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MB 305 - CELL AND TISSUE CULTURE

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BSB3163 PLANT AND MAMALIAN CELL TECHNOLOGY. (PART 2: PLANT TISSUE CULTURE). BY DR TAN SUAT HIAN (TSHIAN@UMP.EDU.MY). ASSIGNMENT NO. 2 (PLANT ...

    BSB3163 Plant and Mamalian Cell Technology

    (PART 2: PLANT TISSUE CULTURE)

    by Dr Tan Suat Hian (tshian@ump.edu.my)

ASSIGNMENT No. 2 (PLANT TISSUE CULTURE):

Learning Outcomes:

    This assignment is designed to assess students’ ability on:

    a) awareness on ethical issue, safety and social responsibility [PO9];

    b) identification of risk and possible problem(s) posted by the risk [PO6]; and

    c) propose a solution towards the problem(s) with the current existing biotech/research

    company available [PO6]

Task / Activities:

    Table 1 shows some contemporary issues in agriculture and in horticulture, whether to improve the yield or quality of the product or to produce new product in the market.

    1) Cultivation of plant 2) Genetic modification 3) Pharmacology: using

    through traditional plant of plant for crop plant to produce edible

    breeding (crop or plantation vaccine

    ornamental plants)

    Working in a group of not more than three (3) students, you are to select one (1) of the contemporary issues given in Table 1 and provide the following in your presentation.

    1. A background of the issue;

    2. Awareness on ethical issue, safety and social responsibility related to the issue;

    3. Identification of risk and possible problem(s) posted by risk;

    4. A possible research solution for the risk identified; and

    5. Propose any suitable research company available in the market to conduct the

    research and identify the apparatus/equipment and method(s) use.

     thththPresentation Date: 29 Nov, 6 Dec and 13 Dec 2011 during class

    Materials and Chemical / Apparatus Equipment

    Methods Biochemical

    Reagents

    2.1 Source ; Half strength ; greenhouse-grown ; Incubator

    tissue MS medium plants (Basella,

     Centella, Withania, ; Laminar

    Amaranthus, air flow ; MS medium

    Catharanthus, bench

     Phlox, ; 2,4

     Petunia) dichloropheno; pH meter

     xyacetic acid

    ; shoot cultures (2,4-D) (1

    (tomato cultivars viz; mg/1)

    Arkavikas, pusa

    Ruby and Punjab ; Kinetin(0?5

    Kesari; chilli, mg/1)

    Nicotiana rustica, N.

    glutinosa and ; 70% ethanol

    N? tabacum cv.

    Thompson) ; Tap water

     ; Cotyledons of ; Liquid

    aseptically- detergent for

    germinated seedlings surface

     (sunflower and niger) sterilization

     ; Placental tissue of

     fruits (tomato)

     ; Friable embryogenic

     callus

     (sandalwood)

     ; Test tubes

     ; Forceps

     ; Spatula

    ; Culture plate

    ; Beakers

    ; Measuring cylinders

    Materials and Chemical / Apparatus Equipment

    Methods Biochemical Reagents 2.2 Explant ; Dilute Labolene ; Leaves of field-; Laminar preparation of Glaxo India grown plants, flow hood prior to enzyme Ltd. fully expanded exposure leaves excised

     from greenhouse- ; 2% sodium

     grown plants hypochlorite

     (Catharanthus,

     Phlox, Petunia)

     ; Protoplast

     ; Shoot cultures isolation

     (Nicotiana rustica, solution

     N. glutinosa and (containing

     N? tabacum cv. digesting

     Thompson) enzymes, the

     osmoticum, a

     protecting ; Cotyledons of

     chemical, CaCl2 sunflower, niger

     (10 mM), all and chilli

     dissolved in seedlings

     distilled water)

     ; Deionized ; Placental tissue of

     water, sterile tomato fruits

     distilled water

     and tap water. ; Embryo genic sandalwood callus ; Spatula ; Sterilized petri plates ; 250mL jar bottles ; Fluorescent light ; Fine scalpel or razor blades ; Scissors

     ; Forceps

     Chemical/

    Material and Biochemical Reagents Equipment Apparatus

    Methods

    2.3 Protoplast isolation, purification and culture

     ; Celluclast and

    (a) Enzymes Pectinex/SP249 ; Laminar flow ; Beaker or test tubes

     hood

     ; Distilled water ; 250mL jar bottles

     ; Fluorescent light

     ; Protoplast

    (b) Osmotic ; Laminar flow isolation solution ; Tissues of conditions hood greenhouse-grown (stabilization) ; Protoplast culture plants ; Cryoscopic solution, osmometer mannitol in ; Tissues of (Osmomat combination with actively-growing 030 Gonotec) either sorbitol or shoot cultures mannitol in combination with ; Cotyledons of magnesium aseptically- sulphate. germinated seedlings ; Placental tissue of tomato ; Sandalwood callus ; Beakers or test tubes ; Pipettes ; Pipette pump ; 250mL jar bottles ; Fluorescent light

     Chemical/

    Material and Biochemical Reagents Apparatus Equipment

    Methods ; Filter sterilized

     (c) Incubation isolation ; Tobacco leaves ; Microscope for protoplast solution release ; Incubation

     room ; Osmotic

     stabilizers eg:

     mannitol as the

     sole

     Osmoticum

     ; Osmotic

     stabilizers eg:

     mannitol in

     combination

     with either

     sorbitol or

     magnesium

     sulphate

     ; Washing

    (d) Protoplast ; Nylon mesh ; Centrifuge mediums (Table

    purification machine 2)

     ; Centrifuge tubes

     ; Hemocyto-; Flotation meter ; 15 ml glass screw medium (Table cap vials 2) ; Pasteur pipette ; Fluorescein diacetate (FDA) ; Pipette tips ; Modified 8E (e) Protoplast ; Incubator liquid culture ; 60 × 15 mm culture medium plastic or glass

    ; Microscope petri plates

     (Table 2) ; Osmotic

    ; Autoclave stabilizer eg:

    machine mannitol

; Growth

regulators

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