Improvement of cold tolerance of the half-high bush Northland blueberry by transformation with the LEA gene from Tamarix androssowii

Abstract

Previous studies have shown that the late embryogenesis abundant (LEA) gene of Tamarix androssowii can enhance the drought tolerance of transgenic tobacco. In this study, the cloned LEA gene was transformed into half-high bush Northland blueberry in order to enhance its ability to tolerate cold stress. The cephalosporin antibiotics ceftriaxone, cefotaxime and cefazolin were used to optimize transformation of leaf explants, and kanamycin sulfate was used to select for transgenic shoots. PCR and Southern blot analysis confirmed 8 transformants with LEA gene copy numbers ranging from 1 to 7. The LEA chimeric gene was found to be normally transcribed in 6 transgenic lines by RT-PCR. The 8 transgenic lines were tested for cold tolerance by measuring the activities of peroxidase (POD) and superoxide dismutase (SOD), malondialdehyde (MDA) content and relative electrolyte leakage (REL). Overexpression of the LEA gene enhanced the activity of both POD and SOD under low temperature stress conditions. Lipid peroxidation in the transgenic lines was significantly lower than in non-transgenic plants after cold stress, as determined by MDA content and REL. Thus, our findings indicate that the LEA gene confers increased cold tolerance in the Northland blueberry and implicate the metabolic pathways through which it exerts this effect.

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

“…The first publications on the application of Timentin date back to 1997 [10]. Ceftriaxone became common in the Agrobacterium-mediated plant transformation rather recently [1,2,4,5].…”

“…It is known that β-lactam antibiotics adversely affect the regeneration of explants cultured in vіtro: Ceftriaxone at a concentration of 400 mg/l completely inhibits blueberry shoots growth [4], 200 mg/l -suppresses leaves, petioles and stems regeneration of ox knee (Achyranthes bidentata) [5]. High concentrations of Timentin (500-1000 mg/l) in culture medium, as well as all β-lactams, decrease the somatic embryoids regeneration in walnut [3].…”

“…or to eliminate any bacterial contamination (in particular during in vіtro Agrobacterium-mediated transformation -Cefo taxime, Carbenicillin, etc.). Recently new generation antibiotics of the β-lactam group, such as Timentin and Ceftriaxone are used more often [1][2][3][4][5][6][7][8][9][10][11].…”

“…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.…”

Effect of antibiotic ceftriaxone on elimination of ABI and GV3101 strains of Agrobacterium tumefaciens

“…Ceftriaxone, a third-generation cephalosporin, characterized by a prolonged half-value period, has the broad-spectrum activity. The successful elimination of various Agrobacterium tumefaciens strains (EHA105, LBA4404, AGL_1) by means of ceftriaxone has been earlier reported [9][10][11][12][13]. However, it has not been yet used for the elimination of ABI Agrobacterium strain.…”