微生物产生的细胞分裂素

微生物产生的细胞分裂素贾小明（浙江农业大学环保系,杭州３１００２９）细胞分裂素（ｃｙｔｏｋｉｎｉｎ）是一类重要的植物激素。它的主要生理功能是刺激细胞分裂,促进侧芽发育,维持蛋白质和核酸的合成,延缓离休叶片衰老等作用。细胞分裂素可用于蔬菜保鲜、防衰和延...     

曼陀罗细胞中6-BA结合蛋白与硝酸还原酶活力的关系

曼陀罗单倍体细胞,在无水层情况下,经紫外线诱变,在氯酸盐培养基上筛选得到一株缺少硝酸还原酶活力的突变细胞株。经3年培养证明突变性状稳定。对它进行生理生化分析,看到它核内细胞分裂素结合蛋白减少。结合蛋白与细胞分裂素结合后能够促进核内 RNA 聚合酶活力,加速基因转绿。在正常细胞中,细胞分裂素能够促进硝酸还原酶诱导活力,而在这些缺少核内细胞分裂素结合蛋白的突变细胞中,外源细胞分裂素对酶活力的诱导没有作用。从而猜测,在突变细胞中,由于激素不能充分与结合蛋白结合,降低了细胞中转绿的总水平,可能是硝酸还原酶活力提不高的原因。本文还讨论了激素与受体结合后在调控基因表达上的可能途径。     

NO对He-Ne激光和增强UV-B辐射小麦幼苗气孔运动的作用机理研究

为探讨NO对He-Ne激光和增强UV-B辐射小麦（Triticum aestivuml）气孔运动的作用机理,采用低剂量（5 mW.mm-2）He-Ne激光和增强（10.08 kJ.m-2.d-1）UV-B辐射并结合药理学实验和激光共聚焦显微技术,对ML7113小麦的叶片及表皮条进行不同的处理,结果显示：（1）UV-B辐射既可诱导小麦叶片气孔关闭,又能够明显增加气孔保卫细胞和叶片的NO水平,且NO清除剂明显抑制了UV-B辐射诱导的小麦叶片气孔关闭,同时气孔保卫细胞和叶片内的NO含量明显减少。（2）一氧化氮合酶（NOS）抑制剂L-NAME对经UV-B辐射诱导的小麦幼苗气孔开度及保卫细胞和叶片内NO含量的抑制程度明显大于硝酸还原酶（NR）抑制剂NaN3对其的抑制程度,说明一氧化氮合酶（NOS）合成途径是小麦叶片经UV-B辐射后NO的主要产生途径。（3）就气孔开度而言,L〉CK〉BL〉B。就小麦叶片及保卫细胞内NO含量而言,B〉BL〉CK〉L。就硝酸还原酶（NR）和一氧化氮合酶（NOS）的活性而言,B组NR活性最低,NOS活性最高,L组NR活性最高,NOS活性最低。表明经He-Ne激光和增强UV-B辐射诱导的小麦气孔开度的变化确实与保卫细胞及叶片中NO含量的多少有关,气孔开度的减小及增大对应于NO含量的增多或减少,同时进一步证实了小麦叶片经He-Ne激光单独辐照后,NO的主要合成途径也来源于NOS途径。     

籽粒满对小麦某些光合特性的影响

籽粒满为近几年来国内研制和开发的植物生长物质,主要成分为细胞分裂素。如众所周知,细胞分裂素能有效地阻滞叶片的衰老。为此,我们在冬小麦扬花期喷施籽粒满,观察对小麦的影响。结果如下: 叶绿素和类胡萝卜素含量 1994年5月9日在小麦扬花期,我们用10×10~(-6)籽粒满水溶液对小麦植株进行喷施,在喷施处理后的第3天、7天、21天和28天时,分别对小麦旗叶的叶绿素和类胡萝卜素的含量进行了测定。结果表     

棉花子房中存在细胞分裂素的专一结合蛋白

棉花子房细胞提取物1000r/min沉淀组分(P_1)和10 000r/min沉淀组分(P_(10))均能与6-BA专一结合,但不能与ABA专一结合。非标记的激动素、玉米素和6-BA等细胞分裂素类物质均可将与P_1结合的[~3H]6-BA取代下来,其它激素IAA和ABA、细胞分裂素的结构类似物cAMP和腺嘌呤均无取代作用。胰蛋白酶处理可明显降低P_1和6-BA的专一结合,说明在棉花子房中存在细胞分裂素的专一结合蛋白。二硫苏糖醇(DTT)、半胱氨酸和还原型谷胱甘肽强烈抑制P_1的专一结合,提示某种可还原基团,很可能是二硫键,定位于结合蛋白,且与其结合活性有关。在子房细胞里存在能钝化细胞分裂素结合蛋白活性的物质,其钝化能力是热不稳定的。     

田间小麦叶片光合作用的光抑制不伴随D1蛋白的净降解

通过测定田间小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ ）叶片Ｄ１蛋白的含量、光合放氧和叶绿素ａ 荧光,探讨了叶片光合作用的光抑制与Ｄ１蛋白净降解的关系。田间的小麦叶片受到晴天中午光照约３ ｈ 以后,表观光合量子效率（Φ）、光系统Ⅱ的光化学效率（Ｆｖ／Ｆｍ ）和初始荧光（Ｆ０）明显下降;若将叶片转入弱光下,这３个指标可在１ ｈ 内基本恢复;强光照射过程中Ｄ１蛋白的含量没有显著变化;Ｄ１蛋白合成抑制剂ＳＭ 使强光下叶片的慢驰豫的非光化学荧光猝灭（ｑＥ－ｓｌｏｗ ）明显增加;在弱光下恢复时引入链霉素（ＳＭ）不影响叶片光合功能的恢复;用二硫苏糖醇（ＤＴＴ）抑制叶黄素循环使中午强光照射后的叶片中Ｄ１蛋白的含量降低３０％ 左右。这些结果都表明,田间小麦叶片光合作用的光抑制不是由于Ｄ１蛋白的净降解,而是由于非辐射能量耗散的增加引起的。     

重金属污染土壤植物修复中细胞分裂素的作用与机制

植物修复因投资成本低、环境扰动少、二次污染易控制、美化环境等优点成为重金属污染土壤修复重要的治理技术。植物内源细胞分裂素调控植物生理活动,外源细胞分裂素对植物生理生态特征产生显著影响,且在植物修复中逐渐受到研究人员的关注。细胞分裂素能够调控植物根茎发育、叶片衰老、激素传递等过程,同时在重金属胁迫下也参与蒸腾、光合、抗性、解毒等系统的运转。以细胞分裂素对植物生理活动的调控作用研究为基础,阐述了细胞分裂素在植物修复中的作用机制。主要包括：增强光合作用,延缓叶片衰老,提升植物抗性能力;调控根茎叶发育,增加植物生物量,强化植物富集效果;增强转运蛋白表达,提高叶面蒸腾作用,促进重金属吸收转运;参与解毒过程,降低重金属毒性,调控重金属体内转化。最后提出了细胞分裂素在重金属污染土壤植物修复中的研究方向,这对促进细胞分裂素在植物修复中的实际应用具有重要意义。     

6-BA延缓大豆叶片衰老的作用与膜蛋白磷酸化状态的关系

蛋白激酶（proteinkinase,PK）和蛋白磷酸酯酶（pIDt6inphOSpha～,PP）是生物体内催化蛋白质磷酸化／脱磷酸化过程的两种重要酶类。目前已有越来越多的实验证据表明：这种可逆的磷酸化／脱磷酸化过程所导致的蛋白质（酶）活性的改变是生物体内信号传导过程中的重要环节（Hunter1995）。已有一些实验系统涉及了植物激素对于植物蛋白磷酸化过程的影响（Mi－zogUchi等1994,Sano和Youssefian1994）,并有一些与此相关的蛋白激酶和蛋白磷酸酯酶的基因被克隆（kleber等1993,temp等1994）。细胞分裂素延缓植物叶片衰老的作用早已被各种实…     

细胞分裂素信号转导研究进展

对应用突变体研究细胞分裂素信号转导、细胞分裂素受体以及参与细胞分裂素信号转导相关的蛋白质作了简要介绍;并且对细胞分裂素信号途径进行了推测：细胞分裂素被受体（CKI1、IBC6或者GCR1）接受后,经传导系统转化形成特定的信息,一方面可能调节基因的表达,从而可能调节受体水平,导致细胞对细胞分裂素浓度应答范围发生改变,另外,基因表达使细胞产生相关的生理反应;另一方面形成的特定信息可能激活MAPK级联途径（mitogen-activated protein kinase cascade）,导致细胞产生相关的生理反应。     

植物体内的芳环细胞分裂素

芳环细胞分裂素包括6-苄基氨基嘌呤（BA）,6-（3-羟苄基氨基）-嘌呤（mT）,6-（2-羟苄基氨基）-嘌呤(oT)和它们的衍生物。它的代谢具有相互转换、羟基化作用、结合作用和氧化降解四个特征。本文从信号的感受,转导和应答阐明细胞分裂素在细胞和分子水平上的 功能。     

Localization of farnesyl diphosphate synthase in chloroplasts.

The subcellular localization of plant farnesyl diphosphate synthase (FPPS) was examined. Immunocytochemical staining using anti-FPPS1 antibody followed by electron microscopy showed that FPPS1 was localized to chloroplasts of rice mesophyll cells. Subcellular fractions from wheat leaves were examined by immunoblot analysis. FPPS was detected in the chloroplast fraction in wheat, and was protected from proteolysis following trypsin treatment of chloroplasts. FPPS was also detected in the chloroplast fraction of a dicot plant, tobacco.     

Vacuolar localization of proteases and degradation of chloroplasts in mesophyll protoplasts from senescing primary wheat leaves

Mesophyll protoplasts isolated from primary leaves of wheat seedlings were used to follow the localization of proteases and the breakdown of chloroplasts during dark-induced senescence. Protoplasts were readily obtained from leaf tissue, even after 80% of the chlorophyll and protein had been lost. Intact chloroplasts and vacuoles could be isolated from the protoplasts at all stages of senescence. All the proteolytic activity associated with the degradation of ribulose bisphosphate carboxylase in the protoplasts could be accounted for by that localized within the vacuole. Moreover, this localization was retained late into senescence. Protoplasts isolated during leaf senescence first showed a decline in photosynthesis, then a decline in ribulose bisphosphate carboxylase activity, followed by a decline in chloroplast number. There was a close correlation between the decline in chloroplast number and the loss of chlorophyll and soluble protein per protoplast, suggesting a sequential degradation of chloroplasts during senescence. Ultrastructural studies indicated a movement of chloroplasts in toward the center of the protoplasts during senescence. Thus, within senescing protoplasts, chloroplasts appeared either to move into invaginations of the vacuole or to be taken up into the vacuole.     

小麦低温种质的器官结构特征

通过对小麦冠层温度的长期观测,发现自然界存在株温持续偏低的低温种质,与其相对应,也有株温持续偏高的高温种质存在。应用光学显微镜和电子显微镜研究了小麦低温种质叶片显微和超微结构,测量统计了叶肉细胞长度、单位面积叶肉细胞数目、单个叶肉细胞中的叶绿体数目、叶肉细胞层数和叶绿体基粒片层数。结果表明,低温小麦种质较高温种质叶肉细胞小,排列紧密,叶肉细胞层数较多;叶绿体数量多,叶绿体基粒片层丰富;叶片维管束密集;随着生育期向成熟趋近,叶肉细胞、叶绿体、籽粒腹沟区有色层细胞等结构衰老缓慢。     

Stochastic dynamics of actin filaments in guard cells regulating chloroplast localization during stomatal movement

Actin filaments and chloroplasts in guard cells play roles in stomatal function. However, detailed actin dynamics vary, and the roles that they play in chloroplast localization during stomatal movement remain to be determined. We examined the dynamics of actin filaments and chloroplast localization in transgenic tobacco expressing green fluorescent protein (GFP)-mouse talin in guard cells by time-lapse imaging. Actin filaments showed sliding, bundling and branching dynamics in moving guard cells. During stomatal movement, long filaments can be severed into small fragments, which can form longer filaments by end-joining activities. With chloroplast movement, actin filaments near chloroplasts showed severing and elongation activity in guard cells during stomatal movement. Cytochalasin B treatment abolished elongation, bundling and branching activities of actin filaments in guard cells, and these changes of actin filaments, and as a result, more chloroplasts were localized at the centre of guard cells. However, chloroplast turning to avoid high light, and sliding of actin fragments near the chloroplast, was unaffected following cytochalasin B treatment in guard cells. We suggest that the sliding dynamics of actin may play roles in chloroplast turning in guard cells. Our results indicate that the stochastic dynamics of actin filaments in guard cells regulate chloroplast localization during stomatal movement.     

In vitro protein import of a putative amino acid transporter from Arabidopsis thaliana into chloroplasts and its suborganellar localization

We identified a gene product of At5g19500 (At5g19500p) from Arabidopsis thaliana that is homologous to EcTyrP, a tyrosine-specific transporter from Escherichia coli. Computational analyses of the amino acid sequence of At5g19500p predicted 11 transmembrane domains (TMDs) and a potential plastid targeting signal at its amino terminus. As a first step toward understanding the possible role of At5g19500p in plant cells, we attempted to determine the localization of At5g19500p by an in vitro chloroplastic import assay using At5g19500p translated in a cell-free wheat germ system (Madin et al., Proc. Natl. Acad. Sci. USA, 97, 559-564 (2000)), followed by subfractionation of the chloroplasts. At5g19500p was successfully imported into chloroplasts, and the newly transported mature form of At5g19500p was recovered from the inner envelope membrane.     

Mg2+对生长在不同光强下的小麦叶绿体光合功能的影响

研究结果表明,Mg~(2+)对生长在不同光强度下的小麦叶绿体光合功能有不同影响。与生长在低光强(2×10~8勒克斯)下的小麦叶绿体相比,Mg~(2+)更加明显地降低从生长在高光强(2×10~4勒克斯)下的小麦所分离的叶绿体的吸收光谱在红区和蓝区的吸收峰值;但它更大幅度地提高后者在低温(77K)下的PSⅡ相对荧光产量(F_(687))与PSⅠ相对荧光产量(F_(742))的比值,PSⅡ活性和PSⅡ原初光能转化效率。实验结果证明,更高的光强度可能有利于叶绿体形成更多可流动的LHC-Ⅱ和LHC-Ⅰ。     

Structure and Photosynthetic Characteristics of Awns of Wheat and Barley

The surface and the cross section of awns of wheat and barley were examined by scanning electron microscopy,ultrastructure of cells were observed under a transmisson electron microscope and the photosynthetic rates were measured with an oxygen, electrode and infra-red CO2 analyser. The main results were as follows :The cross section of wheat awn appeared to be acutely trianglular whereas that of barley awn was obtusely triangular. There were rows of stomota on either side of epidermis in both wheat and barley awns. Under the stomatic band there were green tissues. The green cells in the awn were differentiated from the parenchyma cells . The mature green cells possessed papillae which were rich in chloroplasts and mitochondria. The tamella system in chloroplasts was well developed and contained many starch grains. There were three vascular bundles in each awn. The sheath cells near the green tissues contained chloroplasts. The photosynthate in the green cells might pass through the sheath cells and companion cells to sieve elements. The highest photosynthetic rate of the awn was seen at the flowering stage ,reaching about 20 μmol CO2·m-2·s-1. The light compensation point was 70—80 μE·m-2· s-1. The light saturation point was about 1500 μE·m-2·s-1. The CO2 compensation point was 50—60 ppm and the CO2 saturation point was about 900ppm . The photosynthetic rate and stomatal conductance were easily effected by CO2 concentration, light intensity and the duration of illumination . There was a positive correlation between the photosynthetic rate and the chloro-phyll content in the awns. The CO2-releasing rate in photorespiration of awn was about 4–5 μmol CO2·m-2·s-1.     

Protein synthesis by isolated etioplasts and chloroplasts from pea and wheat and the effects of chloramphenicol and cycloheximide

Etioplasts capable of incorporating ¹⁴C-leucine into protein have been isolated from dark-grown pea and wheat plants. The requirements for leucine incorporation for etioplasts were similar to those for chloroplasts. An ATP-generating system, Mg²⁺, and GTP were required. The amino-acid-incorporation activity of etioplasts from wheat was comparable to that of chloroplasts on an RNA basis, whereas the activity of pea etioplasts was about 50% of the activity of pea chloroplasts. The incorporation of leucine into protein by etioplasts and chloroplasts from pea and wheat was inhibited by chloramphenicol, and to a slight extent by cycloheximide.     

Subcellular localization of dehydrins in wheat plant seedlings subjected to low-temperature adaptation

Subcellular localization of dehydrins (dhn) in stem cell tissues of winter wheat seedlings (Triticulum aestivum L., cult. Irkutskaya ozimaya) was studied by immunoelectron microscopy. It was found that cold hardening at 4°C for 7 days resulted in a duplication of the dhn quantity in the cells as compared with control conditions (22°C). The maximum increase of the dhn content was observed in rough endoplasmic reticulum, mitochondria, cell walls, and intercellular spaces (3.8-, 3.0- and 2.8-fold, respectively); minimum increase was found in chloroplasts (1.4-fold). In the membrane compartments (mitochondria, rough endoplasmic reticulum, chloroplasts) low-temperature stress caused an increase of dhn quantity not only near membranes but also in the intermembrane space. A significant accumulation of dhn (2.5-fold) in the nucleus under low- temperature was found. We conclude that cold hardening of the plant induces accumulation and translocation of dhn to the regions of inter- and intracellular compartments that most require protection during the low-temperature stress.     

四种生态型芦苇叶片结构的研究

The leaf anatomy and uhrastructure of the vascular bundle sheath cells of four ecotypes of reed (Phragmites communis Trin. ) were observed with light and electron microscope. The four ecotypes were the swamp reed, dune reed, heavy salt meadow reed and the light salt meadow reed. It was shown that the intracellular content of the vascular bundle sheath cells of the swamp reed was lower than that of the dune reed; that of the heavy salt meadow reed was between that of the former two reeds and that of the light salt meadow reed and that of the light salt meadow reed was similar to the dune reed. The chloroplasts in the swamp reed were round-shaped with poorly developed grana. Starch grains were not visualized. The uhrastructure of their chloroplasts were similar to those in the typical C3 plants as wheat. By contrast, the chloroplasts in the vascular bundle cells of the dune reed were elliptical with their structure similar to that of the typical C4 plants as maize. These chloroplasts contained some poorly developed grana thykloid. The chloroplasts of the vascular bundle sheath cells in the heavy salt meadow reed were similar to those seen in the dune reed, however, those in the light salt meadow reed appeared to be beth round .and elliptical in shape. The above results suggested a great habitat variation did occur among the four ecotypes of reed in which these exists an obvious evolutional trend from C3 to C4 plants.