Factors influencing Agrobacterium tumefaciens-mediated genetic transformation of Eleusine coracana (L.) Gaertn

Abstract

Agrobacterium-mediated transformation protocol has been developed for Eleusine coracana (var. PR-202) by varying several factors which influence T-DNA delivery. Green nodular regenerative calli with meristematic nodules of seed origin were used as the target tissue for Agrobacterium tumefaciens-mediated gene transfer. The highest frequency of transformation (44.4%) was observed when callus was infected, co-cultivated and incubated at 22°C. Incorporation of higher level of CuSO 4 in the regeneration medium had significantly positive effect on the recovery of transformed plants. PCR analysis of T 0 and T 1 generation plants with nptII-specific primers revealed the amplification of nptII gene. Southern blot analysis of six regenerated plants confirmed selectable marker gene integration in three plants. This is a first report on Agrobacterium-mediated genetic transformation of finger millet and will pave the way for further studies in this and other millet crops.

Agrobacterium-mediated transformation and shoot regeneration in elite breeding lines of western shipper cantaloupe and honeydew melons (Cucumis melo L.)

“…Factors during co-cultivation can enhance transformation efficiency, i.e., temperature (Fullner and Nester 1996;Yasmin and Debener 2010;Sharma et al 2011;Seo et al 2011), lighting conditions (Zambre et al 2003), co-cultivation period (Fang and Grumet 1990;Shilpa et al 2010;Seo et al 2011), addition of acetosyringone (Costa et al 2006;Afroz et al 2010;Sharma et al 2011) and antioxidants (Dan et al 2009;Olhoft and Somers 2001;Ostergaard and Yanofsky 2004;Toldi et al 2002;Zheng et al 2005;Kumar et al 2011;Dutt et al 2011). Co-cultivation periods of 2-6 d were reported to be optimal for melons (Galperin et al 2003;Vallés and Lasa 1994;Akasaka-Kennedy et al 2004;Dong et al 1991).…”

In vitro transformation of pearl millet (Pennisetum glaucum (L). R. BR.): Selection of chlorsulfuron-resistant plants and long term expression of the gus gene under the control of the emu promoter

“…This result confirms the fact that the resistance of selected calli to chlorsulfuron is due to the expression of the ALS transgene. Despite the progress and the new interest in using Agrobacteriummediated methods for in vitro transformation of graminaceous (Jha et al, 2011;Sharma et al, 2011;Plaza-Wüthrich and Tadele, 2012;Hiei et al, 2014), the particles bombardment is still an effective technique for the introduction of transgenes in monocot plant cells with an average transformation frequency of more than 50% (Goldman et al, 2003). Thus, our results indicate that the biolistic method is suitable for in vitro transformation of P. glaucum varieties of Côte d'Ivoire because these varieties are recalcitrant to plants regeneration from protoplasts (Tiécoura et al, 2015).…”

“…However, several transformation systems leading to the introduction of DNA into monocotyledon intact tissues have been developed: for example electroporation into organized plant tissues (Dekeyser et al, 1990), silicon carbide fiber-mediated DNA transfer into maize suspension cells (Kaeppler et al, 1990;Sood et al, 2011) and the microprojectile bombardment or biolistic method (Klein et al, 1987;Prathibha and Sticklen, 2002;Martinez-Trujillo et al, 2003;Singh et al, 2013). Microprojectile bombardment is currently the most used method for in vitro transformation of graminaceous even if improvements of the Agrobacterium-mediated transformation method have been achieved (Liu et al, 2005;Razzaq et al, 2010;Sharma et al, 2011;Plaza-Wüthrich and Tadele 2012). The biolistic method offers an effective alternative for the genetic transformation of cereal crops.…”

Alternative Healthy Food Crops

“…Recently efforts were made to develop highly efficient transformation protocol for finger millets both with Agrobacterium [18] and Biolistic gun to open up new window to fight with the most devastating fungal disease [18,19].…”