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Agrobacterium-Mediated Transformation of Sorghum bicolor Using Immature Embryos

 Last Update: 2020-11-17
 Source: Internet
 Author: User

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heat shock transformation

single use technology

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Successful efforts describing in vitro culturing, regeneration, and transformation of grain sorghum were reported, using particle bombardment, as early as 1993, and with
Agrobacterium tumefaciens
in 2000. Reported transformation efficiencies via
Agrobacterium
routinely range from 1 to 2%. Recently, such efficiencies via
Agrobacterium
in several plant species were improved with the use of heat and centrifugation treatments of explants prior to infection. Here, we describe the successful use of heat pretreatment of immature embryos (IEs) prior to
Agrobacterium
inoculation to increase routine transformation frequencies of a single genotype, P898012, to greater than 7%. This reproducible frequency was calculated as numbers of independently transformed IEs, confirmed by
PCR
, western, and
DNA
hybridization analysis, that produced fertile transgenic plants, divided by total numbers of infected IEs.

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