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Ultrastructure of freeze-substituted pollen tubes ofLilium longiflorum

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Summary

In view of the importance of the lily pollen tube as an experimental model and the improvements in ultrastructural detail that can now be attained by the use of rapid freeze fixation and freeze substitution (RF-FS), we have reexamined the ultrastructure of these cells in material prepared by RF-FS. Several previously unreported details have been revealed: (1) the cytoplasm is organized into axial “slow” and “fast” lanes, each with a distinct structure; (2) long, straight microtubule (MT) and microfilament (MF) bundles occur in the cytoplasm of the fast lanes and are coaligned with every organelle present; (3) the cortical cytoplasm contains complexes of coaligned MTs, MFs, and endoplasmic reticulum (ER); (4) the cortical ER is arranged in a tight hexagonal pattern and individual elements are closely appressed to the plasma membrane with no space between; (5) mitochondria and ER extend into the extreme apex along the flanks of the pollen tube, and vesicles and ER are packed into an inverted cone-shaped area at the center of the apex; (6) MF bundles in the tip region are fewer, finer, and in random orientation in comparison to those of the fast lanes; (7) the generative cell (GC) cell wall complex contains patches of plasmodesmata; (8) The GC cytoplasm contains groups of spiny vesicles that are closely associated with and seem to be fusing with or pinching off from mitochondria, and (9) the vegetative nucleus (VN) contains internal MT-like structures as well as numerous cytoplasmic MTs associated with its membrane and also located between the VN and GC.

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Abbreviations

CF:

chemical fixation

ER:

endoplasmic reticulum

GC:

generative cell

MF:

microfilament

MT:

microtubule

PD:

plasmodesmata

PM:

plasma membrane

RF-FS:

rapid freeze fixation-freeze substitution

VN:

vegetative nucleus

References

  • Arsanto J-P (1982) Observations on P-protein in dicotyledons. Sub-structural and developmental features. Amer J Bot 69: 1220–1212

    Google Scholar 

  • Cresti M, Ciampolini F, Kapil RN (1984) Generative cells of some angiosperms with particular emphasis on their microtubules, J Submicrosc Cytol 16: 317–326

    Google Scholar 

  • —, Lancelle SA, Hepler PK (1987) Structure of the generative cell wall complex after freeze substitution in pollen tubes ofNicotiana andImpatiens. J Cell Sci 88: 373–378

    Google Scholar 

  • Cronshaw J, Esau K (1967) Tubular and fibrillar components of mature and differentiating sieve elements. J Cell Biol 34: 801–815

    PubMed  Google Scholar 

  • Dashek WV, Rosen WG (1966) Electron microscopical localization of chemical components in the growth zone of lily pollen tubes. Protoplasma 61: 192–204

    PubMed  Google Scholar 

  • Derksen J, Pierson ES, Traas JA (1985) Microtubules in vegetative and generative cells of pollen tubes. Eur J Cell Biol 38: 142–148

    Google Scholar 

  • Deshpande BP (1974) On the occurrrence of spiny vesicles in the phloem ofSalix. Ann Bot 38: 865–868

    Google Scholar 

  • Dickinson DB (1965) Germination of lily pollen: respiration and tube growth. Science 150: 1818–1819

    Google Scholar 

  • — (1967) Permeability and respiratory properties of germinating pollen. Physiol Plant 20: 118–127

    Google Scholar 

  • — (1968) Rapid starch synthesis associated with increased respiration in germinating lily pollen. Plant Physiol 43: 1–8

    Google Scholar 

  • Esau K, Gill RH (1970) Observations of spiny vesicles and P-protein inNicotiana tabacum. Protoplasma 69: 373–388

    Google Scholar 

  • Fattah F, Webster JM (1984) Fine structure of the giant cell induced byMeloidogyne javanica in lima bean. Can J Bot 62: 429–436

    Google Scholar 

  • Franke WW, Herth W, VanDerWoude WJ, Morré DJ (1972) Tubular and filamentous structures in pollen tubes: possible involvement as guide elements in protoplasmic streaming and vectorial migration of secretory vesicles. Planta 105: 317–341

    Google Scholar 

  • Herth W (1978) Ionophore A 23187 stops tip growth, but not cytoplasmic streaming, in pollen tubes ofLilium longiflorum. Protoplasma 96: 275–282

    Google Scholar 

  • —, Franke WW, Bittiger H, Kuppel A, Keilich G (1974) Alkali-resistant fibrils of β-1,3- and β-1,4-glucans: structural polysaccharides in the pollen tube wall ofLilium longiflorum. Cytobiologie 9: 344–367

    Google Scholar 

  • Hepler PK, Palevitz BA, Lancelle SA, McCauley MM, Lichtscheidl I (1990) Cortical endoplasmic reticulum in plants. J Cell Sci 96: 355–373

    Google Scholar 

  • Heslop-Harrison J, Heslop-Harrison Y, Cresti M, Tiezzi A, Moscatelli A (1988) Cytoskeletal elements, cell shaping and movement in the angiosperm pollen tube. J Cell Sci 91: 49–60

    Google Scholar 

  • Iwanami Y (1956) Protoplasmic movement in pollen grains and tubes. Phytomorphology 6: 288–295

    Google Scholar 

  • Jaffe LA, Weisenseel MH, Jaffe LF (1975) Calcium accumulations within the growing tips of pollen tubes. J Cell Biol 67: 488–492

    PubMed  Google Scholar 

  • Kohno T, Shimmen T (1987) Ca2+-induced fragmentation of actin filaments in pollen tubes. Protoplasma 141: 177–179

    Google Scholar 

  • — — (1988 a) Accelerated sliding of pollen tube organelles along Characeae actin bundles regulated by Ca2+. J Cell Biol 106: 1539–1543

    PubMed  Google Scholar 

  • — — (1988 b) Mechanism of Ca2+ inhibition of cytoplasmic streaming in lily pollen tubes. J Cell Sci 91: 501–509

    Google Scholar 

  • —, Chaen S, Shimmen T (1990) Characterization of the translocator associated with pollen tube organelles. Protoplasma 154: 179–183

    Google Scholar 

  • Lancelle SA, Hepler PK (1991) Association of actin with cortical microtubules revealed by immunogold localization inNicotiana pollen tubes. Protoplasma 165: 167–172

    Google Scholar 

  • —, Torrey JG, Hepler PK, Callaham DA (1985) Ultrastructure of freeze-substitutedFrankia strain HFPCcI 3, the actinomycete isolated from root nodules ofCasuarina cunninghamiana. Protoplasma 127: 64–72

    Google Scholar 

  • —, Callaham DA, Hepler PK (1986) A method for rapid freeze fixation of plant cells. Protoplasma 131: 153–165

    Google Scholar 

  • —, Cresti M, Hepler PK (1987) Ultrastructure of the cytoskeleton in freeze-substituted pollen tubes ofNicotiana alata. Protoplasma 140: 141–150

    Google Scholar 

  • Li Y-Q, Linkens HF (1983) Neutral sugar composition of pollen tube walls ofLilium longiflorum. Acta Bot Neerl 32: 437–445

    Google Scholar 

  • —, Tsao TH (1985) Covalently bound wall proteins of pollen grains and pollen tubes grown in vitro and in styles after self- and crosspollination inLilium longiflorum. Theor Appl Genet 71: 263–267

    Google Scholar 

  • Miki-Hirosige H, Nakamura S (1982) Process of metabolism during pollen tube wall formation. J Electron Microsc 31: 51–62

    Google Scholar 

  • Newcomb EH (1967) A spiny vesicle in slime producing cells of the bean root. J Cell Biol 35: C17-C22

    PubMed  Google Scholar 

  • Nobiling R, Reiss H-D (1987) Quantitative analysis of calcium gradients and activity in growing pollen tubes ofLilium longiflorum. Protoplasma 139: 20–24

    Google Scholar 

  • Noguchi T (1990) Consumption of lipid granules and formation of vacuoles in the pollen tube ofTradescantia reflexa. Protoplasma 156: 19–28

    Google Scholar 

  • —, Ueda K (1990) Structure of pollen grains ofTradescantia reflexa with special reference to the generative cell and the ER around it, Cell Struct Funct 15: 379–384

    PubMed  Google Scholar 

  • Paulson RE, Webster JM (1970) Giant cell formation in tomato roots caused byMeloidogyne incognita andMeloidogyne hapla (Nematoda) infection. A light and electron microscope study. Can J Bot 48: 271–276

    Google Scholar 

  • Perdue TD, Parthasarathy MV (1985) In situ localization of F-actin in pollen tubes. Eur J Cell Biol 39: 13–20

    Google Scholar 

  • Pierson ES (1988) Rhodamine-palloidin staining of F-actin in pollen after dimethyl sulphoxide permeabilization. Sex Plant Reprod 1: 83–87

    Google Scholar 

  • —, Derksen J, Traas J (1986) Organization of microfilaments and microtubules in pollen tubes grown in vitro or in vivo in various angiosperms. Eur J Cell Biol 41: 14–18

    Google Scholar 

  • —, Kengen HMP, Derksen J (1989) Microtubules and actin filaments co-localize in pollen tubes ofNicotiana tabacum L. andLilium longiflorum Thunb. Protoplasma 150: 75–77

    Google Scholar 

  • —, Lichtscheidl IK, Derksen J (1990) Structure and behaviour of organelles in living pollen tubes ofLilium longiflorum, J Exp Bot 41: 1461–1468

    Google Scholar 

  • Raudaskoski M, Aström H, Perttilä K, Virtanen I, Louhelainen J (1987) Role of the microtubule cytoskeleton in pollen tubes: an immunocytochemical and ultrastructural approach. Biol Cell 61: 177–188

    Google Scholar 

  • Reiss H-D, Herth W (1979) Calcium ionophore A23187 affects localized wall secretion in the tip region of pollen tubes ofLilium longiflorum. Planta 145: 225–232

    Google Scholar 

  • — — (1980) Broad-range effects of ionophore X-537 A on pollen tubes ofLilium longiflorum. Planta 147: 295–301

    Google Scholar 

  • — — (1982) Disoriented growth of pollen tubes ofLilium longiflorum Thunb. induced by prolonged treatment with the calcium chelating antibiotic, chlorotetracycline. Planta 156: 218–225

    Google Scholar 

  • — — (1985) Nifedipine-sensitive calcium channels are involved in polar growth of lily pollen tubes. J Cell Sci 76: 247–254

    PubMed  Google Scholar 

  • Reynolds JD, Dashek WV (1976) Cytochemical analysis of callose localization inLilium longiflorum pollen tubes. Ann Bot 40: 409–416

    Google Scholar 

  • Rosen WG, Gawlik SR (1966) Fine structure of lily pollen tubes following various fixation and staining procedures. Protoplasma 61: 181–191

    PubMed  Google Scholar 

  • — —, Dashek WV, Siegesmund KA (1964) Fine structure and cytochemistry ofLilium pollen tubes. Amer J Bot 51: 61–71

    Google Scholar 

  • Southworth D (1983) PH changes during pollen germination inLilium longiflorum. In: Mulcahy DL. Ottaviano E (eds) Pollen: biology and implications for plant breeding. Elsevier, Amsterdam, pp 61–65

    Google Scholar 

  • Southworth D, Dickinson DB (1981) Ultrastructural changes in germinating lily pollen. Grana 20: 29–35

    Google Scholar 

  • Steer MW, Newcomb EH (1969) Development and dispersal of P-protein the phloem ofColeus blumei Benth. J Cell Sci 4: 155–169

    PubMed  Google Scholar 

  • Tiwari S, Polito V (1988) Organization of the cytoskeleton in pollen tubes ofPyrits communis: a study employing conventional and freeze-substitution electron microscopy, immunofluorescence, and rhodamine-phalloidin. Protoplasma 147: 100–112

    Google Scholar 

  • VanDerWoude WJ, Morré DJ, Bracker CE (1971) Isolation and characterization of secretory vesicles in germinated pollen ofLilium longiflorum. J Cell Sci 8: 331–351

    PubMed  Google Scholar 

  • Weisenseel MH, Nuccitelli R, Jaffe LF (1975) Large electrical currents traverse growing pollen tubes. J Cell Biol 66: 556–567

    PubMed  Google Scholar 

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Lancelle, S.A., Hepler, P.K. Ultrastructure of freeze-substituted pollen tubes ofLilium longiflorum . Protoplasma 167, 215–230 (1992). https://doi.org/10.1007/BF01403385

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  • DOI: https://doi.org/10.1007/BF01403385

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