植物组织培养过程(国外英文资料).doc

植物组织培养过程(国外英文资料) I. principle After the dedifferentiation of plant tissue, the callus is formed. After re differentiation, the calli can be re divided into structural tissues and organs, and finally a complete plant is formed. As early as 1957, Skoog discovered the type and proportion of plant hormones in the culture medium and played an important role in the process of re differentiation. Two reagents and instruments and equipment (I) reagent Ethanol. IAA or 2, 4 - D. HgCl 2 (or sodium hypochlorite). 6- benzyl amino adenine (6-BA) MS medium (see Appendix). (two) instrument and equipment Training room, autoclave, bath, scalpel, flask, beaker (100mL), a Petri dish, cotton, inoculation box or super clean bench, analytical balance, long tweezers, scissors, flask, pipette, kraft paper. Three, experimental steps 1. preparation medium (1) callus induction medium: MS medium (sucrose content was 10 g/L, 2,4 - D content was 2 mg/L, agar 10 g/L). (2) test medium: join IAA and 6 - BA in table 33 - 1 in MS medium. Indole acetic acid was first dissolved with a small amount of 0.1 mol/L NaOH, and 6- benzyl adenine was dissolved with a small amount of 0.1 mol/L HCl, then diluted with distilled water and then added to the medium. Table 33 - 1 test medium IAA and 6 - BA content Serial number IAA (mg/L) 6-BA (mg/L) Relative ratio One Zero Two Two Zero point two Two 1:10 Three Zero point five Two 1:4 Four One Two 1:2 Five Two Two 1:1 Six Two One 2:1 Seven Two Zero point five 4:1 Eight Two Zero point two 10:1 Nine Two Zero 2. medium sterilization Add the medium to the agar, heat it up, dissolve it to pH 5.8, and heat it separately in the 100 mL triangle flask, each of which is about 20 mL. The cooling medium solidified with a layer of a layer of kraft paper and weighing up the bottle (tube), and cotton string, and then in the autoclave at 121 (1 kg/cm 2) under 20 min sterilization. Remove the flask and place it on the table for cooling. All the equipment needed for inoculation (such a