Fig 1.
Schematic representation of A. tumefaciens-mediated transformation of ‘Olathe’ pinto bean using competent cells induced from embryo axes.
Fig 2.
Induction of competent cells from embryo axes for efficient regeneration of ‘Olathe’ pinto bean.
(A) Surface sterilized and soaked seeds for explant preparation. (B) Shoot production from embryo axes after 4-week culture on RM1, on which leaf explants did not survive. A. tumefaciens-mediated transformation of ‘Olathe’ pinto bean. (C, D) Embryo axes after 12-week preculture on RM. All shoots were removed during the subcultures in order to promote callus and bud formation. Bars = 1 cm.
Table 1.
Summary of three transformation experiments of common bean ‘Olathe’.
Fig 3.
A. tumefaciens-mediated transformation of ‘Olathe’ pinto bean.
(A) Explants after 4-day co-cultivation in the dark at 25 °C. (I, II) Half-seeds explants. (III) Precultured embryo axes explants. (IV) Embryo axes explants. (B) Precultured embryo axes explants after 3-week selection on selection RM1. (C) Embryo axes explants after 6-week selection on selection RM3. (D, E) GS-resistant calli and green buds induced from the precultured embryo axes explants after 6-week selection on selection RM3. (F) Rooting of the GS-resistant plants produced from the precultured embryo axes explants. Bars = 1 cm.
Fig 4.
Rooting of T0 ‘Olathe’ pinto bean and T1 seed production.
(A) Rooting of GS-resistant plants on selection MS. (B) Production of T1 seeds at 23–25 °C under a 16 h photoperiod of 55 μmol m-2s-1.
Fig 5.
(A, B) T1 plants of two transgenic lines of ‘Olathe’ pinto bean. Except 2–1*, all the other plants are RT-PCR-positive for the bar gene. (C) Seeds from transgenic 1–1 and nontransgenic (NT) 2–1 and NT plants.
Fig 6.
RT-PCR analysis of bar expression in leaves of T1 transgenic and nontransgenic (NT) plants of ‘Olathe’ pinto bean.
M: size ladder.
Fig 7.
Stable integration of the transgenes confirmed by genome sequencing of a T1 transgenic plant of the transgenic line #2.
(A) Schematic representation of the binary vector pDHB321.1. (B) Detection of the bar in the T-DNA region. (C) Detection of the pRiA4 sequence in the backbone region of the pDHB321.1.