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The transformation frequency is also affected by slight differences in transformation conditions or the physiological state of the recipient cells (Michielse et al. 2005a).

A. tumefaciens C58C1 and a binary vector, pBIG2RHPH2, seem to be appropriate for the AMT of A. aculeatus because the transformation frequency is influenced by differences in the binary vector or bacterial strain used (Mullins et al. 2001).

Marker-free transformation frequency is defined as the percentage of shoots that is PCR positive.

It is known that marker-assisted transformation frequency is highly genotype dependent in potato [ 29].

Transformation efficiency was computed as transformants per microgram of DNA, and transformation frequency was expressed as transformants per microgram of DNA per viable cell.

Transformation frequency was expressed as percentage of transformants against total viable recipient cells per milliliter cell suspension.

The transformation frequency was calculated as the ratio of transformants that exhibited both resistance against paromomycin and the correct aphVIII fragment following PCR and the total number of cells used for each transformation experiment (9 × 10 cells).

Transformation frequency was calculated as the ratio of the transformant number to the recipient (competent) cell number.

Transformation frequency was calculated as the ratio of the transformant number to the estimated recipient cell number, which was regarded as half of the total cell number in each sample.

Unexpectedly, increasing the amount of this mixture did not increase the number of transformants per plate in a dose-dependent manner because the transformation frequency was reduced (Table 4).

Based on 5 independent transformation experiments, the transformation frequency was calculated as 3.7 × 10-7 (± 14%) transformants per cell.