The ultrastructure of chloroplast of a brown alga Sphacelaria sp. III. The replication and segregation of chloroplast genophore

Bisalputra, T.; Bisalputra, A. A.

doi:10.1016/s0022-5320(70)80019-3

Cited by 38 publications

(14 citation statements)

References 9 publications

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“…In cells at or near the apex, DNA rings with hourglass shapes were frequently observed (Fig. 6), reminiscent of chloroplasts pinching in two as described by Bisalputra and Bisalputra (2). Assuming the smallest chloroplasts to be the most newly formed, we observed the chloroplasts and their DNA rings to be smallest in the apical cells, and increasingly larger as one progressed away from the apex .…”

Section: Acetabulariasupporting

confidence: 64%

Use of the fluorochrome 4'6-diamidino-2-phenylindole in genetic and developmental studies of chloroplast DNA.

Coleman¹

1979

The Journal of Cell Biology

104

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Use of the DNA-specific fluorochrome 4'6-diamidino-2-phenylindole (DAPI) makes it possible to examine in situ the structure of chloroplast DNA (chDNA) with the fluorescence microscope . This simplifies the study of genetic and developmental changes in chloroplast DNA . Three examples are presented . (a) Wildtype Euglena gracilis B contains several chloroplast DNA nucleoids per chloroplast . A yellow mutant lacking functional chloroplasts is similar, but such nucleoids are absent in an aplastidic mutant strain known from biochemical studies to have lost its chDNA . (b) In vegetative cells ofthe giant-celled marine algae Acetabularia and Batophora, only about a quarter of the chloroplasts have even one discernible chloroplast DNA particle, and such particles vary in size, showing a 30-fold variation in the amount of DNA-bound DAPI fluorescence detected per chloroplast . By contrast, 98% of chloroplasts in developing Acetabularia cysts contain chDNA, with as many as nine nucleoids per chloroplast. (c) DAPI-stained chloroplasts of chromophyte algae display the peripheral ring of DNA expected from electron microscope studies . However, these rings are not uniform in thickness, but are necklace-like, with the appearance of beads on a string. Since the multiple nucleoids in plastids ofchlorophyte algae also appear to be interconnected throughout the chloroplast, a common structural plan may underlie chDNA morphology in both groups of algae .KEY WORDS Acetabularia algae chloroplast DNA " DAPI " Euglena New techniques can often shed light on longstanding research questions . The recent availability of a new series of fluorescent compounds offers such an opportunity . The compounds in question are 33258 Hoechst (9, 13) and several derivatives of 4'6-diamidine-2-phenylindole (DAPI) (12). These compounds are highly specific in their binding to linear polymers with phosphate backbones, such as polyphosphates and DNA, and they exhibit a much enhanced fluorescence when bound to double-stranded DNA, particularly that with high AT content (10). Hence, aggregates of as little as 10 -16 g of DNA, such as occur in yeast mitochondria, can easily be seen in situ with the fluorescence microscope (17). MATERIALS AND METHODS J . CELL. BIOLOGY

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Section: Acetabulariasupporting

confidence: 64%

Use of the fluorochrome 4'6-diamidino-2-phenylindole in genetic and developmental studies of chloroplast DNA.

Coleman¹

1979

The Journal of Cell Biology

104

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“…As with young wheat chloroplasts the central region of young tobacco and spinach chloroplasts are packed with DAPI staining areas, in contrast to the older chloroplasts in which the areas are more dispersed. Yet another distinctive arrangement of plastid DNA has been described for some algae (3,4,8) in which the cpDNA is associated with the girdle thylakoids and forms a loop around the periphery of the chloroplast. Furthermore, young barley etioplasts have a single polymorphous DNA region located in the centre of the etioplast (22).…”

Section: Discussionmentioning

confidence: 99%

Localization of DNA in Mature and Young Wheat Chloroplasts Using the Fluorescent Probe 4′-6-Diamidino-2-phenylindole

Selldén¹,

Leech²

1981

Plant Physiol.

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The spatial organiation of chloroplast DNA in developing and dividing wheat chloroplasts was studied in the light microscope using the fluorescent probe 4'--diamidino-2-phenylindole, which binds specifically to DNA.The DNA of wheat chloroplasts was localized at the periphery of the plastid, frequently in a discrete band. No relocaization of the DNA was observed during plastid replication. This peripheral location of the DNA was shown to differ from the central random location of DNA in tobacco and spinach chloroplasts. compared with autoradiographic methods since the stain visualizes all the cpDNA, not only the portion which has recently been synthesized. As an alternative to electron microscopy the use of DAPI has the advantage that it avoids digestion of the stroma proteins which conceivably could alter the DNA arrangement within the plastid.This paper reports our observations in young and mature wheat chloroplasts on the nature of the mode of distribution of DNA during chloroplast division. The arrangement of DNA within the chloroplasts of wheat is compared with its arrangement in tobacco and spinach plastids.Chloroplast differentiation in light-grown wheat seedlings is accompanied by a large increase in the number of chloroplasts per cell (7), associated with a decrease in the number of genome copies per plastid (5). Nothing is known about the location of DNA within these developing wheat plastids nor if the DNA becomes redistributed during chloroplast division: indeed there appears to have been no detailed study of the location of DNA within the chloroplast of any monocotyledonous plant. In dicotyledons, electron microscopical and autoradiographic studies have shown that cpDNA3 is located in multiple, interconnected regions associated with the thylakoid membranes (10-12, 14, 15, 17, 18, 21, 25 3Abbreviations: cpDNA, chloroplast DNA; DAPI, 4'-6-diamidino-2-phenylindole; nDNA, nuclear DNA. MATERIALS AND METHODSPlant Material. Seeds of wheat, Triticum aestivum, var. Maris Dove, were soaked in running tap water at 20 C for 17 h, with surface sterilization in NaOCl solution (13% free chlorine) after the first h. The seeds were sown in Levington Universal Compost (Fisons, United Kingdom) at a depth of 1 cm and were grown using a photoperiod of 16 h light at 20 C, 8 h darkness at 15 C, 70%o RH. The light intensity at the level of the seedlings, measured with a solarimeter (Kipp and Zonen), was 4.0mw* cm2. Seedlings were harvested 7 days after sowing, 2 h after the start of the light period; leaves were cut at their bases, and coleoptiles gently pulled off. Tobacco plants, Nicotiana tabacum, were grown in Levington Universal Compost (Fisons, United Kingdom) for 24 days in a photoperiod of 16 h light at 25 C, 8 h darkness at 20 C, 70%o RH. The first of four leaves was used for the isolation of mature plastids. The fourth leaf, which had just emerged and was 0.5 mm long, was used to obtain young plastids. Spinach plants, Spinacia oleracea, were grown for 44 days as previously described (1). Mature plastids ...

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“…Furthermore, by TEM, a genophore, which is the area containing the ring nucleoid, lying just beneath the girdle lamella was not observed in this study. In some algae, cp-DNA has been observed as electron-dense fibrils by TEM (Ris & Plaut 1962;Bisalputra & Bisalputra 1969). In diatoms, however, the fibrils are scarcely recognizable in most TEM photographs published, except for several excellent figures by, for example, Manton and von Stosch (1966) or Manton et al (1969).…”

Section: Discussionmentioning

confidence: 99%

Characterization of linear-oblong pyrenoids with cp-DNA along their sides in Nitzschia sigmoidea (Bacillariophyceae)

Mayama¹,

Mayama²,

Shihira‐lshikawa³

2004

Phycological Res

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SUMMARYIn the past 10 years Nitzschia sigmoidea (Nitzsch) W. Sm. has begun to occur in Japanese rivers in various areas. It is a common diatom in Europe but was previously absent in Japan. Each chloroplast of N. sigmoidea contains many unusual linear-oblong structures. The internal structure of the chloroplast in this species was observed using epifluorescence and electron microscopy with immunolocalization techniques. The linear-oblong structures in the chloroplasts could hardly be observed by conventional light microscopy of living cells, but were obvious in cells stained with propionocarmine. Transmission electron microscopy showed that the cross sections of this structure were lanceolate to fusiform with penetration by a single thylakoid. In cells stained with DAPI, chloroplast DNA was detected along both sides of the linear-oblong structures, and DNA fibrils were detected by electron microscopy. Immunofluorescence microscopy of sectioned cells and also immunoelectron microscopy revealed specific localization of Rubisco between these DNA-containing areas, which divided at the same time as the chloroplast. Our observations confirmed that the linear-oblong structures are pyrenoids. The diversity of localization patterns of chloroplast DNA in diatoms is discussed.

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The ultrastructure of chloroplast of a brown alga Sphacelaria sp. III. The replication and segregation of chloroplast genophore

Cited by 38 publications

References 9 publications

Use of the fluorochrome 4'6-diamidino-2-phenylindole in genetic and developmental studies of chloroplast DNA.

Use of the fluorochrome 4'6-diamidino-2-phenylindole in genetic and developmental studies of chloroplast DNA.

Localization of DNA in Mature and Young Wheat Chloroplasts Using the Fluorescent Probe 4′-6-Diamidino-2-phenylindole

Characterization of linear-oblong pyrenoids with cp-DNA along their sides in Nitzschia sigmoidea (Bacillariophyceae)

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