Citrus Transformation Facility
The Juvenile Tissue Citrus Transformation Facility (JTCTF) has undergone a transition to Educational Business Activity unit in accordance with request made by UF Administration. This transition completely overhauls the operation of the facility and brings into force new set of rules in regard to ordering, pricing, and payments to the JTCTF for the services it provides.
The Juvenile Tissue Citrus Transformation Facility was established to expedite integration of recombinant DNA technologies and genetic engineering into basic research from other related disciplines, including plant pathology, biochemistry, food science/processing, post-harvest physiology, horticulture, and entomology by providing a service that allows researchers to have their genes of interest transferred and tested in the appropriate citrus cultivars. A functional core citrus transformation laboratory benefits nearly all phases of the Florida citrus industry and plays a major role in the long-term improvement of citrus.
The purpose of the JTCTF is to produce genetically modified Citrus plants. The facility is equipped to employ two methods of genetic transformation: Agrobacterium-mediated transformation and biolistic particle bombardment. At this time we are producing transgenic plants only through Agrobacterium-mediated transformation. This method utilizes naturally occurring pathogenic bacterium called Agrobacterium tumefaciens that has the ability to insert the fragment of DNA into the genome of host plant. These bacteria have been manipulated and rendered non-pathogenic although they kept their ability to transfer foreign DNA into host’s genome. The DNA fragment of interest to our client is introduced into Agrobacterium, and such newly created strain is used for treatment of plant tissues that will result in production of plantlets that have their genomes altered by the presence of foreign DNA. Additional tests are performed to confirm that genetic modification took place.
Core Citrus Transformation Facility (CCTF) exists for more than 16 years. Since becoming fully functional 14 years ago, CCTF has succeeded in fulfilling the role of becoming the production site for transgenic Citrus plants. Clients from many states within the US and, from different research organizations and foundations have come to us and requested plants of Citrus cultivars of choice with the desired gene inserted into their genome. Despite detrimental effect of HLB on transformation success rate and exploratory nature of many recent orders, CCTF is still providing transgenic material at satisfactory level. At the moment, capacity of CCTF is to produce about 250 seedlings per year although the final number depends on the cultivars that are ordered/nature of the orders placed.
Most of the plants produced in the CCTF within the last few years were transgenic ‘Duncan’ grapefruit, followed by Carrizo citrange, sweet orange 'Valencia' and ‘Pineapple’ cultivars, Mexican lime, and Swingle citrumelo.
Based on the un-interrupted operation, previous accomplishments, and productivity CCTF has become integral part of many projects that have a goal of production of tolerant/resistant trees to HLB, canker, and CTV.
CCTF is recognized outside of the US as well, with many scientists quoting the publications that were published based on experiments done on transgenic plants made in our facility.
As our major accomplishments I would list the following:
- we were the first lab to produce cis/intragenic Citrus plants;
- we were the first place to produce transgenic Citrus plants with chemically inducible genes;
- we were the first place where transgenic Citrus plants with CRISPR-edited genome were produced.
Citrus Transformation Phases
Sterilized seeds are allowed to germinate in darkness for 4-5 weeks.
Stems of seedlings grown in vitro are cut into 1/2 inch explants, incubated with Agrobacteria and placed on shoot regeneration medium.
Shoots appear in explants (4-5 weeks after treatment with bacteria)
Shoots are collected from explants and allowed to grow for a week.
Tested for reporter genes.
Shoots grafted on rootstocks in vitro.
Two weeks old, soil adapted seedlings
Four months seedlings
Gel with the products of PCR reaction proving that the gene of interest is present within the tissue of transgenic plants
CCTF is located on the campus of CREC. The laboratory has standard equipment including:
- 3 laminar flow hoods
- Sterilizing furnace
- Thermo-controlled shaker
- Zeiss microscopes including inverted stereo microscope and binocular microscope equipped with digital camera which uses SPOT software for fluorescence analysis
Plants and explants are kept in 2 new Percival reach-in growth chambers, and in a greenhouse certified for transgenic material.
Here you will find regularly-updated podcast presentations. You will also be able to read a transcription of the podcast, and view the original articles and relevant materials.
The transition of JTCTF into the EBA unit has changed the way we charge for our services. Please inspect the pricelist below for additional information. The prices marked “internal academic” refer to orders coming from the University of Florida (UF). Those prices marked “external academic” refer to orders coming from other academic institutions, whereas the prices in the column “external market” are for business entities. When hiring the JTCTF, clients will sign the contract that will be supplied by the office of Business Management at UF. Starting with January 1st 2021, JTCTF will bill the clients on a monthly basis. Before we start working on the order, we will provide an approximate total price for the sought service(s). Monthly billing has a purpose of keeping the facility funded throughout the whole year. It also means that clients are assuming the responsibility for the success of the designed experiments. In other words, JTCTF will require payments for the work performed in a certain time frame regardless of the outcome of those efforts. Because of the exploratory nature of these experiments, it will take three months to estimate possible detrimental effect of introduced gene (sensu lato) on regeneration of shoots from explants co-incubated with Agrobacterium and JTCTF will charge clients for the work it performed in that initial period. If the goal of the introduction of transgene into citrus plants is to change phenotypic features that can be recorded by visual inspection, then JTCTF will communicate the success in realized project to clients as the experiments move ahead. Clients will have the chance to stop the work done by JTCTF if there is a lack of progress. In the case where the function of the transgene in transgenic plants needs to be confirmed by additional tests, the responsibility of performing such tests lays on the client except if they wish to hire JTCTF to perform one of the services listed in the Pricelist. It will take 9-12 months, depending on the cultivar, to produce plants that could be tested for the functionality of introduced transgene and JTCTF will require payments for the work done towards production of transgenic plants regardless of the outcome of those tests.
- USDA's Agricultural Biotechnology
- Oklahoma Plant Transformation Facility, Nobel Res Ctr
- Texas A&M Plant Tissue Culture Information Exchange
- Transgenic Crops: Intro and Resource Guide
- UCBiotech - University of California
- USDA Biotechnology Permit Information
- NU Plant Transformation Research Core Facility
- Plant Transformation Core Facility, Univ. of Missouri
- Plant Tissue Culture and Transformation Facility, Cornell
- ISU Plant Transformation Facility
- Plant Biotechnology Core Facility, Univ. of Connecticut
- Plant Biotechnology Facility, University of Wisconsin
Sinn JP, Held JB, Vosburg C, Klee SM, Orbovic V, Taylor EL, Gottwald TR, Stover E, Moore GA, McNellis TW. Flowering Locus T chimeric protein induces floral precocity in edible citrus. Plant Biotechnol J. 2021 Feb;19(2):215-217. https://doi.org/10.1111/pbi.13463. Epub 2020 Sep 16.
Conti G, Gardella V, Vandecaveye MA, Gomez CA, Joris G, Hauteville C, Burdyn L, Almasia NI, Nahirñak V, Vazquez-Rovere C, Gochez AM, Furman N, Lezcano CC, Kobayashi K, García ML, Canteros BI, Hopp HE, Reyes CA. Transgenic Citrange troyer rootstocks overexpressing antimicrobial potato Snakin-1 show reduced citrus canker disease symptoms. J Biotechnol. 2020 Dec 20;324:99-102. https://doi.org/10.1016/j.jbiotec.2020.09.010. Epub 2020 Sep 28.
Corte, L.E., B.M.J. Mendes, F.A.A. Mourao Filho, J.W. Grosser and M. Dutt (2020). Functional characterization of full-length and 5′ deletion fragments of Citrus sinensis-derived constitutive promoters in Nicotiana benthamiana. In Vitro Cellular and Developmental Biology – Plant https://doi.org/10.1007/s11627-019-10044-0
Dutt, M., Mou, Z., Zhang, X. et al. Efficient CRISPR/Cas9 genome editing with Citrus embryogenic cell cultures. BMC Biotechnol 20, 58 (2020). https://doi.org/10.1186/s12896-020-00652-9
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Jardak, R., Boubakri, H., Zemni, H. et al. Establishment of an in vitro regeneration system and genetic transformation of the Tunisian 'Maltese half-blood' (Citrus sinensis): an agro-economically important variety. 3 Biotech 10, 99 (2020). https://doi.org/10.1007/s13205-020-2097-6
Mahmoud, L.M., J.W. Grosser and M. Dutt (2020). Silver compounds regulate leaf drop and improve in vitro regeneration from mature tissues of Australian finger lime (Citrus australasica). Plant Cell Tissue and Organ Culture. https://doi.org/10.1007/s11240-020-01803-8
Pereira W, Takita M, Melotto M, de Souza A. Citrus reticulata CrRAP2.2 Transcriptional Factor Shares Similar Functions to the Arabidopsis Homolog and Increases Resistance to Xylella fastidiosa. Mol Plant Microbe Interact. 2020 Mar;33(3):519-527. https://www.doi.org/10.1094/MPMI-10-19-0298-R. Epub 2020 Jan 23. PMID: 31973654.
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Qiu, W., J. Soares, Z. Pang, Y. Huang, Z. Sun, N. Wang, J.W. Grosser and M. Dutt (2020). Potential Mechanisms of AtNPR1 Mediated Resistance against Huanglongbing (HLB) in Citrus. International journal of molecular sciences 2020, 21, 2009. https://doi.org/10.3390/ijms21062009
Romero-Romero, J.L., Inostroza-Blancheteau, C., Reyes-Díaz, M. et al. Increased Drought and Salinity Tolerance in Citrus aurantifolia (Mexican Lemon) Plants Overexpressing Arabidopsis CBF3 Gene. J Soil Sci Plant Nutr 20, 244–252 (2020). https://doi.org/10.1007/s42729-019-00130-y
Soares, J.M., Weber, K.C., Qiu, W. et al. The vascular targeted citrus FLOWERING LOCUS T3 gene promotes non-inductive early flowering in transgenic Carrizo rootstocks and grafted juvenile scions. Sci Rep 10, 21404 (2020). https://doi.org/10.1038/s41598-020-78417-9
Ying X, Redfern B, Gmitter FG Jr, Deng Z. Heterologous Expression of the Constitutive Disease Resistance 2 and 8 Genes from Poncirus trifoliata Restored the Hypersensitive Response and Resistance of Arabidopsis cdr1 Mutant to Bacterial Pathogen Pseudomonas syringae. Plants (Basel). 2020 Jun 30;9(7):821. https://doi.org/10.3390/plants9070821.
Zhang, F., Rossignol, P., Huang, T., & Irish, V. (2020). Reprogramming of Stem Cell Activity to Convert Thorns into Branches. Current Biology, 30, 2951-2961.e5.
Zhang, XH., Pizzo, N., Abutineh, M. et al. Molecular and cellular analysis of orange plants infected with Huanglongbing (citrus greening disease). Plant Growth Regul 92, 333–343 (2020). https://doi.org/10.1007/s10725-020-00642-z
Ćalović, M., Chen, C., Yu, Q., Orbović, V., Gmitter, F. G., & Grosser, J. W. (2019). New Somatic Hybrid Mandarin Tetraploid Generated by Optimized Protoplast Fusion and Confirmed by Molecular Marker Analysis and Flow Cytometry, Journal of the American Society for Horticultural Science J. Amer. Soc. Hort. Sci., 144(3), 151-163. Retrieved Mar 24, 2021, from https://journals.ashs.org/jashs/view/journals/jashs/144/3/article-p151.xml
Goulin, E.H., dos Santos, P.J.C., Dalio, R.D. et al. In vitro symptom induction of Colletotrichum abscissum infection in detached sweet orange flowers. J Plant Pathol 101, 695–699 (2019). https://doi.org/10.1007/s42161-018-00220-3
Jia H, Orbović V, Wang N. CRISPR-LbCas12a-mediated modification of citrus. Plant Biotechnol J. 2019 Oct;17(10):1928-1937. https://doi.org/10.1111/pbi.13109. Epub 2019 Apr 10.
Peng, A., Zou, X., Xu, L. et al. Improved protocol for the transformation of adult Citrus sinensis Osbeck ‘Tarocco’ blood orange tissues. In Vitro Cell.Dev.Biol.-Plant 55, 659–667 (2019). https://doi.org/10.1007/s11627-019-10011-9
Soriano, Leonardo, Tavano, Eveline Carla da Rocha, Correa, Marcelo Favaretto, Harakava, Ricardo, Mendes, Beatriz Madalena Januzzi, & Mourão Filho, Francisco de Assis Alves. (2019). In vitro organogenesis and genetic transformation of mandarin cultivars. Revista Brasileira de Fruticultura, 41(2), e-116. Epub April 25, 2019.https://doi.org/10.1590/0100-29452019116
Tavano, E.C., Erpen, L., Aluisi, B., Harakava, R., Lopes, J., Vieira, M.C., Piedade, S.M., Mendes, B., & Filho, F.A. (2019). Sweet orange genetic transformation with the attacin A gene under the control of phloem-specific promoters and inoculation with Candidatus Liberibacter asiaticus. The Journal of Horticultural Science and Biotechnology, 94, 210 - 219. https://doi.org/10.1080/14620316.2018.1493361
Wang, L., Chen, S., Peng, A. et al. CRISPR/Cas9-mediated editing of CsWRKY22 reduces susceptibility to Xanthomonas citri subsp. citri in Wanjincheng orange (Citrus sinensis (L.) Osbeck). Plant Biotechnol Rep 13, 501–510 (2019). https://doi.org/10.1007/s11816-019-00556-x
Wu, Hao; Acanda, Yosvanis; Canton, Michel; Zale, Janice. 2019. "Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection" Plants 8, no. 10: 390. https://doi.org/10.3390/plants8100390
Zhu, C., Zheng, X., Huang, Y., Ye, J., Chen, P., Zhang, C., Zhao, F., Xie, Z., Zhang, S., Wang, N., Li, H., Wang, L., Tang, X., Chai, L., Xu, Q. and Deng, X. (2019), Genome sequencing and CRISPR/Cas9 gene editing of an early flowering Mini‐Citrus (Fortunella hindsii). Plant Biotechnol J, 17: 2199-2210. https://doi.org/10.1111/pbi.13132
Chen, L., Li, W., Katin-Grazzini, L. et al. A method for the production and expedient screening of CRISPR/Cas9-mediated non-transgenic mutant plants. Hortic Res 5, 13 (2018). https://doi.org/10.1038/s41438-018-0023-4
Dutt M., F.T. Zambon, L. Erpen, L. Soriano, J.W. Grosser (2018). Embryo-specific expression of a visual reporter gene as a selection system for citrus transformation. PLOS ONE 13 (1):e0190413..pone.0190413. https://doi.org/10.1371/journal.pone.0190413
Dutt, M., L. Erpen, and J.W. Grosser, 2018. Genetic transformation of the ‘W Murcott’ tangor: comparison between different techniques. Scientia Horticulturae, 242:90-94. https://doi.org/10.1016/j.scienta.2018.07.026
Erpen L., H.S. Devi, J.W. Grosser and M. Dutt (2018). Potential use of the DREB/ERF, MYB, NAC and WRKY transcription factors to improve abiotic and biotic stress in transgenic plants. Plant Cell, Tissue and Organ Culture 132 (1):1-25. https://doi.org/10.1007/s11240-017-1320-6
Erpen, L., E.C.R. Tavano, R. Harakava, M. Dutt, J.W. Grosser, S.M.S. Piedade, B.M.J. Mendes and F.A.A. Mourao Filho (2018) Isolation, characterization, and evaluation of three Citrus sinensis-derived constitutive gene promoters. Plant Cell Reports 37(8): 1113–1125. https://doi.org/10.1007/s00299-018-2298-1
Guerra-Lupián MA, Ruiz-Medrano R, Ramírez-Pool JA, Ramírez-Ortega FA, López-Buenfil JA, Loeza-Kuk E, Morales-Galván O, Chavarin-Palacio C, Hinojosa-Moya J, Xoconostle-Cázares B. Localized expression of antimicrobial proteins mitigates huanglongbing symptoms in Mexican lime. J Biotechnol. 2018 Nov 10;285:74-83. https://doi.org/10.1016/j.jbiotec.2018.08.012. Epub 2018 Sep 5.
Hijaz, F., Y., Nehela, S. E., Jones, M. Dutt, J. W., Grosser, J. A., Manthey and N. Killiny. 2018. Metabolically engineered anthocyanin-producing lime provides additional nutritional value and antioxidant potential to juice. Plant Biotechnology Reports, 12(5):329-346. https://doi.org/10.1007/s11816-018-0497-4
Killiny N., S.E. Jones, Y. Nehela, F. Hijaz, M. Dutt, F.G. Gmitter and J.W. Grosser (2018) All roads lead to Rome: Towards understanding different avenues of tolerance to huanglongbing in citrus cultivars. Plant Physiology and Biochemistry129:1-10. https://doi.org/10.1016/j.plaphy.2018.05.005
Levy, A., El-Mohtar, C., Wang, C. et al. A new toolset for protein expression and subcellular localization studies in citrus and its application to citrus tristeza virus proteins. Plant Methods 14, 2 (2018). https://doi.org/10.1186/s13007-017-0270-7
Liu, Z., X. X. Ge, W. Qiu, J. M. Long, H. H. Jia, W. Yang, M. Dutt, X. M. Wu and W Guo. 2018. Overexpression of a B3 transcription factor CsFUS3 promotes somatic embryogenesis in Citrus. Plant Science 277: 121-131. https://doi.org/10.1016/j.plantsci.2018.10.015
Ma, H., Wang, M., Gai, Y., Fu, H., Zhang, B., Ruan, R., Chung, K., & Li, H. (2018). Thioredoxin and glutaredoxin systems required for oxidative stress resistance, fungicide sensitivity and virulence of Alternaria alternata. Applied and Environmental Microbiology, 84(14). https://doi.org/10.1128/AEM.00086-18
Niedz, R.P., Marutani-Hert, M. A filter paper-based liquid culture system for citrus shoot organogenesis—a mixture-amount plant growth regulator experiment. In Vitro Cell.Dev.Biol.-Plant 54, 658–671 (2018). https://doi.org/10.1007/s11627-018-9940-z
Omar, A.A., Murata, M.M., El-Shamy, H.A. et al. Enhanced resistance to citrus canker in transgenic mandarin expressing Xa21 from rice. Transgenic Res 27, 179–191 (2018). https://doi.org/10.1007/s11248-018-0065-2
Vu TX, Ngo TT, Mai LTD, Bui TT, Le DH, Bui HTV, Nguyen HQ, Ngo BX, Tran VT. A highly efficient Agrobacterium tumefaciens-mediated transformation system for the postharvest pathogen Penicillium digitatum using DsRed and GFP to visualize citrus host colonization. J Microbiol Methods. 2018 Jan;144:134-144. https://doi.org/10.1016/j.mimet.2017.11.019. Epub 2017 Nov 23.
Acanda, Y., Canton, M., Wu, H., and Zale, J. (2017). Kanamycin selection in temporary immersion bioreactors allows visual selection of transgenic citrus shoots. Plant Cell Tissue and Organ Culture 129, 351-357.
De Francesco, A., Costa, N., and Garcia, M.L. (2017). Citrus psorosis virus coat protein-derived hairpin construct confers stable transgenic resistance in citrus against psorosis A and B syndromes. Transgenic Res. 26, 225-235.
Noelia Sendin, L., Georgina Orce, I., Liliana Gomez, R., Enrique, R., Grellet Bournonville, C.F., Sergio Noguera, A., Alberto Vojnov, A., Rosa Marano, M., Pedro Castagnaro, A., and Paula Filippone, M. (2017). Inducible expression of Bs2 R gene from Capsicum chacoense in sweet orange (Citrus sinensis L. Osbeck) confers enhanced resistance to citrus canker disease. Plant Mol. Biol. 93, 607-621.
Orbovic, V., Fields, J.S., and Syvertsen, J.P. (2017). Transgenic citrus plants expressing the p35 anti-apoptotic gene have altered response to abiotic stress. Horticulture Environment and Biotechnology 58, 303-309.
Shimada, T., Endo, T., Rodriguez, A., Fujii, H., Goto, S., Matsuura, T., Hojo, Y., Ikeda, Y., Mori, I.C., Fujikawa, T., Pena, L., and Omura, M. (2017). Ectopic accumulation of linalool confers resistance to Xanthomonas citri subsp citri in transgenic sweet orange plants. Tree Physiol. 37, 654-664.
Zhang, Y., Zhang, D., Zhong, Y., Chang, X., Hu, M., and Cheng, C. (2017). A simple and efficient in planta transformation method for pommelo (Citrus maxima) using Agrobacterium tumefaciens. Scientia Horticulturae 214, 174-179.
Zou, X., Jiang, X., Xu, L., Lei, T., Peng, A., He, Y., Yao, L., and Chen, S. (2017). Transgenic citrus expressing synthesized cecropin B genes in the phloem exhibits decreased susceptibility to Huanglongbing. Plant Mol. Biol. 93, 341-353.
Boscariol-Camargo, R.L., Takita, M.A., and Machado, M.A. (2016). Bacterial resistance in AtNPR1 transgenic sweet orange is mediated by priming and involves EDS1 and PR2. Tropical Plant Pathology 41, 341-349.
Dutt, M., Erpen, L., Ananthakrishnan, G., Barthe, G.A., Brlansky, R.H., Maiti, I.B., and Grosser, J.W. (2016). Comparative expression analysis of five caulimovirus promoters in citrus. Plant Cell Tissue and Organ Culture 126, 229-238.
Dutt, M., Barthe, G., Irey, M., and Grosser, J. (2016). Transgenic Citrus Expressing an Arabidopsis NPR1 Gene Exhibit Enhanced Resistance against Huanglongbing (HLB; Citrus Greening) (vol 10, e0137134, 2015). Plos One 11, e0147657.
Hao, G., Pitino, M., Duan, Y., and Stover, E. (2016). Reduced Susceptibility to Xanthomonas citri in Transgenic Citrus Expressing the FLS2 Receptor From Nicotiana benthamiana. Mol. Plant-Microbe Interact. 29, 132-142.
Hao, G., Stover, E., and Gupta, G. (2016). Overexpression of a Modified Plant Thionin Enhances Disease Resistance to Citrus Canker and Huanglongbing (HLB). Frontiers in Plant Science 7, 1078.
Hu, W., Li, W., Xie, S., Fagundez, S., McAvoy, R., Deng, Z., and Li, Y. (2016). Kn1 gene overexpression drastically improves genetic transformation efficiencies of citrus cultivars. Plant Cell Tissue and Organ Culture 125, 81-91.
Jia, H., Orbovic, V., Jones, J.B., and Wang, N. (2016). Modification of the PthA4 effector binding elements in Type I CsLOB1 promoter using Cas9/sgRNA to produce transgenic Duncan grapefruit alleviating XccpthA4:dCsLOB1.3 infection. Plant Biotechnology Journal 14, 1291-1301.
Mcnellis, T., Gottwald, T., Sinn, J., and Orbovic, V. (2016). Phenotypic effects of anti-Candidatus Liberibacter asiaticus antibody expression in grapefruit. Phytopathology 106, 26-26.
Reyes, C.A., De Francesco, A., Ocolotobiche, E.E., Costa, N., and Garcia, M.L. (2016). Uncontrolled Citrus psorosis virus infection in Citrus sinensis transgenic plants expressing a viral 24K-derived hairpin that does not trigger RNA silencing. Physiol. Mol. Plant Pathol. 94, 149-155.
Wu, H., Acanda, Y., Jia, H., Wang, N., and Zale, J. (2016). Biolistic transformation of Carrizo citrange (Citrus sinensis Osb. x Poncirus trifoliata L. Raf.). Plant Cell Rep. 35, 1955-1962.
Yang, L., Hu, W., Xie, Y., Li, Y., and Deng, Z. (2016). Factors affecting Agrobacterium-mediated transformation efficiency of kumquat seedling internodal stem segments. Scientia Horticulturae 209, 105-112.
Zhang, X., Wang, W., Wang, M., Zhang, H., and Liu, J. (2016). The miR396b of Poncirus trifoliata Functions in Cold Tolerance by Regulating ACC Oxidase Gene Expression and Modulating Ethylene-Polyamine Homeostasis. Plant and Cell Physiology 57, 1865-1878.
Alvarez-Gerding, X., Cortes-Bullemore, R., Medina, C., Romero-Romero, J.L., Inostroza-Blancheteau, C., Aquea, F., and Arce-Johnson, P. (2015). Improved Salinity Tolerance in Carrizo Citrange Rootstock through Overexpression of Glyoxalase System Genes. Biomed Research International
Alvarez-Gerding, X., Espinoza, C., Inostroza-Blancheteau, C., and Arce-Johnson, P. (2015). Molecular and physiological changes in response to salt stress in Citrus macrophylla W plants overexpressing Arabidopsis CBF3/DREB1A. Plant Physiology and Biochemistry 92, 71-80.
Cheng Chun-zhen, Yang Jia-wei, Yan Hu-bin, Bei Xue-jun, Zhang Yong-yan, Lu Zhi-ming, and Zhong Guang-yan. (2015). Expressing p20 hairpin RNA of Citrus tristeza virus confers Citrus aurantium with tolerance/resistance against stem pitting and seedling yellow CTV strains. Journal of Integrative Agriculture 14, 1767-1777.
Dutt, M., Barthe, G., Irey, M., and Grosser, J. (2015). Transgenic Citrus Expressing an Arabidopsis NPR1 Gene Exhibit Enhanced Resistance against Huanglongbing (HLB; Citrus Greening). Plos One 10, e0137134.
Orbovic, V., Grosser, J.W. (2015). Citrus Transformation Using Juvenile Tissue Explants. Agrobacterium Protocols, Volume 2, Third Edition 1224, 245-257.
Orbovic, V., Shankar, A., Peeples, M.E., Hubbard, C., and Zale, J. (2015). Citrus Transformation Using Mature Tissue Explants. Agrobacterium Protocols, Volume 2, Third Edition 1224, 259-273.
Peng, A., Xu, L., He, Y., Lei, T., Yao, L., Chen, S., and Zou, X. (2015). Efficient production of marker-free transgenic 'Tarocco' blood orange (Citrus sinensis Osbeck) with enhanced resistance to citrus canker using a Cre/loxP site-recombination system. Plant Cell Tissue and Organ Culture 123, 1-13.
Soler, N., Fagoaga, C., Lopez, C., Moreno, P., Navarro, L., Flores, R., and Pena, L. (2015). Symptoms induced by transgenic expression of p23 from Citrus tristeza virus in phloem-associated cells of Mexican lime mimic virus infection without the aberrations accompanying constitutive expression. Molecular Plant Pathology 16, 388-399.
Wu, H., Acanda, Y., Shankar, A., Peeples, M., Hubbard, C., Orbovic, V., and Zale, J. (2015). Genetic Transformation of Commercially Important Mature Citrus Scions. Crop Sci. 55, 2786-2797.
Gong, X., Zhang, J., and Liu, J. (2014). A stress responsive gene of Fortunella crassifolia FcSISP functions in salt stress resistance. Plant Physiology and Biochemistry 83, 10-19.
Li Ding-li, Xiao Xuan, and Guo Wen-wu. (2014). Production of Transgenic Anliucheng Sweet Orange (Citrus sinensis Osbeck) with Xa21 Gene for Potential Canker Resistance. Journal of Integrative Agriculture 13, 2370-2377.
Ma YuanYuan, Zou XiuPing, Peng AiHong, Xu LanZhen, He YongRui, and Chen ShanChun. (2014). Ectopic expression analysis of Limonoid UDP-glucosyltransferase gene ( citLGT) in transgenic Citrus sinensis 'Jincheng'. Journal of Fruit Science 31, 181-186.
Muniz, F.R., Souza, A., Harakava, R., Alves Mourao Filho,Francisco de Assis, Stach-Machado, D.R., Rezende, J.A.M., Febres, V.J., Moore, G.A., and Mendes, B.M.J. (2014). Reaction of transgenic Citrus sinensis plants to Citrus tristeza virus infection by Toxoptera citricida. Eur. J. Plant Pathol. 139, 151-159.
Pinheiro, T.T., Figueira, A., and Latado, R.R. (2014). Early-flowering sweet orange mutant 'x11' as a model for functional genomic studies of Citrus. BMC Research Notes 7, 511-511.
Rodriguez, A., Shimada, T., Cervera, M., Alquezar, B., Gadea, J., Gomez-Cadenas, A., Jose De Ollas, C., Jesus Rodrigo, M., Zacarias, L., and Pena, L. (2014). Terpene Down-Regulation Triggers Defense Responses in Transgenic Orange Leading to Resistance against Fungal Pathogens. Plant Physiol. 164, 321-339.
Rossignol, P., Orbovic, V., and Irish, V.F. (2014). A dexamethasone-inducible gene expression system is active in Citrus plants. Scientia Horticulturae 172, 47-53.
Sun, L., Zhang, J., Mei, L., and Hu, C. (2014). Molecular cloning, promoter analysis and functional characterization of APETALA 1-like gene from precocious trifoliate orange (Poncirus trifoliata L. Raf.). Scientia Horticulturae 178, 95-105.
Xiao, X., Ma, F., Chen, C., and Guo, W. (2014). High efficient transformation of auxin reporter gene into trifoliate orange via Agrobacterium rhizogenes-mediated co-transformation. Plant Cell Tissue and Organ Culture 118, 137-146.
Zou, X., Song, E., Peng, A., He, Y., Xu, L., Lei, T., Yao, L., and Chen, S. (2014). Activation of three pathogen-inducible promoters in transgenic citrus (Citrus sinensis Osbeck) after Xanthomonas axonopodis pv. citri infection and wounding. Plant Cell Tissue and Organ Culture 117, 85-98.
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