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Capsella Embryogenesis: The Synergids Before and After Fertilization Richardis Schulz; William A. Jensen American Journal of Botany, Vol. 55, No. 5. (May - Jun., 1968), pp. 541-552. Stable URL hitp:/flinks.jstor-org/sicisici=0002-9122% 28 196805% 2F06%2955%3AS%3CS41%3ACETSBA%3E2,0.CO%3B2-V American Journal of Botany is eurrently published by Botanical Society of America, Your use of the ISTOR archive indicates your acceptance of JSTOR's Terms and Conditions of Use, available at hup:/www,jstororglabout/terms.hml. ISTOR’s Terms and Conditions of Use provides, in part, that unless you have obtained prior permission, you may not download an entire issue of a journal or multiple copies of articles, and you may use content in the JSTOR archive only for your personal, non-commercial use. Please contact the publisher regarding any further use of this work. Publisher contact information may be obtained at hupulwww.jstor-org/journals/botsamn. him ch copy of any part of'a JSTOR transmission must contain the same copyright notice that appears on the sereen or printed page of such transmission, ISTOR is an independent not-for-profit organization dedicated to creating and preserving a digital archive of scholarly journals. For more information regarding JSTOR, please contact support @ jstor.org. hupulwww jstor.org/ ‘Thu Jul 13 19:18:06 2006 ‘Ammer. J. Bot, 85(5): SUL-552, 1968 CAPSELLA EMBRYOGENESIS: THE SYNERGIDS BEFORE AND AFTER FERTILIZATION: Sister Ricrarpts Scuuiz? anv WituiaM A. Jensen Department of Botany, University of California, Berkeley ABSTRACT ‘The ultrastructure and composition of the synorgids of Capella burea-pasiois were studied before and after fertilination, The synergide inthe mature embryo sac contain numerous plastids, rmitochondia, dietyosomes and masses of ER and associated ribosomes. Each synergid contains ‘large chalazal vacuole, a nvclens with a single nucleolus and is surrounded by a wall. This wall i thiekest at the mieropyle end of tho ell where it proliferates nto the filiform apparatus, At the ‘halazal end of the oallthe wall thins and may be absent for small distanoes. ‘The pollen tube ‘grows into one of the two synergids through the filiform apparatus and extends one-third the length of the cell before it discharges, Following discharge of the pollen tube, mitochondria and plastids of the tube can be identified in the synergid as can hundreds of 0:5 polysaccharide spheres liberated by the tube. The method by which the sperm or sperm nuclei enter the egg Or central cell s not known although an apparent rupture was found in the wal ofthe egg near the tip of the pollen tube, The second aynergid changes at the timo the pollen tube enters the first synergd. These changes result in the disorganization of the nucleus and loss of the chalazal wall and plasma membrane. Eventually this synergid loses its identity as its cytoplasm merges swith that of the central col ‘Tum, sywmnows are being assigned an ineress- ingly important role in. discussions concerning the entranee and discharge of the pollen tube into the embryo sae (van der Pluijm, 1964; Jensen, 1965; Van Went and Linskens, 1967; Jensen and Fisher, 1968). This report deseribes the synergids of Capsella bursa-pastoris before and after ferti- Tization. Changes. in ‘ultrastructure and com- position of these cells are described from thir ature condition in. the embryo. sae. through their degeneration after fertilization. The data shed additional light on the role of the synergids in the mature megagametophyte and in the events surrounding pollen tube entrance and discharge. Marmarats axp susrions—Plants of Capella iuraa-pasioris L.. “Medic,” the shepherd's purse, were grown in the greenhouse from seeds collected at the Botanieal Garden, University of California, Berkeley. Whole ovules vere dissected from the siliques. and fixed, immediately in 2% KMnO, for 15-25 hr at 4.C. ‘The tips of some of the older ovules were excised before fixation in order to Thliate penetration of fratives and embedding media. Alternately, ovules were placed in 6 lutaraldehyde buffered by 0.06 x phosphate at DIGS fort hr at 4 C-and post-fixated with 2% + Received fr publieation 18 September 1967. sar pied ty NS grant ON 400 a NTL grant S-HOT-CA 03650-4, ‘Research ofthe tefor author conducted while an NSF PMc adgrss: Department of Biology, Rosary “cent addres: Department. of Biology, College, River Foret, 1 6098, mbuifered Os0, containing 4% suerose for 15 hr at 4 C. The tissue was dehydrated in a graded acetone series. Staining with 1% uranyl nitrate was done in the 70% acetone, All material was embedded in. Epon, Sections were stained with lead citrate (Reynolds, 1963) and observed with 1 Zeiss Electron Microscope EM.9. ‘Material prepared for the histochemical locali- zation of nucleic acids was fixed in GA-OsOs, embedded in Epon, sectioned at 2 4 and stained with Azure B (Fensen, 1962). ‘The periodic acid- Schiff (PAS) reaction was used for the localization of insoluble earbyhodrate (Jensen, 1962) and Aniline Blue Black (1% dye in 7% acetic acid, stained for 10 min at 60 C, rinsed in 7% acetic acid) for a, general protein stain (Fisher, un- published), Tissue for the latter procedures was fixed in glutaraldehyde, embedded in Epon and sectioned at 12 u. Permanent mounts were made of material siainod. with Azure B and PAS with Zeiss phase-mounting medium 115..‘Tempor Touts of material stained wth Aniline Blue Black were made with glycerol to which 5% acetic ack had been added. Resvuns—Before Sertilization—At the micro pylar end of the embryo sac the mature egg flanked by two synengids in triangular artange- ‘ment termed the egg apparatus. The synergids are the same length as the xg (254) but ases penetrate deep into the mieropylar apex of the embryo sac whe the base of the ea al tached at the side of the embryo sae approxi- mately 10) back from the tip (Fig. 1). This aL [Vol. 35 AN JOURNAL OF BOTANY ‘May-June, 1968] Fig. 2, SCHULZ AND JENSEN—CAPSELLA EMBRYOGENESIS as —Fig. 2. Longitudinal soction through the two synergids showing the intense staining of the nucleus, nu- leolti, and eytopiasm for protein. The filiform apparatus (PA) and the synergid wall (double arrow) remain unstained. ‘The fused polar nuclei (PN) ean also be seen. Aniline Bl Black, x 2,000-—Fig. 8. Longitudinal rection through the ‘sro synergide (SY) showing the PAS-positive reaction of the wall double arrows) and filiform apparatus for insoluble ‘carbohydrate, The plastids (P) stain intensely for protein, Aniline Blue Black and PAS, X 2,000. causes the chalazal end of the egg to extend beyond the synergids making the exe appear longer. The synergids possess a centrally located nucleus and a large chalazal vacuole (Wig. 1). The micropylar end of both cells is occupied by the complex striated filiform apparatus (Haber- mann, 1906); (Fig. 2, 3). ‘The bulk of the eytoplasm lies between the chalazal vacuole and filiform apparatus. ‘The synergids arise at_ the third mitotic division of megagametogenesis which yields the nucleate embryo sac. They are shortlived cells and become disorganized and disappear soon after fertilization of the egw. ‘A cell wall surrounds each synergid. ‘This wall is thickest toward the micropylar end of the cell where wall synthesis began but becomes thin and irregular over the chalazal end of the cell where it borders the central cell. Often the plasma membranes of the synergid and central cell lie in close contact (Fig. 4), while at other places the two membranes are clearly separated by an clectron-translueent space which contains small patehes of electron-dense material (Fig. ‘The region between the two membranes is PAS. positive (Fig. 3) and is presumed to contain wall material. The synergid wall becomes continuous with the embryo sae wall in the mid-region of the cell. Each synergid also shares. common wall with the egg (Fig, 1) and with its sister synengid (Fig. 2, 3). Plasmodesmata are -seen in all parts of the wail except where it is contin- uous with the embryo sae wall (Fig. 5). At the mieropylar end of each. synergid ‘the wall becomes thickened and folded into the long finger-like projections of the filiform apparatus extending deep into the cytoplasm (Wig. 2, 3, 6). The filiform apparatus appears to consist of ‘two structural phases: a central electron-dense core surrounded by more electron-translucent phase (Fig. 6, 7). The core consists of a. tightly Fig. 1. The micropylar end ofthe embryo sue showing the structure and relative positions of the ogg, synergid (SY), and central ell (CC). GA-0s04, X50. AMERICAN JOURNAL OF BOTANY {Vol. 55 ‘May-June, 1963} packed mass of microfibrils while the peripheral Tegion contains fewer loosely organized, micro- fibrils set in a translucent: matrix (Pig. 7). ‘The filiform apparatus. gives a strong PAS-positive reaction for insoluble earbohydrate (Fig. 8) but is negative for nucleic acids and protein (Fig. 2). ‘The plasma membrane follows the contours of the filiform apparatus which greatly increases its surface area in this region (Fig. 7). ‘The plasma rmerene and th Bare oquently eon in the region of the filiform apparatus (Fig. 6) an ae parallel inthe chalaal Tlf of the cell (ig 4). This parallel relationship gives the plasma- Jesama the sopearance of being » double mem. rane synergid has a large (0 X 11 1) vacuole in the chalazal half of the eell and may contain several smaller randomly distributed vacuoles (1 ») (Fig, 1). “Phe central nucieus is $-6 2 in diam and has single, prominent nucleolus (3.5 a) consisting of a mass of densely packed granules with a few small clear areas in it, Small dense masses of chromatin are frequently seen near the nuclear ‘membrane. The synergid cytoplasm is filled with ribosomes and stains intensely for protein (Fig. 2) and RNA. Characteristic of synergd eytopltam are long profiles of ER frequently appearing in groups of two to four strands oriented parallel to the long axis of the cell (Fig. 5). In general, the cistemal phase of the ER’ is narrow and “uniform except at the tips where it may be swollen (Fig. 8). An exception to this is the ER closely paralleling the plasma membrane which has a ‘vide cisternal phase. At the chalazal end of the synergid the ER often encircles plastids and mitochondria. (Fig. 1) “Phere are relatively few plastids in the synergid compared with their numbers in the egg and central cell (Fig. 1). Most of the plastids are located in. the eytoplasm between the nucleus and the filiform apparatus although a few may ‘ccur at the chalazsl end of the cell. ‘Many cup-shaped plastids are present (Fig. 6). The appearance’ of the eytoplasm within’ the invaginations is altered and contains fewer ribosomes than the neighboring eytoplasm. This form of plastid i also common in the central cell, egg, zygote, and young embryo. ‘The plastids ‘are surromded by a double membrane, and each contains several single lamellae which are invaginations of the inner membrane (Fig. §). Some lamellae are quite vesiculate in appearance and only rarely are two Big. 4, 5.-Fig. 4. The jt SCHULE AND JENSEN—CAPSELLA EMBRYOGENESIS BAS fused. The plastid matrix is very electron-dense and iain intnaly for prota allowing ito be easily seen in light-microscope sections (Fig. 3 ‘There are a few diffuse, light areas in the matric late alo staan of. (ig. 5). The dense, spherical (700A diam) droplets (Fi and ribosome-like partiles (150 A). They cont ery litle starch, even, when stare in the egg and central cell of the same mega- snmetophyte, Mitochondria are much more numerous than plastids. They oceur throughout. the eell but are ‘more concentrated near the, filiform apparatus (Fig. 1, 6). They are rod-shaped (1X 244 u) and contain many short vesicular eristae. Clear ‘areas which contain tiny fibrils (25 A) sometimes ‘oceur in the center of the mitochondria between the eristae (Fig. 6). The mitochondria (Fig. 6) also contain intramitochondrial granules (300 A) (Peachey, 1962) and ribosome-like particles (050 A). ‘The uniform dictyosomes are ubiquitous but fare present in higher numbers in the region between the filiform apparatus and the nucleus (Fig. 1, 5). They have 4-6 flat cisternae (0.7- 0.9.4) which produce vesicles at their tips. ‘There are a few single membrane-bound organelles (0.3 ») randomly distributed in the cell which are spherical or rod-shaped and have dense granular contents (Fig. 6). Crystals are sometimes seen in the BR cisternae. ‘They are long and narrow (0.5 X 0.04) and appear white in GA-Os0, black in KMnO,, They are Believed to be, inorganic beeause they aro pre- served with KMnOi which destroys protein crystals (Jensen, 1965). Microtubules (Ledbetter and Porter, 1963) have also been observed in the synengid cytoplasm, After friization—The pollen tube enters the embryo sae through the flform apparatus. After entering. the ‘synergid_eytoplaam. it continues {fo grov in a chalagal direction close to the com- ron egg-synergid wall until it has traveled one- third the length of the cell. At this point the tp of the pollen tube opens and discharge occurs ‘Thereafter both synergids undergo characteristic changes. "The most immediate and apparent henge oer in the eerie i penetra by the pollen tube (degenerating synergid). ‘The other synergid. (persistent synergid) remains sotive fora short time after fertilization and then ‘becomes disorganized in a manner different from, tion ofthe egg, synergid (SY), and conta ell (CC) showing the close parallel relationship ‘of the ER with the plasma membrane of the synergd (arrows). Gaps which contain a dense material (DM) separate the ‘lasina membranes of the synergid and central eall, but in some areas the two membranes are very close and appear to have little oF no intervening material between them (double arrow). GA-OsO,, x 25,600.—Fig. 6. View of the synergid (SY) and a portion of the egg at the level of the synergid nucleus (N) showing the relative abundance of ER, mito- chondria (M), and dictycsomes (D). Plasmodesmata (PD) are eommon in the wall between the synengid and the exg. Plaatids (P) are relatively undifferentiated and contain dense droplets. GA-OsO,, X 25,600, 546 AMERICAN JOURNAL OF BOTANY [Vol. 55 ‘May-June, 1968] ‘the degenerating synengid. ‘The contents of both synergids are ultimately reabsorbed by the growing embryo sac. Degenerating synergid—'There are _several ryt paso the daca yer fid. and pollen tube oytoplasing. ‘The large chalazal vacuole of the synergid disappears and the upper part of the ell colapecs against the zygote. The cytoplasm is very dark but several structures are still discernible. The nucleus and Icleolus are somewhat flattened and the nuclear membrane has disappeared (Fig. 8). Organelle structure is disorganized and in GA-OsO, material ‘these structures are unrecognizable except where the occasional persistence of a starch grain identifies a plastid (Fig. 8). In KMnO, material some mitochondria can be recognized in the synergid and pollen tube by the presence of long parallel tae (Fig. 10). This type of mito- chondrion is not seen in the synergids before fertlzaton. Dense ribosomes remain inthe synergid and pollen tube. Many are randomly Oren ut ota ar nour i li Gig. 9). The plasma membrane has also dis- {pyar rom che sutfacr of tie syntgié Cg 8) ‘pollen tube liberates hundreds of spherical parte (0102 ) phi. spread dougie id above. the,naintoF rupture (Fig. 3 0 Tn GAa-Os0, these particles have a dense, ular center and are surrounded by a thin Fran of lear material (Pig. 4). In KMnO, they nye‘ coar eutars ant ace. gartouped bys dark band which may be a unit membrane (Fig. What appears to be a rupture i the wall of the egg is seen in Fig. 11. Such structures were repeatedly seen in the egg wall near the tip of the discharged pollen tube “Tho eytgplasm of tho degenerated. synereid stains much more intensely with Aniline Blue Black, Azure B, and PAS than it did before the entrance of the pollen tube. Persistent synergid—After a brief period of normal activity following fertilization the per- sistent synergid begins to show signs of di organization different from those of the do- generating synergid. One of the most notable changes occurs in the nucleus (Fig. 12, 13). ‘The ER becomes oriented in several layers around the nucleus and is continuous. with. the nucl membrane. In some places it is difficult to dis- SCHULZ AND JENSEN—CAPSELLA EMBRYOGENESIS 27 tinguish the nuclear envelope from the ER. ‘The inner nuclear membrane invaginates. to produce small vesicles and several short strands of ER (with ribosomes attached) appear in the center of the nucleus (Fig. 12). The nucleolus breaks up into several large particulate maises which migrate to the periphery of the nucleus (Fig. 12). Small. groups of polyribosomes, pre- sumably originating from the larger mastes of nucleolar material, are seen throughout the nucleus (Fig. 12, 18). These nuclear ribosomes are the same size as the ribosomes in the neighboring cytoplasm. ‘The nucleus gradually takes on the appearance of the. eytoplasm (Fig. 12) and eventually loses its identity. Parts of the wall and plasma membrane at the chalazal end of the synergid disappear and a large vacuole of the central eell makes. contact h the chalazal vacuole of the synengid (Fig. 14). Eventually” the chalazal wall” and’ membrane completely disappear and there is a gradual mixing of the synergid and central cell eytoplasms. As early as the first division of the zygote the large, well-developed plastids of the central cell ean be seen less than 14 from the filiform ap- paratus of the persistent synergid. Discussrox—The fine structural image of th synergids before fertilization is one of metabol cally ‘active cells. ‘The activity of the synergid with its large numbers of dictyosomes and mito- chondria. and abundance of ER is sharply con- trasted with the image of the egg which reflects relatively quiescent eytoplasm. ‘This study sup- ports the conclusion that the synergids function in the absorption. and transport of compounds from the surrounding integuments to the exg and possibly the central cell (Jensen, 1965). ‘The filiform apparatus is a modification of the cell wall which greatly increases the surface ares of the plasma membrane. The position of the filiform apparatus is also important. because it ecupies the micropylar tip of the embryo. sue and comes into intimate contact with metabolite- Jaden. integuments. The proximity of _mito- chondria, offers an available energy supply for active transport across the plasma membrane, and the closely associated ER provides a direct channel to metabolic sites deep in the evtoplasm and ultimately regions bordering, tho gt and contra cals Tho bes parle ap sociation of the ER and plasma membrane in these regions could facilitate the transfer of sub- _ Big. 6,7, in the region traversed by tiny ‘The plastid P) —Fig. 6. View of the filiform apparatus (PA) and associated cytoplasm. There are many mitochondria (M) ‘of the filiform apparatus which have short vesicular erstae, intramitochondtial granules, and clear areas fibrils (F). The ER. is closely associated with the plasma membrane of the flform apparatus. in the upper right corner is wrapped around a portion of the eytoplasm which appears to he inside the organelle, GA-O:0,, X 32,800.—Fig. 7. Enlarged view of the fllform apparatus showing its microuibrillar strusture and the close relationship of the plasma membrane. GA.OsO,, X 45,500. AMERICAN JOURNAL OF BOTANY BAS. ‘May-June, 1963} SCHULZ AND JENSEN—CAPSELEA RMBRYOGENESIS 9 Fig. 10, 11—Fig, 10, The open polle tube (PT) in the degenerating synergid (D SY). Mitochondria with long tubular evistao are frequently seen in the eytoplasm of both the degenersting synergid and the pollen tube, Small spherical particles resembling wall material are lerated from the open end of the pollen tube KMnO,, X 25,600. Fig. 11. View of the open pollen tube (PT) inthe degenerating aynergid and the ruptured sygote (2) wall (W). GA-Os0,, 25,600, Fig. 8. ‘The degenerating synengid containing the pollen tube (PT) which is open at one end, Starch gras (ST) mark the presence of plastids. Hundreds of small particles are liberated from the pollen tube, ‘The mileus (N) and filiform apparstus (PA) of the degenerating synergid can be seen. GA-Os0, X 9,200. ig. 0. Enlarged view of tho eytoplasm of the dogoncrsting synergid. Particles liberated from the pollen tube have & srantiar appearance, Chains of ubosomes and. spherical bodies which may be disorganized mitochondria (MD are fre= tently seen, GA-OsO,, % 59,000. AMERICAN JOURNAL OF BOTANY ‘May-June, 1968] SCHULZ AND JENSEN: A EMBRYOGENESIS Fig. 14. The persistent synergid (PSY) and a portion of the zygote (Z) showing the gap in the chalazal wall (W) fad membrane of the synergid and the cloce relationship hetween the vacuoles of the synergid and central call (CC), GA.Oe04, % 25,00. stances. Numerous plasmodesmata in the walls connecting the synergid with the egg and eéntral cell provide an additional route for the free flow ‘of metabolites. The presence of many active dietyosomes is characteristic of cells engaged in secretion (Mollenhauer and Morré, 1966). The egg and central ecll store metabolites in the form of starch and lipid which are used in the early stages of development, following ferti- lization. The actual amounts of these compounds stored depends in part on the general state of nutrition and metabolism of the plant, but the egg and central coll always have more starch and lipid than the synergids. In contrast. to Capsella the synergids of cotton store large amounts of starch (Jensen, 1965), but this differ ence may be related, in part, to the slower rate of development of the cotton egg. ‘There has been much discussion about the role of the synergids in directing the growth and en- trance of the pollen tube into the embryo. sac (ean der Phun, 1964; Diboll and Larson, 1966; Jensen, 1965; Van Went and Linskens,” 19673 Jensen’ and Pisher, 1968). ‘The pollen tube. in Capella was found to enter and discharge only into a synergid, ‘The pollen tube was never seen to enter the egg or grow through to the central cell. Moreover; the discharge of the pollen tube twas found only’ in the synergid. Thus Capsella fits the pattem established for all the species examined thus far with the electron microscope Torenia fournieri (van der Pluijm, 1964), petunia (Van Went and Linskens, 1967), and cotton Gensen and Fisher, 1968).’‘The data: emphasize the importance of the degenerating synergid in the terminal growth of the pollen tube and pollen tube discharge Ultrastructural studies on the synergids of cotton (Jensen, 1965), Torenia (van der Pluijm, 1964), and maize (Diboll” and Larson, 1966) Fig. 12, 18.—Fig. 12, The disorganization of the persistent synergid nucleus. Polysomes and rough ER (arrow) appear inside the nucleus (N). ‘Tho nuclear mombrane js difieult to distinguish from the surrounding ER and the nucleolus is dispersed into clumps (NU). GA-sO,, X 16,250.—Fig. 18. Hnlarged view of polysomes inside the persistent synergid nutes which have: probabl ‘come from the nucleolus (NU). GA-Os0,, X 59,000. 552 show that a wall is absent at the chalazal end of the cell. ‘The absence of a wall in this position ‘would presumably facilitate the movement of the sperm nuclei into the egg and the central cell. ‘The synergids of Capsella, then, seem unique in possessing’ chalazal wall. In Capsella, how do the two sperm nuclei reach the egg and ‘the polar nuclei once they are discharged from the pollen tube? This is a controversial question because these events happen so quickly that it is difficult tovobserve the material at crtial stages. The present data suggest that the nucleus which fertilizes the exg moves through a rupture in the common egg-synergid wall near the open end of the pollen tube (Fig. 13). This rupture is similar in size to the male gamete and has been seen in the same relation to the pollen tube in material prepared for light and electron microscopy. ire untaual hehavor ofthe persistent synergid in the method by which it becomes disorganized had ‘heen umexpected and stands in sharp con- trast to the way in which the degenerating syner- id is destroyed. This is a good example of how different, mechanisms of destruction may, be employed by cells of the same organism. ‘The sequence of events suggests that before the cyto- plasm of the persistent synergid can mix with that of the central ell the nucleus must. be removed from the eell. The persistent synergidl cotton forms a thiek ‘wall around itself before it finally, undergoes disorganization (Jensen, un- published) AMERICAN JOURNAL OF BOTANY [Vol. 55 Dimou, A. G., axo D. A. Lansox. 1960, An cleetron ‘microscopy study of the mature megagametophyte in Zea mays, Amer. J. Bot, 58: 301-402, Hanuneasy, A. 1006.” Fadenapparat inden Synergiden ‘der Angiospermen. Bot. Centralb. Beih. 20: 300-317. Josep, W. A. 1962. Botanical histochemistry. W. I ‘Froeman and Co, San Francseo, 1965. The ‘ultrastricbiee and histochemistry of the synergds of cotton. Amer. J. Bot. 52: 238-256. sno. B, Fist. 1968, Cotton. embryo. ehesia: ‘The enizance and discharge of the pollen {ie into the embryo sae. Planta 88: 155-183. Luonernen, M.C., aNd IT Porersn, 1963. A “miero- “tubule” in plant ell fine structurd. J. Coll Biol. 19, 239-250. Pusciy, Te D. 1962. Accumulation of divalent fons ‘in mitochondrial grams of intact. ells, p. 00-8. In 8. Broo [ed.)) Fifth International Congress for Eleotron Mieroseopy. II. Academic Press, New York. Puupay J.B vaN DER.” 1964.” An electron microscopic “investigation of the fiform apparatus in. the embryo sae of Torenia fourniri, p. 8-16. In HF. Linskens fed, Pollen. physiology’ and fertilization. North Holland Pub, Co,, Amsterdam, ‘Mousxusvnn, H. 1 ax D. J. Monné. 1966. Golgi ‘pparatus’and plant. secretion. Ann. Rev. Plant Physiol. 17: 27-46. RayNorbs, B.S. 1063. The use of lead citrate at high ‘IL as an electron-opague stain in electron. miero- Seopy. J. Call Biol. 17: 208-212. Van Wav, J, ax HF. Linexess. 1967. Die Ent- ‘wiekhing “des sogensnncen "Fadenapparatae” in Embryoeacke von Petunia hybrila. Genet. Breeding Res. 37 51-56,

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