秋水仙素浓度和处理时间对茉莉腋芽生长发育的影响

为了解秋水仙素处理对茉莉﹝Jasminum sambac (Linn.) Aiton〕腋芽生长发育的影响,以长度为0(未萌发)、3~5和8~10 mm腋芽为实验材料,对500、1000和2000 mg·L-1秋水仙素溶液分别处理24和48 h后腋芽的生长状况以及萌发枝条和叶片的生长和发育状况进行分析,同时,对各处理组花粉母细胞的减数分裂行为进行观察;在此基础上,对秋水仙素处理后茉莉腋芽及萌发枝条的各生物学效应指标进行相关性分析。结果显示：经秋水仙素处理后,各处理组腋芽的生长率和处理指数,以及萌发枝条的长度、最大节间长、最大叶长、最大叶宽、现蕾数、现蕾率、最大蕾长、最大蕾宽和开花率总体上显著(P<0.05)低于对照组(用清水处理未萌发腋芽48 h),而腋芽的抑制率和死亡率显著高于对照组。总体上看,随秋水仙素质量浓度提高,腋芽的生长率和处理指数降低,而抑制率和死亡率升高,萌发枝条的长度和最大节间长缩短,开花率升高,其他指标呈波动变化;随处理时间延长,腋芽的生长率、死亡率和处理指数降低,但抑制率升高,萌发枝条仅现蕾数和开花率降低,其他指标均逐渐提高;随腋芽长度增加,腋芽的抑制率和处理指数降低,但死亡率升高,萌发枝条的现蕾率、现蕾数、最大蕾长和开花率均呈波动变化,其他指标均降低。经秋水仙素处理后,茉莉花粉母细胞在减数分裂过程中存在落后染色体、染色体桥和微核等异常现象,且随秋水仙素质量浓度和腋芽长度增加及处理时间延长,减数分裂后期的细胞异常率逐渐升高。相关性分析结果显示：细胞异常率与腋芽生长率呈显著负相关,与腋芽死亡率呈极显著正相关;腋芽生长率与腋芽抑制率和开花率分别呈显著和极显著(P<0.01)负相关;腋芽抑制率与现蕾率呈显著正相关;处理指数与细胞异常率呈极显著负相关。研究结果表明：秋水仙素浓度和处理时间对茉莉腋芽的生长发育有一定影响;综合考虑各生物学效应指标,茉莉腋芽适宜的诱导条件为用500 mg·L-1秋水仙素溶液处理3~5 mm腋芽24 h。此外,建议将处理指数作为秋水仙素对茉莉腋芽细胞减数分裂影响效应的评价指标之一。     

Somatic chromosome number doubling of selected potato genotypes using callus culture or the colchicine treatment of shoot nodes in vitro

Experiments were carried out to double the somatic cell chromosome numbers of a monoploid and dihaploid of Solanum tuberosum and a genotype of S. circaeifolium subsp. quimense. Colchicine was used in vitro on shoot nodes from which the axillary meristems had been removed. Plants with doubled chromosome numbers were obtained from shoots grown from the tertiary, sub-axillary meristems of all three genotypes. The callus culture of stem and leaf explants was found to produce more shoots with doubled chromosome numbers than the colchicine treatment in the case of the dihaploid and quimense genotypes but no shoots were obtained from callus culture of the monoploid. Fifty-two % of the shoots from the dihaploid and 63% from the quimense clone were ploidy doubled in the case of the best callus culture system. Using a sub-lethal dose of colchicine, the dihaploid yielded 37% ploidy-doubled shoots whereas all the shoots produced from the monoploid were doubled and the quimense clone produced 27% doubled plants. Callus culture was highly dependent upon the type of growth medium and other, unknown, factors. The colchicine treatment, although yielding fewer products, was more reliable for achieving ploidy doubling in selected clones if the number of plants produced is not important.     

Chromosome doubling in sterile Syringa vulgaris × S. pinnatifolia hybrids by in vitro culture of nodal explants

The aim was to produce tetraploid forms of hybrids of Syringa vulgaris × S. pinnatifolia to restore fertility and enable further breeding to be undertaken. Excised nodal sections of three selections were pre-cultured for 5 days then treated with a range of colchicine concentrations for 1, 2 or 3 days, after which they were washed three times in sterile distilled water and cultured on shoot proliferation medium. Frequent movement to fresh medium was beneficial to survival. Three successive experiments established the range of concentrations of colchicine, 0.05 mM to 0.25 mM, at which tetraploids were likely to be produced. Thus a protocol for the production of tetraploids was established. Cytometric analysis showed that tetraploid forms of two selections were produced. This revised version was published online in August 2006 with corrections to the Cover Date.     

Chemically Induced Polyploidization in Spathiphyllum wallisii Regel through Somatic Embryogenesis

Polyploid Spathiphyllum wallisii Regel 'Speedy' plants were obtained from somatic embryos induced on anther filaments exposed to mitosis inhibitors. Primary embryos yielded less polyploid plantlets than secondary embryos. Colchicine could be efficiently replaced by oryzalin or trifluralin (10 μM), resulting in an average yield of 5% polyploids. The mitosis inhibitors were directly added to the induction medium. Morphological differences between diploid and tetraploid S. wallisii hybrids were observed. Throughout our experiments flow cytometry was applied as an unambiguous screening tool. Ploidy breeding within the genus Spathiphyllum holds promises towards the development of tetraploid hybrids with an altered morphology, or triploids with a reduced fertility.     

Methoxy and bromo scans on N-(5-methoxyphenyl) methoxybenzenesulphonamides reveal potent cytotoxic compounds,especially against the human breast adenocarcinoma MCF7 cell line

Thirty seven N-(5-methoxyphenyl)-4-methoxybenzenesulphonamide with methoxy or/and bromo substitutions (series 1-4) and with different substituents on the sulphonamide nitrogen have been synthesised. 21 showed sub-micromolar cytotoxicity against HeLa and HT-29 human tumour cell lines, and were particularly effective against MCF7. The most potent series has 2,5-dimethoxyanilines, especially the 4-brominated compounds 23–25. The active compounds inhibit microtubular protein polymerisation at micromolar concentrations, thus pointing at tubulin as the target. Co-treatment with the MDR inhibitor verapamil suggests that they are not MDR substrates. Compound 25 showed nanomolar antiproliferative potency. It severely disrupts the microtubule network in cells and arrests cells at the G₂/M cell-cycle phase, thus confirming tubulin targeting. 25 triggered apoptotic cell death, and induced autophagy. Docking studies suggest binding in a distinct way to the colchicine site. These compounds are promising new antitumor agents acting on tubulin.     

Modulation of phenotypic expression of fibroblasts by alteration of the cytoskeleton

Several studies indicate that the cytoskeleton may be involved in modulating the cellular response to environmental signals. We have studied the role of the cytoskeleton in regulating glycosaminoglycan (GAG) synthesis and secretion, hyaluronate (HA) endocytosis, the activities of hexoglycosidases, protein synthesis and secretion. Fibroblasts were treated with colchicine (1–8 μM ) and nocodazole (1 or 4 μM ) to alter microtubules or cytochalasin B (0·5–4 μM ) to alter microfilaments. Colchicine inhibited GAG synthesis and secretion in a concentration-dependent manner. It reduced protein and sulphated GAG secretion, while HA secretion was not affected. Concentration-dependent disruption of microtubules from the periphery toward the cellular centre with nocodazole inhibited only the secretion of GAG. Centrosomal microtubles appeared to be required to promote GAG synthesis; intact microtubules promoted the transport of secretory products, intercompatmental transport of lysosomal enzymes and lysosome maturation, but not protein synthesis and HA secretion. Cytochalasin B treatment inhibited, in a concentration-dependent manner, the synthesis and secretion of GAGs and proteins, and the endocytosis of HA. Intact microfilament mesh-works appeared to be required to promote synthesis and secretion of proteins and proteoglycans and to contribute to the transmembrane control of receptor-mediated endocytosis. Drug treatment of concanvalin A (Con A)-stimulated fibroblasts inhibited the stimulation of GAG synthesis. It is probable that this effect may result, in part, from drug-induced effects on Con A-mediated endocytosis.     

甜叶菊同源四倍体离体诱导及鉴定

用不同浓度秋水仙素溶液处理甜叶菊不定芽,诱导同源四倍体,并进行解剖学、染色体鉴定和流式细胞仪鉴定倍性。结果表明:(1)用0.20%的秋水仙素溶液浸泡甜叶菊不定芽12h,同源四倍体诱导率最高,可达32.14%。(2)同源四倍体植株与二倍体(对照)相比,其气孔、叶片等均表现巨大性,且叶片变厚、叶色浓绿、叶片皱缩。(3)对照植株染色体2n=2x=22,四倍体植株染色体2n=4x=44;流式细胞仪倍性鉴定结果显示,对照DNA相对含量为100,四倍体DNA相对含量为200。(4)该研究共鉴定出48株甜叶菊同源四倍体植株,为进行倍性植株的诱导奠定了技术基础,为进一步开展甜叶菊同源四倍体新品种的选育提供了实验材料。     

大花蕙兰四倍体的离体诱导和鉴定

采用组织培养方法研究了秋水仙素处理对大花蕙兰类原球茎增殖、分化和四倍体诱导的效应.结果表明,秋水仙素处理明显抑制类原球茎的增殖和分化,提高类原球茎褐变率和死亡率;在试验浓度和时间范围内,秋水仙素浓度越高,处理时间越长,抑制作用越强,解除抑制作用所需的继代次数越多.所有秋水仙素处理均能诱导出四倍体,但不同处理的四倍体诱导率不同,当处理浓度为0.05%、时间为5 d时,四倍体诱导率最高,为23.7%.二倍体及其四倍体的气孔保卫细胞长度和气孔密度均存在极显著的差异.四倍体植株比二倍体矮,茎部较粗壮,株型紧凑,叶片颜色浓绿、质地变硬.秋水仙素处理和组织培养相结合是创建大花蕙兰多倍体的有效方法.     

Carbamate derivatives of colchicine show potent activity towards primary acute lymphoblastic leukemia and primary breast cancer cells—in vitro and ex vivo study

Colchicine ( COL ) shows strong anticancer activity but due to its toxicity towards normal cells its wider application is limited. To address this issue, a library of 17 novel COL derivatives, namely N‐carbamates of N‐deacetyl‐4‐(bromo/chloro/iodo)thiocolchicine, has been tested against two types of primary cancer cells. These included acute lymphoblastic leukemia (ALL) and human breast cancer (BC) derived from two different tumor subtypes, ER+ invasive ductal carcinoma grade III (IDCG3) and metastatic carcinoma (MC). Four novel COL derivatives showed higher anti‐proliferative activity than COL (IC₅₀ = 8.6 nM) towards primary ALL cells in cell viability assays (IC₅₀ range of 1.1‐6.4 nM), and several were more potent towards primary IDCG3 (IC₅₀ range of 0.1 to 10.3 nM) or MC (IC₅₀ range of 2.3‐9.1 nM) compared to COL (IC₅₀ of 11.1 and 11.7 nM, respectively). In addition, several derivatives were selectively active toward primary breast cancer cells compared to normal breast epithelial cells. The most promising derivatives were subsequently tested against the NCI panel of 60 human cancer cell lines and seven derivatives were more potent than COL against leukemia, non–small‐cell lung, colon, CNS and prostate cancers. Finally, COL and two of the most active derivatives were shown to be effective in killing BC cells when tested ex vivo using fresh human breast tumor explants. The present findings indicate that the select COL derivatives constitute promising lead compounds targeting specific types of cancer.