Shoot regeneration in stem expiants and its amenability to Agrobacterium tumefaciens mediated gene transfer in Brassica carinata

1992 ◽  
Vol 11 (7) ◽  
Author(s):  
S.B. Narasimhulu ◽  
P.B. Kirti ◽  
T. Mohapatra ◽  
Shyam Prakash ◽  
V.L. Chopra
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Shoot Regeneration ◽  
Brassica Carinata

scholarly journals The effect of magnetic field strength on shoot regeneration and Agrobacterium tumefaciens-mediated gene transfer in flax (Linum usitatissimum L.)

2019 ◽  
Vol 55 (No. 1) ◽  
pp. 20-27
Author(s):  
Murat Aycan ◽  
Ramazan Beyaz ◽  
Anzel Bahadir ◽  
Mustafa Yildiz
Keyword(s):  
Magnetic Field ◽  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Shoot Regeneration ◽  
Linum Usitatissimum ◽  
Naphthaleneacetic Acid ◽  
Nptii Gene ◽  
Ms Medium ◽  
Generator System ◽  
Linum Usitatissimum L

This study was conducted to determine the effects of magnetic field (MF) strength on shoot regeneration and Agrobacterium tumefaciens-mediated gene transfer in flax (Linum usitatissimum L.). Seeds of flax cv. Madaras were exposed to different MF strengths (0 – control, 75, 150, and 300 millitesla (mT)) for 24 h by using an electromagnetic generator system fabricated in laboratory conditions. After sterilization, seeds were germinated on MS (Murashige and Skoog) medium in Magenta vessels. Hypocotyl explants excised from 7-days-old seedlings were used for regeneration. GV2260 strain of Agrobacterium tumefaciens was used in transformation studies. Inoculated hypocotyls were cultured on MS medium containing 1 mg/l BAP (6-benzylaminopurine) and 0.02 mg/l NAA (naphthaleneacetic acid) for 2 days by co-cultivation. Then, they were transferred to MS medium containing the same growth regulators, 100 mg/l kanamycin and 500 mg/l Duocid for selection. The presence of the nptII gene was verified by PCR (polymerase chain reaction) analysis in putative transgenic plants. The highest results with respect to shoot regeneration and transformation frequency were obtained from treatments of 75 mT MF strength.  

scholarly journals Strategies to produce T-DNA free CRISPRed fruit trees via Agrobacterium tumefaciens stable gene transfer

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Lorenza Dalla Costa ◽  
Stefano Piazza ◽  
Valerio Pompili ◽  
Umberto Salvagnin ◽  
Alessandro Cestaro ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Genomic Dna ◽  
Fruit Trees ◽  
Plant Genome ◽  
Stable Gene ◽  
Binary Vectors ◽  
Exogenous Dna ◽  
Left Behind ◽  
Dicotyledonous Plants

AbstractGenome editing via CRISPR/Cas9 is a powerful technology, which has been widely applied to improve traits in cereals, vegetables and even fruit trees. For the delivery of CRISPR/Cas9 components into dicotyledonous plants, Agrobacterium tumefaciens mediated gene transfer is still the prevalent method, although editing is often accompanied by the integration of the bacterial T-DNA into the host genome. We assessed two approaches in order to achieve T-DNA excision from the plant genome, minimizing the extent of foreign DNA left behind. The first is based on the Flp/FRT system and the second on Cas9 and synthetic cleavage target sites (CTS) close to T-DNA borders, which are recognized by the sgRNA. Several grapevine and apple lines, transformed with a panel of CRISPR/SpCas9 binary vectors, were regenerated and characterized for T-DNA copy number and for the rate of targeted editing. As detected by an optimized NGS-based sequencing method, trimming at T-DNA borders occurred in 100% of the lines, impairing in most cases the excision. Another observation was the leakage activity of Cas9 which produced pierced and therefore non-functional CTS. Deletions of genomic DNA and presence of filler DNA were also noticed at the junctions between T-DNA and genomic DNA. This study proved that many factors must be considered for designing efficient binary vectors capable of minimizing the presence of exogenous DNA in CRISPRed fruit trees.

scholarly journals Integrative gene transfer in the truffle Tuber borchii by Agrobacterium tumefaciens-mediated transformation

AMB Express ◽  
2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Andrea Brenna ◽  
Barbara Montanini ◽  
Eleonora Muggiano ◽  
Marco Proietto ◽  
Patrizia Filetici ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Tuber Borchii

Development of Auxotrophic Agrobacterium tumefaciens for Gene Transfer in Plant Tissue Culture

2004 ◽  
Vol 20 (3) ◽  
pp. 890-896 ◽  
Author(s):  
J.I. Collens ◽  
D.R. Lee ◽  
A.M. Seeman ◽  
W.R. Curtis
Keyword(s):  
Tissue Culture ◽  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Plant Tissue Culture ◽  
Plant Tissue

scholarly journals Optimization of Agrobacterium tumefaciens-mediated transformation and shoot regeneration after co-cultivation of cabbage (Brassica oleracea subsp. capitata) cv. KY Cross with AtHSP101 gene

2010 ◽  
Vol 124 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Arash Rafat ◽  
Maheran Abd Aziz ◽  
Azmi Abd Rashid ◽  
Siti Nor Akmar Abdullah ◽  
Hossein Kamaladini ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Brassica Oleracea ◽  
Shoot Regeneration
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