Refined glufosinate selection in Agrobacterium -mediated transformation of soybean [ Glycine max (L.) Merrill]

Abstract

Modern genetic analysis and manipulation of soybean ( Glycine max) depend heavily on an efficient and dependable transformation process, especially in public genotypes from which expressed sequence tag (EST), bacterial artificial chromosome and microarray data have been derived. Williams 82 is the subject of EST and functional genomics analyses. However, it has not previously been transformed successfully using either somatic embryogenesis-based or cotyledonary-node transformation methods, the two predominant soybean transformation systems. An advance has recently been made in using antioxidants to enhance Agrobacterium infection of soybean. Nonetheless, an undesirable effect of using these antioxidants is the compromised recovery of transgenic soybean when combined with the use of the herbicide glufosinate as a selective agent. Therefore, we optimized both Agrobacterium infection and glufosinate selection in the presence of L-cysteine for Williams 82. We have recovered transgenic lines of this genotype with an enhanced transformation efficiency using this herbicide selection system.

Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper)

“…DTT is also an efficient antioxidant with potential anti-necrotic property and is least toxic for plant cells (Svabova and Griga 2008). The beneficial role of thiol compounds on greater survival of explants during co-cultivation has been reported in soybean (Olhoft et al 2001(Olhoft et al , 2003Zeng et al 2004), Vigna unguiculata (Popelka et al 2006), Zea maize (Frame et al 2002) and Vitis vinifera (Das et al 2002a, b).…”

Tnt1 Retrotransposon Mutagenesis: A Tool for Soybean Functional Genomics

“…To investigate the feasibility of this approach, we tested the two published methods for soybean regeneration to gauge their ability to reactivate Tnt1 transposition. The cotyledons of the Tnt1-containing T1 plant seeds were used as explants for the first approach (Zeng et al, 2004). The explants were treated by wounding or wounding followed by 12 h of shaking in a 1 M Suc solution.…”

“…To investigate whether the Tnt1 element can transpose in soybean during tissue culture, Agrobacterium tumefaciens-mediated transformation was performed using a modified soybean cotyledonary node transformation protocol (Zeng et al, 2004). The plasmid pSH-Tnt1 containing the Tnt1 element was constructed by inserting the Tnt1 DNA into the binary vector pZY101, which carries a selectable bar gene marker for glufosinate resistance (Fig.…”

Application of somatic embryos to rapid and reliable analysis of soybean seed components by RNA interference-mediated gene silencing

“…Genetic transformation is an essential technique for verification of gene function, and genetic transformation of soybean has become routine with wellestablished methods mediated by Agrobacterium or particle bombardment (Finer and Mcmullen 1991;Kita et al 2007;Olhoft et al 2003;Sato S. et al 1993;Zeng et al 2004). It remains impractical, however, to verify the function of the large number of genes involved in determination of seed components with the use of transgenic soybean seeds, since it takes 3 to 4 months to generate transgenic plants, with transformation efficiencies being relatively low, and it takes several more months to obtain T 1 seeds (Sato H. et al 2007).…”