Transgenic Paulownia elongata S. Y. Hu plants using biolistic-mediated transformation

Abstract

A biolistic protocol for the stable genetic transformation of the hardwood tree Paulownia elongata was developed. Leaf explants were bombarded using the PDS-1000/He system with plasmid pBI121. The introduced DNA contained the b-glucuronidase (GUS) reporter gene and neomycin phosphotransferase (nptII) as a selection marker. Transformed calli were induced and selected on medium supplemented with 50 mg L -1 kanamycin, and transgenic plants were regenerated through indirect organogenesis. Complete plants were successfully transferred to soil and established under greenhouse conditions. Different helium pressures and explant positions were used and the transformation frequency was calculated. Optimal conditions for genetic transformation were bombardment of the abaxial leaf surface at a pressure of 450 psi. The integration of the transgenes in the plant genome and their stable expression was demonstrated by fluorometric GUS assay, determination of NPTII activity and PCR analysis. This method allows the production of transgenic trees of P. elongata in a relatively short time.

Biolistic Transformation of Citrullus vulgaris Schrad (Watermelon)

“…The significance levels (Tukey HSD) between treatments = 0.149 between treatments = 1.00 (Perez-Barranco et al, 2009). However, in a hardwood tree (Paulownia elongata) and garden balsam ( Impatiens balsamina) DNA concentration of 1.66 and 1.0 µg shown to produce the maximum expression for abaxial leaf surface and cotyledonous explants, respectively (Castellanos-Hernandez et al, 2009;Taha et al, 2009). …”

“…2009). In a hardwood tree (Paulownia elongata), 450 psi One of the most important parameters to optimize for was found to be the best helium pressure as compared to biolistic transformation is the target distance between the 900 and 1350 psi when bombarded on abaxial leaf surface macrocarrier to the target tissue in the bombardment (Castellanos-Hernandez et al, 2009). However, in hop chamber.…”

“…This were significantly reduced. This may be due to decrease target distance is similar to the target distance used for in velocity of the microcarriers as a result of increasing sugarcane and hardwood tree (Paulownia elongata) flight distance which consequently reduced penetration (Jain et al, 2007;Castellanos-Hernandez et al, 2009). In force and followed by fewer cells receiving the hop (Humulus lupulus L.) and garden balsam (Impatiens incoming DNA.…”

Histología de la regeneración por organogénesis en Paulownia tomentosa (Paulowniaceae)

“…Paulownia is also useful because of its high transpiration rates, widespread root system, and elevated tolerance to high concentrations of metals in both hydroponic and field studies, and is a good candidate for the phytoremediation of polluted soils (Doumett et al, 2008). The species is also used for the land reclamation of nutrient-poor soils (Marcotrigiano & Jagannathan, 1988), as a fast growing ornamental tree (Castellanos-Hernández et al, 2009), and its leaves and flowers can be used for medicinal purposes, they are rich in nitrogen, serving as good fertilizer and fodder (Zhu, Chao, Lu, & Xiong, 1986).A prerequisite for genetic improvement of this biofuel feedstock plant (Kausch et al, 2010) is the establishment of an efficient transformation system. The use of transgenic tools to improve plant feedstock are required in order to diversify the energy sources, and to obtain plants that produce cellulases or ligninases, plants with lower lignin content or with increased biomass, suitable for producing biofuel (Beltrán, 2008).…”

“…Paulownia is also useful because of its high transpiration rates, widespread root system, and elevated tolerance to high concentrations of metals in both hydroponic and field studies, and is a good candidate for the phytoremediation of polluted soils (Doumett et al, 2008). The species is also used for the land reclamation of nutrient-poor soils (Marcotrigiano & Jagannathan, 1988), as a fast growing ornamental tree (Castellanos-Hernández et al, 2009), and its leaves and flowers can be used for medicinal purposes, they are rich in nitrogen, serving as good fertilizer and fodder (Zhu, Chao, Lu, & Xiong, 1986).…”