Biolistic transformation of wheat using the HMW-GS 1Dx5 gene without selectable markers

Abstract

ABSTRACT. Genetic manipulation using linear DNA was applied to the common wheat variety Xindong No. 26 via particle bombardment with the aim to improve bread-making quality of flour. Initially, 2 biolistic parameters, helium pressure and target distance, were optimized using plasmid pAHC25. We transformed wheat immature embryo scutella with the linear 1Dx5 gene without selectable markers. The highest transient β-glucuronidase expression was obtained when scuttles were bombarded at 1100 psi with a 9-cm target distance. Using the optimized parameters, the transformation of the common wheat variety Xindong No. 26 was carried out using the linear 1Dx5. Three transgenic plants were identified from 1003 transgenic plants, yielding a transformation frequency of 0.4%. A sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis confirmed that the 1Dx5 gene was expressed in 4 T1 seeds of the transgenic plants. These experiments indicate the possibility of obtaining marker-free transgenic wheat plants via particle bombardment using the minimal gene cassettes with the particle bombardment parameters.

Current status and trends of wheat genetic transformation studies in China

“…Cai et al (2013) studied the effects of different media on biolistic transformation efficiency, and obtained highest percentage (65%) of transgenic plantlets by the procedure: the calli were induced on Cu 2+ containing medium and pre-cultured for 4 d before biolistic-mediated transformation, followed by culturing on R medium. Qin et al (2014) obtained transgenic wheat overexpressing 1Dx5 gene without selectable markers from immature embryo scutella by optimizing the biolistic parameters. found that embryo with some endosperm residues after 4°C overnight in soaking could improve the frequency of callus induction and regeneration in wheat mature embryo culture system.…”

Study of transgene expression in Triticum aestivum L. after Agrobacterium-mediated in planta transformation

“…Згідно з опублікованими даними, більшість трансгенних рослин пшениці отримані за допомогою прямого переносу генів -біолістичним методом (Altpeter et al, 1996;Lazzeri, Jones 2009;Vasil et al, 1992;Weeks et al, 1993;Zhou et al, 1995;Qin et al, 2014;Hamada et al, 2017). Тим не менш, така трансформація має ряд обмежень та недоліків, основним з яких є можливі перебудови перенесених ДНК касет та мультикопійність, що зумовлюють «замовчування» відповідних генів (Бончук и др., 2014: пат.…”