Agrobacterium tumefaciens Modified Roy and Sasser Medium for Grapevine Strains

[所属分类:培养基配方] [发布时间:2019-9-23] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Agrobacterium tumefaciens Modified Roy and Sasser
Medium for Grapevine Strains
Composition per 1020mL:
Agar............................................................................................15.0g
Adoniitol.......................................................................................4.0g
H 3 BO 3 ...........................................................................................1.0g
K 2 HPO 4 .........................................................................................0.9g
KH 2 PO 4 .........................................................................................0.7g
NaCl..............................................................................................0.2g
MgSO 4 ·7H 2 O................................................................................0.2g
Yeast extract................................................................................0.14g
Cycloheximide solution...........................................................10.0mL
Triphenyl tetrazolium chloride solution ....................................1.0mL
D -Cycloserine solution...............................................................1.0mL
Trimethoprim solution...............................................................1.0mL
Cycloheximide Solution:
Composition per 10.0mL:
Cycloheximide..........................................................................0.025g
Preparation of Cycloheximide Solution: Add cycloheximide to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Filter sterilize.
Caution:  Cycloheximide is toxic. Avoid skin contact or aerosol for-
mation and inhalation.
Triphenyl Tetrazolium Chloride Solution:
Composition per 10.0mL:
Triphenyltetrazolium chloride ......................................................0.8g
Preparation of Triphenyl Tetrazolium Chloride Solution:
Add triphenyltetrazolium chloride to distilled/deionized water and
bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
D -Cycloserine Solution:
Composition per 10.0mL:
D -Cycloserine................................................................................0.2g
Preparation of  D -Cycloserine Solution: Add  D -cycloserine to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Filter sterilize.
Trimethoprim Solution:
Composition per 10.0mL:
Trimethoprim............................................................................0.025g
Preparation of Trimethoprim Solution: Add trimethoprim to
distilled/deionized water and bring volume to 10.0mL. Add a drop of
dilute HCl. Mix thoroughly. Gently heat while mixing until dissolved.
Filter sterilize.
Preparation of Medium: Add components, except cycloheximide
solution and triphenyl tetrazolium chloride solution,  D -cycloserine so-
lution, and trimethoprim solution to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Adjust pH to 7.2. Distribute 100.0mL
into flasks. Gently heat and bring to boiling. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 50°C. Aseptically add, per 100.0mL of
medium, 0.1 mL sterile triphenyl tetrazolium chloride solution, 0.1mL
sterile  D -cycloserine solution, 0.1mL sterile trimethoprim solution, and
1.0mL cycloheximide solution. Mix thoroughly. Aseptically pour into
sterile Petri dishes.
Use: For the selective cultivation of Agrobacterium tumefaciens biovar 3.