Production of haploid plants that inherit chromosomes from only one parent can greatly accelerate plant breeding1,2,3. Haploids generated from a heterozygous individual and converted to diploid create instant homozygous lines, bypassing generations of inbreeding. Two methods are generally used to produce haploids. First, cultured gametophyte cells may be regenerated into haploid plants4, but many species and genotypes are recalcitrant to this process2,5. Second, haploids can be induced from rare interspecific crosses, in which one parental genome is eliminated after fertilization6,7,8,9,10,11. The molecular basis for genome elimination is not understood, but one theory posits that centromeres from the two parent species interact unequally with the mitotic spindle, causing selective chromosome loss12,13,14. Here we show that haploid Arabidopsis thaliana plants can be easily generated through seeds by manipulating a single centromere protein, the centromere-specific histone CENH3 (called CENP-A in human). When cenh3 null mutants expressing altered CENH3 proteins are crossed to wild type, chromosomes from the mutant are eliminated, producing haploid progeny. Haploids are spontaneously converted into fertile diploids through meiotic non-reduction, allowing their genotype to be perpetuated. Maternal and paternal haploids can be generated through reciprocal crosses. We have also exploited centromere-mediated genome elimination to convert a natural tetraploid Arabidopsis into a diploid, reducing its ploidy to simplify breeding. As CENH3 is universal in eukaryotes, our method may be extended to produce haploids in any plant species.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Dunwell, J. M. Haploids in flowering plants: origins and exploitation. Plant Biotechnol. J. (in the press)
Forster, B. P., Heberle-Bors, E., Kasha, K. J. & Touraev, A. The resurgence of haploids in higher plants. Trends Plant Sci. 12, 368–375 (2007)
Forster, B. P. & Thomas, W. T. B. in Plant Breeding Reviews (ed. Janick, J.) 57–88 (John Wiley & Sons, 2005)
Guha, S. & Maheshwari, S. C. In vitro production of embryos from anthers of Datura . Nature 204, 497 (1964)
Wedzony, M. et al. in Advances in Haploid Production in Higher Plants (eds Touraev, A., Forster, B. P. & Jain, S. M.) 1–33 (Springer, 2009)
Bains, G. S. & Howard, H. W. Haploid plants of Solanum demissum . Nature 166, 795 (1950)
Barclay, I. R. High frequencies of haploid production in wheat (Triticum aestivum) by chromosome elimination. Nature 256, 410–411 (1975)
Burk, L. G., Gerstel, D. U. & Wernsman, E. A. Maternal haploids of Nicotiana tabacum L. from seed. Science 206, 585 (1979)
Clausen, R. E. & Mann, M. C. Inheritance of Nicotiana tabacum. V. The occurrence of haploid plants in interspecific progenies. Proc. Natl Acad. Sci. USA 10, 121–124 (1924)
Hougas, H. W. & Peloquin, S. J. A haploid plant of the potato variety Katahdin. Nature 180, 1209–1210 (1957)
Kasha, K. J. & Kao, K. N. High frequency haploid production in barley (Hordeum vulgare L.). Nature 225, 874–876 (1970)
Bennett, M. D., Finch, R. A. & Barclay, I. R. The time rate and mechanism of chromosome elimination in Hordeum hybrids. Chromosoma 54, 175–200 (1976)
Finch, R. A. Tissue-specific elimination of alternative whole parental genomes in one barley hybrid. Chromosoma 88, 386–393 (1983)
Laurie, D. A. & Bennett, M. D. The timing of chromosome elimination in hexaploid wheat x maize crosses. Genome 32, 953–961 (1989)
Talbert, P. B., Masuelli, R., Tyagi, A. P., Comai, L. & Henikoff, S. Centromeric localization and adaptive evolution of an Arabidopsis histone H3 variant. Plant Cell 14, 1053–1066 (2002)
Henikoff, S. & Dalal, Y. Centromeric chromatin: what makes it unique? Curr. Opin. Genet. Dev. 15, 177–184 (2005)
Henry, I. M. et al. Aneuploidy and genetic variation in the Arabidopsis thaliana triploid response. Genetics 170, 1979–1988 (2005)
Chase, S. S. Monoploids and monoploid-derivatives of maize (Zea mays L.). Bot. Rev. 35, 117–168 (1969)
Jauhar, P. P., Dogramaci-Altuntepe, M., Peterson, T. S. & Almouslem, A. B. Seedset on synthetic haploids of durum wheat: cytological and molecular investigations. Crop Sci. 40, 1742–1749 (2000)
Avetisov, V. A. Production of haploids during in vitro culturing of Arabidopsis thaliana (L.) Heynh anthers and isolated protoplasts. Genetika 12, 17–25 (1976)
Scholl, R. & Amos, J. A. Isolation of doubled-haploid plants through anther culture. Z. Pflanzenphysiol. 96, 407–414 (1980)
Udall, J. A. & Wendel, J. F. Polyploidy and crop improvement. Crop Sci. 46, S3–S14 (2006)
Heppich, S., Tunner, H. G. & Greilhuber, J. Premeiotic chromosome doubling after genome elimination during spermatogenesis of the species hybrid Rana esculenta . Theor. Appl. Genet. 61, 101–104 (1982)
Jin, W. et al. Maize centromeres: organization and functional adaptation in the genetic background of oat. Plant Cell 16, 571–581 (2004)
Coe, E. H. A line of maize with high haploid frequency. Am. Nat. 93, 381–382 (1959)
Hagberg, A. & Hagberg, G. High frequency of spontaneous haploids in the progeny of an induced mutation in barley. Hereditas 93, 341–343 (1980)
Kermicle, J. L. Androgenesis conditioned by a mutation in maize. Science 166, 1422–1424 (1969)
Evans, M. M. The indeterminate gametophyte1 gene of maize encodes a LOB domain protein required for embryo sac and leaf development. Plant Cell 19, 46–62 (2007)
Ori, N., Eshed, Y., Chuck, G., Bowman, J. L. & Hake, S. Mechanisms that control knox gene expression in the Arabidopsis shoot. Development 127, 5523–5532 (2000)
Henikoff, S., Ahmad, K. & Malik, H. S. The centromere paradox: stable inheritance with rapidly evolving DNA. Science 293, 1098–1102 (2001)
Comai, L. & Henikoff, S. TILLING: practical single-nucleotide mutation discovery. Plant J. 45, 684–694 (2006)
Francis, K. E., Lam, S. Y. & Copenhaver, G. P. Separation of Arabidopsis pollen tetrads is regulated by QUARTET1, a pectin methylesterase gene. Plant Physiol. 142, 1004–1013 (2006)
Bushell, C., Spielman, M. & Scott, R. J. The basis of natural and artificial postzygotic hybridization barriers in Arabidopsis species. Plant Cell 15, 1430–1442 (2003)
Paul, W., Hodge, R., Smartt, S., Draper, J. & Scott, R. The isolation and characterisation of the tapetum-specific Arabidopsis thaliana A9 gene. Plant Mol. Biol. 19, 611–622 (1992)
Ross, K. J., Fransz, P. & Jones, G. H. A light microscopic atlas of meiosis in Arabidopsis thaliana . Chromosome Res. 4, 507–516 (1996)
Josefsson, C., Dilkes, B. & Comai, L. Parent-dependent loss of gene silencing during interspecies hybridization. Curr. Biol. 16, 1322–1328 (2006)
We thank L. Comai, D. Melters, J. Maloof and J. Ramahi for comments on the manuscript; and R. McNeilage, R. Clarke, P. Kwong and J. Stewart for technical assistance. This work was supported by a grant from the Hellman Family Foundation to S.W.L.C., and by the University of California, Davis.
Author Contributions M.R. and S.W.L.C. designed the study, performed the experiments, analysed the data and wrote the paper.
The University of California, Davis, has filed a provisional patent application in the USA that is based on results described in the manuscript. M.R. and S.W.L.C. are listed as co-inventors on this application.
About this article
Cite this article
Ravi, M., Chan, S. Haploid plants produced by centromere-mediated genome elimination. Nature 464, 615–618 (2010). https://doi.org/10.1038/nature08842
Site‐directed mutagenesis in bread and durum wheat via pollination by cas9 /guide RNA‐transgenic maize used as haploidy inducer
Plant Biotechnology Journal (2020)
The Plant Journal (2020)
Trends in Genetics (2020)