盐胁迫对玉米叶片叶肉细胞生物膜超微结构的影响

研究了NaCl胁迫对玉米叶肉细胞生物膜超微结构的影响. 结果表明：NaCl胁迫破坏了玉米叶片叶肉细胞生物膜的正常结构,50 mmol·L^-1 NaCl处理胁迫下,玉米叶肉细胞核膜,线粒体膜,细胞膜,叶绿体膜,液泡膜都受到不同程度的破坏,叶绿体基粒类囊体膨胀,间质片层空间增大,片层紊乱。100 mmol·L^-1 NaCl处理胁迫下,质膜,液泡膜,线粒体,叶绿体都受到严重的破坏。细胞质膜破坏,破损的叶绿体充斥在细胞间隙中;叶绿体外膜破坏,甚至解体消失,叶肉细胞中充满膜结构,基粒排列方向改变,垛叠层数减少,基粒和基质片层界限模糊不清,有的基粒解体消失,甚至叶绿体完全解体;核膜破坏、解体,核中的染色质高度凝缩;线粒体的数量增多,线粒体膜破坏,脊的数量减少,甚至整个线粒体破损解体;液泡膜破坏;由于各种生物膜的破坏,使细胞内充满许多囊状小泡、多泡体或斑层小体;叶肉细胞发生严重的质壁分离,严重时发生细胞壁断裂;甚至整个细胞溶解。     

环境胁迫对库拉索芦荟叶片超微结构影响研究

对1年生库拉索芦荟分别用盐(1.8%的NaCl)、低温(10℃)、干旱[25%(w/v)的聚乙二醇-6000]3种胁迫条件处理7d后,对其叶肉细胞超微结构进行观察.结果发现:3种胁迫处理均可使库拉索芦荟细胞膜系统、叶绿体、线粒体、细胞核等结构受到不同程度的破坏,叶绿体周围出现许多小泡,导致细胞内膜系统紊乱,细胞器结构稳定性降低;盐胁迫下高尔基体在细胞质中解体;盐和低温胁迫下均可见线粒体膜与叶绿体膜发生融合、线粒体嵌在叶绿体当中的现象.另外,本研究发现,盐胁迫、低温胁迫比干旱胁迫对库拉索芦荟细胞膜的损伤更严重,而水分胁迫对其的伤害程度较小,表明库拉索芦荟的抗旱性较其抗盐性更强.     

水分胁迫下玉米叶肉细胞超微结构的变化及其与膜脂过氧化伤害的关系

研究ＰＥＧ模拟水分胁迫条件对玉米叶肉细胞超微结构的影响,探讨了超微结构的变化与保护酶活性及膜酯过氧化伤害之间的关系。试验表明,在水分胁迫初期,叶肉细胞超微结构变化较小,此时,叶片的ＳＯＤ及过氧化氢酶活性明显升高,质膜相对透性和丙二醛含量增加缓慢,随着胁迫时间的延长,叶肉细胞超微结构破坏加重,且不同细胞对水分胁迫的敏感性相差很大,叶片ＣＡＴ性下降,质膜透性和ＭＤＡ含量急增;复水后,叶片超微结构,ＳＯ     

Hg2+对海马齿叶肉细胞超微结构的影响

以红树林植物海马齿为材料,将生长一致的海马齿水培苗放到含有不同浓度Hg2+的营养液中进行Hg2+胁迫,用透射电镜观察海马齿叶肉细胞超微结构对不同浓度Hg2+胁迫的响应,以明确重金属汞对海马齿叶肉细胞超微结构的影响,探讨海马齿耐汞机制。结果表明:重金属汞能造成海马齿叶肉细胞不同程度的伤害,主要表现为对叶肉细胞中的叶绿体、线粒体、细胞核以及膜系统的伤害。随着Hg2+浓度不断升高,其叶绿体数目不断减少,形状由船型变成长形以及出现一些巨型叶绿体,类囊体系统受到伤害、基粒片层变得模糊不清。线粒体数目由于Hg2+浓度的不同而不同,形状由棒状变成圆形及椭圆形,线粒体双层膜结构与嵴变得模糊不清。细胞核也受到不同程度的伤害,核仁由一个变成多个,最后消失;同时细胞膜也受到伤害,主要表现为,不断的向胞内形成膜突起再形成空泡。最后在高浓度Hg2+胁迫下,随着叶肉细胞内细胞器的不断减少,最终造成细胞解体死亡。     

水分胁迫下小麦叶肉细胞超微结构变化与抗旱性的关系

本文用电子显微镜观察研究了抗旱性不同的6个小麦品种在不同程度水分胁迫下叶肉细胞超微结构的变化。结果表明:轻度水分胁迫(-0.5MPa)对参试的6个小麦品种叶肉细胞超微结构几乎没有影响。中度(-1.0MPa)和严重(-1.5MPa)水分胁迫下的叶肉细胞超微结构发生了程度不同的变化,且这种变化与品种抗旱性相一致。抗旱性愈弱的品种,对水分胁迫反应愈敏感。但表现在叶肉细胞结构上的变化过程基本一致。胁迫导致叶肉细胞质壁分离,液泡膜破裂。叶绿体变成球形挤入细胞中央,类囊体肿胀。线粒体基质变稀,脊减少。最终叶绿体、线粒体解体。其它细胞器消失,细胞中出现大量的小泡。     

低温锻炼和低温胁迫期间翠南报春叶肉细胞超微结构的适应变化

低温锻炼(10℃)期间翠南报春叶肉细胞线粒体增加并出现质膜内陷现象,低温胁迫(0℃)过程中,低温锻炼植株的细胞超微结构表现相对稳定,叶绿体片层结构和线粒体数目都未发生明显变化。未受低温锻炼植株线粒体数目在低温胁迫第2天增多,第3、4天逐渐减少,并在第3天出现质膜内陷现象。多泡体和囊泡结构有时伴随着质膜内陷出现。这种变化可能是经低温锻炼的翠南报春抗寒性提高的细胞结构基础。     

嫁接对低温弱光下辣椒幼苗膜脂过氧化及抗氧化酶活性的影响

以‘卫士’为砧木,以‘赤峰特选’为接穂进行嫁接,在光照培养箱内对辣椒自根苗(对照)和嫁接苗进行低温 (8 ℃/5 ℃) 弱光(100 μmol·m^-2·s^-1)处理,处理7 d后在正常条件(25 ℃/18 ℃,550～600 μmol·m^-2·s^-1)下恢复3 d,研究低温弱光下辣椒嫁接苗和自根苗电解质渗漏率(EL)、丙二醛(MDA)含量、抗氧化酶活性及根系活力的变化.结果表明：低温弱光胁迫初期,辣椒幼苗叶片与根系的EL、MDA含量和超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性均显著升高,而根系活力大幅降低;1～3 d后EL和MDA含量趋于平稳,SOD、POD、APX、GR活性逐渐降低,根系活力呈上升趋势.恢复3 d后,嫁接苗EL、MDA含量、抗氧化酶活性及根系活力多达到或超过胁迫前水平(根系的MDA含量较胁迫前略高);而自根苗的EL和MDA含量仍显著高于胁迫前.与自根苗相比,嫁接苗在各处理阶段的EL和MDA含量显著降低,而SOD、POD、APX、GR活性及根系活力明显升高,说明嫁接可有效降低辣椒植株的膜脂过氧化,减轻低温弱光对其细胞膜的伤害.     

干旱胁迫下水曲柳苗木细根线粒体的形态及活性变化

根系依赖根细胞内线粒体呼吸代谢产生的能量, 不断从土壤中获取养分。在胁迫条件下, 线粒体的结构和功能会发生一定的变化, 从而影响根系的功能。土壤干旱是最容易引起苗木细根衰老死亡的非生物胁迫因子之一。为了更好地认识干旱胁迫下细根线粒体的结构和功能变化, 对土壤干旱胁迫下水曲柳(Fraxinus mandshurica)不同颜色细根皮层薄壁细胞内线粒体的超微结构(线粒体数量、形态)、线粒体的呼吸功能、线粒体膜脂质氧化(膜透性变化、过氧化氢含量等)情况进行了研究。结果表明: (1)干旱胁迫下, 水曲柳白色及黄色根皮层薄壁细胞内线粒体形状、结构及分布数量与对照相似, 无显著差异。干旱胁迫下产生的褐色根皮层薄壁细胞线粒体数量减少, 分布密度也变小。线粒体内、外膜先后发生不同程度的解体, 最后消失。(2)干旱胁迫显著干扰了线粒体膜的正常呼吸耦联作用, 细根线粒体呼吸控制率(RCR)与磷氧比(无机磷酸/分子氧, P/O)均显著低于对照(p < 0.05)。随着细根颜色加深, 线粒体RCR和P/O值逐渐下降, 白色根﹥黄色根﹥褐色根。褐色根线粒体RCR值最低, 接近极值1。说明褐色根线粒体结构完整性最差, 能量转化效率最低。(3)干旱胁迫下, 不同颜色细根线粒体内的H₂O₂含量、线粒体膜透性、膜脂氧化产物丙二醛(MDA)含量均显著高于对照(p < 0.05)。且随着细根颜色加深, 各个值增加明显。分析可能是由于干旱胁迫导致线粒体内H₂O₂含量升高, 线粒体膜脂质过氧化(MDA含量升高), 膜结构受到破坏(膜透性增加) (电镜下可见部分线粒体内膜电子密度下降及外膜解体)。线粒体膜结构完整性的破坏, 直接影响了线粒体呼吸代谢反应, 使线粒体呼吸功能下降。     

低温对桂花‘状元红’叶肉细胞超微结构的影响

为探讨北引桂花(Osmanthus fragrans)在低温胁迫下叶肉细胞超微结构的变化,揭示桂花于低温胁迫下细胞结构变化规律,该研究以3年生桂花品种‘状元红’(O.fragrans‘Zhuangyuan Hong’)为试材,分别于一系列低温下处理,经制样切片后,用透射电子显微镜观察叶肉细胞超微结构的变化。结果表明:常温(20~25°C)处理时,各细胞器超微结构正常;5°C低温处理时,叶绿体有轻微膨大现象,线粒体结构正常;0°C处理时叶绿体内嗜锇体增多,叶绿体肿胀加剧,线粒体数量增加,淀粉粒出现亮暗相间的轮纹;–10°C处理时,细胞器降解。在同一低温胁迫下不同细胞的叶绿体敏感程度不同,这为遭受低温后植株的恢复生长提供了细胞学基础。叶肉细胞中叶绿体、线粒体、细胞核的稳定性可作为桂花对低温响应的重要参考指标。     

Na2CO3胁迫对星星草叶肉细胞超微结构的影响

利用透射电镜技术对Na2CO3胁迫下星星草叶肉细胞超微结构进行了观察。结果表明：未胁迫的叶肉细胞排列疏松,各种细胞器结构完整,叶绿体含少量淀粉粒和脂质球。轻度盐胁迫（2g/L,4g／LNa2CO3）对叶肉细胞超微结构影响较小。中度盐胁迫（6g／L,8g／L Na2CO3）引起叶肉细胞超微结构的变化,叶绿体类囊体肿胀,基粒紊乱,不含淀粉粒,脂质球数量增加,叶绿体由原来的梭形或椭球形变成圆球状;部分线粒体嵴消失,出现晶体结构;中央大液泡破裂;核逐渐降解。高度盐胁迫（10g／L,12g／LNa2CO3）下,叶绿体片层结构消失,脂质球数量增加,体积变大,被大量的膜片层所包围,叶绿体内、外膜消失,叶肉细胞中看不到叶绿体的存在;膜片层包围线粒体;叶肉细胞中可见大量的泡状结构和膜片层,叶肉细胞死亡。上述结果表明,细胞器特别是叶绿体膜结构的破坏与盐胁迫叶肉细胞最终死亡密切相关。     

Micromanipulation of male and female gametes ofNicotiana tabacum: I. Isolation of gametes

The isolation of male and female gametes is a precondition for the micromanipulation of flowering plant gametes. To reflect their condition at fertilization, isolated gametes need to be physiologically mature and vigorous. Sperm cells are isolated from pollen tubes grown on cut styles using the in vivo/in vitro technique. Embryo sacs are isolated 2 days after anthesis using brief treatments of minimal concentrations of cell-wall-digesting enzymes on ovules of emasculated flowers. Egg cells are then mechanically separated from the embryo sac, allowing unambiguous identification of cells. Two days is usually the minimum required for the pollen tube to penetrate the ovule and effect fertilization in vivo.     

器官发育和程序性细胞死亡中的基因调控——2002年诺贝尔生理学和医学奖工作介绍

2002年的诺贝尔生理学和医学奖授予了在器官发育和程序性细胞死亡研究领域中做出奠基性贡献的三位英美科学家．他们建立了线虫实验模型,完成了其细胞谱图的绘制,而且系统深入地研究了线虫的器官发育和程序性细胞死亡中的基因规则,并在高等哺乳动物中发现了相关的功能基因．这些研究对认识发育过程和揭示人类重大疾病的发病机理具有重要的理论价值和现实意义．     

Development of the Drosophila entero-endocrine lineage and its specification by the Notch signaling pathway

In this paper we have investigated the developmental–genetic steps that shape the entero-endocrine system of Drosophila melanogaster from the embryo to the adult. The process starts in the endoderm of the early embryo where precursors of endocrine cells and enterocytes of the larval midgut, as well as progenitors of the adult midgut, are specified by a Notch signaling-dependent mechanism. In a second step that occurs during the late larval period, enterocytes and endocrine cells of a transient pupal midgut are selected from within the clusters of adult midgut progenitors. As in the embryo, activation of the Notch pathway triggers enterocyte differentiation and inhibits cells from further proliferation or choosing the endocrine fate. The third step of entero-endocrine cell development takes place at a mid-pupal stage. Before this time point, the epithelial layer destined to become the adult midgut is devoid of endocrine cells. However, precursors of the intestinal midgut stem cells (pISCs) are already present. After an initial phase of symmetric divisions which causes an increase in their own population size, pISCs start to spin off cells that become postmitotic and express the endocrine fate marker, Prospero. Activation of Notch in pISCs forces these cells into an enterocyte fate. Loss of Notch function causes an increase in the proliferatory activity of pISCs, as well as a higher ratio of Prospero-positive cells.     

Isolation of gametes and central cells from Oryza sativa L.

In vitro fertilization system of higher plants has been well established using maize gametes and central cells, which can produce embryos and endosperms. In the present study, procedures for isolating gametes and central cells from rice (Oryza sativa L. cv. Nipponbare), a model plant, are reported with the goal of establishing rice in vitro fertilization system. Egg cells and central cells were isolated by manual manipulation of enzyme-treated unpollinated ovules, and an alternative direct isolation method for egg cells that does not use enzymatic treatment was also established. Fluorescent visualization of the granular structures in the cytoplasm of isolated egg cells and the nucleoli in two polar nuclei of isolated central cells suggest that these cells are reliable gametes and central cells. For sperm cell isolation, the contents of rice pollen grains were released by osmotic pressure-induced bursting of the grains. In addition, electrofusion with isolated gametes was successfully conducted.     

植物乳杆菌CGMCC8198干预脂代谢紊乱发挥肝癌防治作用

【背景】脂代谢异常是肝癌发生发展过程中重要的代谢事件,研究发现多种乳酸菌在调节糖脂代谢过程中发挥重要作用。【目的】探究植物乳杆菌CGMCC8198 (TCCC11824)是否会通过调节HMGCR/SMYD3脂代谢通路,进而对肝癌细胞的发生发展产生影响。【方法】采用不同浓度的(5、10、15μg/mL)植物乳杆菌CGMCC8198破碎上清液(Lactobacillus plantarum CGMCC 8198 Crushed Supernatant,LpS)处理HepG2细胞不同时间。利用蛋白免疫印迹(Western Blot)、油红染色以及实时定量荧光PCR(Real Time Quantitative PCR,RT-qPCR)等方法检测LpS对肝癌细胞脂肪变性及HMGCR/SMYD3脂代谢关键通路的影响;通过MTT法、细胞划痕实验、流式细胞术检测LpS对脂代谢紊乱过程中HepG2细胞增殖、迁移和凋亡的影响。【结果】LpS可以抑制脂代谢紊乱肝癌细胞中HMGCR、SMYD3、SREBP-2等基因的表达,同时也可以剂量依赖地抑制细胞增殖、迁移,促进细胞凋亡。【结论】LpS可以通过抑制脂代谢关键转录调控因子SREBP-2和HMGCR的表达来抑制肝癌细胞的脂代谢,进而促进肝癌细胞的内源性凋亡。     

Isolation of third, fourth, and fifth instar larval midgut epithelia of the moth, Trichoplusia ni

A new tissue isolation technique was used to create intact midgut epithelial wholemounts from three Trichoplusia ni (Lepidoptera: Noctuidae) larval instars. The protease, dispase, removed the basal lamina and associated connective tissue and allowed for high resolution light microscopy of entire epithelia. Columnar, goblet, differentiating, and stem cells were characterized by double fluorescent labelling of f-actin and nuclei. A comparison of cell populations by digital image analysis revealed significant regional and temporal changes in the density and number of differentiating and stem cells. Growth of the midgut epithelium from third to fourth instar, and from fourth to fifth instar, was accomplished by both cell differentiation and cell division. Cell division however, was greatly reduced from fourth to fifth instar with a concomitant sharp decrease in the stem cell population.     

Cell structure and reproduction process in the Blue-Green algaChamaesiphon

Ultrathin sections were studied in 2 strains and 2 samples from the nature of the genusChamaesiphon, representing 4 different species. Thylakoids are distributed mainly on the periphery of the cells, the cell-wall is probably 2-layered, and variable multilayered mucilaginous envelopes are developed around the cells. The cell division starts, as well as in otherCyanophyceae, by the invagination of the cytoplasmic membrane and of cell-wall layers into the protoplast; the mucilaginous envelopes—pseudovaginae—do not participate in this process but they form only the firm sheaths around the cells. The way of reproduction is, therefore, essentially the same as that described in other chroococcal Blue-Green algae (e.g.,Synechococcus), and the main difference is the polarized growth of theChamaesiphon cells. The taxonomical position of chamaesiphonoid algae is not as isolated as it was earlier supposed, the similarity withEntophysalidaceae is evident.     

Flow-cytometric cell counting and DNA estimation for the study of plant cell population dynamics

A one-step procedure is presented for simultaneous measurement of cell number and DNA content in cultured plant cells by flow cytometry. In order to obtain nuclei representative of the growth stadium of the culture and of all phases of the cell cycle, cells were carefully sampled and immediately fixed. Next, nuclei were isolated by enzymatic and mechanical maceration, and stained with a DNA-specific fluorescent dye. In the resultant preparation, cells can be counted at relative ease by means of a fluorescence microscope. However, flow-cytometric counting appeared to be superior to manual counting since the time needed for flow-cytometric counting was one-fourth that for manual counting and the variance between counts of the samples was significantly less. In addition, from the same routine, accurate DNA distributions were obtained as a second important parameter of the population dynamics.     

Distribution of cell wall components in<Emphasis Type="Italic"> Sphagnum</Emphasis> hyaline cells and in liverwort and hornwort elaters

Spiral secondary walls are found in hyaline cells of Sphagnum, in the elaters of most liverworts, and in elaters of the hornwort Megaceros. Recent studies on these cells suggest that cytoskeletal and ultrastructural processes involved in cell differentiation and secondary wall formation are similar in bryophytes and vascular plant tracheary elements. To examine differences in wall structure, primary and secondary wall constituents of the hyaline cells of Sphagnum novo-zelandicum and elaters of the liverwort Radula buccinifera and the hornwort Megaceros gracilis were analyzed by immunohistochemical and chemical methods. Anti-arabinogalactan–protein antibodies, JIM8 and JIM13, labeled the central fibrillar secondary wall layer of Megaceros elaters and the walls of Sphagnum leaf cells, but did not label the walls of Radula elaters. The CCRC-M7 antibody, which detects an arabinosylated (16)-linked -galactan epitope, exclusively labeled hyaline cells in Sphagnum leaves and the secondary walls of Radula elaters. Anti-pectin antibodies, LM5 and JIM5, labeled the primary wall in Megaceros elaters. LM5 also labeled the central layer of the secondary wall but only during formation. In Radula elaters, JIM5 and another anti-pectin antibody, JIM7, labeled the primary wall. The distribution of arabinogalactan–proteins and pectic polysaccharides restricted to specific wall types and stages of development provides evidence for the developmental and functional regulation of cell wall composition in bryophytes. Monosaccharide-linkage analysis of Sphagnum leaf cell walls suggests they contain polysaccharides similar to those of higher plants. The most abundant linkage was 4-Glc, typical of cellulose, but there was also evidence for xyloglucans, 4-linked mannans, 4-linked xylans and rhamnogalacturonan-type polysaccharides.Abbreviations AGP Arabinogalactan–protein - Araf Arabinofuranose - Fucp Fucopyranose - GalAp Galacturonopyranose - Galp Galactopyranose - GlcAp Glucuronopyranose - HGA Homogalacturonan - Manp Mannopyranose - RG Rhamnogalacturonan - Rhap Rhamnopyranose - XG Xyloglucan - Xylp Xylopyranose     

Establishment and characterization of ferret cells in culture

Summary Cells derived from the brain of a 6 wk-old ferret have been subcultured over 100 times and have undergone over 400 population doublings in vitro. These cells, referred to as Mpf cells, have an absolute efficiency of colony formation in excess of 45%, exhibit a mean population doubling time of 12.5 h, possess ferret-specific antigens, and have isozymes with electrophoretic properties that are the same as those of isozymes found in ferret liver. The cells exhibit a cytopathic effect and support the synthesis of progeny virus when they are infected with the viruses of lymphocytic choriomeningitis, Newcastle disease, pseudorabies, Sindbis, vaccinia, and vesicular stomatitis. The passage level of the Mpf cells, their elapsed number of population doublings, their possession of ferret-specific antigens, and the comigration of four isozymes obtained from these cells and ferret liver define the cells as an established line of ferret cells.