- Information
- AI Chat
All PDF Maker 20240831 12
Bsc botony (Botony)
University of Calicut
Recommended for you
Preview text
54 INOCULATION INSTRUMENTS USED IN PLANT TISSUE CULTURE The plant tissue culture laboratory should have at least 2 big rooms and a small room. The one room for general laboratory work and the other for keeping cultures under controlled light ture , humidity conditions An ideal tessue culture laboratory should be provided with the following arrangements Washing area it is very important for tissue culture lab It should be provided with large sink. Hot air oven it is necessary for drying washed glass wares. Refrigerator it is essential for storing various thermolabile chemicals vitamins, minerals amino acids. weightng balance used for weighing chemicals sugar, agar etc. Autoclave Used for stirilisation of nutrient media glass ware, Instruments. PH meter Used for measuring and adjusting the p4 of nutrients in the medium. Working table. 53 Heater Laminar air flou chamber. useful for asptic transfer work. culture room Room for keeping or inubbting culture under controlled conditions, temperature and humidity It is filled with double door to make it dust free. To maintain temperature about 25 I 2 coolers are used for light adjustment culture rack is provided with flouresent lamp. Humidity is maintained about Glass wares Measuring cylinder, conual flask, beaker, test tube glass rods and pipettes ... etc. other equipments. a) Forceps Used for transferring explant to culture media. b) scalpel Used to cut the explant in to suitable size for culturing c) Nudles Used to remove seed coat from cotyledons. x 12:13 kinttin AA Hock solution were prepared in such a combination that included in the Stock A. IN order to get 50X solution, required amount of NH4NO3 82 was dessolved in distilled water. Stock B 50X solution was prepared dissolving the requiring amount of KNO3 (45 in distilled water. stock C Required amount of salts such as KHAPO4 (32 H3 BO3 (1) KI (0). cacl2. 6H2O (0) were sequentially added and dissolved in 200 ml of distilled water. 0 of Naz HO 2H2O was dissolved in distilled water seperately second solution was added to the first solution and made up to 12. stock D Required amount of cacl2 2H2O (891k) was dissolved is distilled water stock E Required any of the salt except LUSO4 5H2O were weighed and dissolved in distilled water. CUSO4 5H2O solution was added to the above solution and made up to 12. stock F 2 of FesO4. 7H2O was dissolved in distelled water 3 of Na added EDTA dissolved in distilled water separately Na,EDTA solution was to was Fe SOA along the side. The flask was sinsed with little then amount of water and added to the stock solution. The solution was kept in a magnetic stirue until its colour changes to yellow. It is stored in amber bottlat Vitamins Required qualitity of vitamins (Thiomine Hcl and Pyridoxine Nicotinic acid 0 glycine I ) were dissolved in distilled water seperately and stored in dry bottles seperately. x PREPARATION OF MEDIUM 56 MS medium was prepared from the stock solution (1)) the steps stock A 20 ml stock D to Glycine 1ml stock B 20 ml stock E cml Thymine 0 ml stock C 5 ml stock 7 6 ml Pyridexine 0 ml Nelotinic acid 0 ml 30g of sucrose was in It solution weighed and dissolved 709 of agar was added to 700 ml of this solution and boiled with constant stirring to dissolve others completely. It was allowed to cool and pH was adjusted to The medium was poured into culture tube and kept in standing position. . It was allowed to soldify and store at low temperature under aseptic condition. X 58 INOCULATION TECHNIQUES it is the process of introduction of explant mto the culture medium under aspetic conditions Major steps are Washthe hand throughly with soap and dettol Wipe off the cabinet Switch on the UV for 20 minutes After 20 minutes, switch off the UV and switch on laminar air flow and flourescent lamp Wipe the hands with 95 alcohol Flame the spirit lamp. Fit fresh blade to the bolder. instruments like blade forceps are dipped in alcohol and flamed to the sterilised fitter paper, sterilized explants are transferred Take the test tube containing culture medium in such a way that the mouth of the test tube is in the front of flame. Cap of the tube is removed with right hand twisting slight to the side and mouth of the test tube is flamed. Using the forceps the explant with approximate size is taken and plaud on the medium inthe tube. The cap is replaced after flaming the mouth of tube. The moculated tube is wrapped with cellophane trip The tubes are labelled and placed in test tube rack and kept in culture room. x PROTOCOL FOR PREPARATION OF 59 SYNTHETIC SEED Encapsulation of single somatic embryo. supention of somatic embryos mixed in sodium alginate embryo loaded alginate gel, 100 m M solution of calcium chloride Formation of calcium alginate (Through ion exchange reaction within minutes) Rinsing throughly with soft water 1 tap water (40 minutes) Immerced in pottassium nitrate (200 mm) (60 minutes) Rinsing throughly with water (4 minutes) 8. Beads gradually swell and splitt with in 6 hours of it sown in humid condition K alginate gel, 100 m M solution of calcium chloride Formation of calcium alginate (Through ion exchange reaction within minutes) Rinsing throughly with soft water 1 tap water (40 minutes) Immerced in pottassium nitrate (200 mm) (60 minutes) Rinsing throughly with water (4 minutes) 8. Beads gradually swell and splitt with in 6 hours of it sown in humid condition K APPROACH GRAFTING in this technique two plants ( stock and scion) unite together while both of them are on root systems Pot. grown plants or smckers can be effectively used for the purpose Ater the healing of the wounds the tap of the scion below the graft joint are removed gradually. c) Top of stock and weet of b) Taping with a) making cut on seion WAR removed adhesive stock and seion tape for approach grafting 62 APPROACH GRAFTING in this technique two plants ( stock and scion) unite together while both of them are on root systems Pot. grown plants or sweeks can be effectively used for the purpose. Ater the healing of the wounds the tap of the scion below the graft joint are removed gradually. c) Top of stock and root of b) Taping with a) Making cut on adhesive scion were removed. stock and scion tape for approach grafting BUDDING In this methods a T like cut is made in the bark of the root stock which is about one year old The bask in the cut region is slightly seperated and the bud is inserted into it. The bud is taken from the scion plant in the form of a shill with a bark region. attached to it. The budded region is protected wrapping with a polythine tape or waxed budding tape. D using wrapping care should be taken to see that the bud remains exposed and all the but proper are fully wrapped. CUT SCION STOCK LAYERING AIR LAYERING One of the oldest technique of vegitative propagation ut is also known as chinease layering or Gootee Long shoot of years are suitable for layering Leaves are removed from the base of the shoot A ring of bask of cm long is removed .70 prevent wound healing a thread is tied in the centre. The photo em transport is blocked and hormones build up in upper region of the wound causing root formation In recent times on the cut surfacing rooting hormones like IBA or NAA is smeared The cut surface is then covered some materials which will held water like sphagnum moss saw dust wir pith and this is covered a palythene sheath The two ends are then tid with some threads Rooting takes place with in week time. The rooted portion is seperated from the mothe plant making V shaped cuts The separation is done in days time to reduce the shock of seperation and RING OF BARK REMOVED water sprinkles. Budding and grafting knife Hand sprayer pruning sectateur Hand Troucel Spade
All PDF Maker 20240831 12
Course: Bsc botony (Botony)
University: University of Calicut
- Discover more from:
