Expression of apomixis genes during reproductive development in Urochloa decumbens and Urochloa ruziziensis (Poaceae)
Abstract
This study aimed to correlate the developmental stages of the embryo sac with the morphometric traits of spikelets and to characterize the expression of the BbrizRan, BbrizSti1, and BbrizAGL6 genes in Urochloa ruziziensis (sexual) and U. decumbens (apomictic). Data on the length and diameter of spikelets and anatomical sections were also obtained. Gene expression was determined using RT-qPCR in spikelets at megasporogenesis and megagametogenesis. The spikelet length could be used as a reliable morphological marker to discriminate between the two stages in both species. The embryo sacs observed were of the Polygonum type in U. ruziziensis and the Panicum type in U. decumbens. RT-qPCR analysis revealed that the BbrizRan, BbrizSti1, and BbrizAGL6 genes were differentially expressed in the studied species during reproductive development. The BbrizSti1 gene was more highly expressed in U. decumbens (apomictic) than in U. ruziziensis (sexual) during megasporogenesis, while for BbrizRan, U. decumbens showed higher expression than U. ruziziensis during both megasporogenesis and megagametogenesis. An increase in the expression level of BbrizAGL6 was observed in both species at both developmental stages of the embryo sac.
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Albertini, E., Marconi, G., Barcaccia, G., Raggi, L., & Falcinelli, M. (2004). Isolation of candidate genes for apomixis in Poa pratensis L. Plant Molecular Biology, 56, 879-894. https://doi.org/10.1007/s11103-004-5211-y
Alves, E. R., Carneiro, V. T. C., & Dusi, D. M. A. (2007). In situ localization of three cDNA sequences associated with the later stages of aposporic embryo sac development of Brachiaria brizantha. Protoplasma, 231, 161-171. https://doi.org/10.1007/s00709-007-0253-z
Araújo, A. C. G., Mukhambetzhanov, S., Pozzobon, M. T., Santana, E. F., & Carneiro, V. T. C. (2000). Female gametophyte development in apomictic and sexual Brachiaria brizantha (Poaceae). Revue de Cytologie et Biologie Végetales-Le Botaniste, 23, 13-26.
Araújo, A. C. G., Falcão, R., Simões, K. C. R., & Carneiro, V. T. C. (2004). Identificação de acessos de Brachiaria com interesse ao estudo da apomixia facultativa. Embrapa Recursos Genéticos e Biotecnologia. (Boletim de Pesquisa e Desenvolvimento, 74). https://ainfo.cnptia.embrapa.br/digital/bitstream/CENARGEN/28285/1/bp074.pdf
Chiari, L., Rocha, M., Valle, C. B., & Salgado, L. R. (2008). Variabilidade genética em acessos e cultivares de quatro espécies de Brachiaria estimada por marcadores RAPD. Embrapa Gado de Corte. (Boletim de Pesquisa e Desenvolvimento).
Cruz, R. P., Federizzi, L. C., & Milach, S. C. K. (1998). A apomixia no melhoramento de plantas. Ciência Rural, 28(1), 155-161. https://doi.org/10.1590/S0103-84781998000100028
Boor, S., Knyphausen, P., Kuhlmann, N., Wroblowski, S., Brenig, J., Scislowski, L., Baldus, L., Nolte, H., Krüger, M., & Lammers, M. (2015). Small GTP-binding protein Ran is regulated by posttranslational lysine acetylation. Proceedings of the National Academy of Sciences, 112(28), 3679-3688. https://doi.org/10.1073/pnas.1505995112
Dusi, D. M. A., & Willemse, M. T. M. (1999). Apomixis in Brachiaria decumbens Stapf.: Gametophytic development and reproductive calendar. Acta Biologica Cracoviensia, 41, 151-162.
Fei, X., Shi, J., Liu, Y., Niu, J., & Wei, A. (2019). The steps from sexual reproduction to apomixis. Planta, 249, 1715-1730. https://doi.org/10.1007/s00425-019-03113-6
Ferreira, L. G., Dusi, D. M. A., Irsigler, A. S. T., Gomes, A. C. M. M., Mendes, M. A., Colombo, L., & Carneiro, V. T. C. (2018). GID1 expression is associated with ovule development of sexual and apomictic plants. Plant Cell Reports, 37, 293-306. https://doi.org/10.1007/s00299-017-2230-0
Ferreira, R. C. U., Moraes, A. C. L., Chiari, L., Simeão, R. M., Vigna, B. B. Z., & Souza, A. P. (2021). An overview of the genetics and genomics of the Urochloa species most commonly used in pastures. Frontiers in Plant Science, 12(770461), 1-22. https://doi.org/10.3389/fpls.2021.770461
Gobbe, J., Longly, B., & Louant, B.-P. (1982). Calendrier des sporogénèses et gamétogénèses femelles chez le diploïde et le tétraploïde induit de Brachiaria ruziziensis (Graminée). Canadian Journal of Botany, 60(10), 2032-2036. https://doi.org/10.1139/b82-250
Gounaris, E. K., Sherwood, R. T., Gounaris, I., Hamilton, R. H., & Gustine, D. L. (1991). Inorganic salts modify embryo sac development in sexual and aposporous Cenchrus ciliaris. Sexual Plant Reproduction, 4(3), 188-192.
Guimarães, L. A., Dusi, D. M. A., Masiero, S., Resentini, F., Gomes, A. C. M. M., Silveira, É. D., Florentino, L. H., Rodrigues, J. C. M., Colombo, L. & Carneiro, V. T. C. (2013). BbrizAGL6 is differentially expressed during embryo sac formation of apomictic and sexual Brachiaria brizantha plants. Plant Molecular Biology Reporter, 31(6), 1397-1406. https://doi.org/10.1007/s11105-013-0618-8
Kausar, S., Abbas, M. N., Yang, L., & Cui, H. (2020). Biotic and abiotic stress induces the expression of Hsp70/90 organizing protein gene in silkworm, Bombyx mori. International Journal of Biological Macromolecules, 143, 610-618. https://doi.org/10.1016/j.ijbiomac.2019.12.067
Koehler, A. D., Irsigler, A. S. T., Carneiro, V. T. C., Cabral, G. B., Rodrigues, J. C. M., Gomes, A. C. M. M., Togawa, R. C., Costa, M. M. C., Martinelli, A. P., & Dusi, D. M. A. (2020). SERK genes identification and expression analysis during somatic embryogenesis and sporogenesis of sexual and apomictic Brachiaria brizantha (Syn. Urochloa brizantha). Planta, 252(3), 39. https://doi.org/10.1007/s00425-020-03443-w
Hanna, W. W., & Bashaw, E. C. (1987). Apomixis: Its identification and use in plant breeding. Crop Science, 27(6), 1136-1139. https://doi.org/10.2135/cropsci1987.0011183X002700060010x
Ishigaki, G., Gondo, T., Suenaga, K., & Akashi, R. (2009). Induction of tetraploid ruzigrass (Brachiaria ruziziensis) plants by colchicine treatment of in vitro multiple‐shoot clumps and seedlings. Grassland Science, 55(3), 164-170. https://doi.org/10.1111/j.1744-697X.2009.00153.x
Jank, L., Barrios, S. C., Valle, C. B., Simeão, R. M., & Alves, G. F. (2014). The value of improved pastures to Brazilian beef production. Crop and Pasture Science, 65(11), 1132-1137. https://doi.org/10.1071/CP13319
Lacerda, A. L. M., Dusi, D. M. A., Alves, E. R., Rodrigues, J. C. M., Gomes, A. C. M. M., & Carneiro, V. T. C. (2013). Expression analyses of Brachiaria brizantha genes encoding ribosomal proteins BbrizRPS8, BbrizRPS15a, and BbrizRPL41 during development of ovaries and anthers. Protoplasma, 250(2), 505-514. https://doi.org/10.1007/s00709-012-0433-3
Lamm, C. E., Kraner, M. E., Hofmann, J., Börnke, F., Mock, H.-P., & Sonnewald, U. (2017). Hop/Sti1 a two-faced cochaperone involved in pattern recognition receptor maturation and viral infection. Frontiers in Plant Science, 8(1754), 1-17. https://doi.org/10.3389/fpls.2017.01754
Lituiev, D. S., & Grossniklaus, U. (2014). Patterning of the angiosperm female gametophyte through the prism of theoretical paradigms. Biochemical Society Transactions, 42(2), 332-339. https://doi.org/10.1042/BST20140036
Lutts, S., Ndikumana, J., & Louant, B. P. (1991). Fertility of Brachiaria ruziziensis in interspecific crosses with Brachiaria decumbens and Brachiaria brizantha: meiotic behaviour, pollen viability and seed set. Euphytica, 57, 267-274. https://doi.org/10.1007/BF00039673
Nakamura, A. T., Longhi-Wagner, H. M., & Scatena, V. L. (2009). Desenvolvimento de óvulo, fruto e semente de espécies de Poaceae (Poales). Brazilian Journal of Botany, 32(1), 165-176. https://doi.org/10.1590/S0100-84042009000100016
Naumova, T. N., Hayward, M. D., & Wagenvoort, M. (1999). Apomixis and sexuality in diploid and tetraploid accessions of Brachiaria decumbens. Sexual Plant Reproduction, 12, 43-52. https://doi.org/10.1007/s004970050170
Nicolet, C. M., & Craig, E. A. (1989). Isolation and characterization of STI1, a stress-inducible gene from Saccharomyces cerevisiae. Molecular and Cellular Biology, 9(9), 3638-3646. https://doi.org/10.1128/mcb.9.9.3638
O'Brien, T. P., Feder, N., & McCully, M. E. (1964). Polychromatic staining of plant cell walls by toluidine blue O. Protoplasma, 59, 368-373. https://doi.org/10.1007/BF01248568
Paula, C. M. P., Souza Sobrinho, F., & Techio, V. H. (2017). Genomic constitution and relationship in Urochloa (Poaceae) species and hybrids. Crop Science, 57(5), 2605-2616. https://doi.org/10.2135/cropsci2017.05.0307
Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Research, 29(9), 2003-2007. https://doi.org/10.1093/nar/29.9.e45
Podio, M., Felitti, S. A., Siena, L. A., Delgado, L., Mancini, M., Seijo, J. G., González, A. M., Pessino, S. C., & Ortiz, J. P. A. (2014). Characterization and expression analysis of SOMATIC EMBRYOGENESIS RECEPTOR KINASE (SERK) genes in sexual and apomictic Paspalum notatum. Plant Molecular Biology, 84, 479-495. https://doi.org/10.1007/s11103-013-0146-9
Reinheimer, R., & Kellogg, E. A. (2009). Evolution of AGL6-like MADS box genes in grasses (Poaceae): ovule expression is ancient and palea expression is new. The Plant Cell, 21(9), 2591-2605. https://doi.org/10.1105/tpc.109.068239
Renvoize, S. A., Clayton, W. D., & Kabuye, C. H. S. (1996). Morphology, taxonomy, and natural distribution of Brachiaria (Trin.) Griseb. In J. W. Miles, B. L. Maass, C. B. Valle, & V. Kumble (Eds.), Brachiaria: Biology, agronomy, and improvement (pp. 1-15). CIAT.
R Core Team (2020). R: A language and environment for statistical computing. R Foundation for Computing. https://www.R-project.org
Rodrigo, J. M., Zappacosta, D. C., Selva, J. P., Garbus, I., Albertini, E., & Echenique, V. (2017). Apomixis frequency under stress conditions in weeping lovegrass (Eragrostis curvula). PLoS ONE, 12(4), 1-17. https://doi.org/10.1371/journal.pone.0175852
Rodrigues, J. C., Cabral, G. B., Dusi, D. M. A., Mello, L. V., Rigden, D. J., & Carneiro, V. T. C. (2003). Identification of differentially expressed cDNA sequences in ovaries of sexual and apomictic plants of Brachiaria brizantha. Plant Molecular Biology, 53, 745-757. https://doi.org/10.1023/B:PLAN.0000023664.21910.bd
Röhl, A., Wengler, D., Madl, T., Lagleder, S., Tippel, F., Herrmann, M., Hendrix, J., Richter, K., Hack, G., Schmid, A. B., Kessler, H., Lamb, D. C., & Buchner, J. (2015). Hsp90 regulates the dynamics of its cochaperone Sti1 and the transfer of Hsp70 between modules. Nature Communications, 6, 6655. https://doi.org/10.1038/ncomms7655
Sailer, C., Schmid, B., & Grossniklaus, U. (2016). Apomixis allows the transgenerational fixation of phenotypes in hybrid plants. Current Biology, 26(3), 331-337. https://doi.org/10.1016/j.cub.2015.12.045
Savidan, Y. (2010). Apomixis: genetics and breeding. In Plant breeding reviews (v. 18, pp. 13-86). Wiley-Blackwell. https://doi.org/10.1002/9780470650158.ch2
Schindelin, J., Rueden, C. T., Hiner, M. C., & Eliceiri, K. W. (2015). The ImageJ ecosystem: an open platform for biomedical image analysis. Molecular Reproduction and Development, 82(7-8), 518-529. https://doi.org/10.1002/mrd.22489
Schmidt, A. (2020). Controlling apomixis: shared features and distinct characteristics of gene regulation. Genes, 11(3), 1-29. https://doi.org/10.3390/genes11030329
Silveira, É. D., Alves-Ferreira, M., Guimarães, L. A., Silva, F. R., & Carneiro, V. T. C. (2009). Selection of reference genes for quantitative real-time PCR expression studies in the apomictic and sexual grass Brachiaria brizantha. BMC Plant Biology, 9(84), 1-10. https://doi.org/10.1186/1471-2229-9-84
Silveira, É. D., Guimarães, L. A., Dusi, D. M. A., Silva, F. R., Martins, N. F., Costa, M. M. C., Alves-Ferreira, M., & Carneiro, V. T. C. (2012). Expressed sequence-tag analysis of ovaries of Brachiaria brizantha reveals genes associated with the early steps of embryo sac differentiation of apomictic plants. Plant Cell Reports, 31(2), 403-416. https://doi.org/10.1007/s00299-011-1175-y
Singh, M., Burson, B. L., & Finlayson, S. A. (2007). Isolation of candidate genes for apomictic development in buffelgrass (Pennisetum ciliare). Plant Molecular Biology, 64(6), 673-682. https://doi.org/10.1007/s11103-007-9188-1
Takamori, L. M., Pereira, A. V. C., Souza, G. M., Vieira, L. G. E., & Ribas, A. F. (2017). Identification of endogenous reference genes for RT-qPCR expression analysis in Urochloa brizantha under abiotic stresses. Scientific Reports, 7(8502), 1-11. https://doi.org/10.1038/s41598-017-09156-7
Teo, Z. W. N., Zhou, W., & Shen, L. (2019). Dissecting the function of MADS-box transcription factors in orchid reproductive development. Frontiers in Plant Science, 10(1474), 1-17. https://doi.org/10.3389/fpls.2019.01474
Timbó, A. L. O., Souza, P. N. C., Pereira, R. C., Nunes, J. D., Pinto, J. E. B. P., Souza Sobrinho, F., & Davide, L. C. (2014). Obtaining tetraploid plants of ruzigrass (Brachiaria ruziziensis). Revista Brasileira de Zootecnia, 43(3), 127-131. https://doi.org/10.1590/S1516-35982014000300004
Valle, C. B. D., Miles, J. W., Savidan, Y., Carman, J. G., & Dresselhaus, T. (2001). Breeding of apomictic species. In Y. Savidan, J. G. Carman, & T. Dresselhaus (Eds.), The flowering of apomixis: from mechanisms to genetic engineering (pp. 137-152). CIMMYT.
Valle, C. B. D., & Pagliarini, M. S. (2009). Biology, cytogenetics, and breeding of Brachiaria. In R. J. Singh (Ed.), Genetic resources, chromosome engineering, and crop improvement series: Forage crops (pp. 103-151). CRC Press.
Valle, C. B. D., & Savidan, Y. H. (1996). Genetics, cytogenetics, and reproductive biology of Brachiaria. In J. W. Miles, B. L. Maass, & C. B. D. Valle (Eds.), Brachiaria: Biology, agronomy, and improvement (pp. 147-163). CIAT.
Zhang, Z., Quick, M. K., Kanelakis, K. C., Gijzen, M., & Krishna, P. (2003). Characterization of a plant homolog of hop, a cochaperone of Hsp90. Plant Physiology, 131(2), 525-535. https://doi.org/10.1104/pp.011940
Zorzatto, C., Chiari, L., Bitencourt, G. A., Valle, C. B., Leguizamón, G. O. C., Schuster, I., & Pagliarini, M. S. (2010). Identification of a molecular marker linked to apomixis in Brachiaria humidicola (Poaceae). Plant Breeding, 129(6), 734-736. https://doi.org/10.1111/j.1439-0523.2010.01763.x
Worthington, M., Heffelfinger, C., Bernal, D., Quintero, C., Zapata, Y. P., Pérez, J. G., Vega, J., Miles, J., Dellaporta, S., & Tohme, J. (2016). A parthenogenesis gene candidate and evidence for segmental allopolyploidy in apomictic Brachiaria decumbens. Genetics, 203(3), 1117-1132. https://doi.org/10.1534/genetics.116.190314
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