Agrobacterium tumefaciens-mediated transformation of Campanula carpatica: factors affecting transformation and regeneration of transgenic shoots

2004 ◽  
Vol 23 (1-2) ◽  
Author(s):  
Sridevy Sriskandarajah ◽  
Stefan Frello ◽  
Kirsten J�rgensen ◽  
Margrethe Serek
2000 ◽  
Vol 19 (3) ◽  
pp. 304-310 ◽  
Author(s):  
X. Niu ◽  
X. Li ◽  
P. Veronese ◽  
R. A. Bressan ◽  
S. C. Weller ◽  
...  

2011 ◽  
Vol 49 (No. 6) ◽  
pp. 255-260 ◽  
Author(s):  
J.I. Flores Solís ◽  
P. Mlejnek ◽  
K. Studená ◽  
S. Procházka

Chenopodium rubrum belongs to the plant species in which standard Agrobacterium-mediated transformation procedures remain inefficient. We demonstrate that the employment of sonication-assisted Agrobacterium-mediated transformation (SAAT) effectively enhanced transient expression of GUS gene coding for b-glucuronidase in Chenopodium rubrum. Further the results indicated that the age of seedlings is one of the limiting factors affecting the potency of Agrobacterium tumefaciens infection. Histochemical detection of b-glucuronidase activity revealed that two-days-old seedlings were much more susceptible to infection than ten-days-old ones. According to our results SAAT technology could provide an efficient tool for obtaining stable transformants when applied to two-days-old seedlings of Chenopodium rubrum.


1998 ◽  
Vol 25 (2) ◽  
pp. 207 ◽  
Author(s):  
M. Moralejo ◽  
F. Rochange ◽  
A.M. Boudet ◽  
C. Teuliéres

A procedure for genetic transformation of Eucalyptus globulus Labill. using Agrobacterium tumefaciens is described. Young seedlings obtained from mature seeds were chosen as target material. The most important factors affecting transformation efficiency were the nature of the A. tumefaciens strain, precultivation of the seedlings and wounding of the tissues. The optimised procedure allowed us to obtain the first transgenic plants for E. globulus. On selective medium, 1.2% of the inoculated seedlings regenerated shoots resistant to kanamycin and expressing the gusA gene. Molecular evidence for the presence of stably integrated T-DNA is provided for two plants.


2018 ◽  
Vol 44 (2) ◽  
pp. 323-326
Author(s):  
Nadia Sharif ◽  
Neelma Munir ◽  
Faiza Saleem ◽  
Shugufta Naz

Chlorella vulgarisis Bayerinck widely used as a health food, feed supplement, as well as in the pharmaceutical, biofuel and cosmetics industries. It has been used to determine optimum transformation conditions through Agrobacterium tumefaciens. It has been revealed that bacterial density of OD600 = 1.0, 3 days of co-cultivation at 25°C in pH 5.5, and 100 μM acetosyringone are the optimum conditions to transform C. vulgaris.


2021 ◽  
Vol 22 (8) ◽  
pp. 4181
Author(s):  
Fernando Perez Rojo ◽  
Sumedha Seth ◽  
William Erskine ◽  
Parwinder Kaur

Subterranean clover (Trifolium subterraneum) is the most widely grown annual pasture legume in southern Australia. With the advent of advanced sequencing and genome editing technologies, a simple and efficient gene transfer protocol mediated by Agrobacterium tumefaciens was developed to overcome the hurdle of genetic manipulation in subterranean clover. In vitro tissue culture and Agrobacterium transformation play a central role in testing the link between specific genes and agronomic traits. In this paper, we investigate a variety of factors affecting the transformation in subterranean clover to increase the transformation efficiency. In vitro culture was optimised by including cefotaxime during seed sterilisation and testing the best antibiotic concentration to select recombinant explants. The concentrations for the combination of antibiotics obtained were as follows: 40 mg L−1 hygromycin, 100 mg L−1 kanamycin and 200 mg L−1 cefotaxime. Additionally, 200 mg L−1 cefotaxime increased shoot regeneration by two-fold. Different plant hormone combinations were tested to analyse the best rooting media. Roots were obtained in a medium supplemented with 1.2 µM IAA. Plasmid pH35 containing a hygromycin-resistant gene and GUS gene was inoculated into the explants with Agrobacterium tumefaciens strain AGL0 for transformation. Overall, the transformation efficiency was improved from the 1% previously reported to 5.2%, tested at explant level with Cefotaxime showing a positive effect on shooting regeneration. Other variables in addition to antibiotic and hormone combinations such as bacterial OD, time of infection and incubation temperature may be further tested to enhance the transformation even more. This improved transformation study presents an opportunity to increase the feeding value, persistence, and nutritive value of the key Australian pasture.


HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 875E-875 ◽  
Author(s):  
Jyothi Prakash Bolar ◽  
Susan K. Brown ◽  
John L. Norelli ◽  
Herb S. Aldwinckle

The overall goal of our research is to develop an efficient transformation and regeneration system for `McIntosh' apple. The first objective was to determine the optimum combination of Gelrite (G) and agar (A) to maximize regeneration and minimize vitrification. Treatments included the following combinations of agar (in g–liter–1) and Gelrite (in g–liter–1): 1) 7 and 0; 2) 5.25 and 0.625; 3) 3.5 and 1.875; 4) 1.75 and 1.875; and 5) 0 and 2.5. There were 10 replications, and a single petri plate containing six leaf pieces was the unit of replication. Both 5.25(A) and 0.62(G) and 3.5(A) and 1.25(G) provided high regeneration of healthy, nonvitrified shoots. Since modification of media affects the concentration of antibiotics used in selection due to precipitation of antibiotics, the second objective was to determine the optimal concentration of antibiotic for the selection and regeneration of transformed `McIntosh' on gelrite–agar-based media. Kanamycin was tested at 0, 10, 25, 50, 75, and 100 μg–ml–1 and paromomycin was tested at 0, 50, 100, 150, 200, and 250 μg–ml–1. Antibiotic selection will be discussed relative to optimum concentration and efficiency of selection.


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