Abstract
Angiosperms proliferate through double fertilization mediated by male (pollen) and female (embryo sac) gametophytes. To determine the genes essential for pollen development in Arabidopsis thaliana, we first generated a mutant population using an activation tagging vector with herbicide-resistance gene and screened mature pollen phenotypes. Then, a T-DNA insertional heterozygous line was isolated, initially named AP22-48, which produced high levels of abnormal pollen grains. Reciprocal crosses revealed that the genetic transmission of the mutant allele was completely blocked through the male and was highly limited through the female. Determination of T-DNA flanking sequences and genetic complementation of AP22-48 identified AtCOG8, a subunit of the Conserved Oligomeric Golgi (COG) complex, which is a tethering factor essential for the Golgi architecture and retrograde vesicle trafficking in eukaryotes. We renamed the mutant atcog8-2, with reference to a previous cog8 mutant (atcog8-1). While atcog8-1 induced male-specific defects during pollen tube growth, atcog8-2 mutant failed to produce normal gametophytes in both sexes. Detailed morphological analysis demonstrated aberrant development of the pollen and embryo sac in atcog8-2 mutants. This study, thus, strongly suggests that the COG complex functions are broad and indispensable for accurate gametophyte development, which is a prerequisite for sexual reproduction in Arabidopsis.
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References
Åstrand J, Knight C, Robson J, Talle B, Wilson ZA (2021) Evolution and diversity of the angiosperm anther: trends in function and development. Plant Reprod 34:307–319. https://doi.org/10.1007/s00497-021-00416-1
Becker JD, Boavida LC, Carneiro J, Haury M, Feijo JA (2003) Transcriptional profiling of Arabidopsis tissues reveals the unique characteristics of the pollen transcriptome. Plant Physiol 133:713–725. https://doi.org/10.1104/pp.103.028241
Berger F, Twell D (2011) Germline specification and function in plants. Ann Rev Plant Biol 62:461–484. https://doi.org/10.1146/annurev-arplant-042110-103824
Blackburn JB, D’Souza Z, Lupashin VV (2019) Maintaining order: COG complex controls Golgi trafficking, processing, and sorting. FEBS Lett 593:2466–2487. https://doi.org/10.1002/1873-3468.13570
Boavida L, Becker J, Feijó JA (2005) The making of gametes in higher plants. Int J Dev Biol 49:595–614. https://doi.org/10.1387/ijdb.052019lb
Boavida LC, McCormick S (2007) Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana. Plant J 52:570–582. https://doi.org/10.1111/j.1365-313X.2007.03248.x
Boevink P, Oparka K, Santa Cruz S, Martin B, Betteridge A, Hawes C (1998) Stacks in tracks: the plant Golgi apparatus traffics on an actin/ER network. Plant J 15:441–447. https://doi.org/10.1046/j.1365-313x.1998.00208.x
Borg M, Brownfield L, Twell D (2009) Male gametophyte development: a molecular perspective. J Exp Bot 60:1465–1478. https://doi.org/10.1093/jxb/ern355
Borges F, Gomes G, Gardner R, Moreno N, McCormick S, Feijo JA, Becker JD (2008) Comparative transcriptomics of Arabidopsis sperm cells. Plant Physiol 148:1168–1181. https://doi.org/10.1104/pp108.125229
Christensen CA, King EJ, Jordan JR, Drew GN (1997) Megagametogenesis in Arabidopsis wild type and the Gf mutant. Sex Plant Reprod 10:49–64
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743. https://doi.org/10.1046/j.1365-313x.1998.00343.x
Dresselhaus T, Sprunck S, Wessel GM (2016) Fertilization mechanisms in flowering plants. Curr Biol 26:R125–R139. https://doi.org/10.1016/j.cub.2015.12.032
Drews GN, Koltunow AMG (2011) The female gametophyte. Arabidopsis Book 9:e0155. https://doi.org/10.1199/tab.0155
Gremillion SK, Harris SD, Jackson-Hayes L, Kaminskyj SGW, Loprete DM, Gauthier AC, Mercer S, Ravita AJ, Hill TW (2014) Mutations in proteins of the Conserved Oligomeric Golgi Complex affect polarity, cell wall structure, and glycosylation in the filamentous fungus Aspergillus nidulans. Fungal Genet Biol 73:69–82. https://doi.org/10.1016/j.fgb.2014.10.005
Gu F, Nielsen E (2013) Targeting and regulation of cell wall synthesis during tip growth in plants. J Integr Plant Biol 55:835–846. https://doi.org/10.1111/jipb.12077
Hafidh S, Honys D (2021) Reproduction multitasking: the male gametophyte. Ann Rev Plant Biol 72:581–614. https://doi.org/10.1146/annurev-arplant-080620-021907
Honys D and Twell D (2004) Transcriptome analysis of halpoid male gametophyte development in Arabidopsis. Genome Biol 5:R85. http://genomebiology.com/2004/5/11/R85
Huang J, Dong J, Qu L-J (2021) From birth to function: male gametophyte development in flowering plants. Curr Opin Plant Biol 63:102118. https://doi.org/10.1016/j.pbi.2021.102118
Ishikawa T, Machida C, Yoshioka Y, Ueda T, Nakano A, Machida Y (2008) EMBRYO YELLOW gene, encoding a subunit of the conserved oligomeric Golgi complex, is required for appropriate cell expansion and meristem organization in Arabidopsis thaliana. Genes Cells 13:521–535. https://doi.org/10.1111/j.1365-2443.2008.01186.x
Johnson MA, Harper JF, Palanivelu R (2019) A fruitful journey: pollen tube navigation from germination to fertilization. Annu Rev Plant Biol 70:809–837. https://doi.org/10.1146/annurev-arplant-050718-100133
Kim HJ, Oh SA, Brownfield L, Hong SH, Ryu H, Hwang I, Twell D, Nam HG (2008) Control of plant germline proliferation by SCFFBL17 degradation of cell cycle inhibitors. Nature 455:1134–1137. https://doi.org/10.1038/nature07289
Klodova B, Potesil D, Steinbachova L, Michailidis C, Lindner A-C, Hackenberg D, Becker JD, Zdrahal Z, Twell D, Honys D (2023) Regulatory dynamics of gene expression in the developing male gametophyte of Arabidopsis. Plant Reprod 36:213–241. https://doi.org/10.1007/s00497-022-00452-5
Latijnhouwers M, Hawes C, Carvalho C (2005) Holding it all together? Candidate proteins for the plant Golgi matrix. Curr Opin Plant Biol 8:632–639. https://doi.org/10.1016/j.pbi.2005.09.014
Liu YG, Mitsukawa N, Oosumi T, Whittier RF (1995) Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J 8:457–463. https://doi.org/10.1046/j.1365-313x.1995.08030457.x
Loraine AE, McCormick S, Estrada A, Patel K, Qin P (2013) RNA-seq of Arabidopsis pollen uncovers novel transcription and alternative splicing. Plant Physiol 162:1092–1109. https://doi.org/10.1104/pp/112/211441
McCormick S (1993) Male gametophyte development. Plant Cell 5:1265–1275. https://doi.org/10.1105/tpc.5.10.1265
Miller VJ, Ungar D (2012) Re’COG’nition at the Golgi. Traffic 13:891–897. https://doi.org/10.1111/j.1600-0854.2012.01338.x
Nebenführ A, Gallagher LA, Dunahay TG, Frohlick JA, Mazurkiewicz AM, Meehl JB, Staehelin LA (1999) Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system. Plant Physiol 121:1127–1142. https://doi.org/10.1104/pp.121.4.1127
Nowack MK, Grini PE, Jakoby MJ, Lafos M, Koncz C, Schnittger A (2006) A positive signal from the fertilization of the egg cell sets off endosperm proliferation in angiosperm embyogenesis. Nat Genet 38:63–67. https://doi.org/10.1038/ng1694
Oh SA, Allen T, Twell D (2010) A ticket for the live show: microtubules in male gametophyte development. Plant Signal Behav 5:614–617. https://doi.org/10.4161/psb.11505
Oh SA, Park KS, Twell D, Park SK (2010) The SIDECAR POLLEN gene encodes a microspore-specific LOB/AS2 domain protein required for the correct timing and orientation of asymmetric cell division. Plant J 64:839–850. https://doi.org/10.1111/j.1365-313x.2010.04374.x
Oh SA, Jeon J, Park HJ, Grini PE, Twell D, Park SK (2016) Analysis of gemini pollen 3 mutant suggests a broad function of AUGMIN in microtubule organization during sexual reproduction in Arabidopsis. Plant J 87:188–201. https://doi.org/10.1111/tpj.13192
Pantazopoulou A (2017) The Golgi apparatus: insights from filamentous fungi. Mycologia 108:603–622. https://doi.org/10.3852/15-309
Rotman N, Durbarry A, Wardle A, Yang WC, Chaboud A, Faure J-E, Berger F, Twell D (2005) A novel class of MYB factors controls sperm-cell formation in plants. Curr Biol 15:244–248. https://doi.org/10.1016/j.cub.2005.01.013
Rui Q, Wang J, Li Y, Tan X, Bao Y (2020) Arabidopsis COG6 is essential for pollen tube growth and Golgi structure maintenance. Biochem Biophys Res Commun 528:447–452. https://doi.org/10.1016/j.bbrc.2020.05.189
Schlag M, Hesse M (1992) The formation of the generative cell in Polystachia pubescens (Orchidaceae). Sex Plant Reprod 5:131–137. https://doi.org/10.1007/BF00194872
Scott RJ, Spielman M, Dickinson HG (2004) Stamen structure and function. Plant Cell 16(Suppl):S46–S60. https://doi.org/10.1105/tpc.017012
Steffen JG, Kang IH, Macfarlane J, Drews GN (2007) Identification of genes expressed in the Arabidopsis female gametophyte. Plant J 51:281–292. https://doi.org/10.1111/j.1365-313X.2007.03137.x
Tan X, Cao K, Liu F, Li Y, Li P, Gao C, Ding Y, Lan Z, Shi Z, Rui Q, Feng Y, Liu Y, Zhao Y, Wu C, Zhang Q, Li Y, Jiang L, Bao Y (2016) Arabidopsis COG complex subunits COG3 and COG8 modulate Golgi morphology, vesicle trafficking homeostasis and are essential for pollen tube growth. PLoS Genet 12:e1006140. https://doi.org/10.1371/journal.pgen.1006140
Torutaeva E, Oh S-A, Park SK (2020) Identification of new mutant allelels of Augmin subunits broadens spectrum of augmin function during sexual reproduction in Arabidopsis. J Plant Biol 63:485–494. https://doi.org/10.1007/s12374-020-09276-0
Twell D, Park SK, Lalanne E (1998) Asymmetric division and cell-fate determination in developing pollen. Trends Plant Sci 3:305–310. https://doi.org/10.1016/S1360-1385(98)01277-1
Twell D, Oh SA, Honys D (2006) Pollen development, a genetic and transcriptomic view. Plant Cell Monographs 3:15–45. https://doi.org/10.1007/7089_042
Ungar D, Oka T, Krieger M, Hughson FM (2006) Retrograde transport on the COG railway. Trends Cell Biol 16:113–120. https://doi.org/10.1016/j.tcb.2005.12.004
Vukašinović N, Žarský V (2016) Tethering complexes in the Arabidopsis endomembrane system. Front Cell Dev Biol 4:46. https://doi.org/10.3389/fcell.2016.00046
Weigel D, Ahn JH, Blázquez MA, Borevitz JO, Christensen SK, Fankhauser C, Ferrándiz C, Kardailsky I, Malancharuvil EJ, Nef MM, Nguyen JT (2000) Activation tagging in Arabidopsis. Plant Physiol 122:1003–1014. https://doi.org/10.1104/pp.122.4.1003
Willett R, Ungar D, Lupashin V (2013) The Golgi puppet master: COG complex at center stage of membrane trafficking interactions. Histochem Cell Biol 140:271–283. https://doi.org/10.1007/s00418-013-1117-6
Zuo J, Niu QW, Chua NH (2000) Technical advance: An estrogen receptor-based transactivator XVE mediates highly inducible gene expression in transgenic plants. Plant J 24:265–273. https://doi.org/10.1046/j.1365-313x.2000.00868.x
Acknowledgements
This research was supported by Kyungpook National University Research Fund, 2022. The authors thank Binbin Li, Anchilie Francis-Mangilet, and Jien Jeon for early stage work involved in mutant isolation and vector construction.
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SKP and SAO conceived the research; SKP supervised the experiments; TDN, SAO, MHK, SJ performed the experiments and analyzed the data; SKP, SAO, TDN wrote the manuscript with contributions from MHK and SJ.
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Oh, S.A., Nguyen, T.D., Kim, MH. et al. atcog8-2, A New Mutant Allele of the Conserved Oligomeric Golgi Complex 8, Reveals the Need for the COG Complex for Gametophyte Development in Arabidopsis. J. Plant Biol. 67, 109–121 (2024). https://doi.org/10.1007/s12374-023-09414-4
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DOI: https://doi.org/10.1007/s12374-023-09414-4