PPM plant tissue culture antibacterial protective agent instructions

Item No.: PPM

Storage conditions: 4o C

Transportation conditions: normal temperature

Physical properties: solution

Properties : colorless to amber transparent liquid

pH: 3.8

Validity period : see bottle label description

product description:

1.          PPM ^TM It is a heat-stable antibacterial agent for the prevention and elimination of microbial contamination of plants in plant tissue culture;

2.          At a reasonable dosage, PPM ^TM is a very effective antibacterial and protective agent for plant tissue culture. It does not have any effect on plant growth (such as seed germination, callus growth, regeneration, etc.). Base standard formula ingredients;

3.          PPM ^{TM is} mainly used to prevent and eliminate microbial contamination in plant tissue culture caused by external factors such as air, water or human contact. At the same time, it also has a very good inhibitory effect on endophytic bacteria.

4.          PPM ^{TM is} widely used in most angiosperms and gymnosperms, but is not recommended for aquatic plants such as ferns and algae;

5.          PPM ^TM has thermal stability and can be directly added to the medium for high temperature sterilization without affecting the use effect, and is more convenient to use;

6.          PPM ^TM More cost-effective than conventional antibiotics.

Mechanism of action :

The active ingredient of PPM ^TM penetrates the bacterial or fungal cell wall, inhibits the activity of key enzymes in the tricarboxylic acid cycle and electron transport chain during respiration, and also blocks the absorption and utilization of monosaccharides and amino acids in the culture medium by bacteria and fungi. The process of preventing and eliminating microbial contamination during plant tissue culture.

Advantages of relative antibiotics:

1.          PPM ^TM can widely inhibit and eliminate bacterial and fungal contamination and prevent the germination of fungal spores;

2.          PPM ^TM achieves bacteriostatic effects by inhibiting the activity of various enzymes, thereby avoiding the occurrence of drug-resistant mutants;

3.          PPM ^TM has thermal stability, does not need filtration sterilization, can be directly added to the medium for high temperature sterilization without affecting the use effect, and is more convenient to use;

4.          PPM ^TM is more cost-effective and can be used in a wider range of scientific research, crop breeding, and tissue culture production.

Product patent description:

PPM ^{TM Â}  - Plant Preservative Mixture is a patented product whose product name, formula and method of use are protected by patents, patent number 5,750,402 .

Introduction to general use :

The following usage guidelines are for general use instructions in most cases, and special circumstances can be adjusted as appropriate.

* Description: MS salt solution is a general reference, such as the sample medium salt solution is not MS salt, based on the specific medium salt solution;

Instructions:

The following instructions are for general use instructions in most cases. If necessary, make adjustments according to the specific plant sample. For more guidance, please contact Dr. Assaf Guri at .

1.       Regular dosage:
It is generally recommended to use a concentration of 0.05%-0.2% ( 0.5-0.75ml PPMTM is added to 1L medium ), and the concentration of callus proliferation, organ formation, and embryogenic tissue development is 0.05%-0.075% .

Description:
(a) The medium containing PPMTM does not need to be dispensed in a clean bench. After the medium is solidified, the flask / dish lid is covered.   

Ok. If using an automatic filling system, it is recommended to treat the dispensing tube with autoclaved hot water before and after filling;
(b) If the culture medium is stored in a sterile container and is not contaminated, add PPMTM- prepared medium.

No need to filter again or autoclave. If it is a nutrient medium, such as a medium containing 200 mg/L or higher of amino acid or protein, it is recommended to filter and sterilize.

2.       Plant endophytic contamination treatment :

(a) Seeds: PPMTM is not recommended for direct sterilization of seeds containing large amounts of bacterial or fungal spores. For seed germination, it is recommended to disinfect them by conventional methods and then place them in 2-3% PPMTM . In the culture base broth solution, gently stir for 4-12 hours. In this process, Tween20 should not be added and the pH should be adjusted . After the treatment is completed , the medium containing PPMTM ( 0.05-0.1% of herbaceous plants and 0.2% of woody plants ) can be directly inserted . in;

(b) Explants: Take 1cm explants as an example. Place the explants in a 4-5% PPMTM whole culture base salt solution and gently stir for 4-12 hours. This process must not be added. Tween20 and pH adjustment , after the completion of the treatment, directly into the medium containing PPMTM ( 0.05-0.1% herbaceous plant, 0.2% woody plant ) ;

(c) The ornamental plant specimens tubers, bulbs and squamous thereof: its entirety stirred into the disinfectant after disinfection, rinse with water, the sheet material is cut, placed in 4-5% P PM â„¢ containing the a whole broth culture basal salt mixture, stirred gently for 4-12 hours, this process must not add Tween20 and adjusting the pH, without rinsing after completion of the processing directly into medium containing 0.1-0.2% of PPM â„¢'s cultured.

3.       If thick explants, heavily polluted plants, seeds, etc. are still not ideal after the above treatment, try the following:

(a) Soak the material in water and continue to stir (the soft tissue is stirred for 1 hour, and the hard tissue is stirred for 2 hours ) ;

(b) The material PPM â„¢ containing 50% whole broth a basal salt solution is stirred for 5-10 minutes, this process must not Tween20 added and the pH adjusted;

(c) The treated material can be directly added to the medium without rinsing. For fungal contamination, the appropriate concentration of PPMTM can be added to the medium . Mixed bacterial contamination fungi, recommends adding 0.05% -0.2% in the first month of PPM â„¢ in the culture medium.

4.       Removal and rescue of serious pollution materials (no more than one week of heavy pollution):

(a) The contaminated material is brushed with soft brush in running water and then placed in a sterile aqueous solution containing 50% PPMTM for 5-15 minutes. For bacterial or mixed contamination, 100% PPMTM and 0.6g are recommended. /L citric acid sterile aqueous solution is treated with a 1 : 1 mixed low pH (2.8-3.2) acidic solution;

(b) without washing the treated material directly into PPM â„¢ medium containing 0.05% -0.25 cultured at least one month, wherein the first 10 days of culture required low light;

For some fungal, spore and bacterial contamination hidden deep, PPMTM can not reach the site, it is recommended to cut the material after the initial cleaning, and then put into a sterile aqueous solution containing 50% PPMTM and stir 5-15 Minute processing.

5.       Removal of Agrobacterium:

After co-cultivation, the sample was washed with sterile water, and then the sample was immersed in 100% PPMTM (supplemented with 4X medium salt solution) for 2 minutes, taken out, blotted dry with sterile paper and placed in common resistance. The medium was cultured, and after 3 weeks, it was changed to a medium containing only 0.05% - 0.075% PPMTM ( without common antibiotics ) .

Precautions for use :

1.       In all PPM TM disinfection treatment of explants, try to ensure that the volume of the container is large enough, and that all areas of the explant material are in full contact with the PPM- containing treatment solution to ensure the disinfection effect;

2.       If the explants are highly oxidized, do not throw them away. About 50% of the explants will recover within 4-6 weeks.

3.       The 50% PPMTM treatment solution can be reused but is not recommended because the number of uses and effects are affected by the volume and seeding density of the explant. Store the 50% PPMTM treatment solution at 4oC to prolong its activity, generally no more than 10 times. If necessary, it is recommended to prepare 2 parts of 50% PPMTM solution, one for endophyte disinfection of endophytic bacteria, and the other for eliminating lightly contaminated materials during "culture", the second solution is in each After the treatment, it needs to be filtered with a 0.2mm filter;

4.       If the 50% PPMTM solution is not ideal, it can be treated with 100% PPMTM solution in the same way, but not more than 10 times.

safety instructions:

1.       Although PPM TM is non-toxic, it is also necessary to avoid direct contact with human skin during use. It is recommended to wear gloves to keep the air in circulation. If you inadvertently inhale, immediately seek medical treatment immediately after a large amount of purified water. If you accidentally get into the eyes, immediately rinse your eyes with plenty of water for 15 minutes, and wash the skin parts that PPMTM has touched with soap ;

PPM â„¢ medium containing waste with the same conventional medium processing method.

If you have any questions, please feel free to contact Plant Cell Technology China General Agent - Beijing Ximeijie Technology Co., Ltd.

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