Abstract
Various steps of micropropagation include selection of suitable explant, establishment of adventitious shoot induction cultures, proliferation, rooting, and acclimatization of the resulting plantlets. A systematic protocol is provided for the micropropagation and Agrobacterium tumefaciens-mediated genetic transformation of a fast growing, multipurpose tree, Paulownia elongata. Our studies show that optimum shoot induction is on half leaf with petiole explant on MS medium supplemented with 25 μM thidiazuron and 10 μM indole-3 acetic acid. Micropropagation protocols provided here are applicable to explants collected from the primed in vitro raised seedlings on MS medium containing 2.5 μM 6-benzylaminopurine (BAP) or actively growing shoots collected from greenhouse or field growing plants. We also discuss a possible role of “Python” script guided protocol optimization for higher and consistent multiplication of shoots that can be very helpful for scaled up production in commercial settings. To facilitate future plant improvement and gene editing possibilities, an A. tumefaciens based genetic transformation protocol and molecular identification of transgenic plants using Polymerase Chain Reaction (PCR) and Reverse Transcriptase-PCR (RT-PCR) techniques have also been optimized.
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References
Bing L, Yun-Hong T, Liu S, Olmstead R, Min D-Z, Chen Z-D, Joshee N, Vaidya B, Chung R, Li B (2019) Phylogenetic relationships of Cyrtandromoea and Wightia revisited: a new tribe in Phrymaceae and a new family in Lamiales. J Syst Evol 58(1):1–17. https://doi.org/10.1111/jse.12513
El-Showk S, El-Showk N (2003) The Paulownia tree: an alternative for sustainable forestry. The Farm, 1–8. http://www.cropdevelopment.org/docs/PaulowniaBrochure_print.pdf
Tisserat B, Joshee N, Mahapatra AK, Selling GW, Finkenstadt VL (2013) Physical and mechanical properties of extruded poly(lactic acid)-based Paulownia elongata bio-composites. Ind Crop Prod 44:88–96. https://doi.org/10.1016/j.indcrop.2012.10.030
Tisserat B, Reifschneider L, Joshee N, Finkenstadt VL (2013) Evaluation of Paulownia elongata wood polyethylene composites. J Thermoplastic Comp Mat 28(9):1301–1320. https://doi.org/10.1177/0892705713505856
Tisserat B, Reifschneider L, Joshee N, Finkenstadt VL (2013) Properties of high density polyethylene—Paulownia wood flour composites via injection molding. BioRes 8(3):4440–4458
Stewart WM, Vaidya BN, Mahapatra AK, Terrill TH, Joshee N (2018) Potential use of multipurpose Paulownia elongata tree as an animal feed resource. Am J Plant Sci 9:1212–1227
Vaughn SF, Dinelli FD, Tisserat B, Joshee N, Vaughan MM, Peterson SC (2015) Creeping bentgrass growth in sand-based root zones with or without biochar. Sci Hortic 197:592–596. https://doi.org/10.1016/j.scienta.2015.10.021
Vaughn SF, Kenar JA, Tisserat B, Jackson MA, Joshee N, Vaidya BN, Peterson SC (2017) Chemical and physical properties of Paulownia elongata biochar modified with oxidants for horticultural applications. Ind Crop Prod 97:260–267. https://doi.org/10.1016/j.indcrop.2016.12.017
Yadav NK, Vaidya BN, Henderson K, Lee JF, Stewart WM, Dhekney SA, Joshee N (2013) A review of Paulownia biotechnology: a short-rotation, fast growing multipurpose bioenergy tree. Am J Plant Sci 4:2070–2082. https://doi.org/10.4236/ajps.2013.411259
Bajaj R (2018) Optimization of in vitro propagation and genetic transformation protocols in Paulownia elongata. M.S. Diss., College of Agric., Family Sci., and Technol., Fort Valley State Univ, Fort Valley, GA, p 65
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15(3):473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res 2:208–218
Burrow MD, Chlan CA, Sen P, Murai N (1990) High frequency generation of transgenic tobacco plants after modified leaf disk cocultivation with Agrobacterium tumefaciens. Plant Mol Biol Report 8:24–139
Vaidya BN, Jackson CL, Perry ZD, Dhekney SA, Joshee N (2016) Agrobacterium-mediated transformation of thin cell layer explants of Scutellaria ocmulgee Small: a rare plant with anti-tumor properties. Plant Cell Tissue Org Cult 127(1):57–69. https://doi.org/10.1007/s11240-016-1029-y
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325
Acknowledgments
R.B., L.I., and L.S. would like to thank financial assistance in the form of graduate research assistantship (GRA). We thank Avneesh Srivastava for heat maps. Financial assistance through Evans Allen grant GEOX 5213 and 5220 is appreciated (P.I. N. Joshee).
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Bajaj, R., Irvin, L., Vaidya, B.N., Shahin, L., Joshee, N. (2021). Optimization of Micropropagation and Genetic Transformation Protocols for Paulownia elongata : A Short Rotation Fast Growing Bioenergy Tree. In: Basu, C. (eds) Biofuels and Biodiesel. Methods in Molecular Biology, vol 2290. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1323-8_18
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DOI: https://doi.org/10.1007/978-1-0716-1323-8_18
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