Quantitative Assessment of Ultrastructural Changes in Primary Roots of Corn (Zea mays L.) after Geotropic Stimulation: I. Root Cap

The root cap is the site of gravity perception. In the study of caps of primary roots of corn (Zea mays L.), we compared the ultrastructure of geotropically responding roots that had received a 661 nm (red) irradiation (60 second) with nonresponding dark control roots kept in the dark, at comparable times following geotropic stimulation for a total of 150 minutes. The outstanding differences in the light-exposed root caps at the ultrastructural level were: (a) significantly more Golgi apparatus (dictyosomes) were found in the top than in the bottom of red-exposed cells; a random distribution is seen in the dark control cells; (b) the nucleus preferred the top in a greater number of the red-exposed cells; (c) the pattern of mitochondria localization was identical in both treatments, a greater preference for the top; however, the number of mitochondria was reduced in the bottom of red-treated cap cells as compared to the control cells. A lowering in number in the bottom of the red-treated cells was noted also in the dictyosomes; and (d) in a small percentage of cells that showed a preferential distribution of endoplasmic reticulum (ER), more red-exposed cells than controls, during the period 30 to 135 minutes after stimulation, had less ER in the top; however, a majority of the cells in both treatments showed no preferred position for ER distribution. Commonalities in ultra-structural behavior also existed between the red- and dark-treated root cap cells: (a) sedimentation of amyloplasts, with no difference in total number between treatments; and (b) a close association between amyloplasts and ER in both groups.

Polarization of organelles occurred in both the geotropically responding and nonresponding roots. The differences in dictyosome and nuclear localization, and dictyosome and mitochondrial number could be correlated with the tropic response in the red-exposed roots and no response in the dark roots, which in turn could be related to the reported hormonal events in the geotropism of roots.

     

Distribution and activation of the Golgi apparatus in geotropism

We find a differential distribution of dictyosomes in the avascular tip cells of the oat (Avena sativa) coleoptile upon geostimulation. A differential activation (increased vesicle production) of the dictyosomes with respect to gravity also occurs, but only in the tip cells of the lower tissues. Similar differences in distribution and activation of dictyosomes occur also in cells subjacent to the avascular tip (1st and 5th millimeter from the apex) of both the upper and lower half-tissues. When only the outer epidermal cells below the apex are considered, the differential distribution and activation of dictyosomes occur only in the lower outer epidermis. The changes in distribution of dictyosomes begin at 6 minutes, or sooner, from the start of geostimulation, before the onset of geotropism. The kinetics of amyloplast sedimentation and Golgi movement do not appear to differ in the cells of the avascular tip. We suggest that the Golgi participates in, and possibly initiates, the differential elongation of cells of geotropically stimulated coleoptiles.     

Effects of Hydroxyurea on the Ultrastructure of Giant Cells in Galls Induced by Meloidogyne javanica

Hydroxyurea (HU) at concentrations of 10 or 20 mg/liter was included in a medium on which excised tomato roots infected with the root-knot nematode Meloidogyne javanica were grown. In the HU, treated roots, giant cells were small and contained large vacuoles. Giant cell nuclei were amoeboidal with relatively small nucleoli in treated roots, compared with giant cells of nontreated galls. In treated-root giant cells, the cytoplasm was diffuse and few organelles such as mitochondria, dictyosomes, and endoplasmic reticulum were detected; also, walls of giant cells were thin with less extensive ingrowths than in nontreated roots. We conclude that HU suppressed normal giant cell formation interfering with its function as a feeding cell.     

Effects of Indoleacetic Acid on the Quantity of Mitochondria, Microbodies, and Plastids in the Apical and Expanding Cells of Dark-grown Oat Coleoptiles

We determined the number of mitochondria, microbodies, and plastids in dark-grown oat (Avena sativa) coleoptiles following incubation in indoleacetic acid (IAA) for a period of 60 minutes at 6-minute intervals. In the apical outer epidermis of coleoptiles, the mitochondria increased from 31.4 to 35 per cell section with a 6-minute incubation in IAA, and this trend persisted over the 60-minute incubation. Neither the microbodies, plastids, nor the dicytosomes (Gawlik and Miller 1974 Plant Physiol 54:217-221) responded to the hormone. The apical parenchyma showed no change in quantity of any of the organelles including the dictyosomes during IAA incubation. The quick response of mitochondria in the coleoptile tip could be interpreted as an association of this organelle with hormone transport, growth, or perhaps with gravity perception. In the subapical expansion region, IAA caused significant reductions of mitochondria, microbodies, and dictyosomes in the outer epidermis compared to the control, the timing of which preceded the IAA-induced elongation and of geotropism. The fast response of organelles in the various cells is probably a change in organelle volume rather than number. That microbodies show a response to the plant hormone in the permanently achlorophyllous epidermis indicates that these organelles, in addition to their peroxisomal functions in green leaves, also may have a growth regulation function. IAA treatment was without effect on the quantity of the various types of plastids (including the amyloplasts) in the different oat coleoptile cells.     

ULTRASTRUCTURAL CYTOLOGY OF CALLUS CULTURES OF STREPTANTHUS TORTUOSUS AS AFFECTED BY TEMPERATURE

Tissue culture cells of Streptanthus tortuosus var. orbiculatus (Cruciferae) which have acquired a spherical viruslike particle located in their nucleoli, designated cell line STV, developed supergranal chloroplasts and lost the ability to differentiate vascular tissues. The effect of temperature on the ultrastructural cytology of one line of the STV tissue, STV-I, was compared with the effect of temperature on the ultrastructural cytology of tissue culture cells lacking the viruslike particles (control cell lines). At 4 C, the cellular and ultrastructural appearance of control tissue culture cells differed from that of tissue grown at 22 C by producing increased amounts of endoplasmic reticulum and dictyosomes and by reduction of chloroplast thylakoids. STV-I cells were generally moribund as a result of 4 C treatment. Chloroplast thylakoids were also reduced in control tissue following growth at 10 C and the apparent quantities of endoplasmic reticulum and dictyosomes were similar to those observed in control cells grown at the control temperature (22 C), but less than those observed in tissue subjected to 4 C. STV-I tissue grown at 10 C demonstrated increased endoplasmic reticulum and dictyosomes and reduction of polysomal configurations. The mitochondrial morphology was variable and the cells contained supergranal chloroplasts and proplastids. At the control temperature (22 C), the fine structural appearance of control tissue culture cells was typical of parenchyma cells, but STV-I cells contained mitochondria of variable morphology and two types of chloroplasts— normal and supergranal. Control tissue grown at 30 C also contained proplastids, but these proplastids contained starch in contrast to the proplastids in control tissue grown at low temperatures. The ultrastructural cytology of STV-I cells grown at elevated temperature (30 C) was characterized by enlarged mitochondria containing massive lipid bodies and the presence of protoplastids with starch and supergranal chloroplasts.     

Structure of Columella Cells in Primary and Lateral Roots of Ricinus communis (Euphorbiaceae)

Horizontally oriented primary roots of Ricinus communis aremore graviresponsive than similarly oriented lateral roots.The more pronounced graviresponsiveness of primary roots ispositively correlated with their caps having a more extensivecolumella tissue than caps of lateral roots. Individual columellacells of primary roots contain 2.6 times more protoplasm thando columella cells of lateral roots. Similarly, the absolutevolumes of all cellular components in columella cells of primaryroots are larger than those of lateral roots. However, thereare no statistically significant differences in the relativevolumes of any cellular component in columella cells of primaryvs lateral roots. Endoplasmic reticulum is distributed randomlyin columella cells of both types of roots. Columella cells ofprimary and lateral roots contain numerous sedimented amyloplastswhich do not consistently contact any cellular structure. Nucleitend to be located in the middle thirds of the columella cells,and the vacuole is found in largest concentrations in the middleand upper thirds of columella cells of both types of roots.The largest protoplasmic volumes of mitochondria occur in thelower thirds of columella cells, and dictyosomes are found insimilar concentrations throughout the cells. There is no significantdifference in the intracellular distributions of organellesin columella cells of primary vs lateral roots. We believe thatthe differing graviresponsiveness of primary vs lateral rootsof R. communis is probably due to factors other than the structuresof their individual columella cells. Ricinus communis, columella, graviperception, graviresponsiveness, roots, root cap     

NaCl胁迫对宁夏枸杞叶和幼根显微及超微结构的影响

以宁夏枸杞为材料,采用超薄切片技术制备样品,应用光学显微镜和透射电镜分析了不同浓度NaCl胁迫条件下宁夏枸杞叶和幼根显微及超微结构的变化。结果表明：随着NaCl胁迫的加重,(1)叶片上表皮细胞增厚,栅栏组织细胞出现缩短现象,排列疏松且紊乱;幼根的初生结构无明显变化。(2)叶片栅栏组织中叶绿体不再紧靠在细胞膜上,叶绿体双层膜破坏,基粒片层松散排列,杂乱无章,出现膨胀和空泡现象,淀粉粒和嗜锇颗粒增多,叶肉细胞中线粒体发生轻微变化;幼根中皮层薄壁细胞线粒体形状发生改变,结构破坏,内膜和外膜模糊甚至破裂,大多数嵴模糊,出现空泡现象;细胞核解体,基质外溢。研究表明, 不同浓度的NaCl胁迫对宁夏枸杞叶片和幼根细胞的显微及超微结构影响不同,NaCl浓度大于200 mmol/L时,宁夏枸杞叶片和幼根细胞的显微及超微结构发生了明显变化,且叶肉细胞中线粒体的变化没有叶绿体的变化显著,推测叶肉细胞中线粒体的耐盐性比叶绿体强。     

Immunocytochemical localization of indole-3-acetic acid in primary roots of Zea mays

Colloidal gold-labelled antibody was used to localize indole-3-acetic acid in caps of primary roots of Z. mays cv. Kys. Gold particles accumulated on the nucleus, vacuoles, mitochondria, and some dictyosomes and dictyosome-derived vesicles. This is the first localization of indole-3-acetic acid in dictyosomes and dictyosome-derived vesicles and indicates that dictyosomes and vesicles constitute a pathway for indole-3-acetic acid movement in and secretion from root cap cells. Our findings provide cytochemical evidence to support the hypothesis that indole-3-acetic acid plays an important role in root gravitropism.     

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。     

A role for Ca in the cellular differentiation of root cap cells: A re-examination of root growth control mechanisms

Ultrastructural investigations of root caps (Zea mays) have shown a correlation between the depletion of extracellular Ca²⁺ resources by treatment of intact roots with 50 mM EGTA and changes in the activity of peripheral cap cells, involving reductions in the quantity and changes in the appearance of Golgi apparatus-derived material. In EGTA-treated roots the development of the Golgi apparatus of peripheral cap cells was severely altered and there was no evidence of a granular secretory product, which was encountered in the vesicles of dictyosomes of control roots. Vesicles of dictyosomes of EGTA-treated roots were small and the development of dictyosomes was reminiscent of that encountered in central cap cells of control roots. A decrease in amyloplast numbers brought about by EGTA was concomitant with reduced secretory activity. EGTA treatment was also associated with a redistribution of amyloplasts located in central cap cells. Decreased cap volume arising from EGTA treatment was considered indicative of a decline in overall cap activity. Proposals are made with regard to the integration of intercellular activities in the response of plant roots to stimulus-modulated signals. It is suggested that alterations in peripheral cap cell secretory activity arising from the redistribution of Ca²⁺ may account for the anisotropic growth response of gravireactive roots.     

Intracellular distribution of mitochondria after geotropic stimulation of the oat coleoptile

The number of mitochondria is greater in the bottom than in the top of cells in geotropically stimulated oat (Avena sativa) coleoptiles. In the avascular tip and outer epidermis of subapical regions this difference occurs only in the lower tissues. These inequalities are found both in the KMnO₄⁻ and in the glutaraldehyde-fixed tissues; however, they are significant only in the former. Also, the number of mitochondria scored is consistently lower when the tissues were fixed in KMnO₄. These results suggest that mitochondria undergo a small degree of sedimentation after geostimulation, a redistribution reduced by the slower fixation with glutaraldehyde. Differences in mitochondrial number begin later than those in the amyloplast and the Golgi apparatus after geotropic stimulation. The cells in the avascular-tip region (a region having an important role in geotropism) have two to three times more mitochondria than the subapical cells.     

Ultrastructure of Transfer Cells in Endosperm of Coix lacryma-jobi

Special attention was paid to the ultrastructure of transfer cells (TCs) in different locations of basal endosperm in Coix lacryma-jobi at 10 and 25 days after pollination. At 10 days after pollination. TCs of the outermost layer had long wall ingrowths (WIs) whereas those of the second layer possessed fewer and shorter Wis. In both layers TCs had a lobed nucleus, abundant mitochondria, rough endoplasmic reticulum (RER), ribosomes, and a certain number of dictyosomes and vesicles which contained dense substance connected with plasma membrane of WIs. Mitochondria were located near or between WIs. The distribution of organelles in TCs of the second layer was similar to that of the outermost layer. Mitochondria had well defined cristae and dictyosomes and RER seemed more numerous than in TCs of the outermost layer. At 25 days after pollination, TCs of the outermost and the second layer were almost filled with Wis but the organelles were recognizable. TCs of the fourth layer had branched and network-like WIs, many mitochondria, starch grain within plastids and lipids locating near WIs and in the interstices of WIs. Dictyosomes were frequently found but less RER fragments were seen. TCs of the fifth layer with short WIs contained large starch grains and small protein bodies. Plasmodesmata were not observed in the walls of TCs of the outermost and second layer at both 10 and 25 days after pollination but were found in the walls of TCs of the fourth and upper layers and also in the network-like WIs at 25 days after pollination. The roles of the organelles and functions of TCs of different layers were discussed.     

Ultrastructural investigations of Arbutus unedo-Laccaria amethystea mycorrhiza synthesized in vitro

Summary Anatomy and ultrastructure of the arbutoid mycorrhiza of Arbutus unedo-Laccaria amethystea from axenic culture are described. In comparison to non-inoculated roots, the rhizodermal cells of mycorrhizas are of greater volume, their nuclei are enlarged and show an irregular shape, plasmalemma and cytoplasm with mitochondria, plastids, endoplasmic reticulum and dictyosomes are increased. Several ontogenetical states are documented. The arbutoid mycorrhiza as a connecting link between ectomycorrhiza and ericoid mycorrhiza is discussed.     

ULTRASTRUCTURE OF THE SHOOT APEX OF CHENOPODIUM ALBUM AND CERTAIN OTHER SEED PLANTS

The ultrastructure of cells of the vegetative shoot apices is described for Chenopodium album, Kalanchoë blossfeldiana and K. laxiflora, Bryophyllum daigremontianum, Nicotiana rustica, and N. tabacum (Maryland Mammoth), and Ginkgo biloba. A less intensive study was made of the last three listed. The structures and organelles usually associated with meristematic cells were observed: dictyosomes, plastids (in various stages of development), mitochondria, endoplasmic reticulum (ER), vacuoles, lipid droplets, and plasmalemma. In addition, spherosome-like structures were observed in all zones of the shoot apices. Also, multivesicular bodies were observed in C. album and B. daigremontianum. Ribosome density is greater in cells of the flank meristem. Proplastids, plastids with prolamellar bodies, or grana have a differential distribution in the apex, characteristic for a particular species. Confirmation could not be given to the concept that vacuoles arise as a series of local dilations in long extensions of the so called "smooth ER." The tonoplast and ER are distinguishable at the time of inception of a vacuole, although the tonoplast may arise from the ER. Rapid growth of a vacuole and/or fusion with other vacuoles may result in irregularly shaped prevacuoles. No vacuoles were observed to originate from cisternae of dictyosomes in the species studied.     

ELECTRON MICROSCOPE STUDIES ON THE DICTYOSOMES AND ACROBLASTS IN THE MALE GERM CELLS OF THE CRICKET

The dictyosome (Golgi body) in the secondary spermatocyte of the cricket appears in electron micrographs as a duplex structure composed of (a) a group of parallel double-membraned lamellae and (b) a group of associated vacuoles arranged along the compact lamellae in a chain-like fashion. This arrangement of ultramicroscopic structure for the dictyosomes is strikingly comparable to that described for the Golgi apparatus of vertebrates. Accordingly, the two are considered homologous structures. Associated with the duplex structure of the dictyosomes is a differentiated region composed of small vacuoles. This is thought to represent the pro-acrosome region described in light microscope preparations. In the spermatid the dictyosomes fuse, giving rise to the acroblast. Like the dictyosomes, the acroblasts are made up of double-membraned lamellae and associated vacuoles. In addition, a differentiated acrosome region is present which, in some preparations, may display the acrosome vacuole and granule. Both the dictyosomes and acroblasts are distinct from mitochondria.     

薏苡胚乳传递细胞的超微结构

传粉后１０ ｄ,薏苡（Ｃｏｉｘ ｌａｃｒｙｍ ａ－ｊｏｂｉＬ．）颖果基部胚乳最外层传递细胞具长而多的壁内突,第二层细胞的壁内突较第一层的短而少,均具瓣裂的细胞核、丰富的线粒体、粗糙内质网、核糖体、产生小泡的高尔基体及与壁内突质膜相连的、含深色物质的囊泡。线粒体分布于壁内突附近或其间。授粉后２５ ｄ,第一、二层细胞壁内突发达,几乎充满了细胞,但细胞器可见。第四层传递细胞具树枝状及网状的壁内突,大量线粒体、具质体膜的淀粉粒、脂体存在壁内突附近或壁内突的间隙内。高尔基体常见,仅见很少的片段内质网。第五层传递细胞具短的壁内突、较大的淀粉粒及许多小蛋白质体。两个时期的第一、二层细胞内均未观察到胞间连丝。授粉后２５ ｄ,第四层及以上的传递细胞的细胞壁和呈网状的壁内突均含有胞间连丝。还讨论了各种细胞器的作用及各层传递细胞的功能     

Ultrastructure Changes Induced by Stem Nematodes in Hypocotyl Tissue of Alfalfa

Scarified seeds of Medicago sativa L. ''Ranger'' and ''Lahontan'' alfalfa were allowed to imbibe water for 36 hr and then were inoculated with stem nematodes, Ditylenchus dipsaci Kühn. Seedlings were grown in sterilized Provo sand at 20 C and hypocotyl sections harvested at 1, 3 and 7 days. Evidence from electron micrographs indicated that cells of noninfected control plants contained normally developing chloroplasts bearing stroma, thylakoids, starch grains and plastoglobuli. The cytoplasm contained a nucleus, endoplasmic reticulum, vacuoles, mitochondria, ribosomes and dictyosomes. No morphological symptoms of nematode infection were observed in infected plants of either Ranger of Lahontan alfalfa 1 day after inoculation. Electron micrographs of tissue from the infected plants, however, indicated more osmiophilic bodies (lipid bodies) per cell than did the noninfected control, with more lipid bodies present in Ranger than in Lahontan. Three and 7 days after planting, swollen hypocotyls could be seen; the degree of swelling was greater in Ranger than in Lahontan. Electron micrographs of infected tissues indicated that both cultivars were undergoing the same kind of damage. Injured organelles were endoplasmic reticulum, chloroplasts and the nucleus. Histochemical staining indicated no changes in the middle lamellae.     

Dependence of cytokinin distribution in plants on their physical and chemical properties and transpiration rate

The effect of transpiration on cytokinin accumulation and distribution in 7-day-old wheat (Triticum durum Desf.) seedlings grown on nutrient medium supplemented with zeatin or its riboside was studied. The content of cytokinins in plants and nutrient medium was measured by the immunoenzyme analysis; cytokinin distribution between root cells was assessed immunohistochemically using antibodies against zeatin derivatives. The rate of transpiration was reduced 20-fold by plant placing in humid chamber. At normal transpiration, after 6 h of plant incubation on the solution of zeatin, the level of cytokinins in plant tissues increased stronger than after incubation on the solution of zeatin riboside (by 7.3 and 3.5 times, respectively, as compared with control), although the rates of both cytokinin uptake were equal. Most portions of cytokinins were retained in the roots, which was stronger expressed in the case of free zeatin uptake. A decrease in the rate of transpiration did not affect substantially the zeatin absorption from nutrient medium and the total level of cytokinin accumulation in plants, but these indices were sharply decreased in the case of zeatin riboside. In the zone of absorption of both control roots and roots treated with cytokinins, more intense cytokinin immunostaining was observed in the cells of the central cylinder. The interrelation between cytokinin distribution between the cells and apoplast, their inactivation, and transport over the plant and their form (zeatin or zeatin riboside) used for treatment is discussed.     

Fine structure of protonemal apical cells of the mossPhyscomitrium turbinatum

Summary Elongating caulonemal apical cells of the mossPhyscomitrium turbinatum were cultivatedin vitro and observed during successive stages of cell elongation and division. Actively-growing cells which had completed approximately half of their growth in length were examined by electron microscopy. The distribution of many organelles changes progressively from the cell tip to the distal edge of the large basal vacuole, establishing an apical-basal gradient in organization. Whereas the vacuoles become progressively more extensive in more mature parts of the cell, the dictyosomes, chloroplasts and smooth endoplasmic reticulum are more numerous in younger regions. Some mitochondria in the younger regions of the cell contain localized areas of membrane invagination. Attempts were made to clarify the origin and growth of vacuoles, which become increasingly prominent as the apical cell elongates.Morphological evidence suggests that vacuoles arise in close association with endoplasmic reticulum and dictyosomes as a result of ER dilation and/or cytoplasmic sequestration. The number of vacuolar profiles is highest at the cell tip, decreasing progressively toward the base of the cell, Conversely, the mean area of vacuolar profiles increases progressively toward more basal regions of the cell. These features, along with the increasing number of closely grouped vacuolar profiles along an apical-basal gradient are compatible with the concept of vacuolar growth by coalescence, culminating in their union with the basal vacuole.     

Physio-anatomical responses of tobacco under caffeine stress

Caffeine, a purine alkaloid, is reported to act both as an inducer or inhibitor to plant growth in various species. The aim of this study was to examine the effect of exogenous caffeine on tobacco (Nicotiana tabacum) plants, a plant that does not naturally synthesise caffeine. A hydroponic experiment was carried out in a growth chamber for 14 d using Hoagland’s solution supplemented with 0 (control), 25, 50, 100, 1,000; and 5,000 μM caffeine. None of the investigated caffeine concentrations significantly decreased the net photosynthetic rate except the highest concentrations of 1,000 and 5,000 μM. Light microscopy of thick-sectioned roots showed that 1,000 μM and 5,000 μM caffeine-treated plants possessed deformed epidermal cells, reduced number of cortical cells, and deformed vascular tissues with cells exhibiting thickened xylem walls as compared with control plants. Moreover, transmission electron micrographs of roots revealed that mitochondria and the plasma membrane were affected.