Study On The Tissue Culture Of Clivia Miniata

Clivia miniata Regel also called Clivia is a Amaryllidaceae plant. It is a pe rennial herbaceous flower, which has dignified shape, upstanding body, symmetrical blades, evergreen appearance, lifelong green or dark green color, beautiful flower pos e, brightly colored flowers, and quite spectacular flowering pe riod. Therefore, Clivia miniata is praised as “the queen of Chinese indoor flowers”.The seed propagation can lead to the separa tion of progeny characters, so the excellent properties cannot be reserved. Although the plan t division propagation can reserve the excellent properties, the speed is slow and the random ness is very lar ge. Tissue culture can not only reserve the excellent properties of parents but also reproduce ef ficiently and stably, which is an effective way to produce a lar ge number of identic al plant types. At present, there is still a technical problem, such as the low frequency of in vitro propagation and regeneration, and weak replicability.Through the research on the tissue culture of the four varieties of Clivia miniata Regel such as ‘Monk’, ‘Painter ’, ‘Xiye’ and ‘Crystal’, th is study of fers a suitable way for the rapid propagation of Clivia miniata Regel in order to provide the technical guidance and theoretical support for industrialized breeding. Meanwhile, this study can also provide technical support for the further research such as transgene and cell fusion to i mprove the colors of Clivia flowers. Through the research, Found the sam e culture unde r the condition of dif ferent strains of the callus induction rate and dif ferentiation rate greater im pact, which, ’painter’ strain regeneration rate is the highest.the following findings were obtained.(1). Sampling time of explants: th e current-year immature inflorescence are the best explants; the receptacles in th e young bud stage are the best exp lants, because the callus induction rate is up to 68.8 % ~ 88.9 %.(2). Sterilization method of explants: the expl ants should be soaked with 75 % alcohol in the sterile ultra clean table for 30 seconds, then be sterilized with 0.1 % g/m l mercuric chloride for 8 minutes, and finally be flushed with strerilize d distilled water for 4 times.(3). Suitable culture conditions: the temperature is 25 ± 2 ℃; the light is 24 h/d; the light intensity is 2000 lx; it is not suitable to carry on long dark culture before inoculation; the p H value is 5.8- 6.2; when the p H value is 5.8, the fast growth of tissue culture seedling is conductive to proliferation culture; when the p H value is 6.2, the robust growth of tissue culture seedling is conductive to culturing robust seedling.(4). Callus induction: as for "Monk", the op timal medium is MS + BA 4.0 mg/L + NAA 2.0 mg/L + 2, 4- D 2.0 mg/L, and its induction rate is up to 84.6%; as for "Painter", "Xiye" and "Crystal", the optimal medium is MS + BA 2.0 mg/L + NAA2.0 mg/L + 2, 4-D 3.0 m g/L, and their induction rates are 90.7%, 75.6% and 78.0% respectively; the common optimal medium is MS+BA2.0 mg/+NAA2.0 mg/L+2, 4-D 3.0 m g/L; the p H value is 5.8; after adding 30 g/L sucrose and 4.5 g/L agar strip to the m edium, the induction rate is the highest and the growth status is the best; the average starting rate of the explants is up to 62.96 % after the explants are cultured for 40 days.(5). Tissue differentiation induction: as for "M onk" and "Painter", the optim al medium is MS + NAA 0.5 m g/L + KT 1.0 mg/L, and th eir induction rates are 60.0 % and 91.4 % respectively; as for "Xiye" and "Crystal", the optimal medium is MS + NAA 1.0 mg/L + KT 3.0 mg/L, and their induction rates are up to 73.5 % and 68.7 % respectively; the common optim al medium is MS + NAA 0.5 mg/L + KT 1.0 mg/L, and its average induction rate is up to 66.45 %; the average number of budding is 1 6.7. Enhancing the light intensity, setting the difference of temperature between day and night, and properly increasing the concentration of nutrient elements in the culture m edium are conducive to the cultivation of robust proliferation seedlings.(6). Rooting culture: as for "Monk" and "Xiy e", the 1/2 MS m edium with 25 g/L sucrose and 4.5 g/L agar strip can produce better roo ting effect, and their root formation rates are up to 94.8 % and 100 % respec tively; as for "painter", the 1/2 MS medium with 25 g/L, NAA 0.5mg/L+KT0.25 mg/L sucrose and 4.5 g/L agar strip can produce better rooting ef fect, and its root for mation is up to 97 %; as for "Crystal", the 1/2 m edium with 35 g/L,NAA 0.25 mg/L+KT 0.25 mg/L sucrose and 4. 5 g/L agar strip can produce be tter rooting effect, and its root formation rate is up to 98.9%; the common optimal medium with 25 g/L, NAA 0.5 mg/L + KT 0.25 mg/L sucrose and 4.5 mg/L agar strip can produce better rooting effect, and its average root formation rate is up to 95.9 %.