Skip to main content
Log in

Tip growth of Neurospora crassa upon resource shortage: Disturbances of the coordination of elongation, branching, and septation

  • Published:
Cell and Tissue Biology Aims and scope Submit manuscript

An Erratum to this article was published on 01 January 2017

Abstract

The characteristics of elongation, branching, septation, and nuclear morphology in hyphal tips (of ~400 μm in length) of the mycelial fungus Neurospora crassa isolated from the mycelium and cultivated for several hours have been investigated using intracellular fluorescent markers. The newly formed branches had the following characteristic features: (1) the predefined orientation was conserved, whereas the diameter decreased (from 10–20 to 6.5 ± 0.4 μm), as did the elongation rate (from 24 ± 1 to 6.7 ± 0.5 μm/min); (2) a disturbed branching pattern with abnormally large internodal distances (up to 1471 μm) and developmental arrest of part of the buds of lateral branches; and (3) a conserved septation pattern and a relatively constant length of hyphal segments (68 ± 2 μm). The size of the nucleus-free zone at the tip (5–33 μm) and the distance between the first septum and the growth point (210 ± 15 μm) in the daughter branches of the isolated fragments were almost the same as in hyphae connected to the mycelium, whereas the average distance between the growth point and the first lateral branch (492 ± 127 μm) and the variability of this parameter were higher in the isolated fragments. The morphology of the nuclei and the size of the nucleus-free zone near the growth point did not differ from those reported for normal vegetative hyphae of N. crassa. The experimental model developed may be used for the elucidation of details of molecular genetic mechanisms that underlie the regulation of interactions between the intracellular structures that provide tip growth of the hyphae in N. crassa.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

TG:

tip growth

L :

fragment length

CFW:

calcofluor white

DAPI:

4′,6-diamino-2-phenylindol

E m :

membrane potential

V el :

elongation rate

d :

diameter of a hypha

References

  • Aslanidi, K.B., Aslanidi, O.V., Vachadze, D.M., Mornev, O.M., Potapova, T.V., Chailakhyan, L.M., and Shtemanetian, E.G., Analysis of electrical phenomena accompanying the growth of Neurospora crassa hyphae. Theory and experiment. Membr. Cell Biol., 1997, vol. 11, no. 3, pp. 349–365.

    CAS  PubMed  Google Scholar 

  • Böhm, K.J., Nikolaos, E. Mavromatos, N.E., Michette, A., Stracke, R., and Unger, E., Movement and alignment of microtubules in electric fields and electric-dipole-moment estimates, Electromag. Biol. Med., 2005, vol. 24, pp. 319–330.

    Article  Google Scholar 

  • Borkovich, K.A., Alex, L.A., Yarden, O., Freitag, M., Turner, G.E., Read, N.D., Seiler, S., Bell-Pedersen, D., Paietta, J., Plesofsky, N., Plamann, M., Goodrich-Tanrikula, M., Schulte, U., Mannhaupt, G., Nargang, F.E., Radford, A., Selitrennikoff, C., Galagan, J.E., Dunlap, J.C., Loros, J.J., Catcheside, D., Inoue, H., Aramayo, R., Polymenis, M., Selker, E.U., Sachs, M.S., Marzluf, G.A., Paulsen, I., Davis, R., Ebbole, D.J., Zelter, A., Kalkman, E.R., O’Rourke, R., Bowring, F., Yeadon, J., Ishii, C., Suzuki, K., Sakai, W., and Pratt, R., Lessons from the genome sequence of Neurospora crassa: tracing the path from genomic blueprint to multicellular organism, Microbiol. Mol. Biol. Rev., 2004, vol. 68, pp. 1–108.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brand, A. and Gow, N.A.R., Mechanisms of hypha orientation of fungi, Curr. Opin. Microbiol., 2009, vol. 12, pp. 350–357.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chailakhyan, L.M., Levina, N.N., Belozerskaya, T.A., and Potapova, T.V., Investigation of intercellular interactions in mycelial fungi Neurospora crassa in connection with the photoelectical changes in membranes, Biol. Membr., 1984, vol. 1, no. 1, pp. 44–55.

    CAS  Google Scholar 

  • Davis, R.H. and Perkins, D.D., Neurospora: a model of model microbes, Nature Rev. Genetics, 2002, vol. 3, pp. 7–13.

    Article  Google Scholar 

  • Davis, R.H., Neurospora: Contributions of a Model Organism, Oxford: Oxford Univ. Press., 2000.

    Google Scholar 

  • Delgado-Álvarez, D.L., Bartnicki-García, S., Seiler, S., and Mouriño-Pérez, R.R., Septum development in Neurospora crassa: the septal actomyosin tangle, doi 10.1371/journal.pone.009674410.1371/journal.pone.0096744

  • Dujovne, I., van den Heuvel, M., Shen, Y., de Graaff, M., and Dekker, C., Velocity modulation of microtubules in electric fields, Nano Lett., 2008, vol. 8, pp. 4217–4220.

    Article  CAS  PubMed  Google Scholar 

  • Dunlap, J.C., Borkovich, K.A., Henn, M.R., Turner, G.E., Sachs, M.S., Glass, N.L., McCluskey, K., Plamann, M., Galagan, J.E., Birren, B.W., Weiss, R.L., Townsend, J.P., Loros, J.J., Nelson, M.A., Lambreghts, R., Colot, H.V., Park, G., Collopy, P., Ringelberg, C., Crew, C., Litvinkova, L., DeCaprio, D., Hood, H.M., Curilla, S., Shi, M., Crawford, M., Koerhsen, M., Montgomery, P., Larson, L., Pearson, M., Kasuga, T., Tian, C., Bastürkmen, M., Altamirano, L., and Xu, J., Enabling a community to dissect an organism: overview of the Neurospora, functional genomics project, Adv. Genet., 2007, vol. 57, pp. 49–96.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Fajardo-Somera, R.A., Bowman, B., and Riquelme, M., The plasma membrane proton pump PMA-1 is incorporated into distal parts of the hyphae independentely of the spitzenkorper in Neurospora crassa, Eukaryotic Cell, 2013, vol. 12, pp. 1097–1105.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Freitag, M. and Giuffetti, L.M., Expression and visualization of green fluorescent protein (GFP) in Neurospora crassa, Fungal Genet. Newslett., 2001, vol. 48, pp. 15–19.

    Google Scholar 

  • Freitag, M., Hickey, P.C., Raju, N.B., Selker, E.U., and Read, N.D., GFP as a tool to analyze the organization, dynamics and function of nuclei and microtubules in Neurospora crassa, Fungal Genet. Biol., 2004, vol. 41, pp. 897–910.

    Article  CAS  PubMed  Google Scholar 

  • Fuchs, F., Prokisch, H., Neupert, W., and Westermann, B., Interaction of mitochondria with microtubules in the filamentous fungus Neurospora crassa, J. Cell Sci., 2002, vol. 115, pp. 1931–1937.

    CAS  PubMed  Google Scholar 

  • Harold, F.M., Molecules into cells: specifying spatial architecture, Microbiol. Mol. Biol. Rev., 2005, vol. 69, pp. 544–564. doi 10.1128/MMBR.694.544-564

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Harris, S.D., Branching of fungal hyphae: regulation, mechanisms and comparison with other branching systems, Mycologia, 2008, vol. 100, pp. 823–832.

    Article  PubMed  Google Scholar 

  • Held, M., Edwards, C., and Nicolau, D.V., Temporal and spatial in vivo optical analysis of microtubules in Neurospora crassa, in Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues, San Francisco: SPIE, 2010, vol. VIII. 75680V. doi 101117/12.841328

  • Hickey, P.C., Swift, S.R., Roca, M.G., and Read, N.D., Live-cell imaging of filamentous fungi using vital fluorescent dyes and confocal microscopy, Methods Microbiol., 2005, vol. 34, pp. 63–87.

    Article  Google Scholar 

  • Kato, F., Kurashima, K., Chae, M., Sawada, S., Hatakeyama, S., Tanaka, S., and Inoue, H., Deletion of a novel F-box protein, MUS-10, in Neurospora crassa leads to altered mitochondrial morphology, instability of MtDNA and senescence, Genetics, 2010, vol. 185, pp. 1257–1269. doi 10.1534/genetics.110.117200

    CAS  Google Scholar 

  • Lee, R.R., Comparative analysis of Ca and H flux magnitude and location along growing hyphae of Saprolegnia ferax and Neurospora crassa, Eur. J. Cell Biol., 1999, vol. 78, pp. 892–902.

    Article  Google Scholar 

  • Levina, N.N. and Lew, R.R., The role of tip-localized mitochondria in hyphal growth, Fungal Genet. Biol., 2006, vol. 43, pp. 65–74.

    Article  CAS  PubMed  Google Scholar 

  • Minke, P.F., Lee, I.H., and Plamann, M., Microscopic analysis of Neurospora crassa mutants defective in nuclear distribution, Fungal Biol. Rev., 1999a, vol. 28, pp. 55–67.

    Article  CAS  Google Scholar 

  • Minke, P.F., Lee, I.H., Tinsley, J.H., Bruno, K.S., and Plamann, M., Neurospora crassa ro-10 and ro-11 genes encode novel proteins required for nuclear distribution, Mol. Microbiol., 1999b, vol. 32, pp. 1065–1076.

    Article  CAS  PubMed  Google Scholar 

  • Money, N.P., Functions and evolutionary origin of hyphal turgor pressure, in Cell Biology of Plant and Fungal Tip Growth, NATO Science Series, Amsterdam etc.: IOS Press., 2001, vol. 328, pp. 161–170.

    Google Scholar 

  • Mouriño-Pérez, R.R., Septum development in filamentous ascomycetes, Fungal Biol. Rev. 2013, vol. 27, pp. 1–9. doi 10.1016/j.fbr.2013.02.002

    Article  Google Scholar 

  • Mouriño-Pérez, R.R., and Riquelme, M., Recent advances in septum biogenesis in Neurospora crassa, Adv. Genet., 2013, vol. 83, pp. 99–134. doi 10.1016/b978-0-12-407675-4.00003-1

    PubMed  Google Scholar 

  • Mourino-Perez, R.R., Robertson, R.W., and Bartnicki-Garcia, S., Microtubule dynamics and organization during hyphal growth and branching in Neurospora crassa, Fungal Gen. Biol., 2006, vol. 43, pp. 389–400.

    Article  CAS  Google Scholar 

  • Potapova, T.V., Intercellular interactions in the Neurospora crassa hyphae–twenty years later, Biol. Membr., 2004, vol. 21, no. 3, pp. 163–191.

    Article  CAS  Google Scholar 

  • Potapova, T.V., Cell-to-cell communication in the tip growth of mycelial fungi, in Biocommunication of Fungi, Berlin-Heidelberg: Springer-Verlag, 2012, pp. 103–114.

    Chapter  Google Scholar 

  • Potapova, T.V., Structural and functional organization of growing tips of Neurospora crassa hyphae, Biochemistry (Moscow), 2014, vol. 79, no. 7, pp. 593–607.

    Article  CAS  Google Scholar 

  • Potapova, T.V. and Boitzova, L.Yu., Structure, function, regulation: experimental analysis in groups of non-excitable cells coupled via permeable junctions, Membr. Cell Biol., 1998, vol. 11, no. 6, pp. 817–829.

    CAS  PubMed  Google Scholar 

  • Potapova, T.V., Levina, N.N., and Belozerskaya, T.A., Investigation of electrophysiological responses of Neurospora crassa to blue light, Arch. Microbiol., 1984, vol. 137, pp. 262–265.

    Article  CAS  Google Scholar 

  • Potapova, T.V., Aslanidi, K.B., Belozerskaya, T.A., and Levina, N.N., Transcellular ionic currents studied by intracellular potential recordings in Neurospora crassa hyphae, (transfer of energy from proximal to apical cells), FEBS Lett., 1988, vol. 241, pp. 173–176.

    Article  CAS  PubMed  Google Scholar 

  • Potapova, T.V., Boitzova, L.Yu., and Golyshev, S.A., Problem of interactions between intracellular structures during the Neurospora crassa tip growth, Dokl. Biochem. Biophys., 2011, vol. 436, pp. 44–48.

    Article  CAS  PubMed  Google Scholar 

  • Potapova, T.V., Boitzova, L.Yu., Golyshev, S.A., and Popinako, A.V., The organization of mitochondria in growing hyphae of Neurospora crassa, Cell Tissue Biol., 2013, vol. 8, no. 2, pp. 166–174.

    Article  Google Scholar 

  • Prokisch, H., Neupert, W., and Westermann, B., Role of MMM1 in maintaining mitochondrial morphology in Neurospora crassa, Mol. Biol. Cell., 2000, vol. 11, pp. 2961–2971.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ramos-Garcia, S.L., Roberson, R.W., Freitag, M., Bartnicki-Garcia, S., and Mourino-Perez, R.R., Cytoplasmic bulk flow propels nuclei in mature hyphae of Neurospora crassa, Eukaryot. Cell, 2009, vol. 8, pp. 1880–1890. doi 10.1128/EC.00062-09

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Riquelme, M., Robetson, R.W., McDaniel, D.P., and Bartnicki-Garcia, S., The effect of ropy-1 mutation on cytoplasmic organization and intracellular motility in mature hyphae of Neurospora crassa, Fung. Genet. Biol., 2002, vol. 37, pp. 171–179.

    Article  CAS  Google Scholar 

  • Riquelme, M., Freitag, M., Leon-Hing, E.S., and Bowman, B., Live imaging of the secretory pathway in hyphae of Neurospora crassa, Fungal Genet. Newslett., 2005, vol. 52 (suppl.), p. 53.

    Google Scholar 

  • Riquelme, M., Yarden, O., Bartnicki-Garcia, S., Bowman, B., Castro-Longoria, E., Free, S.J., Fleibner, A., Freitag, M., Lew, R., Mourino-Perez, R., Plamann, M., Rasmussen, C., Richthammer, C., Roberson, R.W., Sanchez-Leon, E., Seiler, S., and Watters, M.K., Architecture and development of the Neurospora crassa hypha—a model cell for polarized growth, Fungal Biol., 2011, vol. 115, pp. 446–474. doi 10.1016/j.funbio.2011.02.008

    Article  PubMed  Google Scholar 

  • Roca, M.G., Kuo, H-Ch., Lichius, A., Freitag, M., and Read, N.D., Nuclear dynamics, mitosis and the cytoskeleton during the early stages of colony initiation in Neurospora crassa, Eukar. Cell, 2010, vol. 9, pp. 1171–1183. doi 10.1128/EC.00329-09

    Article  CAS  Google Scholar 

  • Roper, M., Ellison, C., Taylor, J.W., and Glass, N.L., Nuclear and genome dynamics in multunucleate ascomycete fungi, Curr. Biol. 2011, vol. 21, pp. R786–R793. doi 10.1016/j.cub.2011.06.042

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roper, M., Simonin, M., Hickey, P.C., Leeder, A., and Glass, N.L., Nuclear dynamics in a fungal chimera, Proc. Natl. Acad. Sci. U. S. A., 2013, vol. 110, pp. 12875–12880.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schnedl, W., Roscher, U., van der Ploeg, M., and Dann, O., Cytofluorometric analysis of nuclei and chromosomes by DAPI staining, Cytobiologie, 1977, vol. 15, pp. 337–362.

    Google Scholar 

  • Seiler, S. and Plamann, M., The genetic basis of cellular morphogenesis in the filamentous fungus Neurospora crassa, Mol. Biol. Cell., 2003, vol. 14, pp. 4352–4364. doi 0.1091/mbc.E02-07-0433

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Slayman, C.L., The plasma membrane ATPase of Neurospora: a proton-pumping electroenzyme, J. Bioenerget. Biomemb., 1987, vol. 19, pp. 1–20.

    CAS  Google Scholar 

  • Slayman, C.L. and Slayman, C.W., Measurements of membrane potential in Neurospora, Science, 1962, vol. 136, pp. 876–877.

    Article  CAS  PubMed  Google Scholar 

  • Smolianinov, V.V., and Potapova, T.V., Estimation of the critical length of the growing Neurospora crassa hyphae, Biol. Membr., 2003, vol. 20, no. 4, pp. 304–312.

    Google Scholar 

  • Sugden, K.E.P., Evans, M.R., Poon, W.C.K., and Read, N.D., Model of hyphal tip growth involving microtubule- based transport, Phys. Rev. E Stat. Nonlin. Soft Matter. Phys. Rev., 2007, vol. E75, p. 031909. doi http://dx.doi.org/10.1103/PhysRevE.75.031909

    Article  Google Scholar 

  • Takeuchi, Y., Schmid, J., Caldwell, J.H., and Harold, F.M., Transcellular ion currents and extension of Neurospora crassa hyphae, J. Membr. Biol., 1988, vol. 101, pp. 33–41.

    Article  CAS  PubMed  Google Scholar 

  • Thompson-Cofee, C., and Zicker, D., How the cytoskeleton recognizes and sorts nuclei of opposite mating type during the sexual cycle of filamentous ascomycetes, Dev. Biol., 1994, vol. 165, pp. 257–271.

    Article  Google Scholar 

  • Trinci, A.P.J., The duplication cycle and vegetative development in moulds, in The Filamentous Fungi, London: Edward Arnold. III, 1978, pp. 132–163.

    Google Scholar 

  • Trinci, A.P.J., The duplication cycle and branching, in Fungal Wall and Hyphal Growth, London: Cambridge Univ. Press, 1979, pp. 319–358.

    Google Scholar 

  • Trinci, A.P.J., Wiebe, M.G., and Robson, G.D., The mycelium as an integrated entity, in The Mycota I. Growth, Differentiation and Sexuality, Berlin–Heidelberg: Springer–Verlag, 1994, pp. 175–193.

    Chapter  Google Scholar 

  • Westermann, B., Molecular machinery of mitochondrial fusion and fission, J. Biol. Chem., 2008, vol. 283, pp. 13501–13505.

    Article  CAS  PubMed  Google Scholar 

  • Wideman, J.G., Go, N.E., Klein, A., Redmond, E., Lackey, S.W.K., Tao, T., Kalbacher, H., Rapaport, D., Neupert, W., and Nargang, F.E., Roles of the Mdm1, Tom7, Mdm12, and Mmm1 proteins in the assembley of mitochondrial outer membrane proteins in Neurospora crassa, Mol. Biol. Cell, 2012, vol. 21, pp. 1725–1736. doi 10.1091/mbc.E09–10–084

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to T. V. Potapova.

Additional information

Original Russian Text © T.V. Potapova, L.Yu. Boitsova, S.A. Golyshev, A.Ya. Dunina-Barkovskaya, 2016, published in Tsitologiya, 2016, Vol. 58, No. 8, pp. 634–645.

An erratum to this article is available at http://dx.doi.org/10.1134/S1990519X17010138.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Potapova, T.V., Boitsova, L.Y., Golyshev, S.A. et al. Tip growth of Neurospora crassa upon resource shortage: Disturbances of the coordination of elongation, branching, and septation. Cell Tiss. Biol. 10, 486–499 (2016). https://doi.org/10.1134/S1990519X16060055

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1990519X16060055

Keywords

Navigation