The effect of temperature on Agrobacterium tumefaciens‐mediated gene transfer to plants

Abstract

Agrobacterium tumefaciens is generally used to achieve genetic transformation of plants. The temperatures that have been used for infection with Agrobacterium in published transformation protocols differ widely and, to our knowledge, the effect of temperature on the efficiency of T-DNA transfer to plants has not been investigated systematically. Agrobacterium tumefaciens strains harbouring a binary vector with the beta-glucuronidase (uidA) gene and either a nopaline-, an octopine- or an agropine/succinamopine-type helper plasmid were tested in two transformation systems at temperatures between 15 and 29 degrees C. One system involved cocultivation of Phaseolus acutifolius callus whereas in the other system Nicotiana tabacum leaves were vacuum-infiltrated. In both situations, irrespective of the type of helper plasmid, the levels of transient uidA expression decreased notably when the temperature was raised above 22 degrees C. Expression was low at 27 degrees C and undetectable at 29 degrees C. We anticipate that the efficiency of many published transformation protocols can be improved by reconsidering the factor of temperature

Agrobacterium tumefaciens Mediated Genetic Transformation in Coker-312 (Gossypium hirsutum L.) Using Hypocotyls Explants

“…Jin et al, (2005) suggested that co-cultivation at 19 0 C for 48 hours was effective for the efficient transformation which was confirmed earlier by Dillen et al, (1997) and Sunilkumar and Rathore (2001).…”

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

“…In the present study, transformation frequency was highest when co-cultivation was performed at 24°C. Co-cultivation at low temperature has been reported in several instances Kapila et al 1997;Dillen et al 1997;Popelka et al 2006). It has also been found that the temperature dependence profile of T-DNA transfer does not parallel with that of vir gene induction, indicating that another factor possibly the formation of a conjugal pilus is causing temperature sensitivity of T-DNA transfer .…”

Transgenic Indian Cotton (Gossypium hirsutum) Harboring Rice Chitinase Gene (Chi II) Confers Resistance to Two Fungal Pathogens

“…Although cotton transformation rates have been significantly improved, increasing its efficiency on plant recovery is still needed [30] . Many factors such as plant genotype, explants type, pH, co-cultivation media, On growth response and transformation frequency of shoot tip explants temperature and period influence the gene transfer efficiency from Agrobacterium to plant cells and the above external factors play a critical role in T-DNA transfer mechanism in particularly vir gene activation [31] .…”