Anthurium is a perennial evergreen plant in the Araceae family. Its leaf leather is bright, and its single flower is at its top. The flowering period is about one and a half months. The use of conventional sowing and ramets to propagate cut flowers and red palms has a very low reproductive rate, requires long time for sowing, and is prone to variation. The use of tissue culture to rapidly propagate cut flowers can meet the large demand of the market.
Materials and Methods
The source of the material is the high-quality Anthurium cut flower varieties bred by Henan Puyang Shijin Company.
The explants were sterilized. The young leaves and petioles of the cut hibiscus were taken as the test materials. First, they were rinsed under running water for 1 hour. At the same time, they were brushed gently with a brush. Before the disinfection, the leaves were cut into 1.5 cm 1.5 cm The petiole was cut into small segments about 1.5 cm long, placed in sterile jars, soaked in 75% alcohol for 30 seconds, and soaked in 0.1% HgCl2 for 8 minutes. Sterile water was used. Rinse 5 times, cut the leaves into pieces of about 1cm1cm on sterile paper, cut the petioles into small pieces of about 1cm, inoculate on the sterile medium, all the aseptic operation must be carried out in the clean bench.
Medium and culture conditions Basic medium MS plus sucrose 30g/L, agar 6g/L, pH 5.8. Different types and concentrations of plant growth regulators were added at different stages of culture. The culture temperature was 25° C. and 1° C., and the light intensity was 2000-3000 lux. The light was lighted for 14 hours per day.
results and analysis
The degree of dedifferentiation of different explants was compared. Two different types of explants were inoculated on the same medium of MS+6-BA1.5+NAA0.7. After 3 weeks of culture, the explants of the leaves first expanded significantly and grew out. Callus tissue pieces, 5 weeks later, leaf explants grow callus pieces (see Table 1). It was observed that two different explants were able to induce callus, but the leaves were more easily dedifferentiated.
Effect of 6-BA on Callus Induction The sterile leaves were inoculated on a medium supplemented with 6-BA with different concentrations of MS+6-BA+NAA0.7, and observed for different concentrations of 6-BA on leaves. The effect of explant callus induction (see Table 2).
It can be seen from Table 2 that different concentrations of 6-BA have distinctly different effects on the induction of leaves of C. chinensis. In the absence of 6-BA medium, callus could not be induced. When the concentration of 6-BA was lower than 1.5 mg/L, the callus induction rate and proliferation rate increased with the increase of 6-BA concentration, while the higher concentrations of 6-BA inhibited the callus. Induction and proliferation of tissues. When 6-BA was 1.5 mg/L, callus induction rate and proliferation rate reached the maximum, and the best effect was obtained. Therefore, MS+6-BA1.5+NAA0.7 is an ideal induction medium for cut flower Anthurium.
The effect of 6-BA on the differentiation and proliferation of callus The successful callus pieces were transferred to differentiation medium containing different concentrations of 6-BA, respectively, and cultured to observe the effects of different concentrations of exogenous hormone 6-BA on callus. The effect of tissue differentiation budding (see Table 3).
As can be seen from Table 3, different concentrations of 6-BA have distinctly different effects on the differentiation of callus of Cutflower Anthurium. With the increase of the concentration of low concentration of 6-BA, the bud differentiation rate and bud yield increased. When 6-BA reached 1.0 mg/L, bud differentiation was significantly inhibited and the growth of callus was accelerated. When 6-BA was 0.5 mg/L, the highest bud differentiation rate was 91.43%. The fastest speed. The calluses covered with buds were cut into small pieces, subcultured, and subcultured once every 30 days to achieve the purpose of factory production.
Rooting culture A single bud of 2 to 3 cm in length was inoculated from the bud clusters and inoculated in a rooting medium. After 15 days of cultivation, the growth in different media was observed (see Table 4).
From Table 4, it can be seen that NAA and IBA 0.2 mg/L have a good promotion effect on the rooting of cut flowers. The shape of the former root is thick and short, while the latter is slender, but the NAA price is lower than that of IBA. Therefore, 1/2MS+NAA 0.2mg/L is the first choice for the rooting medium for cutting Anthurium andraeanum.
The test-tube seedlings were transplanted to take good test-tube seedlings that were rooted, and the medium at the base of the seedlings was washed clean with water, planted in a 1:1 matrix of peat and perlite, and irrigated with water. The temperature was maintained at 18 in a shaded greenhouse. Between °C and 25°C, the surrounding humidity is maintained at about 90%. After careful management, after 2 weeks, the survival rate of plantlets in vitro is generally 90%. After several transplanting experiments, it has been found that keeping the air around the seedlings moist, well-ventilated and well-hydrated cultivation substrates and the appropriate temperature are the key to the success of transplanting the test tube seedlings.
discuss
During the process of tissue culture of cut flower Anthurium, high-quality callus is firstly induced, and fine clones with more buds are differentiated as fast-producing materials, so that a large number of high quality seedlings can be produced in a short time.
Transferring soil from the medium to the soil is a transition from heterotrophic to autotrophic. Test tube heterotrophic conditions grow in a sterile, high-humidity, low-light, and temperature-stable environment, whereas greenhouses are relatively dry, strong light, and have a large temperature difference. Therefore, in the period of transplanting and acclimation of test-tube seedlings, the cut flower red should be used. The physiological characteristics of the palm and the characteristics of the test-tube seedlings were selected to be suitable for the transplanting substrate with good ventilation and heat preservation and suitable for cutting the growth of Anthurium andraeanum. It is necessary to ensure the proper temperature and humidity, create a good growth environment, reduce the pests and diseases, and improve the survival rate of transplanting.
It is well known that high concentrations of cytokinins can cause unpredictable variation in plants. Although the concentration of mitogen used in the tissue culture rapid propagation technique described above does not cause significant variation, the concentration must be appropriate.
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