Abstract
Transgenesis through biolistic of immature embryos is the most convenient way to introduce artificially new genes in bread wheat (Triticum aestivum L.). However, only a few genotypes can be efficiently transformed. To improve the transformation of wheat varieties, we stored immature seeds at room temperature or 4 °C during 4 or 7 days and extracted immature embryos prior to transformation. Shelling stops the embryo’s growth and almost all the embryos formed a callus on selective media when stored at 4 °C for 4 or 7 days (respectively 87% and 99%). We also used hybrid immature embryos derived from a cross between a transformable line (Courtot) and a non-transformable line (Chinese Spring) for biolistic transformation. Hybrid embryos showed the same response to biolistic than the responsive parent. All together, these results improve significantly the biolistic protocol for wheat transformation by reducing the number of mother plants in the greenhouse, and improve the transformation of additional genotypes through hybrid transformation.
Similar content being viewed by others
References
Altpeter F, Springer NM, Bartley LE et al (2016) Advancing crop transformation in the era of genome editing. Plant Cell 28:1510–1520. https://doi.org/10.1105/tpc.16.00196
Amer IB, Worland AJ, Korzun V, Börner A (1997) Genetic mapping of QTL controlling tissue-culture response on chromosome 2B of wheat (Triticum aestivum L.) in relation to major genes and RFLP markers. TAG Theor Appl Genet 94:1047–1052. https://doi.org/10.1007/s001220050513
Bailey SF (1935) Thrips as vectors of plant disease. J Econ Entomol 28:856–863. https://doi.org/10.1093/jee/28.6.856
Birchler JA, Yao H, Chudalayandi S et al (2010) Heterosis. Plant Cell 22:2105–2112. https://doi.org/10.1105/tpc.110.076133
Bolibok H, Rakoczy-Trojanowska M (2006) Genetic mapping of QTLs for tissue-culture response in plants. Euphytica 149:73–83. https://doi.org/10.1007/s10681-005-9055-6
Breitler J-C, Labeyrie A, Meynard D et al (2002) Efficient microprojectile bombardment-mediated transformation of rice using gene cassettes. TAG Theor Appl Genet 104:709–719. https://doi.org/10.1007/s00122-001-0786-z
Carlos Popelka J, Altpeter F (2003) Agrobacterium tumefaciens-mediated genetic transformation of rye (Secale cereale L.). Mol Breed 11:203–211. https://doi.org/10.1023/A:1022876318276
Cheng M, Fry JE, Pang S et al (1997) Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol 115:971–980. https://doi.org/10.1104/pp.115.3.971
Choulet F, Alberti A, Theil S et al (2014) Structural and functional partitioning of bread wheat chromosome 3B. Science 345:1249721. https://doi.org/10.1126/science.1249721
Darrier B, Rimbert H, Balfourier F et al (2017) High-resolution mapping of crossover events in the hexaploid wheat genome suggests a universal recombination mechanism. Genetics 206:1373–1388. https://doi.org/10.1534/genetics.116.196014
Dennis ES, Peacock WJ (2009) Vernalization in cereals. J Biol 8:57. https://doi.org/10.1186/jbiol156
Fábián A, Jäger K, Rakszegi M, Barnabás B (2011) Embryo and endosperm development in wheat (Triticum aestivum L.) kernels subjected to drought stress. Plant Cell Rep 30:551–563. https://doi.org/10.1007/s00299-010-0966-x
Fehér A (2015) Somatic embryogenesis—stress-induced remodeling of plant cell fate. Biochim Biophys Acta 1849:385–402. https://doi.org/10.1016/j.bbagrm.2014.07.005
Florentin A, Damri M, Grafi G (2013) Stress induces plant somatic cells to acquire some features of stem cells accompanied by selective chromatin reorganization. Dev Dyn 242:1121–1133. https://doi.org/10.1002/dvdy.24003
Geard A, Spurr CJ, Brown PH (2007) Embryo development and time of cutting in cool temperate carrot seed crops. In: Adkins SW, Ashmore S, Navie SC (eds) Seeds: biology, development and ecology. Proceedings of the eighth international workshop on seeds. CABI, Wallingford, pp 120–129
Golovina EA, Hoekstra FA, Van Aelst AC (2001) The competence to acquire cellular desiccation tolerance is independent of seed morphological development. J Exp Bot 52:1015–1027. https://doi.org/10.1093/jexbot/52.358.1015
Grafi G, Barak S (2015) Stress induces cell dedifferentiation in plants. Biochim Biophys Acta 1849:378–384. https://doi.org/10.1016/j.bbagrm.2014.07.015
Gu HH, Hagberg P, Zhou WJ (2004) Cold pretreatment enhances microspore embryogenesis in oilseed rape (Brassica napus L.). Plant Growth Regul 42:137–143. https://doi.org/10.1023/B:GROW.0000017488.29181.fa
Hiei Y, Ishida Y, Komari T (2014) Progress of cereal transformation technology mediated by Agrobacterium tumefaciens. Front Plant Sci 5:628. https://doi.org/10.3389/fpls.2014.00628
Hodges TK, Kamo KK, Imbrie CW, Becwar MR (1986) Genotype specificity of somatic embryogenesis and regeneration in maize. Nat Biotechnol 4:219–223. https://doi.org/10.1038/nbt0386-219
International Wheat Genome Sequencing Consortium (IWGSC) (2014) A chromosome-based draft sequence of the hexaploid bread wheat (Triticum aestivum) genome. Science 345:1251788. https://doi.org/10.1126/science.1251788
International Wheat Genome Sequencing Consortium (IWGSC) (2018) Shifting the limits in wheat research and breeding using a fully annotated reference genome. Science 361:7191. https://doi.org/10.1126/science.aar7191
Jia H, Yi D, Yu J et al (2007) Mapping QTLs for tissue culture response of mature wheat embryos. Mol Cells 23:323–330
Jones HD (2005) Wheat transformation: current technology and applications to grain development and composition. J Cereal Sci 41:137–147. https://doi.org/10.1016/j.jcs.2004.08.009
Kim D, Alptekin B, Budak H (2018) CRISPR/Cas9 genome editing in wheat. Funct Integr Genomics 18:31–41. https://doi.org/10.1007/s10142-017-0572-x
Kiviharju E, Pehu E (1998) The effect of cold and heat pretreatments on anther culture response of Avena sativa and A. sterilis. Plant Cell Tissue Organ Cult 54:97–104. https://doi.org/10.1023/A:1006167306638
Li B, Caswell K, Leung N, Chibbar RN (2003) Wheat (Triticum aestivum L.) somatic embryogenesis from isolated scutellum: days post anthesis, days of spike storage, and sucrose concentration affect efficiency. In Vitro Cell Dev Biol 39:20–23. https://doi.org/10.1079/IVP2002356
Liang Z, Chen K, Zhang Y et al (2018) Genome editing of bread wheat using biolistic delivery of CRISPR/Cas9 in vitro transcripts or ribonucleoproteins. Nat Protoc 13:413–430. https://doi.org/10.1038/nprot.2017.145
Lublin A, Sela S (2008) The impact of temperature during the storage of table eggs on the viability of Salmonella enterica serovars enteritidis and virchow in the eggs. Poult Sci 87:2208–2214. https://doi.org/10.3382/ps.2008-00153
Luo J, Jiang S, Pan L (2003) Cold-enhanced somatic embryogenesis in cell suspension cultures of Astragalus adsurgens Pall.: relationship with exogenous calcium during cold pretreatment. Plant Growth Regul 40:171–177. https://doi.org/10.1023/A:1024295901808
Machii H, Mizuno H, Hirabayashi T et al (1998) Screening wheat genotypes for high callus induction and regeneration capability from anther and immature embryo cultures. Plant Cell Tissue Organ Cult 53:67–74. https://doi.org/10.1023/A:1006023725640
Malabadi RB, van Staden J (2006) Cold-enhanced somatic embryogenesis in Pinus patula is mediated by calcium. S Afr J Bot 72:613–618. https://doi.org/10.1016/j.sajb.2006.04.001
Montalbán IA, García-Mendiguren O, Goicoa T et al (2015) Cold storage of initial plant material affects positively somatic embryogenesis in Pinus radiata. New Forest 46:309–317. https://doi.org/10.1007/s11056-014-9457-1
Özgen M, Türet M, Altınok S, Sancak C (1998) Efficient callus induction and plant regeneration from mature embryo culture of winter wheat (Triticum aestivum L.) genotypes. Plant Cell Rep 18:331–335. https://doi.org/10.1007/s002990050581
Pastori GM, Wilkinson MD, Steele SH et al (2001) Age-dependent transformation frequency in elite wheat varieties. J Exp Bot 52:857–863. https://doi.org/10.1093/jexbot/52.357.857
Pescitelli SM, Johnson CD, Petolino JF (1990) Isolated microspore culture of maize: effects of isolation technique, reduced temperature, and sucrose level. Plant Cell Rep 8:628–631. https://doi.org/10.1007/BF00270070
Przetakiewicz A, Karaś A, Orczyk W, Nadolska-Orczyk A (2004) Agrobacterium-mediated transformation of polyploid cereals. The efficiency of selection and transgene expression in wheat. Cell Mol Biol Lett 9:903–917
Rasco-Gaunt S, Riley A, Cannell M et al (2001) Procedures allowing the transformation of a range of European elite wheat (Triticum aestivum L.) varieties via particle bombardment. J Exp Bot 52:865–874. https://doi.org/10.1093/jexbot/52.357.865
Saintenac C, Falque M, Martin OC et al (2009) Detailed recombination studies along chromosome 3B provide new insights on crossover distribution in wheat (Triticum aestivum L.). Genetics 181:393–403. https://doi.org/10.1534/genetics.108.097469
Samarah NH (2005) Effects of drought stress on growth and yield of barley. Agron Sustain Dev 25:145–149. https://doi.org/10.1051/agro:2004064
Sánchez-León S, Gil-Humanes J, Ozuna CV et al (2018) Low-gluten, nontransgenic wheat engineered with CRISPR/Cas9. Plant Biotechnol J 16:902–910. https://doi.org/10.1111/pbi.12837
Shrawat AK, Lörz H (2006) Agrobacterium-mediated transformation of cereals: a promising approach crossing barriers. Plant Biotechnol J 4:575–603. https://doi.org/10.1111/j.1467-7652.2006.00209.x
Stoykova P, Stoeva-Popova P (2011) PMI (manA) as a nonantibiotic selectable marker gene in plant biotechnology. Plant Cell Tissue Organ Cult 105:141–148. https://doi.org/10.1007/s11240-010-9858-6
Tassy C, Barret P (2017) Biolistic transformation of wheat. In: Bhalla P, Singh M (eds) Wheat biotechnology. Methods in molecular biology, vol 1679. Humana Press, New York, pp 141–152
Tassy C, Partier A, Beckert M et al (2014) Biolistic transformation of wheat: increased production of plants with simple insertions and heritable transgene expression. Plant Cell Tissue Organ Cult 119:171–181. https://doi.org/10.1007/s11240-014-0524-2
Vasil V, Castillo AM, Fromm ME, Vasil IK (1992) Herbicide resistant fertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus. Nat Biotechnol 10:667–674. https://doi.org/10.1038/nbt0692-667
Wang Y, Cheng X, Shan Q et al (2014) Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew. Nat Biotechnol 32:947–951. https://doi.org/10.1038/nbt.2969
Wright M, Dawson J, Dunder E et al (2001) Efficient biolistic transformation of maize (Zea mays L.) and wheat (Triticum aestivum L.) using the phosphomannose isomerase gene, pmi, as the selectable marker. Plant Cell Rep 20:429–436. https://doi.org/10.1007/s002990100318
Yao Q, Cong L, He G et al (2007) Optimization of wheat co-transformation procedure with gene cassettes resulted in an improvement in transformation frequency. Mol Biol Rep 34:61–67. https://doi.org/10.1007/s11033-006-9016-8
Zhang K, Liu J, Zhang Y et al (2015) Biolistic genetic transformation of a wide range of Chinese elite wheat (Triticum aestivum L.) varieties. J Genet Genomics 42:39–42. https://doi.org/10.1016/j.jgg.2014.11.005
Zong Y, Wang Y, Li C et al (2017) Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol 35:438–440. https://doi.org/10.1038/nbt.3811
Acknowledgements
Members of the team CPCC are greatly acknowledged for taking care of the plants. Members of the team ValFon are also acknowledged for helpful discussions and for providing with all facilities for biolistic transformation. RM is funded by ANRT CIFRE Grant No. 2014/1020.
Author information
Authors and Affiliations
Contributions
RM, CT, MCD, AL and MB conducted the experiments; RM analyzed all data; RM, PB and PS conceived this work and wrote the paper. GB and AN reviewed the paper. All authors contributed in the writing of this paper.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Communicated by Francisco de Assis Alves Mourão Filho.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Michard, R., Batista, M., Debote, MC. et al. Cold-conserved hybrid immature embryos for efficient wheat transformation. Plant Cell Tiss Organ Cult 136, 365–372 (2019). https://doi.org/10.1007/s11240-018-1521-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-018-1521-7