Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

Abstract

Abstract:Soybean transformation by ovary-drip was improved by optimizing the length of the transformation pathway by cutting the styles. These modifications facilitated soybean transformation manipulation and improved transformation reproducibility and efficiency. Using a linear minimal gus gene cassette as the foreign DNA, a maximum transformation frequency of 11% was obtained in flowers of the soybean cultivar 'Liaodou 14' with their styles mostly removed, whereas removal of only the stigma, partial style cutting and partial ovary cutting gave transformation frequencies of 0%, 1%, and 2%, respectively. An average transformation frequency of 8.2% was obtained when 619 flowers from three soybean cultivars ('Liaodou 14', 'Liaodou 13', and 'Tiefeng 29') were transformed by this optimized method. Southern blotting analysis showed that the gus reporter gene (encoding β-glucuronidase) was stably inherited with a simple pattern. Reverse transcription-polymerase chain reaction (RT-PCR) and GUS staining confirmed the expression of the gus gene in transgenic plants.

Transgene integration complexity and expression stability following biolistic or Agrobacterium-mediated transformation of sugarcane

“…To date, MC technology has been used to introduce genes of interest into several plant species such as rice (Oryza sativa; Fu et al 2000;Breitler et al 2002;Loc et al 2002;Agrawal et al 2005;Zhao et al 2007), corn (Zea mays; Lowe et al 2009;Prakash et al 2009), wheat (Triticum aestivum; Yao et al 2006Yao et al , 2007, creeping bentgrass (Agrostis stolonifera; Jayaraj et al 2008), bahiagrass (Paspalum notatum, Sandhu and Altpeter 2008), sugarcane (Saccharum spp. hybrids, Taparia et al 2012a;Taparia et al 2012b;Jackson et al 2013), soybean (Glycine max; Vianna et al 2004;Gao et al 2008;Liu et al 2009), grapevine (Vitis vinifera; Vidal et al 2006), potato (Solanum tuberosum; Romano et al 2003), and common bean (Phaseolus vulgaris; Vianna et al 2004).…”

Female reproductive system of <i>Amaranthus</i> as the target for <i>Agrobacterium</i>-mediated transformation

“…In order to reduce the amount of AGL1 solution needed, infiltration solution containing transformed AGL1 was pipetted rather than dipping the whole plant [22]. This was preferred as flowering Amaranthus reaching up to 90 cm tall are difficult to be dipped.…”

Plant Transformation via Pollen Tube-Mediated Gene Transfer

“…For instance, even though PTT has yielded transformants in soybean (Glycine max L.) (Hu and Wang 1999;Liu et al 2009b), it has not been universally successful (Li et al 2002). Where PTT was unsuccessful in soybean, the expression of beta glucuronidase (GUS) activity of seed recovered from some treated plants was endogenous (i.e., exogenous DNA detected in the cytoplasm but not integrated into the recipient's genome) (Li et al 2002).…”