Rewiring carotenoid biosynthesis in plants using a viral vector

[EN] Plants can be engineered to sustainably produce compounds of nutritional, industrial or pharmaceutical relevance. This is, however, a challenging task as extensive regulation of biosynthetic pathways often hampers major metabolic changes. Here we describe the use of a viral vector derived from Tobacco etch virus to express a whole heterologous metabolic pathway that produces the health-promoting carotenoid lycopene in tobacco tissues. The pathway consisted in three enzymes from the soil bacteria Pantoea ananatis. Lycopene is present at undetectable levels in chloroplasts of non-infected leaves. In tissues infected with the viral vector, however, lycopene comprised approximately 10% of the total carotenoid content. Our research further showed that plant viruses that express P. ananatis phytoene synthase (crtB), one of the three enzymes of the heterologous pathway, trigger an accumulation of endogenous carotenoids, which together with a reduction in chlorophylls eventually result in a bright yellow pigmentation of infected tissues in various host-virus combinations. So, besides illustrating the potential of viral vectors for engineering complex metabolic pathways, we also show a yellow carotenoid-based reporter that can be used to visually track infection dynamics of plant viruses either alone or in combination with other visual markers.We thank Veronica Aragones and M. Rosa Rodriguez-Goberna for excellent technical assistance. This research was supported by Spanish Ministerio de Economia y Competitividad (MINECO) grants BIO2014-54269-R to J.-A.D., and BIO2014-59092-P and BIO2015-71703-REDT to M. R.-C. Financial support from the Generalitat Valenciana (PROMETEOII/2014/021), the Programa Iberoamericano de Ciencia y Tecnologia para el Desarrollo (Ibercarot 112RT0445), and the Generalitat de Catalunya (2014SGR-1434) is also acknowledged. E.M. is the recipient of a pre-doctoral fellowship (AP2012-3751) from the Spanish Ministerio de Educacion, Cultura y Deporte. B.L. is supported by a postdoctoral fellowship (FPDI-2013-018882) from MINECO.Majer, E.; Llorente, B.; Rodríguez-Concepción, M.; Daros Arnau, JA. (2017). Rewiring carotenoid biosynthesis in plants using a viral vector. Scientific Reports. 7. https://doi.org/10.1038/srep41645S7O’Connor, S. E. Engineering of secondary metabolism. Annu. Rev. Genet. 49, 71–94 (2015).Sainsbury, F. & Lomonossoff, G. P. Transient expressions of synthetic biology in plants. Curr. Opin. Plant Biol. 19, 1–7 (2014).Gleba, Y. Y., Tusé, D. & Giritch, A. Plant viral vectors for delivery by Agrobacterium. Curr. Top. Microbiol. 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High expression of tumour-associated trypsin inhibitor correlates with liver metastasis and poor prognosis in colorectal cancer

Increased expression of tumour-associated trypsin inhibitor (TATI) in tumour tissue and/or serum has been associated with poor survival in various cancer forms. Moreover, a proinvasive function of TATI has been shown in colon cancer cell lines. In this study, we have examined the prognostic significance of tumour-specific TATI expression in colorectal cancer, assessed by immunohistochemistry (IHC) on tissue microarrays (TMAs) with tumour specimens from two independent patient cohorts. Kaplan–Meier analysis and Cox proportional hazards modelling were used to estimate time to recurrence, disease-free survival and overall survival. In both cohorts, a high (>50% of tumour cells) TATI expression was an independent predictor of a significantly shorter overall survival. In cohort II, in multivariate analysis including age, gender, disease stage, differentiation grade, vascular invasion and carcinoembryonal antigen (CEA), high TATI expression was associated with a significantly decreased overall survival (HR=1.82; 95% CI=1.19–2.79) and disease-free survival (HR=1.56; 95% CI=1.05–2.32) in curatively treated patients. Moreover, there was an increased risk for liver metastasis in both cohorts that remained significant in multivariate analysis in cohort II (HR=2.85; 95% CI=1.43–5.66). In conclusion, high TATI expression is associated with liver metastasis and is an independent predictor of poor prognosis in patients with colorectal cancer

Relocation of the NIb gene in the tobacco etch potyvirus genome

Supplemental material for this article may be found at http://dx.doi.org/10.1128/JVI.03336-13[EN] Potyviruses express most of their proteins from a long open reading frame that is translated into a large polyprotein processed by three viral proteases. To understand the constraints on potyvirus genome organization, we relocated the viral RNA-dependent RNA polymerase (NIb) cistron to all possible intercistronic positions of the Tobacco etch virus (TEV) polyprotein. Only viruses with NIb at the amino terminus of the polyprotein or in between P1 and HC-Pro were viable in tobacco plants.This research was supported by grant BIO2011-26741 from the Spanish Ministerio de Economia y Competitividad (MINECO) to J.-A.D., grant PROMETEO/2010/019 from Generalitat Valenciana to S. F. E. and J.-A.D., and grants BFU2012-30805 and 22371 from MINECO and the John Templeton Foundation, respectively, to S. F. E. E. M. was supported by a predoctoral fellowship (AP2012-3751) from the Spanish Ministerio de Educacion, Cultura y Deporte. M.P.Z. was supported by a Juan de la Cierva postdoctoral contract (JCI-2011-10379) from MINECO and a Rubicon grant from the Netherlands Organization for Scientific Research (www.nwo.nl).Majer, E.; Salvador, Z.; Zwart, MP.; Willemsen, A.; Elena Fito, SF.; Daros Arnau, JA. (2014). Relocation of the NIb gene in the tobacco etch potyvirus genome. Journal of Virology. (88):4586-4590. https://doi.org/10.1128/JVI.03336-13S4586459088Roossinck, M. J. (2011). The big unknown: plant virus biodiversity. Current Opinion in Virology, 1(1), 63-67. doi:10.1016/j.coviro.2011.05.022Chirico, N., Vianelli, A., & Belshaw, R. (2010). Why genes overlap in viruses. Proceedings of the Royal Society B: Biological Sciences, 277(1701), 3809-3817. doi:10.1098/rspb.2010.1052Belshaw, R., Pybus, O. G., & Rambaut, A. (2007). The evolution of genome compression and genomic novelty in RNA viruses. 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Proceedings of the National Academy of Sciences, 92(2), 457-461. doi:10.1073/pnas.92.2.457Bedoya, L., Martínez, F., Rubio, L., & Daròs, J.-A. (2010). Simultaneous equimolar expression of multiple proteins in plants from a disarmed potyvirus vector. Journal of Biotechnology, 150(2), 268-275. doi:10.1016/j.jbiotec.2010.08.006Bedoya, L. C., Martínez, F., Orzáez, D., & Daròs, J.-A. (2012). Visual Tracking of Plant Virus Infection and Movement Using a Reporter MYB Transcription Factor That Activates Anthocyanin Biosynthesis. Plant Physiology, 158(3), 1130-1138. doi:10.1104/pp.111.192922Majer, E., Daròs, J.-A., & Zwart, M. (2013). Stability and Fitness Impact of the Visually Discernible Rosea1 Marker in the Tobacco etch virus Genome. Viruses, 5(9), 2153-2168. doi:10.3390/v5092153Bedoya, L. C., & Daròs, J.-A. (2010). Stability of Tobacco etch virus infectious clones in plasmid vectors. Virus Research, 149(2), 234-240. doi:10.1016/j.virusres.2010.02.004Chen, C.-C., Chen, T.-C., Raja, J. A. J., Chang, C.-A., Chen, L.-W., Lin, S.-S., & Yeh, S.-D. (2007). Effectiveness and stability of heterologous proteins expressed in plants by Turnip mosaic virus vector at five different insertion sites. Virus Research, 130(1-2), 210-227. doi:10.1016/j.virusres.2007.06.014Merits, A., Runeberg-Roos, P., Rajamäki, M.-L., Puustinen, P., Mäkeläinen, K., Saarma, M., … Kekarainen, T. (2002). Proteolytic processing of potyviral proteins and polyprotein processing intermediates in insect and plant cells. Journal of General Virology, 83(5), 1211-1221. doi:10.1099/0022-1317-83-5-1211Parks, T. D., Howard, E. D., Wolpert, T. J., Arp, D. J., & Dougherty, W. G. (1995). Expression and Purification of a Recombinant Tobacco Etch Virus NIa Proteinase: Biochemical Analyses of the Full-Length and a Naturally Occurring Truncated Proteinase Form. Virology, 210(1), 194-201. doi:10.1006/viro.1995.1331KIM, D.-H., PARK, Y. S., KIM, S. S., LEW, J., NAM, H. G., & CHOI, K. Y. (1995). Expression, Purification, and Identification of a Novel Self-Cleavage Site of the NIa C-Terminal 27-kDa Protease of Turnip Mosaic Potyvirus C5. Virology, 213(2), 517-525. doi:10.1006/viro.1995.0024Dolja, V. V., McBride, H. J., & Carrington, J. C. (1992). Tagging of plant potyvirus replication and movement by insertion of beta-glucuronidase into the viral polyprotein. Proceedings of the National Academy of Sciences, 89(21), 10208-10212. doi:10.1073/pnas.89.21.10208Fernández-Fernández, M. R., Mouriño, M., Rivera, J., Rodríguez, F., Plana-Durán, J., & García, J. A. (2001). Protection of Rabbits against Rabbit Hemorrhagic Disease Virus by Immunization with the VP60 Protein Expressed in Plants with a Potyvirus-Based Vector. Virology, 280(2), 283-291. doi:10.1006/viro.2000.0762Sánchez, F., Sáez, M., Lunello, P., & Ponz, F. (2013). Plant viral elongated nanoparticles modified for log-increases of foreign peptide immunogenicity and specific antibody detection. 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Antagonist of potato common scab pathogen increases microbial diversity in rhizosphere

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Maan taudinestokykyä voi edistää

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First report of Streptomyces europaeiscabiei causing common scab on potato in Finland

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