Table 5.
Heat-shocked and regenerated plants from different HSP::TRANSPOSASE/ATDs double transgenic aspen lines without any phenotypic alterations (anonymous approach) grown in the greenhouse were randomly selected and tested for ATDs transposition with the primer pair 16/37.
| Transgenic line | Tested plants | PCR-positive (16/37) (%) | TAIL-PCR and positive BLAST hits | Transcript annotation |
|---|---|---|---|---|
| N82-2 | 22 | 3 | 0 | --- |
| N82-5 | 26 | 11 | 5 | 3 |
| N82-7 | 24 | 4 | 0 | --- |
| N82-11 | 20 | 4 | 1 | 0 |
| N82-14 | 13 | 4 | 2 | 0 |
| N82-15 | 25 | 14 | 8 | 6 |
| N92-1 | 19 | 16 | 6 | 4 |
| N92-2 | 19 | 10 | 0 | --- |
| N92-3 | 20 | 0 | 0 | --- |
| N92-4 | 10 | 6 | 1 | 0 |
| N95-1 | 23 | 2 | 0 | --- |
| N95-2 | 19 | 2 | 0 | --- |
| N95-3 | 19 | 9 | 1 | 1 |
| N95-4 | 10 | 0 | 0 | --- |
| N95-5 | 10 | 0 | 0 | --- |
| N95-6 | 21 | 10 | 2 | 1 |
| SUM (%) | 300 | 95 (32%) | 26 (8.7%) | 15 (5.0%) |
Positive candidates were subjected to TAIL-PCR and sequencing to determine the new ATDs genomic insertion locus. Obtained sequences were blasted against the publicly available genome sequence of P. trichocarpa v2.0 (Phytozome v7.0; http://www.phytozome.net/poplar). Numbers of positive BLAST hits and, if applicable, of transcript annotations are given.
In bold the three lines with highest numbers of positive ATDs transposition and BLAST hits.