PlantTC Assignment HanMeng

8/9/2019 PlantTC Assignment HanMeng

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Title:

Question 2: Design and plan the operation of the laboratory

Introduction:

Plant tissue culture is a propagation technique widely used in modern agriculture that

allows a complete plant to be grown from a single plant cell. Producing a plant from a

single cell in the laboratory is not a simple process. In the process, we need to use

more techniques and strict environments. In order to get more accurate result from

plant tissue culture, a good laboratory is needed. In the laboratory, there are more

instructors and equipments in it that we should set them up in a proper position.

Therefore, we can work well in the lab and provide a good surrounding.

Bodies:

A.

Culture room

Media preparation room

A

C

C

CH

DE

F

F

E

E

E

E

G

B

Comment [cyn1]: Media prep room should be

isolated from the laminar floor chambers, and

there is just one laminar floor chamber/hood?


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A: washing bath

B: autoclave

C: work table

D: teansference (transfer) chamber

E: incubate shelves

F: main lab bench

G: Laminar airflow chamber

H: refrigerator-freezer

B.

1. Autoclave

2.water bench

3. heat-dry sterilization unit

4. Laminar airflow chamber

5. refrigerator-freezer

6. Weighing scales

7. A hot plate with an automatic stirrer

8. The pH meter

9. Aspirator

10. Drying oven

11. Gyratory shaker or a reciprocal shaker

12. Glass atomizer

Comment [cyn2]: No description of the design

is provided


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13. Pipette washers

14. Centigram balances

15. Small transfer instruments

16. Hot air oven

17. Culture racks

18. Draining-boards

19. Optional equipment

C.

An estimation of the number of staff that you will require to produce 3 million tissue

cultured plants per year (assuming that each culture vessel can house 15 plants and

each person working 8 hours a day can handle 200 culture vessels a day) .

We can calculate in this way:

3000000/365/ (200*15) 3

To be frugal, we just required 2 persons a year that we can produce 3 million tissue

cultured plants per year.

D.

The preparation of media should be in the media preparation room. The media

preparation area is the room where material is sterilized, so it requires a good cleaning

to avoid media contamination before it is used. This area must be maintained in

isolation to protect it from dust and other air-borne contaminants. The media may be

Comment [cyn3]: What are these supposed to

be?

Comment [cyn4]: What about the preparation

of media? Stage 4 work? Cleaning/washing?

Also you have not factored in the

multiplication of cultures from Stage 1

4.time frame for initiation, how many

sub-culture you are allowed etc.


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purchased readymade or may be prepared in the laboratory, the latter being cheaper, if

needed in large quantities. The washing and sterilization facilities (autoclave, etc.) are

often kept outside the tissue culture laboratory, since they would generate moisture

(increased humidity) and heat. Facilities should also be available for the storage of the

following: 1. sterile liquids (media, serum, etc.) at 4C, - 20C or - 70C. 2. Sterile

glassware and plastic ware (bottles, flasks, pipettes, Petri dishes, syringes). 3. Screw

caps, filter tubes, stoppers, etc. 4. Gloves, disposable bags, etc. 5.other equipment

(filters, large receiver flasks). Media preparation area should be equipped with both

tap and purified water. An appropriate system for water purification must be selected

and fitted after careful consideration of the cost and quality. A number of electrical

appliances are required for media preparation; hence, it is essential to have safety

devices like fire extinguisher, fire blanket and a first aid kit in the media preparation

room. A variety of glassware, plastic ware and stainless steel apparatus is required for

measuring, mixing, and media storage. These should be stored in the cabinets built

under the worktables and taken out for use as and when required. This would save the

cost and space for building storage shelves. The use of glassware should be kept at a

minimum, as it will help in reducing losses due to breakage. As far as possible, plastic

ware and stainless steel vessels should be used, as they are much cheaper and more

durable than glassware. The water source and glassware storage area should be

convenient to the media preparation area. Benches, suitable for comfortable working

while standing and deep enough to hold equipment listed below are essential. Their

tops should be made with molded plastic laminate surfaces that can tolerate frequent

Comment [cyn5]: This was copied directly

from:

http://www.molecular-plant-biotechnology.inf

o/animal-biotechnology-genomics/laboratory-

facilities-culture-media-procedures/laboratory

-facilities-for-tissue-culture.html

Comment [cyn6]: This was copied directly

from:

http://dbtncstcp.nic.in/downloads/Plant%20Ti

ssue%20Culture.%20Techno-Commercial%20F

easibility.pdf.
http://dbtncstcp.nic.in/downloads/Plant%20Tissue%20Culture.%20Techno-Commercial%20Feasibility.pdfhttp://dbtncstcp.nic.in/downloads/Plant%20Tissue%20Culture.%20Techno-Commercial%20Feasibility.pdfhttp://dbtncstcp.nic.in/downloads/Plant%20Tissue%20Culture.%20Techno-Commercial%20Feasibility.pdfhttp://dbtncstcp.nic.in/downloads/Plant%20Tissue%20Culture.%20Techno-Commercial%20Feasibility.pdfhttp://dbtncstcp.nic.in/downloads/Plant%20Tissue%20Culture.%20Techno-Commercial%20Feasibility.pdfhttp://dbtncstcp.nic.in/downloads/Plant%20Tissue%20Culture.%20Techno-Commercial%20Feasibility.pdfhttp://dbtncstcp.nic.in/downloads/Plant%20Tissue%20Culture.%20Techno-Commercial%20Feasibility.pdf


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cleanings. The area for washing should have a big washbasin (of stainless steel, and

be acid and alkali resistant), tap water, tables that allow stand-up work and shelves to

dry and keep the washed materials. The media preparation area must be equipped with

a refrigerator to keep the chemicals and solutions used in the media culture, scales, a

potentiometer, a kitchen, a media mixer, a water distiller, and an autoclave or pressure

pot. The last two must be located as close as possible to the washbasin. The stove may

be used to dry the materials.

E.

Because we will produce a large amount of plants, we should depends on the

production of the plants it might be worth it to invest more money in a well-equipped

glasshouse. Such a glasshouse would include a mistbed or plastic tunnels to keep the

humidity high and a very good temperature control system. A method to control the

light intensity would also be needed, as plantlets should be gradually exposed to an

increase in light intensity during hardening off. In vitro Plantlets, when in vitro shootsare first rooted in tissue culture, the plantlets must be healthy and well proportioned.

No callus must be present between the root and the shoot. If sugar was included in the

rooting medium, it is necessary to wash away the agar. Sucrose found on the surface

of the roots can cause the plantlets to be infected with disease causing

micro-organisms. Otherwise, plantlets must be treated with fungicide and nutrient

solution, and rooted in growth medium as previously described for ex vitro rooting. It

is essential to work as quickly as possible with the plantlets as soon as they have been

removed from the tissue culture vessels. They may desiccate in a couple of minutes

Comment [cyn7]: This was copied from here:

https://www.msu.edu/course/css/451/LabSkill

s/Setting%20Up%20a%20Tissue%20Culture%2

0Lab.pdf

Comment [cyn8]: this was copied from here:

http://www.ipps.org/Papers/SouthAfrican/thi

art.pdf


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under hot, dry conditions. Often, it is easier to work with the plantlets when the roots

are still quite short. The longer the roots are, the more likely they are to be damaged.

When using a seedling tray or container where a lot of plants are planted in the same

container, the plants must be sprayed with water regularly. As soon as the leaves start

to curl up, the plant is under stress. A spray bottle with distilled water works very

well.

Conclusion:

Therefore, a good laboratory needs a perfect design of laboratory. Because it is to

design a lab that should know more knowledge about all the process and the

instruments we required.

Comment [cyn9]: this was also copied from

the same article above (Comment 8):


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References:

1.Ammirato, P.V. 1983. Embryogenesis. In: Handbook of Plant Cell Culture.

Evans, D., Sharp,W.R., Ammirato, P.V. and Yamada, Y. (Eds.) MacMillan,New

York. Pp.82-123.

2. http://www.cipotato.org/csd/materials/Tissue/Capitulo1.pdf

3.

https://www.msu.edu/course/css/451/LabSkills/Setting%20Up%20a%20Tissue%20C

ulture%20Lab.pdf

4.http://www.ipps.org/Papers/SouthAfrican/thiart.pdf

Marks allocation

No. Criterion Weightage Marks Allocated Remark

1 Adherence to format 10% 7%

2 Referencing 10% 4%

ref. used not

cited

3 Coverage of topic 40% 30%

4 Depth of research 20% 15%5 Flow of writing 10% 5%

6 Language usage 5% 4%

7 Conclusion 5% 2%

8

Bonus mark for early

submission 5% -45%

+5% for early

submission, -

50% for

plagiarism

Total 105% 22%
http://www.cipotato.org/csd/materials/Tissue/Capitulo1.pdfhttp://www.cipotato.org/csd/materials/Tissue/Capitulo1.pdfhttps://www.msu.edu/course/css/451/LabSkills/Setting%20Up%20a%20Tissue%20Culture%20Lab.pdfhttps://www.msu.edu/course/css/451/LabSkills/Setting%20Up%20a%20Tissue%20Culture%20Lab.pdfhttps://www.msu.edu/course/css/451/LabSkills/Setting%20Up%20a%20Tissue%20Culture%20Lab.pdfhttps://www.msu.edu/course/css/451/LabSkills/Setting%20Up%20a%20Tissue%20Culture%20Lab.pdfhttp://www.cipotato.org/csd/materials/Tissue/Capitulo1.pdf

PlantTC Assignment HanMeng

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