IMPROVEMENT OF INNER SOYBEAN DISEASE RESISTANCE BY GENETIC ENGINEERING OF THE PHENYLPROPANOID CYCLE: MOLECULAR DETECTION OF TRANSGENIC PLANTS
Main Article Content
Authors
О.I. Kershanskaya
Institute Plant Biology and Biotechnology, 45 Timiryazev str., 050040, Almaty, Kazakhstan
М.А. Abdulzhanova
Institute Plant Biology and Biotechnology, 45 Timiryazev str., 050040, Almaty, Kazakhstan
G.L. Esenbaeva
Institute Plant Biology and Biotechnology, 45 Timiryazev str., 050040, Almaty, Kazakhstan
D.S. Nelidova
Institute Plant Biology and Biotechnology, 45 Timiryazev str., 050040, Almaty, Kazakhstan
O.V. Zernova
University of Illinois at Urbana-Champaign, Edward R. Madigan Laboratory, 1201 W. Gregory Drive, Urbana, 61801, Il, USA
V.V. Lozovaya
University of Illinois at Urbana-Champaign, Edward R. Madigan Laboratory, 1201 W. Gregory Drive, Urbana, 61801, Il, USA
J.M. Widholm
University of Illinois at Urbana-Champaign, Edward R. Madigan Laboratory, 1201 W. Gregory Drive, Urbana, 61801, Il, USA
Abstract
Soybean diseases in Kazakhstan are a serious problem that reduces soybean yield up to 15%. However, information is lacking on these diseases. Plant resistance is an economical and sustainable disease management option. Genetic engineering of the key metabolic pathway components, which result in a broad range of products, including improving complex plant resistance to stress and increasing yield, is needed. One of the most important metabolic pathways of secondary metabolism in the plant is the phenylpropanoid cycle, which involves, in particular, the formation of lignin and flavonoids. Efforts to increase the strength of the innate defense systems, including lignin biosynthesis, would help limit the colonization of these pathogens. Lignin is the most significant polymer on Earth after cellulose. The use of lignin biosynthesis genes for genetic transformation is only developing now, and it is unknown for soybeans. The objective of this study was to establish approaches to improve the innate resistance of soybeans to biotic stresses and to create resistant soybeans through genetic engineering of the phenylpropanoid pathway to increase the biosynthesis of lignin, which is a natural antimicrobial compound, in order to improve management of micro-pathogens-caused diseases. The main results and novelty of the study were that genetic constructions of valuable genes, including transcription factor Cs/MYB4sens and the main genes of the lignification process (35S/PAL, С4Н/F5H, and the antioxydative stress anti-ROX gene FeSOD) have been optimized and used for soybean genetic transformation. Transgenic soybean plants of the first T1 and second T2 generations with lignification genes integrated into the genome were confirmed by PCR and RT-PCR methods with a transformation efficiency of 5.63% in the first T1 and 75% in the second T2 generations.
Keywords
diseases, lignin, germ-line genetic transformation, molecular detection, soybean
Article Details
References
Kliebenstein D.J. Secondary metabolites and plant/environment interactions: a view through Arabidopsis thaliana tinged glasses. Plant Cell and Environ., 2004, vol. 27, pp. 675-684.
Ahuja I., Kissen R., Bones A. Phytoalexins in defense against pathogens. Trends in Plant Science, 2012, vol. 17, pp. 73-90. doi: 22209038. Available at: Crossref.
Grobinsky D.K., van der Graaff E., Roitsch T. Phytoalexintransgenics in crop protection – Fairy tale with a happy end? Plant Sci., 2012, vol. 195, pp. 54-70. doi: 22920999. Available at: Crossref.
Lozovaya V.V., Lygin A.V., Zernova O.V., Widholm J.M. Genetic engineering of plant; disease resistance by modification of thephenylpropanoid pathway. PlantBiosystems, 2005, vol. 139, pp. 20-23.
Aharoni A., Galili G. Metabolic engineering of the plant primary-secondary metabolism interface.Current Opin in Biotechnol., 2011, vol. 22, pp. 239-244. doi: 21144730. Available at: Crossref.
Jeandet P., Clement C., Courot E., Cordelier S. Modulation of PhytoalexinBiosyntesis in Engineered Plants for Disease Resistance. Int. J. Mol. Sci., 2013, vol. 14, pp. 14136-14170. doi: 23880860. Available at: Crossref.
Lozovaya V.V., Lygin A.V., Li S., Hartman G.L., Widholm J.M. Biochemical response of soybean roots to Fusariumsolani f. sp. glycinesinfection. Crop Sci., 2004, vol. 44, pp. 819-826.
Lozovaya V.V., Lygin A.V., Zernova O.V., Ulanov A.V., Li S., Hartman G.L., Widholm J.M. Modification of phenolic metabolism in soybean hairy roots through down regulation of chalcone synthase or isoflavone synthase. Planta,2007, vol. 225, pp. 665-679. doi: 16924535. Available at: Crossref.
Lygin A.V., Li S., Vital R., Widholm J.M., Hartman G.L., Lozovaya V.V. The importance of phenolic metabolism to limit the growth of Phakopsorapachyrhizi.Phytopathol., 2009, vol. 99, pp. 1412-1420.
Lygin A.V., Hill C.B., Zernova O.V., Crull L., Widholm J.M., Hartman G.L., Lozovaya V.V. Response of Soybean Pathogens to Glyceollin. Phytopathol., 2010, vol. 100, pp. 897-903. doi: 20701487. Available at: Crossref.
Lygin A.V., Zernova O.V., Hill C.B., Kholina N.A., Widholm J.M., Hartman G.L., Lozovaya V.V. Glyceollin is an important component of soybean plant defense against Phophthorasojae and Macrophominaphaseolina. Phytopathol., 2013, vol. 103, pp. 984-994. doi: 23617338. Available at: Crossref.
Lozovaya V.V., Lygin A.V., Zernova O.V., Li S., Hartman G.L., Widholm J.M. Isoflavonoid accumulation in soybean hairy roots upon treatment with Fusariumsolani. Plant Physiol. Biochem., 2004, vol. 42, pp. 671-679. doi: 15331097. Available at: Crossref.
Zernova O.V., Lygin A.V., Widholm J.M., Lozovaya V.V. Modification of isoflavones in soybean seeds via expression of multiple phenolic biosynthetic genes.Plant Physiol. Biochem, 2009, vol. 47, pp. 769-777. doi: 19539487. Available at: Crossref.
Zernova O.V., Lygin A.V., Pawlowski M.L., Kershanskaya O.I., Hill C.B., Hartman G.L., Widholm J. M., Lozovaya V.V. Regulation of Plant Immunity Through Modulation of Phytoalexin Synthesis. Phytopathology, 2014, vol. 104, pp. 843-850. doi: 24914895. Available at: Crossref.
Wilson R.F. The role of genomics and biotechnology in achieving global food security for high-oleic vegetable oil.Functional and Integrative Genomics, 2012, pp. 447-462. doi: 22790166. Available at: Crossref.
Hall R.D., Hardy N.W. Practical Applications of Metabolomics in Plant Biology.In Book: Plant Metabolomics: Methods and Protocols. Series: Methods in Molecular Biology, 2012, vol. 860, pp. 1-10. doi: 22351167. Available at: Crossref.
Iwase A., Matsui K., Ohme-Takagi M. Manipulation of plant metabolic pathways by transcription factors. Plant Biotechnology, 2009, vol. 26, pp. 29-38.
Perez-Clemente R.M., Vives V., Zandalinas S.I., Climent M.F.L. Biotechnological approaches to study plant responses to stress. BioMed Research International, 2013, vol. 2013, p.10. doi: 23509757. Available at: Crossref.
Tran L.S., Mochida. K. Functional genomics of soybean for improvement of productivity in adverse conditions. Journal of Oleo Science, 2010, vol. 61, pp. 357-367. doi: 20582712. Available at: Crossref.
Morgenthal K., Wienkoop S., Wolschin F., Weckwerth W. Integrative Profiling of Metabolites and Proteins. In Metabolomics. Methods in Molecular Biology, 2007, vol. 358, pp. 57-75. doi: 17035680. Available at: Crossref.
Li Z., Nelson R.L., Widholm J.M., Bent A. Soybean transformation via the pollen tube pathway. Soybean Genet Newslett, 2002, vol. 29, pp. 1-11.
Dixon R.A., Lamb C.J., Masoud S., Sewalt V. J.H., Paiva N.L. Metabolic engineering: prospects for crop improvement through the genetic manipulation of phenylpropanoid biosynthesis and defense responses. A review. Gene, 1996, vol. 179, pp. 61-71. doi: 8955630. Available at: Crossref.
Fumiko H., Shinya K., Tomonori S., Mikiko U., at el. Manipulation of phenylpropanoid-biosynthetic pathway by genetic engineering with bacterial genes.Proceedings of the Lignin Symposium, 2001, vol. 46, pp. 115-118.
Chapple C.C.S. Manipulation of lignin composition in plants using a tissue-specific promoter / Patent USA, N 6610908, 2003.
Harakava R. Genes encoding enzymes of the lignin biosynthesis pathway in Eucalyptus. Genetics and Molecular Biology, 2005, vol. 28, p. 12.