The Role of Druse and Raphide Calcium Oxalate Crystals in Tissue Calcium Regulation in Pistia stratiotes Leaves

Abstract: Ca oxalate crystal formation was examined in Pistia stratiotes L. leaves during excess Ca and Ca-deficient conditions. Pistia produces druse crystal idioblasts in the adaxial mesophyll and raphide idioblasts in the abaxial aerenchyma. Raphide crystals were previously found to grow bidirectionally, and here we show that Ca is incorporated along the entire surfaces of developing druse crystals, which are coated with membrane-bound microprojections. Leaves formed on plants grown on 0 Ca medium have fewer and smaller druse crystals than leaves formed under 5 mM Ca ("control") conditions, while raphide crystal formation is completely inhibited. When plants were moved from 0 to 15 mM ("high") Ca, the size and number of crystals in new leaves returned to (druse) or exceeded (raphide) control levels. High Ca also induced formation of druse, but not raphide, crystals in differentiating chlorenchyma cells. When plants were transferred from 15 mM Ca to 0 Ca, young druse crystals were preferentially partially dissolved. Oxalate oxidase, an enzyme that degrades oxalate, increased during Ca deficiency and was localized to the crystal surfaces. The more dynamic nature of druse crystals is not due to hydration form as both crystal types are shown to be monohydrate. Part of the difference may be because raphide idioblasts have developmental constraints that interfere with a more flexible response to changing Ca. These studies demonstrate that excess Ca can be stored as Ca oxalate, the Ca can be remobilized under certain conditions, and different forms of Ca oxalate have different roles in bulk Ca regulation.     

Twinned Raphides of Calcium Oxalate in Grape (Vitis): Implications for Crystal Stability and Function

Among the higher plants that accumulate crystalline calcium oxalate, many taxa characteristically produce raphides, or needle-shaped crystals. Substantial evidence has accumulated indicating that raphides function in plant defenses against herbivory and that their acicular shape is a critical component in proposed mechanisms for these defenses. Previous observations have shown that raphides in members of the Vitaceae are twinned crystals. In this study, the fine structure of raphides in Vitis was examined in order to determine the nature of twinning. Rotation of isolated raphides under cross-polarized light revealed extinction patterns that indicated that raphides are twinned along their length. In cross sections of raphides, the twin plane extends across the raphides, parallel to their surfaces. The dissolution patterns observed in etched crystals indicate that the type of twinning is rotational. Parallels in other biomineralization systems indicate that the rotational nature of the twinning may increase mechanical strength. In addition, because twinning may affect crystal growth or enhance stability and persistence of crystals, it could be an important factor in allowing plant cells to produce the raphide morphology.     

Histochemical Localization of Calcium Oxalate Crystals in Starch Grains of Yams (Dioscorea)

The bulbils and/or tubers of seven species of yams (Dioscorea)were examined for crystal content using light microscopy andhistochemistry. Calcium oxalate crystals in the form of raphide bundles werelocalized in the parenchymatous tissues. Within starch grains,crystals of various shapes and sizes were observed. The variationin shape and sizes of the intra-amylar crystals could be exploitedfor taxonomic purposes. Calcium oxalate crystals appear to serve a storage functionin these starch grains. Yams, Dioscorea, raphides, oxalate crystals, histochemistry     

Calcium oxalate formation is a rapid and reversible process inLemna minor L.

Summary Lemna minor root tips form raphide Ca oxalate crystals in both the root cap and root proper. An in vivo system was developed to examine raphide crystal bundle formation in the root of intact plants. By increasing the exogenous Ca concentration, crystal bundle formation could be induced. Entire new crystal bundles could be formed within 30 minutes of an inductive stimulus. The process was reversible with recently formed crystal bundles being dissolved over a period of about 3 hours. Older, previously existing bundles were more resistant to dissolution. The calmodulin antagonists, chlorpromazine and trifluoperazine (300 M), prevented crystal formation and caused dissolution of some crystal bundles, even in the presence of exogenous Ca. When the antagonists were flushed out and replaced with fresh medium, crystals were formed in cells where dissolution had occurred under the influence of the antagonists. The Ca ionophore A 23187 (20 M) caused slow dissolution of crystal bundles, even in the presence of exogenous Ca. A model describing the control of and physiological significance of Ca oxalate formation in plants is presented and discussed with respect to the results obtained in this study.     

Isolation of a crystal matrix protein associated with calcium oxalate precipitation in vacuoles of specialized cells

The formation of calcium (Ca) oxalate crystals is considered to be a high-capacity mechanism for regulating Ca in many plants. Ca oxalate precipitation is not a stochastic process, suggesting the involvement of specific biochemical and cellular mechanisms. Microautoradiography of water lettuce (Pistia stratiotes) tissue exposed to 3H-glutamate showed incorporation into developing crystals, indicating potential acidic proteins associated with the crystals. Dissolution of crystals leaves behind a crystal-shaped matrix "ghost" that is capable of precipitation of Ca oxalate in the original crystal morphology. To assess whether this matrix has a protein component, purified crystals were isolated and analyzed for internal protein. Polyacrylamide gel electrophoresis revealed the presence of one major polypeptide of about 55 kD and two minor species of 60 and 63 kD. Amino acid analysis indicates the matrix protein is relatively high in acidic amino acids, a feature consistent with its solubility in formic acid but not at neutral pH. 45Ca-binding assays demonstrated the matrix protein has a strong affinity for Ca. Immunocytochemical localization using antibody raised to the isolated protein showed that the matrix protein is specific to crystal-forming cells. Within the vacuole, the surface and internal structures of two morphologically distinct Ca oxalate crystals, raphide and druse, were labeled by the antimatrix protein serum, as were the surfaces of isolated crystals. These results demonstrate that a specific Ca-binding protein exists as an integral component of Ca oxalate crystals, which holds important implications with respect to regulation of crystal formation.     

虎掌（天南星科）中草酸钙晶体的形态和分布

在光学显微镜下对虎掌（Pinellia pedatisecta）营养器官和繁殖器官中晶体的类型和分布进行了观察和分析,探讨晶体的功能与作用机制。结果表明：（1）虎掌各个器官中都发现有晶体,且晶体类型有针晶、簇晶、砂晶和柱晶4种形态,其中针晶最为常见。（2）虎掌叶中的晶体大多以针晶状分布在叶片上表皮下的叶肉中,少数分布在叶下表皮下的叶肉中,其次砂晶和星芒状簇晶也在叶中较常见,叶中也有少量的柱晶。（3）虎掌的块茎中分布有大量的针晶束,在输导组织附近还有一些大的簇晶;虎掌的不定根中分布有不整齐的针晶和排列不规则的针晶束以及少量大的簇晶。（4）虎掌的佛焰苞中分布有针晶、簇晶和砂晶,且在佛焰苞中的针晶主要分布于上、下表皮之下的叶肉中,砂晶多分布在佛焰苞的上、下表皮上。（5）虎掌的花药壁中分布有针晶束,其方向和花药壁表面垂直,而花粉囊中只有小的簇晶。（6）虎掌的果皮和种皮上分布有大量的针晶。根据晶体在酸中的溶解性,虎掌体内所有晶体的化学成分都为草酸钙。研究认为,虎掌各个器官中的各种草酸钙晶体对于保护虎掌免受食草动物取食具有重要的作用。     

Calcium Oxalate Crystals in Eucalypt Ectomycorrhizae: Morphochemical Characterization

Ectomycorrhizal fungi are ubiquitous in forest ecosystems, benefitting plants principally by increasing the uptake of water and nutrients such as calcium from the soil. Previous work has demonstrated accumulation of crystallites in eucalypt ectomycorrhizas, but detailed morphological and chemical characterization of these crystals has not been performed. In this work, cross sections of acetic acid-treated and cleared ectomycorrhizal fragments were visualized by polarized light microscopy to evaluate the location of crystals within cortical root cells. Ectomycorrhizal sections were also observed by scanning electron microscopy (SEM) coupled with energy dispersive x-ray (EDS) microprobe analysis. The predominant forms of crystals were crystal sand (granules) and concretions. Calcium, carbon and oxygen were detected by EDS as constituent elements and similar elemental profiles were observed between both crystal morphologies. All analyzed crystalline structures were characterized as calcium oxalate crystals. This is the first report of the stoichiometry and morphology of crystals occurring in eucalypt ectomycorrhizas in tropical soils. The data corroborates the role of ectomycorrhizae in the uptake and accumulation of calcium in the form of calcium oxalate crystals in hybrid eucalypt plants.     

Insolubilization of potassium chloride crystals in Tradescantia pallida

Summary. Calcium oxalate crystals are by far the most prevalent and widely distributed mineral deposits in higher plants. In Tradescantia pallida, an evergreen perennial plant widely used as an ornamental plant, calcium oxalate crystals occur in the parenchymal tissues of stem, leaf, and root, as well as in flower organs, in the form of either raphides or tetragonal prismatic crystals or both. Energy-dispersive X-ray analysis revealed that C, O, and Ca were the main elements; and K, Cl, and Si, the minor elements. Infrared and X-ray analyses of crystals collected from these tissues detected the coexistence of two calcium oxalate chemical forms, i.e., whewellite and weddellite, as well as calcite, opal, and sylvite. Here, we show for the first time the occurrence of epitaxy in mineral crystals of plants. Epitaxy, which involves the oriented overgrowth of one crystal onto a second crystalline substrate, might explain how potassium chloride (sylvite) – one of the most water-soluble salts – stays insoluble in crystal form when coated with a calcium oxalate epilayer. The results indicate the potential role of crystals in regulating the ionic equilibrium of both calcium and potassium ions. Correspondence and reprints: Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pabellón 2, Ciudad Universitaria, C1428EGA, Ciudad de Buenos Aires, Argentina.     

五种C4荒漠植物光合器官中含晶细胞的比较分析

 为了探讨荒漠植物适应干旱环境的机理, 选择光合器官发生很大变化的5种C₄荒漠植物进行了解剖结构的对比研究。结果表明, 这5种 植物中含晶细胞的数量、大小、形态和分布位置等存在差异。白梭梭(Haloxylon persicum)和梭梭(H. ammodendron)的同化枝普遍具有含晶细 胞; 沙拐枣(Calligonum mongolicum)的含晶细胞很少, 一般只分布在贮水组织或靠近栅栏组织处; 木本猪毛菜(Salsola arbuscula)的含晶细 胞也不多, 主要分布在栅栏组织和表皮细胞之间; 猪毛菜(S. collina)的含晶细胞更少, 仅在贮水组织中偶尔可见晶簇。比较梭梭、白梭梭和 沙拐枣同化枝不同部位的解剖结构发现, 梭梭同化枝基部含晶细胞最多, 中部次之, 顶部最少; 白梭梭同化枝顶部的含晶细胞数量较多, 中部 及基部较少; 沙拐枣同化枝顶部与基部的粘液细胞较多, 中部较少, 基部几乎没有栅栏组织, 而其维管组织较为发达。综合晶体的酸碱溶解性 及硝酸银组化分析结果, 并参照能谱仪的分析结果得知, 梭梭、白梭梭、沙拐枣和木本猪毛菜的叶片或同化枝中所含晶体的主要成分为草酸钙 。通过比较解剖结构发现, 梭梭和白梭梭的同化枝中含晶细胞最多, 其它3种植物的同化器官中含晶细胞较少, 而沙拐枣同化枝中有粘液细胞存 在。     

Cell and calcium oxalate crystal growth is coordinated to achieve high-capacity calcium regulation in plants

Summary Crystal idioblasts are cells which are specialized for accumulation of Ca²⁺ as a physiologically inactive, crystalline salt of oxalic acid. Using microautoradiographic, immunological, and ultrastructural techniques, the process of raphide crystal growth, and how crystal growth is coordinated with cell growth, was studied in idioblasts ofPistia stratiotes. Incorporation of⁴⁵Ca²⁺ directly demonstrated that, relative to surrounding mesophyll cells, crystal idioblasts act as high-capacity Ca²⁺ sinks, accumulating large amounts of Ca²⁺ within the vacuole as crystals. The pattern of addition of Ca²⁺ during crystal growth indicates a highly regulated process with bidirectional crystal growth. In very young idioblasts,⁴⁵Ca²⁺ is incorporated along the entire length of the needle-shaped raphide crystals, but as they mature incorporation only occurs at crystal tips in a bidirectional mode. At full maturity, the idioblast stops Ca²⁺ uptake, although the cells are still alive, demonstrating an ability to strictly regulate Ca transport processes at the plasma membrane. In situ hybridization for ribosomal RNA shows young idioblasts are extremely active cells, are more active than older idioblasts, and have higher general activity than surrounding mesophyll cells. Polarizing and scanning electron microscopy demonstrate that the crystal morphology changes as crystals develop and includes morphological polarity and an apparent nucleation point from which crystals grow bidirectionally. These results indicate a carefully regulated process of biomineralization in the vacuole. Finally, we show that the cytoskeleton is important in controlling the idioblast cell shape, but the regulation of crystal growth and morphology is under a different control mechanism.Abbreviation SEM scanning electron microscopy     

Herbivory and Calcium Concentrations Affect Calcium Oxalate Crystal Formation in Leaves ofSida (Malvaceae)

Calcium oxalate crystals have potential roles in plants as partof a defence mechanism against herbivores and/or in accumulatingexcess calcium. To date, these potential roles have been studiedindependently. In this experimental study the effects of calciumlevels and herbivory on the production of calcium oxalate crystals(i.e. druse, spherical crystal aggregates) were examined inseedlings of Sida rhombifolia. Seedlings were subjected to threecalcium levels (low, normal or high) and an artificial herbivorytreatment. Calcium levels and herbivory both affected densityof crystals in leaves. Leaves from seedlings grown in low calciumhad a greater crystal density than those grown in high calcium.Leaves from seedlings subjected to herbivory had a greater crystaldensity than those from seedlings not subjected to herbivory.This study provides additional evidence that calcium oxalatecrystal production depends not only on calcium levels but canalso be influenced by external pressures such as herbivory.In addition to their physiological role in plants, these resultssuggest that calcium oxalate crystals can also act as a defencemechanism against herbivores. Copyright 2001 Annals of BotanyCompany Calcium concentrations, calcium oxalate crystals, herbivory, Malvaceae, Sida rhombifolia     

Calcium oxalate crystals in plants

Calcium (Ca) oxalate crystals occur in many plant species and in most organs and tissues. They generally form within cells although extracellular crystals have been reported. The crystal cells or idioblasts display ultrastructural modifications which are related to crystal precipitation. Crystal formation is usually associated with membranes, chambers, or inclusions found within the cell vacuole(s). Tubules, modified plastids and enlarged nuclei also have been reported in crystal idioblasts. The Ca oxalate crystals consist of either the monohydrate whewellite form, or the dihydrate weddellite form. A number of techniques exist for the identification of calcium oxalate. X-ray diffraction, Raman microprobe analysis and infrared spectroscopy are the most accurate. Many plant crystals assumed to be Ca oxalate have never been positively identified as such. In some instances, crystals have been classified as whewellite or weddellite solely on the basis of their shape. Certain evidence indicates that crystal shape may be independent of hydration form of Ca oxalate and that the vacuole crystal chamber membranes may act to mold crystal shape; however, the actual mechanism controlling shape is unknown. Oxalic acid is formed via several major pathways. In plants, glycolate can be converted to oxalic acid. The oxidation occurs in two steps with glyoxylic acid as an intermediate and glycolic acid oxidase as the enzyme. Glyoxylic acid may be derived from enzymatic cleavage of isocitric acid. Oxaloacetate also can be split to form oxalate and acetate. Another significant precursor of oxalate in plants is L-ascorbic acid. The intermediate steps in the conversion of L-ascorbic acid to oxalate are not well defined. Oxalic acid formation in animals occurs by similar pathways and Ca oxalate crystals may be produced under certain conditions. Various functions have been attributed to plant crystal idioblasts and crystals. There is evidence that oxalate synthesis is related to ionic balance. Plant crystals thus may be a manifestation of an effort to maintain an ionic equilibrium. In many plants oxalate is metabolized very slowly or not at all and is considered to be an end product of metabolism. Plant crystal idioblasts may function as a means of removing the oxalate which may otherwise accumulate in toxic quantities. Idioblast formation is dependent on the availability of both Ca and oxalate. Under Ca stress conditions, however, crystals may be reabsorbed indicating a storage function for the idioblasts for Ca. In addition, it has been suggested that the crystals serve purely as structural supports or as a protective device against foraging animals. The purpose of this review is to present an overview of plant crystal idioblasts and Ca oxalate crystals and to include the most recent literature.     

Renal tubular cell membranes inhibit growth but promote aggregation of calcium oxalate monohydrate crystals

Cell membranes have been proposed to serve as promoters for calcium oxalate monohydrate (COM) kidney stone formation. However, direct evidence to demonstrate the modulatory effects of renal tubular cell membranes on COM crystals does not currently exist. We thus examined the effects of intact MDCK cells and their fragmented membranes on COM crystal growth, aggregation and transformation. COM crystals were generated in the absence (control) or presence of intact MDCK cells or their membrane fragments. Intact MDCK cells and their membrane fragments significantly inhibited COM crystal growth (22.6% and 25.2% decreases in size, respectively) and significantly reduced COM total crystal mass (23.1% and 25.6% decreases, respectively). In contrast, both of them markedly promoted crystal aggregation (1.9-fold and 3.2-fold increases, respectively). Moreover, both intact cells and membrane fragments could transform COM to calcium oxalate dihydrate (COD) crystals. Finally, COM crystal growth inhibitory activities of both membrane forms were successfully confirmed by a spectrophotometric oxalate-depletion assay. Our data provide the first direct evidence to demonstrate the dual modulatory effects of MDCK membranes on COM crystals. Although growth of individual COM crystals was inhibited, their aggregation was promoted. These findings provide additional insights into the mechanisms of COM kidney stone formation.     

Cellular ultrastructure and crystal development in Amorphophallus (Araceae)

Background and Aims: Species of Araceae accumulate calcium oxalate in the form ofcharacteristically grooved needle-shaped raphide crystals andmulti-crystal druses. This study focuses on the distributionand development of raphides and druses during leaf growth inten species of Amorphophallus (Araceae) in order to determinethe crystal macropatterns and the underlying ultrastructuralfeatures associated with formation of the unusual raphide groove. Methods: Transmission electron microscopy (TEM), scanning electron microscopy(SEM) and both bright-field and polarized-light microscopy wereused to study a range of developmental stages. Key Results: Raphide crystals are initiated very early in plant development.They are consistently present in most species and have a fairlyuniform distribution within mature tissues. Individual raphidesmay be formed by calcium oxalate deposition within individualcrystal chambers in the vacuole of an idioblast. Druse crystalsform later in the true leaves, and are absent from some species.Distribution of druses within leaves is more variable. Drusesinitially develop at leaf tips and then increase basipetallyas the leaf ages. Druse development may also be initiated incrystal chambers. Conclusions: The unusual grooved raphides in Amorphophallus species probablyresult from an unusual crystal chamber morphology. There aremultiple systems of transport and biomineralization of calciuminto the vacuole of the idioblast. Differences between raphideand druse idioblasts indicate different levels of cellular regulation.The relatively early development of raphides provides a defensivefunction in soft, growing tissues, and restricts build-up ofdangerously high levels of calcium in tissues that lack theability to adequately regulate calcium. The later developmentof druses could be primarily for calcium sequestration.     

The intravacuolar organic matrix associated with calcium oxalate crystals in leaves of Vitis

In Vitis (grape) calcium oxalate crystals form in a needle-like morphology unique to plants, presenting an intriguing system of biological control over mineral formation. Crystals develop within an organic matrix which appears to provide control over the sites and forms of crystal deposition; however, little is known about the chemical nature of the matrix. A procedure has been developed to isolate crystals along with their associated intravacuolar matrix from leaves of grape, and studies have been initiated into the chemical composition of the matrix by characterizing elemental content, carbohydrates, and protein. The isolated matrix consisted of two structural phases, membrane chambers enclosing developing crystals, and a water-soluble phase surrounding the crystal chambers. Elemental analysis detected substantial calcium and potassium, as well as some iron in the water-soluble phase. Analysis of the water-soluble matrix by GC-MS showed that it contained an unusual polymer with novel glucuronic acid linkages. In addition, linkage analysis indicated 5-linked arabinans, arabinogalactan, and various mannosyl units typical of complex carbohydrates of N-linked glycoproteins. SDS—PAGE analysis of the water-soluble matrix and crystal chambers showed that each had distinct banding profiles in silver-stained gels, with prominent 60 and 70 kDa polypeptides in crystal chamber extracts. Demineralization studies provided direct evidence that the isolated matrix promotes crystal nucleation. The findings about the organic matrix associated with calcium oxalate crystals in grape are discussed in relation to crystal nucleation and growth and features shared with animal and microbial biomineralization systems.     

Engineering calcium oxalate crystal formation in Arabidopsis

Many plants accumulate crystals of calcium oxalate. Just how these crystals form remains unknown. To gain insight into the mechanisms regulating calcium oxalate crystal formation, a crystal engineering approach was initiated utilizing the non-crystal-accumulating plant, Arabidopsis. The success of this approach hinged on the ability to transform Arabidopsis genetically into a calcium oxalate crystal-accumulating plant. To accomplish this transformation, two oxalic acid biosynthetic genes, obcA and obcB, from the oxalate-secreting phytopathogen, Burkholderia glumae were inserted into the Arabidopsis genome. The co-expression of these two bacterial genes in Arabidopsis conferred the ability not only to produce a measurable amount of oxalate but also to form crystals of calcium oxalate. Biochemical and cellular studies of crystal accumulation in Arabidopsis revealed features that are similar to those observed in the cells of crystal-forming plants. Thus, it appears that at least some of the basic components that comprise the calcium oxalate crystal formation machinery are conserved even in non-crystal-accumulating plants.     

The Isolation and Properties of Oxalate Crystals from Plants

A method of isolation of crystalline inclusions of plant cellsis described. The crystals consist mainly of calcium oxalatein plants grown under normal conditions, but when calcium isreplaced by magnesium, barium, or strontium in the culture solutionthese elements substitute for calcium in the crystals; evenunder normal conditions magnesium occurs in the crystals tothe extent of about 2 per cent. The crystal morphology vanesin the species examined from raphides to complex conglomeratesand X-ray diffraction demonstrates an association of raphideswith calcium oxalate monohydrate whilst other solitary formsand conglomerates are associated with calcium oxalate 2.25H₂O.On this basis the species examined can be divided mto threegroups.     

香樟叶肉含晶细胞季节变化研究

为探讨香樟(Cinnamomum camphora)叶肉含晶细胞超微结构的季节变化,阐明香樟叶肉中草酸钙晶体在春夏秋冬的变化规律。该研究以多年生香樟(C. camphora)叶片为材料,分别于春夏秋冬四个季节露地取样,制作超薄切片,用透射电子显微镜(TEM)观察叶肉含晶细胞超微结构的变化。结果表明:春季时香樟叶肉中只有少数细胞有草酸钙晶体,数量较少,晶体结构多为柱状晶、方晶; 夏季时香樟叶肉细胞中随机分布于液泡的草酸钙晶体明显比春季的数量多、体积大、形态丰富,晶体多为柱状晶、方晶、针晶、簇晶; 秋季时香樟叶肉细胞草酸钙晶体和夏季的类似,数量较多,形态多样,以方晶和柱状晶针晶为主,伴有晶簇; 冬季时香樟叶肉含晶细胞晶体形态为柱状晶、方晶、针晶,数量比夏季和秋季的数量略有减少。该研究结果表明在一年四季中香樟叶肉细胞液泡中均有草酸钙晶体结构存在。     

Cytoplasmic streaming of calcium oxalate crystals in Callipsygma wilsonis (Bryopsidales, Chlorophyta)

Light microscopic study of the giant‐celled, marine green alga Callipsygma wilsonis J. Agardh (Udoteaceae, Bryopsidales) revealed numerous birefringent crystalline inclusions in the terminal segments of the assimilatory axes. The inclusions were thin plates with a triangular shape in face view, a base up to 75 μm in length, and a height that was one‐seventh the length of the base. Crystals of various sizes commonly were stacked face‐to‐face with one or more edges coinciding, but removal of organic material by treatment in sodium hypochlorite resulted in disaggregation. The crystals were soluble in dilute hydrochloric acid without effervescence but were insoluble in acetic acid. These diagnostic chemical solubility tests and a positive reaction to the Yasue staining reaction indicated that the crystals were composed of calcium oxalate. Scanning electron microscopy showed that most crystals had smoothly curving edges, but some had truncate or beveled margins. Calcium oxalate crystals have been reported to occur in the large central vacuoles of several bryopsidalean species, but the crystals in C. wilsonis were present in the parietal cytoplasm, which was evident from the presence of crystals in streaming cytoplasm. Calcium oxalate crystals, amyloplasts, chloroplasts, and other cytoplasmic constituents moved along cytoskeletal cables at rates of approximately 2.8 μm s⁻¹. These findings add to a growing body of evidence that calcium oxalate crystals in diverse algae may be present in cellular compartments other than the central vacuole.