Synthesis of a Wheat/Maize Hybrid CENH3 Gene, the Genetic Transformation of Wheat, Its Chromosomal Localization and Effects on Chromosome Behaviors in Wheat/Maize Somatic Hybrids — Oak Academic Publishing
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Synthesis of a Wheat/Maize Hybrid CENH3 Gene, the Genetic Transformation of Wheat, Its Chromosomal Localization and Effects on Chromosome Behaviors in Wheat/Maize Somatic Hybrids
Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, China
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School of Life Sciences, State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Hong Kong, China
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Guangdong Provincial Key Laboratory for Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen University, Shen-zhen, China
1 Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, China
2 School of Life Sciences, State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Hong Kong, China
3 Guangdong Provincial Key Laboratory for Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen University, Shen-zhen, China
Centromere-specific histone H3 (CENH3) replaces the canonical histone H3 in nucleosomes of functional centromeres, and plays important roles in faithful chromosome segregation during cell division. CENH3 is also important in the recognition of alien centromeres and determines the accommodation or elimination of alien chromosomes in interspecific or intergenic hybridization. In this study, a maize full length CENH3 with a yellow fluorescent protein ( YFP ) tag at C-terminus ( Zm CENH3- YFP ) and a synthetic hybrid wmCENH3 with the N-terminus from wheat CENH3 and the histone fold domain (HFD) from maize tagged with a red fluorescent protein ( RFP ) at the C-terminus (wmCENH3- RFP ) were transformed to wheat by biolistics transformation. Transgenic wheat plants with both Zm CNEH3- YFP and wmCENH3- RFP genes were identified by PCR. The expression of ZmCENH3-YFP was not observed, while the expression of wmCENH3-RFP could be detected by RT-PCR, direct fluorescence microscopy, and immunostaining with anti-RFP antibody. The expressed wmCENH3-RFP was localized to nuclei as dotted patterns, indicating its targeting to wheat centromeres. Somatic hybridization was performed between wmCENH3- RFP transgenic wheat and transgenic maize that expressed a Zm CENH3- YFP gene to investigate chromosome behaviors in somatic hybrids. Cytological and FISH analyses of somatic hybrid cells showed the formation of micronuclei and lagging chromatin in both somatic hybridizations with or without the wmCENH3- RFP transgene, indicating that ectopically expressed wmCENH3 could not overcome chromosome elimination in wheat/maize somatic hybrids. Immunostaining of wmCENH3-RFP and ZmCENH3-YFP in early stage somatic hybrid cells indicated that both wmCENH3-RFP and ZmCENH3-YFP proteins were expressed, but their binding patterns changed from the commonly observed dotted patterns to diffused ones, suggesting that the inactivation of CENH3 might be a factor for chromosome elimination in wheat/maize somatic hybridization.
Bloom, K. (2007) Centromere Dynamics. Current Opinion in Genetics & Development, 17, 151-156. https://doi.org/10.1016/j.gde.2007.02.009
Ravi, M. and Chan, S.W.L. (2010) Haploid Plants Produced by Centromere-Mediated Genome Elimination. Nature, 464, 615-618. https://doi.org/10.1038/nature08842
Maheshwari, S., Tan, E.H., West, A., Franklin, F.C., Comai, L. and Chan, S.W. (2015) Naturally Occurring Differences in CENH3 Affect Chromosome Segregation in Zygotic Mitosis of Hybrids. PLOS Genetics, 11, e1004970. https://doi.org/10.1371/journal.pgen.1004970
Sanei, M., Pickering, R., Kumke, K., Nasuda, S. and Houben, A. (2011) Loss of Centromeric Histone H3 (CENH3) from Centromeres Precedes Uniparental Chromosome Elimination in Interspecific Barley Hybrids. Proceedings of the National Academy of Sciences, 108, E498-E505. https://doi.org/10.1073/pnas.1103190108
Jin, W., Melo, J.R., Nagaki, K., Talbert, P.B., Henikoff, S., Dawe, R.K. and Jiang, J. (2004) Maize Centromeres: Organization and Functional Adaptation in the Genetic Background of Oat. Plant Cell, 16, 571-581. https://doi.org/10.1105/tpc.018937
Malik, H.S. and Henikoff, S. (2002) Conflict Begets Complexity: The Evolution of Centromeres. Current Opinion in Genetics & Development, 12, 711-718. https://doi.org/10.1016/S0959-437X(02)00351-9
Talbert, P.B., Masuelli, R., Tyagi, A.P., Comai, L. and Henikoff, S. (2002) Centromeric Localization and Adaptive Evolution of an Arabidopsis Histone H3 Variant. Plant Cell, 14, 1053-1066. https://doi.org/10.1105/tpc.010425
Rieseberg, L.H. and Carney, S.E. (1998) Plant Hybridization. New Phytologist, 140, 599-624. https://doi.org/10.1046/j.1469-8137.1998.00315.x
Brock, M. (2009) Prezygotic Barriers to Gene Flow between Taraxacum ceratophorum and the Invasive Taraxacum officinale (Asteraceae). Oecologia, 161, 241-251. https://doi.org/10.1007/s00442-009-1383-0
Schiestl, F.P. and Schlüter, P.M. (2009) Floral Isolation, Specialized Pollination, and Pollinator Behavior in Orchids. Annual Review of Entomology, 54, 425-446. https://doi.org/10.1146/annurev.ento.54.110807.090603
Swanson, R., Edlund, A.F. and Preuss, D. (2004) Species Specificity in Pollen-Pistil Interactions. Annual Review of Genetics, 38, 793-818. https://doi.org/10.1146/annurev.genet.38.072902.092356
Kranz, E. and Scholten, S. (2008) In Vitro Fertilization: Analysis of Early Post-Fertilization Development Using Cytological and Molecular Techniques. Sexual Plant Reproduction, 21, 67-77. https://doi.org/10.1007/s00497-007-0060-x
Stebbins, G.L. and Demerec, M. (1958) The Inviability, Weakness, and Sterility of Interspecific Hybrids. Advances in Genetics, 9, 147-215. https://doi.org/10.1016/S0065-2660(08)60162-5
Wu, C.I. (1992) A Note on Haldane’s Rule: Hybrid Inviability versus Hybrid Sterility. Evolution, 46, 1584-1587. https://doi.org/10.1111/j.1558-5646.1992.tb01152.x
Johansen-Morris, A.D. and Latta, R.G. (2006) Fitness Consequences of Hybridization between Ecotypes of Avena barbata: Hybrid Breakdown, Hybrid Vigor, and Transgressive Segregation. Evolution, 60, 1585-1595. https://doi.org/10.1111/j.0014-3820.2006.tb00503.x
Bomblies, K. and Weigel, D. (2007) Hybrid Necrosis: Autoimmunity as a Potential Gene-Flow Barrier in Plant Species. Nature Reviews Genetics, 8, 382-393. https://doi.org/10.1038/nrg2082
Gernand, D., Rutten, T., Varshney, A., Rubtsova, M., Prodanovic, S., Bruss, C., Kumlehn, J., Matzk, F. and Houben, A. (2005) Uniparental Chromosome Elimination at Mitosis and Interphase in Wheat and Pearl Millet Crosses Involves Micronucleus Formation, Progressive Heterochromatinization, and DNA Fragmentation. Plant Cell, 17, 2431-2438. https://doi.org/10.1105/tpc.105.034249
Gernand, D., Rutten, T., Pickering, R. and Houben, A. (2006) Elimination of Chromosomes in Hordeum vulgare × H. bulbosum Crosses at Mitosis and Interphase Involves Micronucleus Formation and Progressive Heterochromatinization. Cytogenetic and Genome Research, 114, 169-174. https://doi.org/10.1159/000093334
Baum, M., Lagudah, E.S. and Appels, R. (1992) Wide Crosses in Cereals. Annual Review of Plant Physiology and Plant Molecular Biology, 43, 117-143. https://doi.org/10.1146/annurev.pp.43.060192.001001
Sharma, D.R., Kaur, R. and Kumar, K. (1996) Embryo Rescue in Plants—A Review. Euphytica, 89, 325-337.
Barclay, I.R. (1975) High Frequencies of Haploid Production in Wheat (Triticum aestivum) by Chromosome Elimination. Nature, 256, 410-411. https://doi.org/10.1038/256410a0
Laurie, D.A. and Bennett, M.D. (1986) Wheat × Maize Hybridization. Canadian Journal of Genetics and Cytology, 28, 313-316. https://doi.org/10.1139/g86-046
Laurie, D.A. and Bennett, M.D. (1988) The Production of Haploid Wheat Plants from Wheat × Maize Crosses. Theoretical and Applied Genetics, 76, 393-397. https://doi.org/10.1007/BF00265339
Laurie, D.A. and Bennett, M.D. (1989) The Timing of Chromosome Elimination in Hexaploid Wheat × Maize Crosses. Genome, 32, 953-961. https://doi.org/10.1139/g89-537
Xu, C., Xia, G., Zhi, D., Xiang, F. and Chen, H. (2003) Integration of Maize Nuclear and Mitochondrial DNA into the Wheat Genome through Somatic Hybridization. Plant Science, 165, 1001-1008. https://doi.org/10.1016/S0168-9452(03)00287-5
Mochida, K., Tsujimoto, H. and Sasakuma, T. (2004) Confocal Analysis of Chromosome Behavior in Wheat × Maize Zygotes. Genome, 47, 199-205. https://doi.org/10.1139/g03-123
Chen, W., Zhu, Q., Wang, H., Xiao, J., Xing, L., Chen, P., Jin, W. and Wang, X.E. (2015) Competitive Expression of Endogenous Wheat CENH3 May Lead to Suppression of Alien ZmCENH3 in Transgenic Wheat × Maize Hybrids. Journal of Genetics and Genomic, 42, 639-649. https://doi.org/10.1016/j.jgg.2015.05.006
Christensen, A.H. and Quail, P.H. (1996) Ubiquitin Promoter-Based Vectors for High-Level Expression of Selectable and/or Screenable Marker Genes in Monocotyledonous Plants. Transgenic Research, 5, 213-218. https://doi.org/10.1007/BF01969712
Redway, F.A., Vasil, V., Lu, D. and Vasil, I.K. (1990) Identification of Callus Types for Long-Term Maintenance and Regeneration from Commercial Cultivars of Wheat (Triticum aestivum L.). Theoretical and Applied Genetics, 79, 609-617. https://doi.org/10.1007/BF00226873
Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. (1987) GUS Fusions: Beta-Glucuronidase as a Sensitive and Versatile Gene Fusion Marker in Higher Plants. The EMBO Journal, 6, 3901. https://doi.org/10.1002/j.1460-2075.1987.tb02730.x
Bimboim, H.C. and Doly, J. (1979) A Rapid Alkaline Extraction Procedure for Screening Recombinant Plasmid DNA. Nucleic Acids Research, 7, 1513-1523. https://doi.org/10.1093/nar/7.6.1513
Altpeter, F., Vasil, V., Srivastava, V., Stöger, E. and Vasil, I.K. (1996) Accelerated Production of Transgenic Wheat (Triticum aestivum L.) Plants. Plant Cell Reports, 16, 12-17. https://doi.org/10.1007/BF01275440
Vasil, I.K. and Vasil, V. (2006) Transformation of Wheat via Particle Bombardment. Methods in Molecular Biology, 318, 273. https://doi.org/10.1385/1-59259-959-1:273
Edwards, K., Johnstone, C. and Thompson, C. (1991) A Simple and Rapid Method for the Preparation of Plant Genomic DNA for PCR Analysis. Nucleic Acids Research, 19, 1349. https://doi.org/10.1093/nar/19.6.1349
Kanai, R. and Edwards, G.E. (1973) Separation of Mesophyll Protoplasts and Bundle Sheath Cells from Maize Leaves for Photosynthetic Studies. Plant Physiology, 51, 1133-1137. https://doi.org/10.1104/pp.51.6.1133
Xia, G. and Chen, H. (1996) Plant Regeneration from Intergeneric Somatic Hybridization between Triticum aestivum L. and Leymus chinensis (Trin.) Tzvel. Plant Science, 120, 197-203. https://doi.org/10.1016/S0168-9452(96)04492-5
Grosser, J., Calović, M. and Louzada, E. (2010) Protoplast Fusion Technology-Somatic Hybridization and Cybridization. In: Plant Cell Culture, John Wiley & Sons, Hoboken, 175-198. https://doi.org/10.1002/9780470686522.ch10
Ross, K., Fransz, P. and Jones, G. (1996) A Light Microscopic Atlas of Meiosis in Arabidopsis thaliana. Chromosome Research, 4, 507-516. https://doi.org/10.1007/BF02261778
Storey, W.B. and Mann, J.D. (1967) Chromosome Contraction by O-Isopropyl-N-Phenylcarbamate (IPC). Biotechnic & Histochemistry, 42, 15-18. https://doi.org/10.3109/10520296709114976
Alfenito, M.R. and Birchler, J.A. (1993) Molecular Characterization of a Maize B Chromosome Centric Sequence. Genetics, 135, 589-597.
Ananiev, E.V., Phillips, R.L. and Rines, H.W. (1998) Chromosome-Specific Molecular Organization of Maize (Zea mays L.) Centromeric Regions. Proceedings of the National Academy of Sciences, 95, 13073-13078. https://doi.org/10.1073/pnas.95.22.13073
Wiegant, J., Verwoerd, N., Mascheretti, S., Bolk, M., Tanke, H.J. and Raap, A.K. (1996) An Evaluation of a New Series of Fluorescent dUTPs for Fluorescence in Situ Hybridization. Journal of Histochemistry & Cytochemistry, 44, 525-529. https://doi.org/10.1177/44.5.8627009
Kato, A., Lamb, J.C. and Birchler, J.A. (2004) Chromosome Painting Using Repetitive DNA Sequences as Probes for Somatic Chromosome Identification in Maize. Proceedings of the National Academy of Sciences, 101, 13554-13559. https://doi.org/10.1073/pnas.0403659101
Kato, A. (1999) Air Drying Method Using Nitrous Oxide for Chromosome Counting in Maize. Biotechnic & Histochemistry, 74, 160-166. https://doi.org/10.3109/10520299909047968
Cui, H., Sun, Y., Deng, J., Wang, M. and Xia, G. (2015) Chromosome Elimination and Introgression Following Somatic Hybridization between Bread Wheat and Other Grass Species. Plant Cell, Tissue and Organ Culture, 120, 203-210. https://doi.org/10.1007/s11240-014-0594-1
Kohli, A., Leech, M., Vain, P., Laurie, D.A. and Christou, P. (1998) Transgene Organization in Rice Engineered through Direct DNA Transfer Supports a Two-Phase Integration Mechanism Mediated by the Establishment of Integration Hot Spots. Proceedings of the National Academy of Sciences, 95, 7203-7208. https://doi.org/10.1073/pnas.95.12.7203
Pawlowski, W.P. and Somers, D.A. (1998) Transgenic DNA Integrated into the Oat Genome Is Frequently Interspersed by Host DNA. Proceedings of the National Academy of Sciences, 95, 12106-12110. https://doi.org/10.1073/pnas.95.21.12106
Waterworth, W.M., Drury, G.E., Bray, C.M. and West, C.E. (2011) Repairing Breaks in the Plant Genome: The Importance of Keeping It Together. New Phytologist, 192, 805-822. https://doi.org/10.1111/j.1469-8137.2011.03926.x
Svitashev, S., Ananiev, E., Pawlowski, W.P. and Somers, D.A. (2000) Association of Transgene Integration Sites with Chromosome Rearrangements in Hexaploid Oat. Theoretical and Applied Genetics, 100, 872-880. https://doi.org/10.1007/s001220051364
Moraes, I., Lermontova, I. and Schubert, I. (2011) Recognition of A. thaliana Centromeres by Heterologous CENH3 Requires High Similarity to the Endogenous Protein. Plant Molecular Biology, 75, 253-261. https://doi.org/10.1007/s11103-010-9723-3
Maheshwari, S., Ishii, T., Brown, C.T., Houben, A. and Comai, L. (2017) Centromere Location in Arabidopsis Is Unaltered by Extreme Divergence in CENH3 Protein Sequence. Genome Research, 27, 471-478. https://doi.org/10.1101/gr.214619.116
Heckmann, S., Macas, J., Kumke, K., Fuchs, J., Schubert, V., Ma, L., Novak, P., Neumann, P., Taudien, S., Platzer, M. and Houben, A. (2012) The Holocentric Species Luzula elegans Shows Interplay between Centromere and Large-Scale Genome Organization. The Plant Journal, 73, 555-565. https://doi.org/10.1111/tpj.12054
Nagaki, K., Terada, K., Wakimoto, M., Kashihara, K. and Murata, M. (2010) Centromere Targeting of Alien CENH3s in Arabidopsis and Tobacco Cells. Chromosome Research, 18, 203-211. https://doi.org/10.1007/s10577-009-9108-0