How do you make acetosyringone stock solution?
Acetosyringone (AS) – 100mM Stock (mw. 196.20 g/mol): • Dissolve in 95% ethanol and adjust volume with water. Dissolve 0.3924 g in 12 ml 95% ethanol, then add 8 ml of sterile milli-Q water to equal 20 ml. Filter Sterilize and store at -20°C.
Can we autoclave acetosyringone?
Acetosyringone is a naturally occurring compound secreted from wounded dicot plant tissues and is a chemotractant for Agrobacterium sp. It is generally used at 5 uM to 200 uM to enhance transformation in monocots. Acetosyringone should NOT be autoclaved. This is a sterile solution of Acetosyringone in DMSO.
How do you store acetosyringone?
Storage instructions Store at Room Temperature. Store under desiccating conditions. The product can be stored for up to 12 months.
How do you dissolve acetosyringone?
Acetosyringone is soluble in organic solvents such as ethanol, DMSO, and dimethyl formamide (DMF), which should be purged with an inert gas. The solubility of acetosyringone in ethanol is approximately 2 mg/ml and approximately 20 mg/ml in DMSO and DMF.
What is acetosyringone used for?
Acetosyringone is known to improve transformation efficiency by up-regulating the virulence of Agrobacterium. In previous studies using Eustoma, the effect of this compound has only been tested during inoculation.
Why is acetosyringone used?
What is the function of acetosyringone?
Acetosyringone is a phenol secreted by wounded plant tissues. It induces expression of virulence A genes and chemotaxis in A. tumefaciens strains that contain a tumor-inducing plasmid used to transfer genetic information to plant cells.
What is Acetosyringone used for?
How do you make Agrobacterium competent cells?
Distribute culture into 8 x 50 ml tubes and pellet by centrifugation at 3000 – 4000 x g for 20-30 minutes at 4°C. Discard supernatant and resuspend each pellet in 10 ml ice cold H2O. Adjust volume to 50 ml each with ice cold H2O. Pellet cells by centrifugation at 3000 – 4000 x g for 20-30 minutes at 4°C.
How do you dissolve Acetosyringone?
What is Agrobacterium used for?
Uses in biotechnology. The ability of Agrobacterium to transfer genes to plants and fungi is used in biotechnology, in particular, genetic engineering for plant improvement. Genomes of plants and fungi can be engineered by use of Agrobacterium for the delivery of sequences hosted in T-DNA binary vectors.
What is vacuum infiltration?
The term “vacuum infiltration”, as used herein, relates to a method to allow the penetration of pathogenic bacteria, e.g. Agrobacterium, into the intercellular or interstitial spaces and in that way study the interaction between plants and pathogenic bacteria.
What is acetosyringone?
Acetosyringone (AS) is a phenol secreted by wounded plant tissues.Acetosyringone is a phenolic natural product with analgesic, antipyretic and anti-inflammatory actions. 0-4℃ for short term (days to weeks), or -20℃ for long term (months).
How to prepare acetosyringone in autoclave?
I’ve prepared my acetosyringone by dissolving 196.2 mg of it in both 10 ml of DMSO and ethanol separately. Acetosyringone prepared in DMSO precipitated out when I added it into my autoclaved MS medium. As for ethanol-dissolved acetosyringone, it evaporated as soon as I added it into my autoclaved medium.
What is the maximum um of acetosyringone?
It is generally used at 5 uM to 200 uM to enhance transformation in monocots. Acetosyringone should NOT be autoclaved. This is a sterile solution of Acetosyringone in DMSO.
How does acetosyringone activate Virg in Agrobacterium?
Acetosyringone, which attracts the bacteria to the wounded plant, also induces the virulence genes, thus facilitating the transfer of the T-DNA region ( Fig. 23.25 ). Acetosyringone binds to VirA protein in the Agrobacterium membrane. This activates VirG, which in turn switches on the other vir genes, including virD and virE2.