Plant regeneration and Agrobacterium-mediated transformation of Achyranthes bidentata using cotton EREBP gene

Abstract

The aim of this work was to study the plant regeneration and Agrobacterium-mediated transformation of Achyranthes bidentata using cotton EREBP gene. Results showed that the callus induction rate of stems from A. bidentata was the highest (100%) and bud was in approximately 70% of calli from stems. However bud differentiation rate of the callus from leaves and petioles was very low. Compared with ceftriaxone, 200mg/L cefotaxime could completely control Agrobacterium tumefaciens and had relatively less toxic action on the stems of A. bidentata. In addition, the induction rate of callus resistant to hygromycin was the highest when infected for 3 min and co-cultivated for 3 d. Six positive transgenic plants transformed with pCAMBIA1304-GhEREB2 expression vector were obtained and confirmed by PCR. The expression of target gene GhEREB2 was detected in five transgenic plants by RT-PCR. In brief, an efficient system of genetic transformation and plant regeneration was established for A. bidentata.

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].…”

“…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.…”

Piercing and incubation method of in planta transformation producing stable transgenic plants by overexpressing DREB1A gene in tomato (Solanum lycopersicum Mill.)

“…Incubation period is a critical factor that incredibly influences explants survival and transformation efficiency because it is directly related with the sensitivity of explants and virulence of bacterial strain (Carvalho et al 2004;Orzaez et al 2006). Shorter time period is not favorable for attachment and T-DNA transfer by Agrobacterium into plant genome, while long period of infection might injure the explants resulting in necrosis, due to which competence of T-DNA transfer would be limited (Duan et al 2013). Co-cultivation period is another important factor that significantly affects transformation efficiency because T-DNA transfer into plant genome via Agrobacterium is a lengthy phenomenon that takes a few hours to some days depending on plant species, type of explants and culturing environment (Mayavan et al 2013).…”

Roles of plant growth substance in callus induction of Achyranthes bidentata

“…As is well known, the growth and development of plant need water, mineral and organic matter, and also are regulated by plant growth substance which is very critical to growth and differentiation of cell and tissue, induction and formation of bud and root (Melissa et al, 2006;Randy et al, 2009;Duan et al, 2013;Dragicevic et al, 2013;Gerszberg et al, 2015). At present, 2,4-dichlorophenoxy acetic acid (2,4-D), Naphthylacetic acid (NAA), 6-benzylaminopurine (6-BA) and other plant growth substances are often applied in plant tissue culture, in which NAA, 2,4-D and Indole butyric acid (IBA) belong to auxin and mainly play a role in callus formation, embryoid regeneration, and so on, yet 6-BA, Kinetin (KT) and Zeatin (ZT) 7 belong to cytokinin, and principally act on stimulants of cell division and induction of bud differentiation.…”