Acute morphological changes of palisade cells ofSaintpaulia leaves induced by a rapid temperature drop

Early changes in fine structures of intracellular organelles ofSaintpaulia leaves exhibiting chilling-induced degeneration of photosynthetic activities were investigated by electron microscopy. As early as 3 min after exposure to 10C water, thylakoid lamellae in the palisade cells showed extensive disarrangement, especially in intergrana lamellae. Simultaneously, other organelles showed drastic morphological changes, including the condensation of chromatin in the nucleus.     

Low temperature-induced modifications in cell ultrastructure and localization of phenolics in winter oilseed rape (Brassica napus L. var. oleifera L.) leaves

Acclimation of winter oilseed plants in the cold (i.e. at temperatures >0 degrees C) followed by short exposure to sub-lethal freezing temperatures resulted in pronounced ultrastructural changes of leaf epidermal and mesophyll cells. The following major changes were observed upon acclimation at 2 degrees C: increased thickness of cell walls; numerous invaginations of plasma membranes; the appearance of many large vesicles localized in the cytoplasm in close proximity to the central vacuole; the occurrence of abundant populations of microvesicles associated with the endoplasmic reticulum (ER) cisternae or located in the vicinity of dictyosomes; and the occurrence of paramural bodies and myelin-like structures. In addition, large phenolic deposits were observed in the vicinity of the plasma membrane and membrane-bound organelles such as chloroplasts, large vesicles or cytoplasm/tonoplast interfaces. Transient freezing (-5 degrees C for 18 h) of the cold-acclimated leaves led to reversible disorganization of the cytoplasm and to pronounced structural changes of the cellular organelles. Chloroplasts were swollen, with the stroma occupying one half of their volume and the thylakoid system being displaced to the other half. Large phenolic aggregates disappeared but distinct layers of phenolic deposits were associated with mitochondrial membranes and with chloroplast envelopes. In frost-thawed cells recovered at 2 degrees C for 24 h, dictyosomes and dictyosome- or ER-derived small vesicles reappeared in the ribosome-rich cytoplasm. Aberrations in the structure of chloroplasts and mitochondria were less pronounced. Few phenolic deposits were seen as small grains associated with chloroplast envelopes and vesicle membranes. These observations demonstrate that plants undergo different changes in cell ultrastructure depending on whether they are subjected to chilling or freezing temperatures. Results are discussed in relation to membrane recycling and the possible role of phenolics during the first and second stages of plant acclimation at low temperature.     

EFFECTS OF 2,3,5-TRIIODOBENZOIC ACID ON THE STRUCTURE OF SOYBEAN LEAVES

The effects of 2,3,5-triiodobenzoic acid (TIBA) on soybean leaves (Glycine max. [L.] Merrill ‘Harosoy‘) include thickening with intensification of color and some raised intercostal regions, giving a wrinkled appearance. These effects are not restricted to early stages of leaf development but are pronounced during and after unfolding of the leaf. Proliferation of tracheary elements, increased procambial activity, and hypertrophy of bundle sheath extension cells occurred in the leaflet midvein of the youngest expanded leaf treated with 50 ppm or 100 ppm of TIBA. The youngest treated leaves exhibited differential growth rates and expansion within the palisade and spongy layers. Hypertrophy of spongy cells in these leaves occurred independently or simultaneously with elongation of the upper and lower palisade layers. The palisade and spongy tissues had undergone cell division and expansion at a greater pace than the epidermal layers. This, along with hypertrophy in the bundle sheath extension cells, would explain the wrinkled appearance of the lamina. The treated leaf became thicker than the control as a result of the increased number of cells in the spongy layer and elongation in the palisade layer. The observed aberrations in leaf structure suggest that TIBA interferes with some auxin-translocating system within the plant.     

贵州西北喀斯特区古茶树叶片解剖结构及抗旱性评价

该研究以贵州西北喀斯特区茶树分布集中的亮岩镇为采样点,选取亮岩镇8个村30个点的古茶树(bj-1～bj-30)叶片为试验材料,采用石蜡切片技术观察其15项叶片解剖结构指标;通过描述性及方差分析、相关性分析和聚类分析,利用相关指数大小筛选出4项典型指标;运用隶属函数综合评价了30个点古茶树的抗旱性。结果表明：(1)亮岩镇古茶树叶片由上角质层、上表皮、栅栏组织、海绵组织、下表皮、下角质层、气孔以及叶脉组成。栅栏组织大多2层,极少数叶片排列有3层栅栏组织。(2)15个叶片解剖结构指标在不同样点间均存在显著差异,且各指标的变异系数存在较大差异,其中以栅栏组织厚度变异系数最大(41.21%),栅海比变异系数次之(35.45%),叶脉厚度变异系数最小(7.31%)。(3)采用隶属函数法筛选出栅栏组织厚度、下表皮厚度、下角质层厚度以及叶脉突起度作为评价亮岩古茶树抗旱性的典型指标;综合分析发现bj-15、bj-16、bj-29、bj-10和bj-24的抗旱性较强, bj-22、bj-20、bj-2、bj-11以及bj-6的抗旱性较弱。     

蒙古莸叶片解剖结构的地理种源变异及其对环境变化响应的意义

长期受到生长环境影响而形成的遗传变异对植物生长发育有着显著的影响。叶片是植物对环境变化最敏感的器官, 了解叶片解剖结构在不同环境中产生的适应性变异是探索植物对环境适应的基础。同质园试验是研究遗传与环境因素对植物生长代谢等影响的一种有效方法, 该研究利用同质园试验排除了环境梯度的影响, 通过常规石蜡切片、多重比较、相关性分析、一般线性模型分析等方法, 对7个不同种源地的蒙古莸(Caryopteris mongholica)叶片解剖结构及其影响因素进行了定量比较。结果表明, 7个种源地的蒙古莸叶片均为等面叶, 无海绵组织分化, 其上表皮细胞较下表皮细胞厚, 上栅栏组织较下栅栏组织厚; 叶片各解剖结构参数间存在显著的自相关性, 不同种源叶片解剖结构存在显著差异: 随种源地年平均气温升高, 叶厚度、栅栏组织厚度呈增大趋势, 其中, 最西南部的阿左旗种源蒙古莸叶片的上下栅栏组织、叶厚度及叶片结构紧密度值均最大, 表现出明显的抗旱特征。种源地经纬度、气温、降水等对解剖结构指标有显著的影响, 其解释程度为34.09%-81.43%。同质园试验说明, 种源地气候差异驱动的遗传变异是引起不同种源叶片解剖结构差异的重要因素。     

Cellular fine structures and histochemical reactions in the tissue of a cypress twig preserved in Baltic amber

A twig of a cypress plant preserved for ca. 45 Myr in Baltic amber was analysed by light and electron microscopy. Cross-sections of the whole plant showed an almost intact tissue of the entire stem and leaves, revealing, to our knowledge, the oldest and most highly preserved tissue from an amber inclusion reported so far. The preparations are based on a new technique of internal imbedding, whereby the hollow spaces within the inclusion are filled with synthetic resin which stabilizes the cellular structures during the sectioning procedure. Cytological stains applied to the sections reacted with cell walls and nuclei. A strong green auto-fluorescence of the cuticle and the resin canals in the leaves was observed. Transmission electron micrographs revealed highly preserved fine structures of cell walls, membranes and organelles. The results were compared with taxonomically related recent Glyptostrobus and Juniperus plants.     

TISSUE PREPARATION AND FINE STRUCTURE OF THE RADICLE APEX FROM COTTON SEEDS

A comparative methodological study was made of the fine structure of apical cortical cells in excised radicles from cotton (Gossypium hirsutum L. var M-8) seeds. Radicles from dry seed had 12% moisture content and were prepared for electron microscopy using several different techniques. These included different methods of chemical fixation or freeze-fracture and etching of unfixed tissue for transmission electron microscopy (TEM) and cryofracturing of fixed and dehydrated radicles for scanning electron microscopy (SEM). Cortical cells had a similar appearance regardless of the method used in tissue preparation. Cell walls had a pronounced waviness which was particularly evident in SEM images of cells lining the elongated intercellular air spaces. The plasma membrane (PM) delimited the cytoplasm of each cell as an intact unit membrane. Single layers of tightly-packed lipid bodies (LB) were apposed to the PM and protein bodies (PB). Distension of cells, membranous organelles and LB was observed in radicles fixed by immersion in aqueous solutions, suggesting that a certain amount of hydration occurred during fixation. This interpretation was supported by the compact appearance of cells and organelles in tissue prepared by freeze-etch or vapor fixation. We conclude that freeze-fracture and etching of unfixed tissue provided the best information for cell morphology and structure of membranes and organelles in dry tissue. Complementary data on the fine details of nuclei and cytoplasmic organelles were best observed with TEM of fixed tissue. These data when viewed collectively indicate the advantage of using several techniques to obtain analogous and complementary information essential for establishing a baseline level of information on the fine structure of cells in dry tissue.     

金叶连翘叶片色素含量和解剖结构研究

金叶连翘不同冠层的成熟叶片呈现为不同颜色。以朝鲜连翘深绿色叶为对照,观察金叶连翘冠层上、中、下位叶色,测定其叶片大小和叶绿素a、叶绿素b、总叶绿素及类胡萝卜素含量,同时观察分析叶片横切面解剖结构,旨在阐明叶片色素含量和解剖结构对叶色的影响。研究表明:上层黄色、中层黄绿色、下层浅绿色,黄、黄绿、浅绿色叶总叶绿素含量分别是对照组的0.51%、4.44%和66.47%,均极显著低于对照(P <0.01),但黄绿叶的叶绿素a/b比值显著升高(P <0.05),黄、黄绿叶的总叶绿素/类胡萝卜素比值极显著降低(P <0.01)。黄、黄绿叶的叶绿体发育停滞于单片层时期,类囊体分化程度低,浅绿叶类囊基粒片层肿胀;黄叶细胞器降解,栅栏组织细胞形状难以辨别,黄绿叶上表皮细胞凸起。金叶连翘属于总叶绿素及叶绿素b合成减少型突变体,表现为叶绿素严重缺失,类胡萝卜素相对含量升高;其叶绿体发育停滞,类囊体结构异常,是金叶连翘叶片呈现不同颜色的主要因素,与其叶片解剖显微结构无关。     

The paraveinal mesophyll of soybean leaves in relation to assimilate transfer and compartmentation

Nitrogen and carbohydrate assimilates were temporally and spatially compartmented among various cell types in soybean (Glycine max L., Merr.) leaves during seed filling. The paraveinal mesophyll (PVM), a unique cell layer found in soybean, was demonstrated to function in the synthesis, compartmentation and remobilization of nitrogen reserves prior to and during the seed-filling stages. At anthesis, the PVM vacuoles contain substantial protein which completely disappears by two weeks into the seed filling. Distinct changes in the PVM cytoplasm, tonoplast and organelles were correlated with the presence or absence of the vacuolar material. Microautoradiography following the accumulation of several radiolabeled sugars and amino acids demonstrated the glycoprotein nature of the vacuolar material. Incorporation of methionine, leucine, glucose, and glucosamine resulted in heavy labelling of the PVM vacuole, in contrast to galactose, proline, and mannose which resulted in a much reduced labelling pattern. In addition, starch is unequally compartmented and degraded among the various leaf cells during seed filling. At the end of the photoperiod at the flowering stage, the highest starch accumulation was in the second palisade layer followed by the spongy mesophyll and the first (uppermost) palisade layer. Starch in the first palisade layer was completely degraded during the dark whereas the starch in the second palisade and spongy mesophyll was not remobilized to any appreciable extent. By mid-podfilling (approximately five weeks postanthesis) starch was absent in the first palisade layer at the end of the photoperiod while the second palisade and spongy mesophyll layers contained substantial starch. Starch was remobilized from these latter cells during the remainder of seed filling when current photosynthetic production is low. Structural changes associated with cell senescence first appear in the upper palisade layer and then progress (excluding the PVM) to the second palisade and spongy mesophyll layer. The PVM and phloem appear to retain their structural integrity into the leaf yellowing stage. Reducing sink capacity by pod removal resulted in a continued accumulation of vacuolar protein, an increase in cytoplasmic volume, and fragmentation of the vacuole in the PVM. Pod removal also resulted in an increased amount of accumulated starch (which did not turn over) in all mesophyll layers, and an increase in cell size and cell-wall thickness.     

Ultrastructural features of chilling-injury in Episcia reptans

Abstract. Chilling the leaves of the extremely chill-sensitive plant Episcia reptans for only a few hours at 5°C causes ultrastructural changes such as swelling and disorganization of chloroplasts and mitochondria, and vesiculation of cytoplasmic membranes. Rewarming plants after 5–6 h chilling causes further deterioration in chloroplast and mitochondrial structure and the appearance of fibrous material in the cytoplasm of epidermal, mesophyll and vascular cells. The nature of the deposit is not known but it may be the cause of the rapid development of chilling-injury on return of plants to the warmth.     

家蚕消化管内桑叶的银染法研究

通过银染法对家蚕整体染色,结果表明:家蚕消化管内的桑叶由叶表皮、叶肉和叶脉组成。叶表皮包括上表皮和下表皮。上表皮细胞可分为三种:钟乳体细胞、绿色表皮细胞和黄色表皮细胞;下表皮内含有气孔;叶肉组织内含有晶体,其中海绵组织内的最多。家蚕消化管由前向后可分为前肠、中肠和后肠,由外向内依次为肌层、底膜、上皮细胞层、内膜。中肠最为发达,其发达的上皮细胞向内表面突起形成许多大的指突形皱褶;上皮细胞层内有圆筒形细胞、杯形细胞两种细胞,两者在形状、功能以及嗜银性等方面有所差异。家蚕消化管对桑叶不同组织的消化吸收效率有差异,上表皮吸收效率最高,下表皮和栅栏组织次之,最低的是海绵组织。采用动物细胞染色方法对植物细胞进行染色,并与常规植物学染色方法进行了比较;依据细胞嗜银性的不同,可将桑叶的上表皮细胞分为两种亚型。     

枫香变红过程中叶片组织结构、光合特性及色素含量变化研究

枫香(Liquidambar formosana)因其叶片入秋后逐渐变红而极具观赏价值,是优良的景观生态树种。为了解枫香叶片结构变化与叶色的关系,该文通过连续监测枫香叶片变红过程中组织结构、光合特性及色素含量的变化,分析叶片结构与其光合特性和色素的关系。结果表明：(1)叶片变色过程中,表皮细胞均为椭圆形,紧密排列,未观察到明显的细胞变异,表面未附着绒毛和蜡质,且上表皮细胞与栅栏组织细胞间排列紧密,未出现较大的气室。(2)随着叶片逐渐变红,叶片结构变化显著,其中叶片、上表皮、栅栏组织和海绵组织厚度及气孔开度均逐渐减小,而气孔器长和宽、单个气孔器面积则逐渐增大。(3)随着叶片结构的变化,其叶绿素含量逐渐减少,致使净光合速率逐渐减小,在出现光破坏时,叶片通过在栅栏组织细胞液泡内合成花色苷来自我保护,而大量的花色苷致使叶片表面呈现红色。综上认为,叶绿素含量降低,花色素苷大量积累是导致枫香叶片变红的直接原因,而枫香叶色变红则是其一系列生理结构特征综合作用的结果。     

Production of volatiles by degradation of lipids during manufacture of black tea

During manufacture of black tea, lipids are degraded to volatile constituents. Cis-3-hexenal was present in appreciable amounts in the various parts of fresh shoots and decreased in the second leaves during manufacture. There was a simultaneous increase in trans-2-hexenal. Linalol and methyl salicylate also increased appreciably during rolling and fermentation. Most of the volatiles were lost during the firing process. The above trend was borne out by the ‘potential’ of the leaves for the production of volatiles as indicated by the increased amounts of volatiles produced by homogenizing the tissue in water against controls homogenized in 0.1 N acid. The C₆-aldehydes present in the headspace of withered shoots increased significantly following mechanical damage. The major fatty acids of the lipids in the various parts of the shoots were linolenic, linoleic, palmitic, oleic and stearic acids. The ratio of linoleic to linolenic acid in the stems was much higher than that of the leaves or buds and this was reflected in its higher 'potential for formation of hexanal. During withering and rolling of the second leaves, the unsaturated fatty acids showed substantial losses compared with the saturated acids. It is suggested that the enzymic breakdown of membrane lipids initiate the formation of volatile carbonyl compounds which are partly responsible for the flavour of black tea.     

Effect of water stress on cotton leaves : I. An electron microscopic stereological study of the palisade cells

Palisade cells from fully expanded leaves from irrigated and nonirrigated, field grown cotton (Gossypium hirsutum L. cv. Paymaster 266) were subjected to a microscopic examination to evaluate the effect of water stress on subcellular structures. The water potential difference between the two treatments was 13 bars at the time of sampling. The dimensions of the palisade cells and their density per unit leaf area were determined by light microscopy. Palisade cells from stressed plants had the same diameter, but were taller than their counterparts in irrigated plants. The density of the palisade cells was the same in both treatments as was the fractional volume of the intercellular space. It was concluded that the reduced leaf area observed in the stressed plants resulted primarily from a mitotic sensitivity to water stress. Further, expansion of palisade cells was not inhibited by the stress imposed in this study.

Morphometric analysis of electron micrographs was used to evaluate the subcellular structure of palisade cells from nonstressed and stressed plants. The fractional volumes of cell walls, total cytoplasm, chloroplasts, starch granules, intrachloroplast bodies, mitochondria, peroxisomes, and central vacuoles were determined. The surface densities of grana and stroma lamellae, outer chloroplast membranes, mitochondrial cristae, endoplasmic reticulum and Golgi cisternae were also measured. The number of chloroplasts, mitochondria, and peroxisomes were determined. These data were expressed as actual volumes, areas, and numbers per palisade cell for each treatment. Palisade cells from stressed plants had thinner cell walls, larger central vacuoles and approximately the same amount of cytoplasm compared to cells from nonstressed plants. Within the cytoplasm, stressed plants had more but smaller chloroplasts with increased grana and stroma lamellae surfaces, larger mithchondria with reduced cristae surfaces, smaller peroxisomes and reduced membrane surfaces of endoplasmic reticulum and Golgi cisternae.

     

SUBEPIDERMAL AIR SPACES: BASIS FOR THE PHENOTYPIC EXPRESSION OF THE ARGENTEUM MUTANT OF PISUM

The Argenteum (Arg) mutant of pea is characterized by a gray-green, silvery cast of the leaves and stipules. Light and electron microscopy showed that the nature of the silvery cast was due to extensive intercellular air space between the epidermis and palisade parenchyma. Adaxial epidermal strips from mutant leaves were free of palisade cell wall fragments, whereas similar strips from normal leaves had remnants of the torn palisade cells attached to the epidermal cell walls. The periphery of the epidermal-palisade cell wall interfaces showed a more electron-opaque middle lamella in the normal leaves than in the Arg mutant leaves.     

Cytological studies on rust fungi. (vi) fine structures of infection process of Kuehneola japonica (diet.) dietel

The behavior of rust fungi in their host plants has been elucidated by electron microscopy. However, most of the ultrastructural studies on rust fungi have focused on the uredial stage. In order to elucidate the features of the sporidial stage, we studied the fine structure of Kuehneola japonica, a short-cycle rust, in rose leaves. Infection pegs arising from appressoria penetrated the host walls. Papillae formed at the time of penetration against the outer epidermal cell walls. The papillae which had formed at the penetration sites grew extensively and partially surrounded the intracellular hyphae which were connected with the infection pegs. The intracellular hyphae in the epidermal cells developed further and entered adjacent parenchyma cells. Walls of parenchyma cells either invaginated or thin papillae formed at penetration sites and the invaginated walls or papillae surrounded the necks of the intracellular hyphae. Intracellular hyphae in both epidermal and parenchyma cells were not enveloped by the sheath before 20 days after inoculation. In specimens prepared 20 days after inoculation, some of the intracellular hyphae were enveloped by a sheath in both palisade and spongy parenchyma cells. The sheathed hyphae resembled haustoria of other rust fungi which had been described previously. Teliospore initials were formed in mycelial masses in intercellular spaces between the epidermal cells and palisade parenchyma cells 20 days after inoculation. Uninucleate teliospores developed from teliospore initials 30 days after inoculation.Contribution No. 32.     

Expression and location of caffeine synthase in tea plants

Tea caffeine synthase (CS) is a key enzyme in tea plant for synthesis of caffeine. We firstly systematically investigated the gene expression of CS. Northern blot analysis showed that CS gene was not expressed in stems and roots but efficiently expressed in leaves of tea plants. The expression level of CS gene in summer-grown leaves was much higher than in spring leaves. Its expression level in leaves of the shaded or fertilized tea plants was significantly higher. Along the same shoot, CS gene was expressed at much higher level in the young leaves (bud and the first leaf) than in the more mature second and third leaves. RNA in situ hybridization indicated that tea CS gene was mainly expressed in the palisade parenchyma and the epicuticle of leaves but less expressed in the spongy parenchyma and the hypoderm. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 5, pp. 786–789. The text was submitted by the authors in English.     

Fine structure of isolated mesophyll protoplasts of tobacco

Summary Protoplasts of palisade cells isolated enzymatically from mature leaves of tobacco were studied with the electron microscope. A cell wall was completely absent, and the chloroplasts contained large inclusion bodies which were believed to be a crystalline form of fraction I protein. The fine structure of the protoplasts was otherwise that of healthy mesophyll cells, indicating that they are in a good physiological state. Some protoplasts were multinucleate as a result of fusion during the isolation process.     

6种含笑属植物抗寒性分析与综合评价

以6种含笑属植物为试材,进行田间冻害调查,拟合Logistic方程计算其低温半致死温度,采用石蜡切片技术,测定9项叶片解剖结构指标,运用隶属函数法对各树种的抗寒性进行综合分析与评价.结果表明: 6种含笑属植物叶片的相对电导率与0～25 ℃低温处理3 h的半致死温度之间呈显著正相关.6种含笑属植物低温半致死温度在-20.48～-8.67 ℃,高低顺序为深山含笑>红花深山含笑>峨眉含笑>杂交11-8>阔瓣含笑>六瓣含笑;叶片表皮细胞1～2层,垂周壁略呈波浪状,栅栏组织细胞1～3层,9项叶片解剖结构指标差异显著,栅栏组织厚度、栅海比和主脉厚度是影响抗寒性的主要叶片解剖结构指标.电导法和隶属函数法对6种含笑属植物的抗寒性评价结果与田间调查结果基本一致.6种含笑属植物抗寒性强弱顺序为:六瓣含笑>阔瓣含笑>杂交11-8>峨眉含笑>红花深山含笑>深山含笑.     

银杏贮粉室发生部位的珠心细胞程序性死亡的形态学观察

银杏（Ginkgo bilobaL)贮粉室的发生涉及位于珠孔端的珠心细胞的程序性死亡（PCD）,本研究观察了贮粉室发生过程中发生PCD的珠心细胞的形态学变化。这些珠心细胞在PCD过程中形态变化显著,细胞组分有序地降解,液泡在此起关键作用。在液泡化过程中,细胞质基质和一些细胞器被液泡所吞噬,此时的细胞器结构完整。当液泡膜破裂,细胞质基质消失之后。细胞器才逐步解体。最终,这些珠心细胞仅具有残留的细胞壁,随着胚珠的生长,细胞壁也被破坏,在整个PCD过程中,内膜系统发生明显改变;细胞质膜出泡,产生多泡体;形成多环膜结构;出现由膜包围的小体,其中含有细胞质基质和一些细胞器;液泡膜破裂;细胞器解体;细胞中出现大量的小膜泡。珠孔端的珠心表皮开裂形成贮粉室的开口有两种方式：一种为专一细胞的自溶,而另一种是在两个邻接细胞的中胶层处分离,没有发生细胞的自溶破裂。贮粉室开口位置的特定表皮细胞在开裂发生前就死亡,从而提前标示出表皮开裂的发生位置。这些细胞形态的变化反映出银杏珠心细胞的死亡是受发育调控的PCD过程。