Nuclear crystalloids in sieve elements of Boraginaceae: a protein digestion study

1983 ◽  
Vol 64 (1) ◽  
pp. 37-47
Author(s):  
J. Thorsch ◽  
K. Esau

Nuclear crystalloids have been found in sieve elements of several Boraginaceae. Nuclei of differentiating sieve elements of Echium and other genera except Amsinckia contain one or more crystalloids composed of thin rods densely packed in parallel arrangement. After the nuclei disintegrate in the maturing sieve element the crystalloids are released into the cell lumen where they persist intact. In Amsinckia the crystalloid consists of two components: a dense component, similar to the crystalloid in the other genera and a loosely arranged paracrystalline component. The proteinaceous nature of the nuclear crystalloids and their possible similarity to P-protein was investigated by enzyme digestion techniques. Three proteolytic enzymes were employed in this study: protease, pepsin and trypsin. Successful digestion of the dense crystalloid in both Echium and Amsinckia was obtained with each enzyme tested. P-protein plugging the sieve plate pores was also digested. The loose component in Amsinckia and the aggregated and dispersed P-protein were not affected by the enzyme digestion procedures. These results seemed to indicate that the density or compactness of the proteinaceous inclusions may play a role in the differential response.

1967 ◽  
Vol 34 (3) ◽  
pp. 801-815 ◽  
Author(s):  
James Cronshaw ◽  
Katherine Esau

An ontogenetic study of the sieve element protoplast of Nicotiana tabacum L. by light and electron microscopy has shown that the P-protein component (slime) arises as small groups of tubules in the cytoplasm. These subsequently enlarge to form comparatively large compact masses of 231 ± 2.5 (SE)A (n = 121) tubules, the P-protein bodies. During subsequent differentiation of the sieve element, the P-protein body disaggregates and the tubules become dispersed throughout the cell. This disaggregation occurs at about the same stage of differentiation of the sieve elements as the breakdown of the tonoplast and nucleus. Later, the tubules of P-protein are reorganized into smaller striated 149 ± 4.5 (SE)A (n = 43) fibrils which are characteristic of the mature sieve elements. The tubular P-protein component has been designated P1-protein and the striated fibrillar component P2-protein. In fixed material, the sieve-plate pores of mature sieve elements are filled with proteinaceous material which frays out into the cytoplasm as striated fibrils of P2-protein. Our observations are compatible with the view that the contents of contiguous mature sieve elements, including the P-protein, are continuous through the sieve-plate pores and that fixing solutions denature the proteins in the pores. They are converted into the electron-opaque material filling the pores.


1979 ◽  
Vol 38 (1) ◽  
pp. 11-22
Author(s):  
K. Esau ◽  
A.C. Magyarosy

The compounds crystalloids formed in sieve element nuclei of Amsinckia douglasiana A. DC. (Boraginaceae) during differentiation of the cell become disaggregated during the nuclear breakdown characteristic of a maturing sieve element. The phenomenon occurs in both healthy and virus-infected plants. The crystalloid component termed cy, which is loosely aggregated, separates from the densely aggregated component termed cx and disperses. The cx component may become fragmented, or broken into large pieces, or remain intact after the cell matures. After their release from the nucleus both crystalloid components become spatially associated with the dispersed P-protein originating in the cytoplasm, but remain distinguishable from it. The component tubules of P-protein are hexagonal in transections and are somewhat wider than the 6-sided cy tubules. The cx tubules are much narrower than the P-protein or the cy tubules and have square transections. Both the P-protein and the products of disintegrated crystalloids accumulate at sieve plates in sieve elements subjected to sudden release of hydrostatic pressure by cutting the phloem. The question of categorizing the tubular components of the nuclear crystalloid of a sieve element with reference to the concept of P-protein is discussed.


1979 ◽  
Vol 38 (1) ◽  
pp. 1-10
Author(s):  
K. Esau ◽  
A.C. Magyarosy

The presence of usually single, elongated, compound crystalloids in nuclei of sieve elements is an outstanding characteristic of the phloem of Amsinckia douglasiana A. DC. (Boraginaceae). The crystalloid consists of two components forming alternating panels, or blocks, that extend through the entire length of the crystalloid and radiate from the centre where one of the components predominates. Three to seven panels for each component were recorded. One component consists of 4-sided tubules closely packed in highly ordered aggregates, the other of wider 6-sided tubules rather loosely arranged in paracrystalline aggregates. The crystalloid arises at the beginning of sieve element differentiation. Aggregates of 4-sided tubules appear first. In plants infected with the curly top virus, the crystalloids do not differ from those in non-infected controls in structure and conformation. But because the phloem in infected plants is hyperplastic, with most of the cells differentiating as sieve elements, the crystalloids are far more abundant in diseased than in healthy plants.


1982 ◽  
Vol 54 (1) ◽  
pp. 149-160
Author(s):  
KATHERINE ESAU ◽  
JENNIFER THORSCH

Ultrastructural studies of differentiating phloem of the midvein in 15 species of Echium (Boraginaceae) have shown that in every species one or more crystalloids develop in sieve-element nuclei. These inclusions appear as soon as the cell begins to differentiate from a procambial derivative. Small at first, they enlarge later, particularly in length. The crystalloid may extend end-to-end in the correspondingly elongated nucleus. When the nucleus disintegrates in the maturing cell the crystalloid is released into the cell lumen where it remains intact or becomes fractured into large pieces. No dissociation into subunits was observed. The crystalloids are composed of tightly packed narrow rods (tubules) forming a two-dimensional lattice of squares in transections and a system of parallel striations in longitudinal sections. The similarities of these inclusions to the crystalline form of P-protein in Fabaceae raises the question of terminological delimitation of P-protein, a common component of angiospermous sieve-element protoplasts. The occurrence of nuclear crystalloids in another previously investigated genus in the Boraginaceae, Amsinckia, suggests that this inclusion should be explored in boraginaceous representatives for its possible value as a systematic character.


1989 ◽  
Vol 67 (12) ◽  
pp. 3608-3617 ◽  
Author(s):  
Deborah D. Fisher ◽  
Jennifer Thorsch ◽  
Katherine Esau

A survey of 68 species representing 28 genera in the family Boraginaceae was conducted at the ultrastructural level to determine presence of nuclear crystalloids in sieve elements and occurrence of crystalline structures in chloroplasts of phloem parenchyma cells. Nuclear crystalloids were identified in 55 of the species examined, and 25 of the species contained chloroplast crystals. The nuclear crystals were mainly composed of thin rods densely packed in parallel arrangement. Their sizes, shapes, and numbers varied, but they were basically prismatic or possibly cubical. During maturation, the sieve element nuclei disintegrated and the crystalloids were released into the cell lumen where they remained intact. Loosely arranged paracrystalline components associated with the dense nuclear crystalloids were found only in the genus Amsinckia. Crystalline inclusions in the parenchyma chloroplasts were made up of fibers loosely aligned in a herringbone pattern. The lability of the chloroplast crystals to the proteolytic enzyme, protease, was tested on 10 species, and only Onosma stellulatum Waldst. & Kit. chloroplast crystals were routinely digested. The high percentage of nuclear crystalloids found in this family suggests that these inclusions could be valuable as a systematic character. Key words: sieve elements, nuclear inclusions, plastid crystals, Boraginaceae.


Author(s):  
Lidija Murmanis

At the earliest stage, P-protein in red oak sieve elements can be identified with the fine filaments embedded in the cytoplasm (Fig. 1), and they appear similar to the P-protein component in Cucurbita maxima at an early differentiation stage (1). At this time ribosomes (in a spiral configuration) and the rough ER are numerous and in intimate association with the filaments. As differentiation proceeds P-protein bodies are formed in which the filaments are randomly arranged (Fig. 2). Later the filaments become oriented parallel to one another (Fig. 1, at right) and are transformed structurally into tubules. Tubules aggregate into crystalline bodies (Figs. 3, 4) which resemble extruded nucleoli reported previously (2, 3). Individual tubules have a diameter of about 235A and a tripartite structure--60-70A thick electron-dense walls separated by an electron-transparent core. During the disintegration of sieve element protoplast the crystalline bodies disperse and tubules gradually change into fibrils.


1969 ◽  
Vol 17 (3) ◽  
pp. 441 ◽  
Author(s):  
S Zee

The fine structure of the sieve elements of the primary phloem of the epicotyl of Vicia faba is described. The cytoplasm of the young sieve element contains four distinct forms of "slime" body: amorphous, crystalline, tubular (each tubule measuring about 140 Å in diameter), and short fibrillar (each fibril measuring about 350 Å in diameter). At the very early stages of differentiation, polysome helices are prevalent often in close association with the amorphous but not the other forms of "slime" bodies. At the early stages of development of the sieve element the tubular form of "slime" is closely associated with the endoplasmic reticulum, which suggests their possible origin. The plastids of the sieve element lack a well-developed internal membrane system but contain two characteristic types of inclusion bodies, starch granules and crystalloids identical to those revorted for the secondarv vhloem of the root of Pisum sativum. Mitochondria - remain apparently unchanged throughout sieve element development. Microtubules are present during the early stages of sieve element development but become scarce at later stages. Dictyosomes, coated vesicles, ribosomes, polysomes, and nucleus disappear as the sieve element matures. The fine structure of the sieve plate pore initial is complex. It consists of an outer electron-dense ring ("desmotubule") which encloses a central dark core. The developmental pattern of the sieve plate pore has been traced from the very young to the mature stages.


1968 ◽  
Vol 38 (2) ◽  
pp. 292-303 ◽  
Author(s):  
James Cronshaw ◽  
Katherine Esau

During maturation of sieve elements in Cucurbita maxima Duchesne, the P-protein bodies (slime bodies) usually disperse in the tonoplast-free cell. In some sieve elements the P-protein bodies fail to disperse. The occurrence of dispersal or nondispersal of P-protein bodies can be related to the position of the sieve elements in the stem or petiole. In the sieve elements within the vascular bundle the bodies normally disperse; in the extrafascicular sieve elements the bodies often fail to disperse. Extrafascicular sieve elements showing partial dispersal also occur. The appearance of the sieve plate in fixed material is related to the degree of dispersal or nondispersal of the P-protein bodies. In sieve elements in which complete dispersal occurs the sieve plate usually has a substantial deposit of callose, and the sieve-plate pores are filled with P protein. In sieve elements containing nondispersing P-protein bodies the sieve plate bears little or no callose, and its pores usually are essentially "open." The dispersed P-protein components may aggregate into loosely organized "strands," which sometimes extend vertically through the cell and continue through the sieve-plate pores; but they may be oriented otherwise in the cell, even transversely.


Author(s):  
James Cronshaw

Long distance transport in plants takes place in phloem tissue which has characteristic cells, the sieve elements. At maturity these cells have sieve areas in their end walls with specialized perforations. They are associated with companion cells, parenchyma cells, and in some species, with transfer cells. The protoplast of the functioning sieve element contains a high concentration of sugar, and consequently a high hydrostatic pressure, which makes it extremely difficult to fix mature sieve elements for electron microscopical observation without the formation of surge artifacts. Despite many structural studies which have attempted to prevent surge artifacts, several features of mature sieve elements, such as the distribution of P-protein and the nature of the contents of the sieve area pores, remain controversial.


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