An Efficient Agrobacterium-Mediated Transformation of Strawberry cv. Camarosa by a Dual Plasmid System

Abstract

An Agrobacterium-mediated transformation method was applied to introduce the luciferase reporter gene under the control of the CaMV35S promoter in the pGreen0049 binary vector into strawberry cv. Camarosa. The in vitro regeneration system of strawberry leaves to be used in the transformation was optimized using different TDZ concentrations in MS medium. TDZ at 16 µM showed the highest percentage (100%) of shoot formation and the highest mean number of shoots (24) produced per explant. Studies on the effects of different antibiotics, namely timentin, cefotaxime, carbenicillin and ampicillin, on shoot regeneration of strawberry leaf explants showed the best shoot regeneration in the presence of 300 mg/L timentin and 150 mg/L cefotaxime. Assessment of the different factors affecting Agrobacterium mediated-transformation of strawberry with the luciferase gene showed the highest efficiency of putative transformant production (86%) in the treatment with no preculture, bacterial OD600 of 0.6 and the addition of 150 mg/L cefotaxime in the pre-selection and selection media. The presence of the luciferase gene in the plant genome was verified OPEN ACCESSMolecules 2015, 20 3648 by the luciferase reporter gene assay, nested PCR amplification and dot blot of genomic DNA isolated from the young leaves of each putatively transformed plantlet.

Transformasi Gen Proteolisis 6 (PRT6) Berperantarakan Agrobacterium tumefaciens ke dalam Kotiledon Tomato kultivar Micro Tom

“…Tempoh masa ko-penanaman selama 48 jam meningkatkan peluang pemindahan transgen daripada A. tumefaciens ke dalam eksplan (Tan et al 2017). Kajian lepas oleh Sun et al (2006) (Haddadi et al 2015). Antibiotik turut digunakan bagi merencat pertumbuhan A. tumefaciens dalam proses penyaringan transgenik putatif.…”

Effect of Ceftriaxone and Timentin antibiotics on morphogensis in the in vitro culture of bread wheat Triticum aestivum L.

“…In some cases Timentin (250 mg/l) has no negative effect on the callus growth, for example, in rice (Oryza sativa L.) [6]. There are reports that Timentin stimulates morphogenetic processes in plants [7][8][9][10][11][12][13]. It increases organogenesis from leaf explants of tobacco Nicotiana tabacum (150 mg/l) [10] and cotyledon of tomato explants (300 mg/l) [11], callus formation of mature wheat germ (50 mg/l) [9], shoots regeneration from strawberry leaf explants (300 mg/l) [13] and the regeneration of white ash (Fraxinus americana) (500 mg/l) [7], efficient plant regeneration from protoplasts of three carrots cultivars (400-500 mg/l) [12] and the shoot induction in tomato (400 mg/l) [8].…”

“…All researchers, which apply antibiotics when working with plant culture, report on their effects on the morphogenetic processes. In some cases the antibiotics Timentin and Ceftriaxone inhibit plant regeneration [3][4][5], in others -vice versa, increase it [6][7][8][9][10][11][12][13]. Accordingly, the objective of our study was to find out how antibiotics Timentin and Ceftriaxone affect morphogenic callus ISSN 1993-6842 (on-line); ISSN 0233-7657 (print) Biopolymers and Cell.…”

Establishment of a Rapid and Stable Transgenic System by Agrobacterium-Mediated Transformation of the Germination Seeds in Diploid Strawberry.

“…Strawberry (Fragaria), as a distinct member of Rosaceae, is not only an important fruit crop worldwidely, but is also regarded as a model for studying non-climacteric fruit ripening [1]. In the past years, a large of genes related to strawberry fruit ripening have been identi ed via traditional stable and transient transgenic systems in strawberry; however, more or less, the limitation is existed [2][3][4]. Thus, development of a rapid, e cient, and stable transformation system is required for strawberry research and breeding.…”

“…It is known that strawberry leaf tissue has been extensively studied to develop e cient transformation and regeneration methods through somatic tissue or callus formation [5][6][7][8][9][10][11][12][13][14]. However, the regeneration of transgenic strawberry plants not only remains di cult, but also takes a long time from transformation to identi cation [3,15,16]. Such as, it takes at least 15 months from transformation to gaining transgenic fruits for gene function analysis in ripening [17,18].…”