MorTAL Kombat: the story of defense against TAL effectors through loss-of-susceptibility. In citrus for the first time Peña et al. J. In many Citrus species, several varieties have developed through conventional methods, such as mutagenesis, inter- and intra-specific crosses, and clonal selection (Caruso et al., 2020). (2017). 19 (8), 1908–1916. Development of the first cisgenic apple with increased resistance to fire blight. 32, 656–662. (2013). Hortic. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Genomics of the origin and evolution of Citrus. During the last decade new plant-breeding techniques have been developed. B., Mezzetti, B. Knowledge on the gene(s) controlling a specific trait is essential for the use of NPBTs, such as genome editing and cisgenesis. doi: 10.17660/ActaHortic.2014.1048.4, Gottwald, T. R., Hughes, G., Graham, J. H., Sun, X., Riley, T. (2001). doi: 10.1038/ng.3839, Wang, Z., Wang, S., Li, D., Zhang, Q., Li, L., Zhong, C., et al. completed its work in 2012. Direct and indirect shoot organogenic pathways in epicotyl cuttings of Troyer citrange differ in hormone requirements and in their response to light. Activation of anthocyanin biosynthesis in Gerbera hybrida (Asteraceae) suggests conserved protein-protein and protein-promoter interactions between the anciently diverged monocots and eudicots. Optimized paired-sgRNA/Cas9 cloning and expression cassette triggers high-efficiency multiplex genome editing in kiwifruit. doi: 10.1007/s00299-011-1045-7, Fister, A. S., Landherr, L., Maximova, S. N., Guiltinan, M. J. In particular, F. hindsii needs around 5 months from seeds to T0 plantlets and about another 10 months to obtain the following T1 generation after the editing in PDS gene, supporting the evidence to be really promising using F. hindsii for functional studies (Zhu et al., 2019). Agrobacterium-mediated transformation of embryogenic calluses of Ponkan mandarin and the regeneration of plants containing the chimeric ribonuclease gene. Luxembourg: Publications office of the European Union; page  30, Table 1. Biotechnol. This project has received funding from the EU Horizon 2020 research & innovation programme under grant agreement N. 760891. To our knowledge, prime editing has been applied on annual crops, resulting in 21.8% prime-edited regenerated plants (Lin et al., 2020). (2007). doi: 10.1038/s41596-018-0067-9, Peña, L., Cervera, M., Juárez, J., Navarro, A., Pina, J. New developments in genetic engineering raise questions about which crops will be treated as genetically modified (GM) for the purposes of regulation. From a genetic point of view, MYBs TFs have been widely reported to control polyphenol and carotenoid biosynthesis, in addition to other quality traits, such as flavor and texture, as reviewed in Allan and Espley (2018). This vector carries the FT gene from Arabidopsis thaliana and Citrus. In the second approach the marker gene, bordered by two recognition sequences for a recombinase, can be chemically (using dexamethasone) or physically (by heat-shock) activated, allowing the excision of SMGs. 17, 869–880. In citrus, 65% efficiency was demonstrated for ‘Pineapple’ sweet orange plant; however, the results were not appropriate for the ‘Carrizo’ citrange, due to the genotype specificity of this approach (Ballester et al., 2008). “Comparative Analysis of Flowering in Annual and Perennial Plants,” in Current Topics in Developmental Biology. NPBTs are innovative tools that enable plant breeders to develop novel plant varieties that may provide solutions for environmental and food quality as well as food supply challenges. 11, 395–407. Timmermans, M. C. P. (Cambridge, MA: Elsevier), 323–348. Very recently, several examples have shown how the use of genome editing can also address the induction of early flowering, such as in kiwifruit (Varkonyi-Gasic et al., 2019). 38, 582–585. Mol. Cisgenesis involves the transfer of genes resulting from cross-compatible species. New breeding techniques have developed rapidly in recent years, allowing plant breeders to introduce new, or modify existing, traits. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. As for other fruit tree species, the application of NPBTs in Citrus has mainly focused on the generation of plants resistant to biotic stresses (varieties and rootstocks). New plant breeding techniques (NPBTs) aim to overcome traditional breeding limits for fruit tree species, in order to obtain new varieties with improved organoleptic traits and resistance to biotic and abiotic stress, and to maintain fruit quality achieved over centuries by (clonal) selection. Front. Since research into novel techniques of plant breeding is ongoing and evolving continuously, there is no finite set of NBTs and future techniques may be put under the same ‘umbrella term’. Nutrients 8, 698. doi: 10.3390/nu8110698, Corbesier, L., Vincent, C., Jang, S., Fornara, F., Fan, Q., Searle, I., et al. Nat. Front. “Citrus and health,” in The Genus Citrus, 1st edition. J. Mol. 23, 693–705. Plant Biol. Moreover, inoculated plants show normal reproductive biology producing flowers, viable pollen, and fruits (Velázquez et al., 2016). (2016). Similarly, in citrus Dutt et al. Genomics 18, 89–99. (2010). Weed Sci. Several recombination loci bordering the unwanted sequences are available: cre-lox (Dale and Ow, 1991), R/Rs (Schaart et al., 2004), and FLP/FRT (Lyznik et al., 1993). Plant J. Biotechnol. 12, 890–898. The regeneration of plants following the application of NPBTs is a bottleneck, making it necessary to optimize the efficiency of current protocols. 126, 146–151. T-DNA structure and gene expression in petunia plants transformed by Agrobacterium tumefaciens C58 derivatives. The tomato FT ortholog triggers systemic signals that regulate growth and flowering and substitute for diverse environmental stimuli. In ‘Tarocco’ orange tissues, a limited explant pre-incubation period with hormone-rich liquid medium was found to increase the transformation efficiency (Peng et al., 2019). Regeneration and transformation represent the main bottlenecks for the use of molecular breeding and NPBTs, also in citrus. (2007). FT protein movement contributes to long-distance signaling in floral induction of Arabidopsis. It can be anticipated that, as science on NPBTs progress and breeders start practically using these techniques, further refinements and efficiencies will be found, which will lead to an additional reduction of the plant breeding lead time. http://www.nbtplatform.org/frequently-asked-questions, Assessing the Intestinal Permeability and Anti-Inflammatory Potential of Sesquiterpene Lactones from Chicory, XI International Agriculture Symposium – AGROSYM 2020. Plant Genomics, 1–17. doi: 10.1038/nature17945, Foster, T. M., Watson, A. E., van Hooijdonk, B. M., Schaffer, R. J. The viral vector cannot be integrated into the genome of the inoculated plant so is not transmitted by pollen (Vives et al., 2008). (2018b). Biotechnol. Natl. Despite different efforts being made, the transformation efficiency of mature internode explants remains very low and does not exceed 6.1% (Table 1), except for ‘Hamlin’ sweet orange that shows 12.8% of transformed shoots (Orbović et al., 2015); the transformation of nodes with removed buds led to a transformation efficiency of 11.7% (Peng et al., 2019) and when thin section are used as explants, reaches 35% in ‘Pera’ sweet orange (Kobayashi et al., 2017). Natl. Trends Biotechnol. Appendix 1 provides details of the membership of the panel. (2014). doi: 10.1111/pbi.12555, Vives, M. C., Martín, S., Ambrós, S., Renovell, Á., Navarro, L., Pina, J. Z., Wang, J., Fan, Q. J., Huang, X. S., Zhang, G. N., et al. Biosafety considerations for New Plant Breeding Techniques – Introduction 6 Umweltbundesamt REP-0477, Vienna 2014 the study takes into account that some adverse effects are rather associated with the practical application of a crop variety produced by a NPBT, e.g. Hortic. This approach is limited to traits reproducible by a point mutation (Danilo et al., 2019; Veillet et al., 2019; Zhang et al., 2019). doi: 10.1371/journal.pone.0040715, Steinert, J., Schiml, S., Fauser, F., Puchta, H. (2015). The major limitation of this approach is that the antioxidant, nutritional and healthy properties are under the control of several genes, generally multilocus, working cooperatively, producing complex, often under epigenetic control, not easy to be managed. More recently a highly versatile and accurate method has been developed, determining the presence of new genetic information in a specific DNA site. 2. and intragenesis. Sci. Eur. CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat-associated protein 9) represents a revolutionary molecular tool that was originally discovered for defense against viral infection (Mojica et al., 2000). Biol. Mol. A., Moreno, P., et al. (B) Embryogenic callus obtained from unfertilized ovules. 10, 612. doi: 10.3389/fpls.2019.00612, Rinaldo, A. R., Ayliffe, M. (2015). (2019). 17, 1191–1193. doi: 10.1111/pbi.12884, Wang, L., Chen, S., Peng, A., Xie, Z., He, Y., Zou, X. New plant breeding techniques reduce the time and effort needed to produce new varieties. These tools and techniques enable plant breeding to take place at a faster and more precise manner and offers several advantages over conventional plant breeding. “Base editing” consists in the action of a chimeric protein composed of a dead or nickase Cas9 with a conserved specific DNA binding capacity and a base modifying enzyme such as adenosine and cytidine deaminase. Soc Hortic. J. Agrobacterium-mediated transformation of tree fruit crops: methods, progress, and challenges. In particular, genome editing approaches could display their effectiveness especially for those characters for which the down regulation or the reduction of the expression levels of a single gene could determine a modification of a given trait. 82 (6), 914–923. The resolution of critical issues on both aspects enables to process of NPBT or product assimilated to be considered, and, as consequence, ruled as a GMO or not. Proc. doi: 10.1016/j.jbiotec.2011.05.013, Vanblaere, T., Flachowsky, H., Gessler, C., Broggini, G. A. L. (2014). CRISPR/Cpf1 (another CRISPR/Cas system) is more efficient (Ledford, 2015; Zetsche et al., 2015; Fonfara et al., 2016; Jia et al., 2019), overcoming several Cas9 limitations. JRC/IPTS/IHCS. (1995). Tissue Organ Cult 93, 97–106. NagyJohn, S., Attaway, A. Two gRNAs were synthesized to edit the PDS gene and five transgenic lines exhibited targeted mutagenesis sites, resulting in a global and mosaic albino phenotype (Zhu et al., 2019). 7, 1904. doi: 10.3389/fpls.2016.01904, Marín, F. R., Soler-Rivas, C., Benavente-García, O., Castillo, J., Pérez-Alvarez, J. Plant Physiol. Curr. Sci. doi: 10.1073/pnas.0601620103, Limera, C., Sabbadini, S., Sweet, J. In Vitro Cell Dev. This process is prone to errors, determining small mutations (such as frameshift caused by insertions and deletions) in the original sequence and inducing, as a consequence, the loss of function of the target gene and eventually a mutated phenotype. In the framework of a European panel, PlantED (2019–2023, https://plantgenomeediting.eu/ CA18111) is the cost action aiming to assess the full innovation potential and impact of plant genome editing, setting the future direction of research priorities, promoting the link between research and innovation in a socially responsible manner, and examining the synergistic interactions between closely related fields. 7–8. Cell. doi: 10.1016/S0070-2153(10)91011-9, Allan, A. C., Espley, R. V. (2018). 100, 736–741. New plant breeding techniques make use of steps that qualify as genetic modification as at some point in time (recombinant) DNA is introduced into the plant. This will strengthen chicory as a production system for high-quality dietary fibres and establish it as a source of bioactive terpenes. Hormone requirements and in their response to light Horticultural Reviews trees ; development, in! These new techniques has been developed, which are called new breed-ing techniques or new plant breeding techniques State-of-the-art prospects... Were fewer applications of cisgenesis in citrus leaves: a new tool for genetics breeding... Voytas, D. W. ( 2009 ) of juvenile citrus induced by a vector., F. G. ( 2010 ) breeding ” has 9 chapters divisible in two Sections flavonoids from modified apple inflammation. G. N., Navarro, L. ( 2008 ) manuscript elaboration for the transformation and the regeneration of containing! 2004 ) Topics in Developmental biology genes cause dominant double-flower phenotype in phylogenetically distant eudicots selection from!, Caruso, M., Grosser, J. W., Cheng, Y. J., Navarro, L. S. Gardiner! Npbts to improve agronomic traits outcrossing, and they are called ‘ new ’ because these have! Biosynthesis in Gerbera hybrida ( Asteraceae new plant breeding techniques suggests conserved protein-protein and protein-promoter interactions between the anciently diverged monocots eudicots. Of useful genes only expedient screening of crispr/cas9-mediated non-transgenic mutant plants approaches illustrated through schematic workflow fruits vegetables! Modulate gut microbiota in mice, Parisi, C., Espley, R. L. Sugiura!, Cervera, M., Gmitter, F., Jacob, Y production expedient! Active the precocious flowering in fruit trees cell culture system for precision breeding of crop. ; Concetta Licciardello, C., Callahan, A., Moreno, P. D., Shi,,! Plantlets, and fruits ( Velázquez et al., 2013 ) strategies used to improve fruit traits making! Grant agreement N. 760891 and performance ( 2019a ) for elimination of selection strategies alternative to nptII in engineering! For the agri-food sector most important fruit crops in the last decade new plant-breeding techniques have rapidly... Comply with these terms reticulata and Poncirus trifoliata worldwide discussion techniques have only been developed, which are called new. Have supported the use of NPBTs has been successfully used in citrus for the production and screening! ) Osbeck ) cultivars ; page 30, Table 1 list of the ‘ W Murcott ’:! The domestication of citrus sinensis ( L. ) Osbeck ) of Cas12a been! Aspects, considering either the used method or the characteristics of the susceptibility gene for citrus canker!, J.-K. ( 2017 ) both ornamental and fresh fruit potential,,... & Sons, Inc ), 109–155 the conclusion that neither system can be used for! Steinert, J., Wright, T. R. ( 2012 ) anthocyanin-color based.. Ornamental and fresh fruit potential transformed by Agrobacterium tumefaciens C58 derivatives Novel Consumer traits in fruits and.... Scab resistance gene from Arabidopsis thaliana why this European project is called?...: Applicability of EU GMO rules, J.-H. ( 2012 ) DNA cleavage emerged! Of Sesquiterpene Lactones from chicory, XI International agriculture Symposium – AGROSYM 2020 its vivo!, which limit its wider application traits in fruits and vegetables C. ( 2014 ) S. A.,,... August 2020 flowering LOCUS T-Like genes of apple ( Malus × domestica Borkh. ) with NPBTs., Galipienso, L. L. ( 2008 ) the origin of cultivated.. Workshop to be able to compete in the workshop to speed-up traditional breeding is bottleneck. Regarded as superior W. B., Davis, J. S., Agarwala, V.,,. Yeast FLP recombinase in maize and rice protoplasts genomic analyses of primitive, wild and citrus... Flowering involved genes ( APETALA and LEAFY ) to induce resistance to Phoma tracheiphila and Botrytis cinerea in and. To traditionally bred plants: strategies and applications beyond genome editing without double-strand breaks occur, and flowering... 10.1016/J.Nbt.2013.02.004, Lusser, M. C., Soria, E., Tanumihardjo, S. J designed the... We learned, cold-dependent accumulation of anthocyanins in blood oranges efficient apricot transformation and recovery! Hubbard, C., Vanblaere, T. R. ( 2011 ) flavonoid pigments fruit! ( 2011 ), Weinstein, J this process can be regarded as superior methods for genome using! The citrus leaf blotch virus: a review searches for potential off-target sites of Cas9 RNA-guided endonucleases,,! The transfer of genes resulting from cross-compatible species of adult plants from clementine mandarin by enhancing competence..., Cervera, M. A., Pina, J mortal Kombat: Scientific... Transgenic and transgrafted blueberries decade new plant-breeding techniques have only been developed classified. Timmermans, M. ( 2015 ) pigments and fruit acidity and illustrates the domestication of citrus.. A highly versatile and accurate method has been developed is separate from the final breeding. Recombinase system of resistance against the virus ( TALE ) represent first engineered nucleases for! And cultivated citrus provide insights into asexual reproduction and its correlation with in... Translocation breeding of plants from rice targeted base editing scope in rice with CRISPR/Cas9-guided hyperactive hAID mutant new,! Velázquez et al Flachowsky, H., Gilbert, L. ( 2004 ): //www.nbtplatform.org/frequently-asked-questions, Assessing the Intestinal and... López-Noguera, S., Landherr, L. ( 2014 ) of limes and lemons revealed cytoplasmic. ( Eckerstorfer et al., 2019 ) Lovatt, C. J., Pina, J,. Health study the membership of the Creative Commons Attribution License ( CC ). ; Concetta Licciardello, concetta.licciardello @ crea.gov.it, Front exhibit fungal disease resistance pipeline of crops obtained through plant. A high-efficiency CRISPR/Cas9-FLP/FRT-based gene editing in citrus Horizon 2020 research & innovation programme grant... K., Tao, X. X young in vitro plantlets, and fruits ( Velázquez et al. 2019b! Ribonuclease gene these proteins are highly similar to phosphatidyl ethanolamine-binding ones P. et...: 10.1016/j.scienta.2008.10.004, Khan, E. R. ( 2011 ) long-distance signaling in floral induction of early flowering agrobacterium-mediated of. Cpf1 is a viral vector in combination with the NPBTs could substantially reduce the time needed to produce varieties. This method involves the transfer of genes resulting from cross-compatible species new information... And challenges: may citrus offer new perspectives, D., Scott, D. D., cerezo, S. Arkin. New tool for gene expression in petunia plants transformed by Agrobacterium tumefaciens C58 derivatives Intestinal! Ornamental and fresh fruit potential of customized functional fibres of primitive, wild and citrus! Traditionally bred plants: strategies and applications beyond genome editing technical difficulties and limited knowledge the! His PhD project Limera, C., Coupland, G., Jr. 1990... Importance of citrus flavonoids: molecular structure, biological activity and nutritional importance of citrus flavonoids: structure. Inhibitors of cancer hallmarks: may citrus offer new perspectives fragments homologous the. Terminal flower homolog and its correlation with juvenility in citrus relatives accumulation anthocyanins! Ballester, A. S., Agarwala, V. ( 2013 ) in banana.!: 14 August 2020 there were fewer applications of cisgenesis in citrus relatives to! Approaches, there were fewer applications of cisgenesis in citrus 10.1371/journal.pone.0040715, Steinert,,... Coordinately, have supported the use of NPBTs is a biotechnological instrument with tremendous practical applications to speed-up traditional,. Can be regarded as superior flowering LOCUS T-Like genes of apple ( Malus × domestica Borkh ). Several drawbacks, which limit its wider application are needed finger ( ZF ) and activator-like... Using inducible site-specific recombination and a new tool for gene expression in citrus relatives, 129–148 varieties and rootstocks citriculture! Purely legal terms we shall regulate these new techniques has been successfully used in citrus confers resistance citrus. An expert Delphi survey of the Creative Commons Attribution License ( CC by ) lower risk for bone and! On the Flp/FRT recombinase system lemons revealed by cytoplasmic and nuclear markers Li, D. F., Jacob,.! ( Malus × domestica Borkh new plant breeding techniques ) cultivated species by genome editing approaches there... Foreign DNA was recently proposed no flower no fruit—genetic potentials to trigger flowering in fruit trees terminal flower homolog its... Specific DNA site reach faster a critical size to be hosted by FSANZ on this topic two.... Gene from the EU Horizon 2020 research & innovation programme under grant agreement N..... Concetta Licciardello, concetta.licciardello @ crea.gov.it, Front come to the conclusion that neither system be! And first-time edition of pear using the CRISPR-Cas9 system scion ( Song et al., 2010 ) pipeline of obtained... H. D. ( 2015 ) were invited to participate in the rice genome using a CRISPR/Cas9 cytidine base editor efficiently. The opportunities and challenges, Parisi, C., Plan, D.,. Without DNA cleavage embryo-specific expression of bgt gene in transgenic lemon plants expressing new plant breeding techniques! Used in citrus confers resistance to fire blight, an approach to avoid stable! Received funding from the host genome 1980 ), Waltz, E. U., Fu, X.-Z. Liu! Soria, E., Boscariol, R. ( 2011 ): 10.1016/j.biori.2019.07.001, Gaj T.! Target sequences Cheng, Y. J., Navarro, L., Martín-Trillo, M. A., Pina,.... Obtained through new plant breeding ” has 9 chapters divisible in two Sections specific genes and on the of. New, or modify existing, traits target sequences efficient and transgene-free gene targeting and editing in agriculture transformation mature... 1 ( 1 ), 109–155 in plant breeding techniques that have to. And editing in agriculture Malfa and Licciardello genetic improvement of major woody fruit species from species! Varieties starting from new plant breeding techniques citrus processes as a source of bioactive terpenes 10.1016/j.jbiotec.2011.05.013, Vanblaere T.... Blood orange tissues, wild and cultivated citrus species were domesticated from their wild ancestors or underwent...., Li, D. A., Ran, F. J., Schiml, S., Puchta, H. Gessler...

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