Abstract
Callose, a β-1,3-glucan, lines the pollen tube cell wall except for the apical growing region, and it constitutes the main polysaccharide in pollen tube plugs. These regularly deposited plugs separate the active portion of the pollen tube cytoplasm from the degenerating cell segments. They have been hypothesized to reduce the total amount of cell volume requiring turgor regulation, thus aiding the invasive growth mechanism. To test this, we characterized the growth pattern of Arabidopsis callose synthase mutants with altered callose deposition patterns. Mutant pollen tubes without callose wall lining or plugs had a wider diameter but grew slower compared to their respective wildtype. To probe the pollen tube's ability to perform durotropism in the absence of callose, we performed mechanical assays such as growth in stiffened media and assessed turgor through incipient plasmolysis. We found that mutants lacking plugs had lower invading capacity and higher turgor pressure when faced with a mechanically challenging substrate. To explain this unexpected elevation in turgor pressure in the callose synthase mutants we suspected that it is enabled by feedback-driven increased levels of de-esterified pectin and/or cellulose in the tube cell wall. Through immunolabeling we tested this hypothesis and found that the content and spatial distribution of these cell wall polysaccharides was altered in callose-deficient mutant pollen tubes. Combined, the results reveal how callose contributes to the pollen tube's invasive capacity and thus plays an important role in fertilization. In order to understand, how the pollen tube deposits callose, we examined the involvement of the actin cytoskeleton in the spatial targeting of callose synthases to the cell surface. The spatial proximity of actin with locations of callose deposition and the dramatic effect of pharmacological interference with actin polymerization suggest a potential role for the cytoskeleton in the spatial control of the characteristic wall assembly process in pollen tubes.
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References
Abou-Saleh RH, Hernandez-Gomez MC, Amsbury S, Paniagua C, Bourdon M, Miyashima S, Helariutta Y, Fuller M, Budtova T, Connell SD (2018) Interactions between callose and cellulose revealed through the analysis of biopolymer mixtures. Nat Commun 9:1–13. https://doi.org/10.1038/s41467-018-06820-y
Barratt DP, Kölling K, Graf A, Pike M, Calder G, Findlay K, Zeeman SC, Smith AM (2011) Callose synthase GSL7 is necessary for normal phloem transport and inflorescence growth in Arabidopsis. Plant Physiol 155:328–341. https://doi.org/10.1104/pp.110.166330
Benkert R, Obermeyer G, Bentrup F-W (1997) The turgor pressure of growing lily pollen tubes. Protoplasma 198:1–8. https://doi.org/10.1007/BF01282125
Blake AW, Marcus SE, Copeland JE, Blackburn RS, Knox JP (2008) In situ analysis of cell wall polymers associated with phloem fibre cells in stems of hemp, Cannabis sativa L. Planta 228:1–13. https://doi.org/10.1007/s00425-008-0713-5
Brown GD, Gordon S (2003) Fungal β-glucans and mammalian immunity. Immunity 19:311–315. https://doi.org/10.1016/S1074-7613(03)00233-4
Brownfield L, Wilson S, Newbigin E, Bacic A, Read S (2008) Molecular control of the glucan synthase-like protein NaGSL1 and callose synthesis during growth of Nicotiana alata pollen tubes. Biochem J 414:43–52. https://doi.org/10.1042/BJ20080693
Brownfield L, Doblin M, Fincher GB, Bacic A (2009) Biochemical and molecular properties of biosynthetic enzymes for (1, 3)-β-glucans in embryophytes, chlorophytes and rhodophytes. Chem, Biochem Biol 1–3 Beta Glucans Related Polysacch https://doi.org/10.1016/B978-0-12-373971-1.00008-X
Cai G, Moscatelli A, Del Casino C, Cresti M (1996) Cytoplasmic motors and pollen tube growth. Sex Plant Reprod 9:59–64. https://doi.org/10.1007/BF02153052
Cai G, Faleri C, Del Casino C, Emons AMC, Cresti M (2011) Distribution of callose synthase, cellulose synthase, and sucrose synthase in tobacco pollen tube is controlled in dissimilar ways by actin filaments and microtubules. Plant Physiol 155:1169–1190. https://doi.org/10.1104/pp.110.171371
Carreño NL, Barbosa AM, Noremberg BS, Salas M, Fernandes S, Labidi J (2017) Advances in nanostructured cellulose-based biomaterialsAdvances in nanostructured cellulose-based biomaterials. Springer, pp. 1–32
Çetinbaş-Genç A, Conti V, Cai G (2022) Let’s shape again: the concerted molecular action that builds the pollen tube. Plant Reprod. https://doi.org/10.1007/s00497-022-00437-4
Chebli Y, Kaneda M, Zerzour R, Geitmann A (2012) The cell wall of the Arabidopsis pollen tube—spatial distribution, recycling, and network formation of polysaccharides. Plant Physiol 160:1940–1955. https://doi.org/10.1104/pp.112.199729
Chen X-Y, Kim J-Y (2009) Callose synthesis in higher plants. Plant Signal Behav 4:489–492. https://doi.org/10.4161/psb.4.6.8359
Derksen J (1996) Pollen tubes: a model system for plant cell growth. Botanica Acta 109:341–345. https://doi.org/10.1111/j.1438-8677.1996.tb00582.x
Dong X, Hong Z, Sivaramakrishnan M, Mahfouz M, Verma DPS (2005) Callose synthase (CalS5) is required for exine formation during microgametogenesis and for pollen viability in Arabidopsis. Plant J 42:315–328. https://doi.org/10.1111/j.1365-313X.2005.02379.x
Enns LC, Kanaoka MM, Torii KU, Comai L, Okada K, Cleland RE (2005) Two callose synthases, GSL1 and GSL5, play an essential and redundant role in plant and pollen development and in fertility. Plant Mol Biol 58:333–349. https://doi.org/10.1007/s11103-005-4526-7
Fayant P, Girlanda O, Chebli Y, Aubin C-E, Villemure I, Geitmann A (2010) Finite element model of polar growth in pollen tubes. Plant Cell 22:2579–2593. https://doi.org/10.1105/tpc.110.075754
Ferreira AR, Alves VD, Coelhoso IM (2016) Polysaccharide-based membranes in food packaging applications. Membranes 6:22. https://doi.org/10.3390/membranes6020022
Geitmann A (2010) How to shape a cylinder: pollen tube as a model system for the generation of complex cellular geometry. Sex Plant Reprod 23:63–71. https://doi.org/10.1007/s00497-009-0121-4
Geitmann A, Cresti M (1998) Ca2+ channels control the rapid expansions in pulsating growth of Petunia hybrida pollen tubes. J Plant Physiol 152:439–447. https://doi.org/10.1016/S0176-1617(98)80261-7
Geitmann A, Steer M (2006) The architecture and properties of the pollen tube cell wallThe pollen tube. Springer, pp. 177–200
Geitmann A, Wojciechowicz K, Cresti M (1996) Inhibition of intracellular pectin transport in pollen tubes by monensin, brefeldin A and cytochalasin D. Botanica Acta 109:373–381. https://doi.org/10.1111/j.1438-8677.1996.tb00586.x
Geitmann A (1999) The rheological properties of the pollen tube cell wallFertilization in higher plants. Springer, pp. 283–302
Ghanbari M, Packirisamy M, Geitmann A (2018) Measuring the growth force of invasive plant cells using Flexure integrated Lab-on-a-Chip (FiLoC). Technology 6:101–109. https://doi.org/10.1142/S2339547818500061
Gossot O, Geitmann A (2007) Pollen tube growth: coping with mechanical obstacles involves the cytoskeleton. Planta 226:405–416. https://doi.org/10.1007/s00425-007-0491-5
Green PB (1962) Mechanism for plant cellular morphogenesis. Science 138:1404–1405
Greenberg JT (1996) Programmed cell death: a way of life for plants. Proc Natl Acad Sci 93:12094–12097. https://doi.org/10.1073/pnas.93.22.12094
Guyon V, Tang Wh, Monti MM, Raiola A, Lorenzo GD, McCormick S, Taylor LP (2004) Antisense phenotypes reveal a role for SHY, a pollen-specific leucine-rich repeat protein, in pollen tube growth. Plant J 39:643–654. https://doi.org/10.1111/j.1365-313X.2004.02162.x
Hepler PK, Vidali L, Cheung AY (2001) Polarized cell growth in higher plants. Annual Rev Cell Develop Biol 17:159. https://doi.org/10.1146/annurev.cellbio.17.1.159
Herrero M, Hormaza J (1996) Pistil strategies controlling pollen tube growth. Sex Plant Reprod 9:343–347. https://doi.org/10.1007/BF02441953
Hiscock SJ, Bown D, Gurr SJ, Dickinson HG (2002) Serine esterases are required for pollen tube penetration of the stigma in Brassica. Sex Plant Reprod 15:65–74. https://doi.org/10.1007/s00497-002-0143-7
Kapoor K, Geitmann A (2023) Invasive processes in the life cycle of plants and fungi. In: Jensen K, Forterre Y (eds.) Soft matter in plants: from biophysics to biomimetics
Li H, Bacic A, Read SM (1999) Role of a callose synthase zymogen in regulating wall deposition in pollen tubes of Nicotiana alata Link et Otto. Planta 208:528–538. https://doi.org/10.1007/s004250050590
Lockhart JA (1965) An analysis of irreversible plant cell elongation. J Theor Biol 8:264–275. https://doi.org/10.1016/0022-5193(65)90077-9
Lord SJ, Velle KB, Mullins RD, Fritz-Laylin LK (2020) SuperPlots: communicating reproducibility and variability in cell biology. J Cell Biol. https://doi.org/10.1083/jcb.202001064
Lovy-Wheeler A, Wilsen KL, Baskin TI, Hepler PK (2005) Enhanced fixation reveals the apical cortical fringe of actin filaments as a consistent feature of the pollen tube. Planta 221:95–104. https://doi.org/10.1007/s00425-004-1423-2
Malhó R, Read ND, Trewavas AJ, Pais MS (1995) Calcium channel activity during pollen tube growth and reorientation. Plant Cell 7:1173–1184. https://doi.org/10.1105/tpc.7.8.1173
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Nishikawa S-i, Zinkl GM, Swanson RJ, Maruyama D, Preuss D (2005) Callose (β-1, 3 glucan) is essential for arabidopsispollen wall patterning, but not tube growth. BMC Plant Biol 5:1–9. https://doi.org/10.1186/1471-2229-5-22
Obermeyer G, Weisenseel MH (1991) Calcium channel blocker and calmodulin antagonists affect the gradient of free calcium ions in lily pollen tubes. Eur J Cell Biol 56:319–327
Palanivelu R, Brass L, Edlund AF, Preuss D (2003) Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABA levels. Cell 114:47–59. https://doi.org/10.1016/S0092-8674(03)00479-3
Parre E, Geitmann A (2005) More than a leak sealant. The mechanical properties of callose in pollen tubes. Plant Physiol 137:274–286. https://doi.org/10.1104/pp.104.05077
Parrotta L, Faleri C, Del Casino C, Mareri L, Aloisi I, Guerriero G, Hausman J-F, Del Duca S, Cai G (2022) Biochemical and cytological interactions between callose synthase and microtubules in the tobacco pollen tube. Plant Cell Rep 41:1301–1318. https://doi.org/10.1007/s00299-022-02860-3
Reimann R, Kah D, Mark C, Dettmer J, Reimann TM, Gerum RC, Geitmann A, Fabry B, Dietrich P, Kost B (2020) Durotropic growth of pollen tubes. Plant Physiol 183:558–569. https://doi.org/10.1104/pp.19.01505
Richmond TA, Somerville CR (2000) The cellulose synthase superfamily. Plant Physiol 124:495–498. https://doi.org/10.1104/pp.124.2.495
SanatiNezhad A, Geitmann A (2013) The cellular mechanics of an invasive lifestyle. J Exp Bot 64:4709–4728. https://doi.org/10.1093/jxb/ert254
SanatiNezhad A, Naghavi M, Packirisamy M, Bhat R, Geitmann A (2013) Quantification of cellular penetrative forces using lab-on-a-chip technology and finite element modeling. Proc Natl Acad Sci 110:8093–8098. https://doi.org/10.1073/pnas.1221677110
Sanati Nezhad A, Geitmann A (2015) Tip growth in walled cells: cellular expansion and invasion mechanismsCells, forces, and the Microenvironment. Pan Stanford, pp. 335–356
Thiele K, Wanner G, Kindzierski V, Jürgens G, Mayer U, Pachl F, Assaad FF (2009) The timely deposition of callose is essential for cytokinesis in Arabidopsis. Plant J 58:13–26. https://doi.org/10.1111/j.1365-313X.2008.03760.x
Töller A, Brownfield L, Neu C, Twell D, Schulze-Lefert P (2008) Dual function of Arabidopsis glucan synthase-like genes GSL8 and GSL10 in male gametophyte development and plant growth. Plant J 54:911–923. https://doi.org/10.1111/j.1365-313X.2008.03462.x
Turner A, Bacic A, Harris PJ, Read SM (1998) Membrane fractionation and enrichment of callose synthase from pollen tubes of Nicotiana alata Link et Otto. Planta 205:380–388. https://doi.org/10.1007/s004250050334
Verma DPS, Hong Z (2001) Plant callose synthase complexes. Plant Mol Biol 47:693–701
Wang Y, Li X, Fan B, Zhu C, Chen Z (2021) Regulation and function of defense-related callose deposition in plants. Int J Mol Sci 22:2393. https://doi.org/10.3390/ijms22052393
Wu S-W, Kumar R, Iswanto ABB, Kim J-Y (2018) Callose balancing at plasmodesmata. J Exp Bot 69:5325–5339. https://doi.org/10.1093/jxb/ery317
Xie B, Hong Z (2011) Unplugging the callose plug from sieve pores. Plant Signal Behav 6:491–493. https://doi.org/10.4161/psb.6.4.14653
Xie B, Wang X, Zhu M, Zhang Z, Hong Z (2011) CalS7 encodes a callose synthase responsible for callose deposition in the phloem. Plant J 65:1–14. https://doi.org/10.1111/j.1365-313X.2010.04399.x
Zerzour R, Kroeger J, Geitmann A (2009) Polar growth in pollen tubes is associated with spatially confined dynamic changes in cell mechanical properties. Dev Biol 334:437–446
Zhang R, Edgar KJ (2014) Properties, chemistry, and applications of the bioactive polysaccharide curdlan. Biomacromol 15:1079–1096. https://doi.org/10.1021/bm500038g
Zhang D, Wengier D, Shuai B, Gui C-P, Muschietti J, McCormick S, Tang W-H (2008) The pollen receptor kinase LePRK2 mediates growth-promoting signals and positively regulates pollen germination and tube growth. Plant Physiol 148:1368–1379. https://doi.org/10.1104/pp.108.124420
Acknowledgements
This study was supported by Natural Sciences and Engineering Research Council of Canada (NSERC) and by the Canada Research Chair Program. Sample preparation and image acquisition were performed at the McGill University Multi-Scale Imaging Facility, Sainte-Anne-de-Bellevue, Québec, Canada. We thank Dr. Giampiero Cai from the University of Siena for providing us with anti-callose synthase antibodies.
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Kapoor, K., Geitmann, A. Pollen tube invasive growth is promoted by callose. Plant Reprod 36, 157–171 (2023). https://doi.org/10.1007/s00497-023-00458-7
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DOI: https://doi.org/10.1007/s00497-023-00458-7