Transgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system

Abstract

After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.

Integration and expression of Bluetongue VP2 gene in somatic embryos of peanut through particle bombardment method

“…for checking the transformation efficiency [11]. Forty milligrams per liter concentration of 2,4-D used in the induction medium had good proliferation amongst all concentrations tested.…”

“…1A). It is known that osmosis can alleviate the damage caused after bombardment [11], and incubation in mannitol helps in decreasing such damages after bombardment. Maximum survival was observed when the 5 days precultured embryos were used with 3 h of incubation in osmosis medium prior to bombardment (Table 1).…”

“…Each plate had 7-12 precultured embryos that were incubated in osmotic medium for 1-3 h. The acceleration with which the microcarriers were bombarded with the explants was 1200 psi and the sample platform was positioned 5 cm below the launch assembly in all experiments. Each bombardment delivered approximately 1 mg DNA and 300 mg tungsten particle 1.0 mm in diameter [7,11]. Selection for Kanamycin resistant somatic embryos was initiated on the 12th day of post-bombardment on M.S medium containing 30-50 mg/L Kanamycin.…”

“…The effects of pre culture and osmosis treatment on transformation were checked as previously described [11]. After 1-7 days of pre-culture in induction medium, part of the pre-cultured embryos were treated for 1-3 h by osmosis medium (induction medium containing 0.4 M mannitol) and the embryos were used for particle bombardment.…”

Effects of osmoticum treatments and shooting chances on the improvement of particle gun-mediated transformation in Phalaenopsis

“…그러나 대부분 아그로박테리 움을 이용한 논문이고 10여 편 중 유전자총을 이용한 연 구는 3편에 불과하다 (Roh et al 2011). (Denchev et al 1997), 옥수수 (Vain et al 1993), peanut (Deng et al 2001)과 난과식물에서는 온시디움 (Li et al 2005 …”