The protection of Taxus chinensis has been highly valued internationally, but the strong market demand still brings long-term and lasting pressure to the survival of endangered Taxus chinensis. According to reports, Taxus plants in China. Meili Forest Park will do its best to protect wild resources and cultivate artificial seedlings. At present, the local government will cooperate with Fudan University and other relevant scientific research institutions on introduction, propagation and seedling raising. At present, more than 70 mu of seedling base has been initially built, and nearly 20,000 seedlings of Taxus chinensis have been artificially cultivated, which will exceed 654.38+10,000 in three years. According to the person in charge, the base will take artificial cultivation measures to develop Taxus resources in the early stage, and then use the artificially developed Taxus resources to extract paclitaxel after expanding the planting area to occupy the international taxol market.
The State Forestry Administration has a clear instruction on the current "Taxus chinensis Project": "Taxus chinensis is a first-class protected tree species in China, with good ecological benefits and extremely high economic value. The state supports the vigorous construction of yew forests in appropriate areas and encourages all sectors of society to participate in afforestation and greening in various forms such as contracting, leasing, auction and joint-stock cooperation. " At present, the development goal of the yew industry in Weihe Forestry Bureau is: the resource reserve is the first in the world, and the seedling output is the first in the world. Therefore, the "yew project" is not only an urgent life-saving project, but also a great sunshine cause that benefits mankind.
Compared with other countries in the world, China's yew resources account for more than half, but the world's demand for paclitaxel is far from enough. Since the US Food and Drug Administration (FAD) approved paclitaxel as a drug for the treatment of advanced cancer in 1992, the excessive logging of wild yew resources in China has become increasingly serious, and the wild yew resources have been seriously damaged. Moreover, many biological characteristics of wild yew restrict the development of natural communities. According to statistics, the annual incidence of cancer in the world is more than 654.38+million. If 0.006%~0.06% paclitaxel is extracted, it will consume 700 ~ 654.38+0000 tons of yew bark every year. Even if all the bark of yew in the world is collected completely, it can only maintain short-term needs. Moreover, the methods of synthetic or chemical taxol are still in the laboratory stage. Therefore, it is urgent to expand and develop Taxus resources artificially and establish a timber forest base for Taxus medicinal raw materials.
Jiangsu Hodo Group Co.,Ltd. Co., Ltd. began to cultivate Taxus cuspidata artificially on 1997, and obtained the national patent. Now, more than 30 million strains of Taxus cuspidata have been planted artificially.
Several propagation methods of Taxus seedlings at present
1. Pre-propagation planting method
The preservation of Taxus resources and rapid propagation of seedlings are the basis to solve the problem of taxol timber forest construction, and the key to solve the problem of taxol raw materials is to build a large-scale Taxus plantation.
When raising seedlings of Taxus chinensis seeds, it is necessary to maintain the storage mode of seeds, and store sand seeds together or control the temperature, which has a good effect on budding after wintering and breaking the dormancy habit. Rub the seed coat before sowing, soak it in warm water and treat it with drugs and hormones. Shading after emergence is the key to raising seedlings. It can prevent the seedlings from being burned at high temperature, keep them moist and have a light transmittance of 40%. The emergence rate of Taxus cuspidata and Taxus mairei can reach 70~80%, and the emergence temperature should be higher than 65438 05 degrees.
The dynamic growth of seedlings is as follows: the plant height and stem diameter of seedlings grow slowly two years before emergence, and the plant height generally grows about 10CM in one year, and grows faster after transplanting for one year. The annual growth can reach 20~30CM in 3~5 years.
2. Tissue culture and breeding of Taxus chinensis
Tissue culture makes use of the totipotency and clonality of plant cells. The tender stems, needles, bark, cambium, aril and embryo of Taxus plants are used as plants for cultivation and research.
(1). Fine varieties of Taxus cuspidata, such as fine organs of Taxus cuspidata (with high taxol content), were selected as explants, inoculated into culture medium, and after callus formation, rooting and bud formation, a large number of tissue culture seedlings were obtained in the laboratory. After substrate transplanting, seedling hardening and epidemic prevention, it became a production seedling.
(2) Different species and different plants of the same species have different proportions of callus formation in the morning and evening. At the same time, it is related to explant type, sampling place and season, lighting conditions, culture medium types and other factors. Taxus cuspidata, Taxus mairei and Taxus yunnanensis can all form callus on the induction medium. However, due to the different species and plants of Taxus chinensis, the formation situation is also different.
The results showed that the hydrolyzed complex protein could promote the growth of callus of Taxus cuspidata and Taxus mairei to some extent, but the concentration greater than 0.5% was not conducive to the formation and accumulation of paclitaxel.
When the concentration is greater than 1%, it can promote callus healing and form new growth points without affecting the accumulation of paclitaxel.
Comparatively speaking, 2 days and 4 days are also beneficial to the formation of callus and can also improve the induction rate. Callus in 2-day and 4-day culture medium is bright, large and soft, which is easy to induce callus of Taxus chinensis. However, other medium additives, such as cell suspension culture, B5 medium and carbon source sucrose, can also increase the content of paclitaxel. However, glucose in high-yield sucrose will inhibit the synthesis of paclitaxel. At the same time, galactose is of great significance to promote cell growth.
GIBSON's research shows that whether it is cell suspension culture or inducing callus cell growth and taxol content formation, it is better in the dark than in light.
No matter what species and parts are used for tissue culture, we should choose the cell components with strong proliferation ability in the middle (the surface cells contain a lot of starch granules, the central cells are seedless, and tubular molecules are easy to differentiate). This is conducive to the formation of cell filaments between cells, thus forming a large cell mass, which is conducive to the signal transmission of small substances between cells. Therefore, the cells of Taxus chinensis exist in the form of cell clusters, producing functional cell bodies, promoting the regionalization of cell clusters and cell functional differentiation, which is the premise for Taxus chinensis to produce metabolites.
3. Artificial cutting propagation of Taxus chinensis
The cutting propagation of Taxus chinensis is better in spring and autumn. Generally, low shed shading treatment should be done when cutting. Generally, the survival rate of cutting can reach more than 70%. Conventional cutting is only about 3%~20%. Shading rate is not less than 60%. Cutting media should be carefully rooted. At the beginning, the humidity should be kept at about 75~85% intermittently. Avoid water loss caused by strong wind.
The factors affecting the survival rate of cutting are generally: tree age, temperature, drug concentration, substrate, season, humidity, variety and other human factors. According to the data, the highest survival rates of several kinds of yew are: Taxus cuspidata 95%, Taxus mairei 95%, Taxus yunnanensis 90% and Taxus China 86%.
In the process of rooting in the first year, the aboveground part of the cuttings grew slowly, but took root quickly and developed lateral roots. After transplanting in the second year, further shading treatment and seedbed management are needed. Ensure the corresponding environmental conditions and accelerate the growth. The growth rate is the fastest in summer and slower in winter and spring. Pay attention to the change of temperature at this time. Long-term drying can lead to growth inhibition or sudden death. However, after the seedlings of Taxus chinensis are strong in the seedbed, the survival rate of afforestation is very high. It rarely dies and grows rapidly.
Cutting is the most effective way to solve the shortage of Taxus resources and accelerate the construction of Taxus medicinal forest.
Protection knowledge of Taxus bonsai
1, "Maintenance of newly purchased bonsai" is best to spray 800 times 1000 times potassium dihydrogen phosphate solution on the leaves every three or four days, usually before 10 am or around 4 pm. After three or four consecutive sprays, it is gradually reduced to once a month.
2, "lighting requirements" Taxus chinensis is a shade-loving plant, suitable for indoor display, but it should be noted that it should be properly shaded in summer and should not be placed in a room with western photos.
3. "Soil selection" Taxus chinensis should be planted in loose, humus-rich, fertile and slightly acidic soil (PH=5~6.5).
4. "Water requirement" The surface of the basin soil is slightly yellow and white, and the leaves are slightly curled. Pot soil doesn't need watering, just spraying leaves. When the soil in the basin turns white, it is necessary to water it. Pay attention to one-time watering to make the basin soil fully absorb water.
5. The three elements of "soil fertility" plant growth are nitrogen, phosphorus and potassium fertilizer, and nitrogen fertilizer can promote the growth of flower branches; Phosphate fertilizer can promote the formation of flowers and fruits; Potassium fertilizer can promote the growth of stems and roots. Because the newly bought Taxus bonsai has already used the prepared nutrient soil, it is not necessary to apply fertilizer within 3 months, and it can be applied every 2 to 3 months in the future. Cake fertilizer is the best fertilizer. Pay attention to the operation along the basin edge when applying fertilizer to avoid touching the root of bonsai.
"Selection of flowerpot" The flowerpot should be appropriately large, and the bottom of the flowerpot should be perforated more, mainly to enhance the water permeability and air permeability of the flowerpot.
6. "Requirements for Transplanting and Changing Pots" After purchasing bonsai for half a year, the saplings gradually grew up and the roots were developed. In order to ensure its normal growth, users are advised to transplant and change pots. When changing pots, it is best to break the old pots without destroying the original soil balls, slowly move them into new pots, and water them to take root to ensure survival.
7, "bonsai pruning" If the dry yellow leaves at the lower part of the bonsai fall off seriously, it may be that the lower branches are too dense, and it is necessary to prune a few branches that are too dense at the lower part. Pruning can be arbitrary, so it can be trimmed into umbrella, tower, circle and other shapes. The newly bought Taxus bonsai above No.3 will damage part of the root system when transplanted from the planting base to the flowerpot, resulting in the imbalance of nutrient supply between the root system and the branches and leaves. When the leaves are still curly and dry under the condition of moist soil, it is best to trim the branches and leaves of bonsai properly to reduce the excessive consumption of nutrients by branches and leaves and restore the growth of roots.
8. "Pest control" In the hot and dry season, leaf blight and bacterial blight will occur in individual young yew trees, which can be controlled by spraying 1% Bordeaux solution.
9. Artificial seedlings are being cultivated locally.
The protection of Taxus chinensis has been highly valued internationally, but the strong market demand still brings long-term and lasting pressure to the survival of endangered Taxus chinensis. According to reports, Taxus plants in China. Meili Forest Park will do its best to protect wild resources and cultivate artificial seedlings. At present, the local government will cooperate with Fudan University and other relevant scientific research institutions on introduction, propagation and seedling raising. At present, more than 70 mu of seedling base has been initially built, and nearly 20,000 seedlings of Taxus chinensis have been artificially cultivated, which will exceed 654.38+10,000 in three years. According to the person in charge, the base will take artificial cultivation measures to develop Taxus resources in the early stage, and then use the artificially developed Taxus resources to extract paclitaxel after expanding the planting area to occupy the international taxol market.
The State Forestry Administration has a clear instruction on the current "Taxus chinensis Project": "Taxus chinensis is a first-class protected tree species in China, with good ecological benefits and extremely high economic value. The state supports the vigorous construction of yew forests in appropriate areas and encourages all sectors of society to participate in afforestation and greening in various forms such as contracting, leasing, auction and joint-stock cooperation. " At present, the development goal of the yew industry in Weihe Forestry Bureau is: the resource reserve is the first in the world, and the seedling output is the first in the world. Therefore, the "yew project" is not only an urgent life-saving project, but also a great sunshine cause that benefits mankind.
Compared with other countries in the world, China's yew resources account for more than half, but the world's demand for paclitaxel is far from enough. Since the US Food and Drug Administration (FAD) approved paclitaxel as a drug for the treatment of advanced cancer in 1992, the excessive logging of wild yew resources in China has become increasingly serious, and the wild yew resources have been seriously damaged. Moreover, many biological characteristics of wild yew restrict the development of natural communities. According to statistics, the annual incidence of cancer in the world is more than 654.38+million. If 0.006%~0.06% paclitaxel is extracted, it will consume 700 ~ 654.38+0000 tons of yew bark every year. Even if all the bark of yew in the world is collected completely, it can only maintain short-term needs. Moreover, the methods of synthetic or chemical taxol are still in the laboratory stage. Therefore, it is urgent to expand and develop Taxus resources artificially and establish a timber forest base for Taxus medicinal raw materials.
Jiangsu Hodo Group Co.,Ltd. Co., Ltd. began to cultivate Taxus cuspidata artificially on 1997, and obtained the national patent. Now, more than 30 million strains of Taxus cuspidata have been planted artificially.
Several propagation methods of Taxus seedlings at present
1. Pre-propagation planting method
The preservation of Taxus resources and rapid propagation of seedlings are the basis to solve the problem of taxol timber forest construction, and the key to solve the problem of taxol raw materials is to build a large-scale Taxus plantation.
When raising seedlings of Taxus chinensis seeds, it is necessary to maintain the storage mode of seeds, and store sand seeds together or control the temperature, which has a good effect on budding after wintering and breaking the dormancy habit. Rub the seed coat before sowing, soak it in warm water and treat it with drugs and hormones. Shading after emergence is the key to raising seedlings. It can prevent the seedlings from being burned at high temperature, keep them moist and have a light transmittance of 40%. The emergence rate of Taxus cuspidata and Taxus mairei can reach 70~80%, and the emergence temperature should be higher than 65438 05 degrees.
The dynamic growth of seedlings is as follows: the plant height and stem diameter of seedlings grow slowly two years before emergence, and the plant height generally grows about 10CM in one year, and grows faster after transplanting for one year. The annual growth can reach 20~30CM in 3~5 years.
2. Tissue culture and breeding of Taxus chinensis
Tissue culture makes use of the totipotency and clonality of plant cells. The tender stems, needles, bark, cambium, aril and embryo of Taxus plants are used as plants for cultivation and research.
(1). Fine varieties of Taxus cuspidata, such as fine organs of Taxus cuspidata (with high taxol content), were selected as explants, inoculated into culture medium, and a large number of tissue culture seedlings were obtained in the laboratory after callus formation, rooting and seedling formation. After substrate transplanting, seedling hardening and epidemic prevention, it became a production seedling.
(2) Different species and different plants of the same species have different proportions of callus formation in the morning and evening. At the same time, it is related to explant type, sampling place and season, lighting conditions, culture medium types and other factors. Taxus cuspidata, Taxus mairei and Taxus yunnanensis can all form callus on the induction medium. However, due to the different species and plants of Taxus chinensis, the formation situation is also different.
The results showed that the hydrolyzed complex protein could promote the growth of callus of Taxus cuspidata and Taxus mairei to some extent, but the concentration greater than 0.5% was not conducive to the formation and accumulation of paclitaxel.
When the concentration is greater than 1%, it can promote callus healing and form new growth points without affecting the accumulation of paclitaxel.
Comparatively speaking, 2 days and 4 days are also beneficial to the formation of callus and can also improve the induction rate. Callus in 2-day and 4-day culture medium is bright, large and soft, which is easy to induce callus of Taxus chinensis. However, other medium additives, such as cell suspension culture, B5 medium and carbon source sucrose, can also increase the content of paclitaxel. However, glucose in high-yield sucrose will inhibit the synthesis of paclitaxel. At the same time, galactose is of great significance to promote cell growth.
GIBSON's research shows that whether it is cell suspension culture or inducing callus cell growth and taxol content formation, it is better in the dark than in light.
No matter what species and parts are used for tissue culture, we should choose the cell components with strong proliferation ability in the middle (the surface cells contain a lot of starch granules, the central cells are seedless, and tubular molecules are easy to differentiate). This is conducive to the formation of cell filaments between cells, thus forming a large cell mass, which is conducive to the signal transmission of small substances between cells. Therefore, the cells of Taxus chinensis exist in the form of cell clusters, producing functional cell bodies, promoting the regionalization of cell clusters and cell functional differentiation, which is the premise for Taxus chinensis to produce metabolites.
3. Artificial cutting propagation of Taxus chinensis
The cutting propagation of Taxus chinensis is better in spring and autumn. Generally, low shed shading treatment should be done when cutting. Generally, the survival rate of cutting can reach more than 70%. Conventional cutting is only about 3%~20%. Shading rate is not less than 60%. Cutting media should be carefully rooted. At the beginning, the humidity should be kept at about 75~85% intermittently. Avoid water loss caused by strong wind.
The factors affecting the survival rate of cutting are generally: tree age, temperature, drug concentration, substrate, season, humidity, variety and other human factors. According to the data, the highest survival rates of several kinds of yew are: Taxus cuspidata 95%, Taxus mairei 95%, Taxus yunnanensis 90% and Taxus China 86%.
In the process of rooting in the first year, the aboveground part of the cuttings grew slowly, but took root quickly and developed lateral roots. After transplanting in the second year, further shading treatment and seedbed management are needed. Ensure the corresponding environmental conditions and accelerate the growth. The growth rate is the fastest in summer and slower in winter and spring. Pay attention to the change of temperature at this time. Long-term drying can lead to growth inhibition or sudden death. However, after the seedlings of Taxus chinensis are strong in the seedbed, the survival rate of afforestation is very high. It rarely dies and grows rapidly.
Cutting is the most effective way to solve the shortage of Taxus resources and accelerate the construction of Taxus medicinal forest.
Protection knowledge of Taxus bonsai
1, "Maintenance of newly purchased bonsai" is best to spray 800 times 1000 times potassium dihydrogen phosphate solution on the leaves every three or four days, usually before 10 am or around 4 pm. After three or four consecutive sprays, it is gradually reduced to once a month.
2, "lighting requirements" Taxus chinensis is a shade-loving plant, suitable for indoor display, but it should be noted that it should be properly shaded in summer and should not be placed in a room with western photos.
3. "Soil selection" Taxus chinensis should be planted in loose, humus-rich, fertile and slightly acidic soil (PH=5~6.5).
4. "Water requirement" The surface of the basin soil is slightly yellow and white, and the leaves are slightly curled. Pot soil doesn't need watering, just spraying leaves. When the soil in the basin turns white, it is necessary to water it. Pay attention to one-time watering to make the basin soil fully absorb water.
5. The three elements of "soil fertility" plant growth are nitrogen, phosphorus and potassium fertilizer, and nitrogen fertilizer can promote the growth of flower branches; Phosphate fertilizer can promote the formation of flowers and fruits; Potassium fertilizer can promote the growth of stems and roots. Because the newly bought Taxus bonsai has already used the prepared nutrient soil, it is not necessary to apply fertilizer within 3 months, and it can be applied every 2 to 3 months in the future. Cake fertilizer is the best fertilizer. Pay attention to the operation along the basin edge when applying fertilizer to avoid touching the root of bonsai.
"Selection of flowerpot" The flowerpot should be appropriately large, and the bottom of the flowerpot should be perforated more, mainly to enhance the water permeability and air permeability of the flowerpot.
6. "Requirements for Transplanting and Changing Pots" After purchasing bonsai for half a year, the saplings gradually grew up and the roots were developed. In order to ensure its normal growth, users are advised to transplant and change pots. When changing pots, it is best to break the old pots without destroying the original soil balls, slowly move them into new pots, and water them to take root to ensure survival.
7, "bonsai pruning" If the dry yellow leaves at the lower part of the bonsai fall off seriously, it may be that the lower branches are too dense, and it is necessary to prune a few branches that are too dense at the lower part. Pruning can be arbitrary, so it can be trimmed into umbrella, tower, circle and other shapes. The newly bought Taxus bonsai above No.3 will damage part of the root system when transplanted from the planting base to the flowerpot, resulting in the imbalance of nutrient supply between the root system and the branches and leaves. When the leaves are still curly and dry under the condition of moist soil, it is best to trim the branches and leaves of bonsai properly to reduce the excessive consumption of nutrients by branches and leaves and restore the growth of roots.
8. "Pest control" In the hot and dry season, leaf blight and bacterial blight will occur in individual young yew trees, which can be controlled by spraying 1% Bordeaux solution.
9. Because the climate in the north is dry and there is little rain, you can water the soil surface when it is dry. Pay attention to a thorough watering, especially in summer. When the air in the north is dry, the leaves are prone to water shortage and drooping, and the appearance of the leaves is not full, it is necessary to spray water on the leaves from bottom to top with a small watering can (it can be sprayed every day in summer). Taxus chinensis is a shade-loving plant, which is suitable for indoor display, but it should not be placed at the air outlet of air conditioner or next to heating, otherwise it will increase the evaporation of water in the leaves of Taxus chinensis, which will easily dehydrate the branches and leaves of Taxus chinensis quickly, resulting in curly and dry leaves.
10, Taxus bonsai should not be watered with tea. In particular, the water quality in Beijing is hard and high in alkali, so it must be used for watering and spraying at the end of the day. Because the climate in the north is dry and there is little rain, you can water the soil when it is dry, and pay attention to watering it once, especially in summer. When the air in the north is dry, the leaves are prone to water shortage and drooping, and the appearance of the leaves is not full, it is necessary to spray water on the leaves from bottom to top with a small watering can (it can be sprayed every day in summer). Taxus chinensis is a shade-loving plant, which is suitable for indoor display, but it should not be placed at the air outlet of air conditioner or next to heating, otherwise it will increase the evaporation of water in the leaves of Taxus chinensis, which will easily dehydrate the branches and leaves of Taxus chinensis quickly, resulting in curly and dry leaves.
10, Taxus bonsai should not be watered with tea. In particular, the water quality in Beijing is hard and high in alkali, so it must be used for watering and spraying at the end of the day.