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G. B E N J A M I N BOUCK
523
plants were then rinsed rapidly in distilled water, chloroplasts or join a peripheral band which fol-
dehydrated gradually with methyl alcohol, and lows the contours of the chloroplast (Berkaloff,
embedded in Epon (Luft, 1961) without a propylene 1961). The area just within this terminal band
oxide intermediate step (of Bouck, 1964). The transfer and at the terminus (or beginning) of the other
from absolute methyl alcohol to the Epon mixture chloroplast bands is usually less electron-opaque
was done gradually and the tissue remained at room (see also Leyon and yon Wettstein, 1954; yon
temperature for 24 hours to insure adequate in-
Wettstein, 1954) and appears to contain fine fibers
filtration. The blocks were polymerized at 60°C.
Sections were cut with a diamond knife on a Porter- (Fig. 8) similar to the 25-A DNA fibers found in
Blum uhramicrotome and either stained in Millonig's the chloroplast of Chlamydomonas (Ris and Plaut,
(1961) lead stain alone or double stained with 1962). The usual number of discs per band is
uranyl acetate in 50 per cent alcohol (Watson, 1958) three, although four and two are not uncommon.
followed by lead staining. A J E M 6C electron Discs in chloroplasts from cells near the meriste-
microscope was utilized for examining and photo- marie region of Fucus are often single or paired
graphing the sectioned material. (Fig. 11). Electron-opaque "globuli" are scattered
throughout the matrix, but starch is absent.
Key to Labels
B, chloroplast band N~ nucleus
Ch, chloroplast NE, nuclear envelope
CE, chloroplast envelope NL, nucleolus
CER, caloroplast endoplasmic reticulum P, pyrenoid
ER, endoplasmic reticulum PS, pyrenoid sac
F, formative face of Golgi apparatus T, tubule
G, Goigi apparatus Ira, vacuole
M, mitochondrion V, vesicle
Ma, maturation face of Golgi apparatus
FIa~RE 1 Portion of an outer assimilatory cell in Chordafilum. Note the large Golgi
apparatus (G) in close association with tile nuclear envelope (NE). The latter envelope
is seen to be continuous with the chloroplast ER (CER), but distinct and separate from
the chloroplast envelope (CE). Ch, chloroplast; M, mitochondrion; N, nucleus; Nl, nu-
cleolus. X 88,000.
525
G. BENJAMIN BOUCK OrganelleAssociations in Brown Algae
seemlngiy unattached tubules may, in part, arise the pyrenoid likewise is usually free of bands or
through evaginations from the chloroplast E R discs.
(Fig. 7). Surrounding the pyrenoid are three separate
3-he chloroplast E R is present also in Giffordia double membranes or envelopes (Fig. 2). The
(Fig. 6) and Futus (Fig. 11), but in neither genus innermost of these envelopes marks the continua-
have direct connections with the nuclear envelope tion of the chloroplast envelope around the margin
been observed. In both plants, there are many of the pyrenoid. Outside this chloroplast envelope
chloroplasts per cell, as distinguished from Chorda lies the chloroplast ER which comprises the mid-
in which there are one or two large chloroplasts dle of the three envelopes and is continuous with
per cell. In the latter case, because of the small the chloroplast E R surrounding the rest of the
size of the cell the nucleus is usually forced up chloroplast. However, in contrast to the rather
against the chloroplast, while in Fucus or Giffordia loose association of chloroplast E R to the chloro-
there may be much greater distances separating plast envelope in adjacent regions, around the
the chloroplasts and nucleus. In Giffordia, indi- pyrenoid the two envelopes become closely and
vidual chloroplasts can often be seen linked to one very regularly appressed. Finally, the outermost
FIGURE ~ A section through the pyrenoid (P) of Chorda.filum. Note that the matrices
of the pyrenoid (P) and chloroplast (Ch) are continuous. An electron-transparent pyrenoid
sac (PS) is seen extending around the pyrenoid, but not over the remainder of the chloro-
plast. In the inset, the various envelopes (double membranes) of the pyrenoid are shown.
The chloroplast envelope (CE) limits the pyrenoid, the chloroplast ER (CER) consti-
tutes the middle pair of membranes, and the pyrenoid sac (PS) is formed from the outer-
most pair of membranes. M, mitochondrion. )< ~7,000. Inset, )< 5~,000.
FIGURE 3 A section through an outer assimilatory cell of Chorda filum showing tile nu-
cleus (N) in close association with the chloroplast and the Golgi apparatus (G). A por-
tion of the nuclear envelope can be seen extending into the Golgi apparatus (at single
arrow). A bifurcation of the nuclear envelope which is continuous with the chloroplast
ER is seen at the double arrow. × 25,000.
527
G° B~JA~N BOUCK OrganelleAs~iations in Brown Algae
appears in m a n y cases highly irregular and always associated with either the nuclear envelope
convoluted while the inner membrane seems fairly or a portion of the endoplasmic reticulum (Fig.
regular. The irregular outer surface often appears I0). In the latter case, blebs may be formed
to be giving rise to blebs, especially in the region from the cytoplasmic E R cisternae, but not from
of the Golgi apparatus. Continuity of the nuclear the membranes of the chloroplast ER.
envelope with the endoplasmic reticulum may be
seen in m a n y regions, and an extension of the Mitochondria
nuclear envelope around the chloroplasts is espe- Mitochondria are limited by the usual double-
cially well seen in Chorda where the nucleus is membraned envelope. Characteristically, the in-
closely associated with the surface of the chloro- vaginati0ns of the inner membrane which form
plasts (Fig. 1). Direct continuity of the outermost the cristae are tubular or villus-like rather than
envelope of the chloroplast with the nuclear en- plate-like (Figs. l, 2, and 14). Electron-opaque
velope has been established with certainty only in globuli (of Peachey, 1964) appear to be present in
Chordafilum. small quantities in those genera examined. The
shape of the mitochondria is highly variable, and
FIGURE 4, Portion of a cell of Giffordia showing the relation of nucleus (N) and Golgi
apparatus (G). A blebbing of the outer of the two nuclear membranes is apparent at ar-
rows in the inset. X S9,000. Inset, X 50,000.
529
G. BENJAMIN BOUCK OrganelleAssociations in Brown Algae
crystal-containing bodies of fungi and higher discs. Near the ends of the chloroplasts electron-
plants (Thornton and Thimann, 1964; Cronshaw, transparent areas often contain fine fibers, and in
1964). some genera pyrenoids arise from the chloroplast
Vacuoles are limited by a single membrane and surface. Mitochondria are found to possess villus-
may contain a variety of materials varying in their like rather than plate-like cristae, and are, there-
electron opacity. In Fucus especially, the contents fore, suggestive of the mitochondrial structure
of the vacuoles frequently appear heavily electron- reported for protozoa (cf Sedar and Rudzinska,
opaque (Fig. 14) and resemble the accumulations 1956). Crystal-containing bodies similar to those
of tannins found in other plant tissues (Politis, found in higher plants and fungi are seen in small
1961). Also in Fucus, spherical membrane-limited quantities in brown algae, and thereby extend
vesicles about 800 m# in diameter are often abun- the range of these anomalous structures. Paired
dant in the outer cells. In some sections, these centrioles and associated spindle-like fibers appear
vesicles appear to protrude from the cytoplasm structurally identical to their counterparts in ani-
into the wall region, suggesting that there is a mals and lower plants. In the brown algae, how-
release of material to the wall (Fig. 10). Many of ever, the membranes surrounding the pyrenoid,
FIGURE 6 Portion of a cell of Giffcrdia either not sectioned through the neck of the
pyrenoid (P) or sectioned through a pyrenoid not attached to the chloroplast. The chloro-
plast ER (CER) cleaily can be traced around the pyrenoid. B, chloroplast band; CE,
chloroplast envelope. X 30,000.
Fmvu~. 8 A region near the end of the chloroplast which is usually less electron-opaque
than other parts of the chloroplast matrix. Note appearance of fine fibers (arrows). X
70,000.
531
G. BENJAMINBovcK OrganelleAesoclations in Brown Algae
envelope, but does form a loosely fitting sac sur- maintains contact with the regions blebbing ves-
rounding the chloroplast. Four features of this icles to the Golgi apparatus. In the brown algae,
sac are worthy of notice and may provide clues to the Golgi apparatus is consistently oriented adja-
its function: (1) around the pyrenoid and in con- cent to the nuclear envelope except in the order
trast to the remainder of the chloroplast surface Fucales in which the Golgi apparatus may be
the chloroplast E R is very regularly and closely associated with a portion of the cytoplasmic E R
appressed to the pyrenoid (i.e., chloroplast) en- as well as the nuclear envelope. The connection
velope; (2) the space between the chloroplast between Golgi apparatus and nuclear envelope
envelope and the chloroplast E R frequently con- appears to be indirect. That is vesicles derived
tains projections or villi from the chloroplast ER; from the nuclear membrane are first seen as free
(3) the chloroplast ER may be continuous with spheres and are then incorporated into the form-
the nuclear envelope; and (4) the chloroplast E R ative region of the Golgi apparatus.
lacks ribosomes on its inner surface. Considering The dynamic nature of the Golgi apparatus has
that the E R may serve as a channeling or conveyor been frequently noted, and, in general, materials
system in other kinds of cells (e.g., Porter, 1961; incorporated into the formative region are be-
FmuaE 10 A section through two Golgi bodies (G) in Fucus showing the association of
the formative face with a portion of the ER. Membrane-bounded electron-opaque vesicles
(V) appear protruding on the wall (CW). At the arrow is a vesicle which may have re-
leased its contents. )< 35,000.
FIGURE 11 A portion of a cell of Fucus veslculosus sectioned near the apical growing
region. Chloroplasts appear to have an outer chloroplast ER (CER) envelope. CE, chloro-
plast envelope; CW, cell wall; NE e nuclear envelope. X 31,000.
533
G. BENJAMINBOUCK OrganeUeAssociation~ in Brown Algae
ciated with the Golgi apparatus in the hornwort, Golgi apparatus, and ultimately extruded from
Anthoceros. However, in higher plants, no convinc- the cell. Thus, the expansion of the nuclear en-
ing evidence of an association of the Golgi appa- velope and the chloroplast E R system to include
ratus with the E R or nuclear envelope has yet the Golgi apparatus, at least in brown algae, seems
been recorded (e.g. Whaley, Mollenhauer, and appropriate. It provides a structural channeling
Kephart, 1959), and there the mechanisms of system from the site of carbon fixation and sugar
acquisitions by the Golgi apparatus remain ob- synthesis to the probable site of the sequestering,
scure. possible polymerization, and secretion of a poly-
In summary, in the brown algae it would seem saccharide (Fig. 16).
that materials manufactured within the pyrenoid
are passed across the surrounding E R system into The technical assistance of Miss Joan E. Ncuchal is
a collecting sac and perhaps stored there for in- gratefully acknowledged. This investigation was
ternal use. Conceivably, other materials synthe- supported by funds from grants NSF GB 279, N I I t
sized within the chloroplast are passed through GB 111 00-01, and N I H GB 111 00-02.
the surrounding E R system, sequestered by the Receivedfor publication, December 2, 1964.
FIGURE 12 Transverse section through a centriole of Fucus showing the typical nine
skewed triplet fibers. X 85,000.
Fmtrag 18 A section through the long axis of a centriole of Fucus. Note the depression
in the nuclear envelope (NE) in which the rod-like centriole is placed. Spindle fiber-like
elements radiate from the lower portion of the centriole over the nucleus as well as
towards the cell surface. X 89,000.
FIGURE 14 A cell of Fucus illustrating some of the variety of vacuoles and membrane-
limited vesicles. A vacuole at lower right appears to be filled with electron-opaque tubules.
Some of these vacuoles probably correspond to the "physodes" of light microscopists.
Ch, chloroplast; M, mitochondrion. X 19,000.
FmURE 15 Single membrane-limited body of Fueus showing a lattice of regularly spaced,
electron-opaque lines or dots. Spacings are about 175 A. X 56,000.
535
G. BENJAMIN BOUCK OrganelleAssociations in Brown Algae
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FIGURE 16 Diagram of a hypothetical brown alga cell illustrating some of the organelle associations
found to occur within the group. The chloroplast envelope (CE) limits the chloroplast (Ch) and pyrenoid
(P). On the cytoplasmic side of the chloroplast envelope lies the chloroplast E R (CER) which is seen,
at least in Ctwrda filum, to be continuous with the nuclear envelope (NE) and also follows the contours
of the pyrenoid. Golgi bodies (G) are closely associated with the nuclear envelope surface or a portion
of the ER. It is believed that vesicles originating from either the nuclear envelope or the E R are incor-
porated into the formative region of the Golgi apparatus. The pyrenoid sac (PS) forms a distinct and
often dilated envelope which may contain reserve carbohydrates synthesized and/or polymerized within
the pyrenoid. M, mitochondrion; N, nucleus; N1, nueleolus.
BIBLIOGRAPHY
BERKALOFF, C., 1961, l~tude au microscope 61ec- CHADEFAUD, ~¢~., 1936, Le cytoplasme des algues
tronique des plastes de Laminaria saccharina L., vertes et des algues brunes, scs 616ments figures et
Compt. rend., Aead. sc. (Paris), 252, 2747. ses inclusions, Rev. Algol., 8, 1.
BERKALOFF, C., 1963, Les cellules m6rist6matiques CRONSHAW, J., 1964, Crystal containing bodies of
d'Himanthalia Lorea (L.), S. F. Gray, l~tude au plant ceils, Protoplasma, 59, 318.
microscope ~lectronique, J. Micr., 2, 213. CRONSHAW, J., MYERS, A., and PRESTON, R. D.,
BOUCK, G. B., 1962, Chromatophore development, 1958, A chemical and physical investigation of
pits, and other fine structure in the red alga, the cell wails of some marine algae, Biochim. et
Lomentaria baileyana (Harv.) Farlow, or. Cell Biol., Biophysica Acta, 27, 89.
12, 553. DAWES, C. J., SCOTT, F. M., and BOWLER, E., 1961,
BOUCK, G. B., 1964, Fine structure in Acetabularia A light and electron-microscopic survey of algal
and its relation to protoplasmic streaming, in cell walls. I. Phaeophyta and Rhodophyta, Am.
Primitive Motile Systems, (R. D. Allen and N. d. Bot., 48, 925.
Kamiya, editors), New York, Academic Press, FRIEND, O. S., 1964, The fine structure of Brunner's
Inc. gland in the mouse, J. Cell Biol., 23, No. 2, Pt. 2,
CARO, L. G., and PAEADE, G. E., 1964, Protein 32A.
synthesis, storage, and discharge in the pancreatic FRITSCH, F. E., 1945, The structure and reproduction
exocrine cell. An autoradiographic study, J. of the algae, 2, Cambridge University Press.
Cell Biol., 20, 473. GIBBS, S. P., 1962 a, The ultrastructure of the