together, our results suggest that changes in size, phenotype, as well as the expression of NCC and NKCC cotransporters of MR cells in the tilapia are microtubule‐dependent. J. Morphol. 277:1113–1122, 2016. © 2016 Wiley Periodicals, Inc.     

Plastid microtubule-like structures in wheat are insensitive to microtubule inhibitors

The effects of microtubule inhibitors on the spectral properties of leaves of wheat ( Triticum aestivum L. cv. Walde) and on the presence of plastid microtubule–like structures (MTLS) during etioplast to chloroplast transformation were examined. Amiprophos-methyl (APM, 0.1 m M ), fed to leaf sections of 7-day-old dark-grown wheat, reduced the ration of phototransformable to non-phototransformable proto-chlorophyllide (PChlide), decreased the rate of the Shibata shift, and inhibited chlorophyll accumulation and grana stacking. The spectral properties of isolated etioplasts were not affected by APM. Colchicine (10 m M ), fed to leaf sections, inhibited greening but had no effect on the PChlide ratio or the Shibata shift. MTLS were still visible on electron micrographs after treatment with APM or colchicine at frequencies similar to controls. A third inhibitor, vinblastine, had no effect on the spectral properties of non-irradiated or irradiated etiolated leaves except at concentrations that produced visible tissue damage before the irradiation. The effects of APM and colchicine may reflect inhibitions of respiration and protein synthesis, respectively. It is concluded that MTLS are insensitive to microtubule inhibitors and thus are probably not composed of tubulin.     

PREMATURE CHROMOSOME CONDENSATION OF THE KARYOGAMY-BLOCKED SPERM PRONUCLEUS IN THE FERTILIZATION OF FUCUS DISTICHUS (FUCALES,PHAEOPHYCEAE)1

Mitosis of egg and sperm pronuclei of Fucus distichus subsp. evanescens (C. Agardh)Powell was examined by fluorescence and electron microscopy when migration of the sperm pronucleus and, as a result, karyogamy were blocked by colchicine treatment after plasmogamy. Chromosome condensation was obsewed in both pronuclei Microspectrophotometric studies after staining the nuclei with mithramycin A clearly showed that DNA synthesis ocurred in the egg pronucleus but not in the sperm pronucleus. This means that chromosomes condensed prematurely in the sperm pronucleus (premature chromosome condensation). In some cases, the egg chromosomes became arranged on a metaphase plate, whereas the sperm chromosomes lay scattered near the egg pronucleus. Immuno fluorescence microscopy using anti-β-tubulin antibody confirmed that a normal spindle was formed at the egg pronucleus. A pair of centrioles existed at the two poles of this spindle. The sperm nuclear membrane disappeared, and microtubules radiated to the sperm chromosomes from one pole of the egg spindle.     

DIATOM MINERALIZATION OF SILICIC ACID. VI. THE EFFECTS OF MICROTUBULE INHIBITORS ON SILICIC ACID METABOLISM IN NAVICULA SAPROPHILA1

The role of microtubules in silicon metabolism leading to valve formation was investigated in the pennate diatom Navicula saprophila Lange-Bertalot & Bonik. By using synchronized cells blocked after mitosis and cytokinesis but prior to cell wall formation, effects due to inhibition of mitosis were eliminated. Cells were treated with three anti-microtubule drugs to assess the role of microtubules. Chemical analogs to two of the drugs provided controls for inhibition not related to microtubule disruption. Although all three anti-microtubule drugs reduced cell separation at high concentrations (1 × 10^?3 M), podophyllotoxin was the only drug which reduced cell separation at concentrations lower than 1 × 10^?5 M. None of the drugs at any concentration tested affected cell viability. There was no differential inhibitory effect between the active and inactive drugs on silicic acid transport, total uptake, incorporation, or pool formation. There was no qualitative difference between silica incorporated in treated and untreated cells. A colchicine binding component was isolated from N. saprophila. The characteristics of colchicine binding suggest this component may be tubulin. Microtubules do not appear to be involved in any of the steps of silicon metabolism leading to valve formation and yet they have profound influence on the symmetry and pattern of the mineralized product, the siliceous valve.