A method for tissue culture and rapid propagation of Ilex red fruit superior strain

Abstract

The invention discloses a tissue culture and rapid propagation method for an ilex purpurea superior plant and belongs to the technical field of plant tissue culture. The method comprises the followingsteps: obtaining an explant; selecting an annual tender shoot tip of the ilex purpurea superior plant, and cleaning, disinfecting and sterilizing the same; induced culture: transferring the explant into an induced culture medium to induce new buds; enrichment culture: transferring the new buds to an enrichment medium to be cultured for 30 days to obtain the proliferated shoot buds; rooting culture: slicing the shoot tips of the shoot buds and transferring the shoot tips into a rooting culture medium to be cultured for 15 days to obtain rooting seedlings; and acclimatization and transplant: sealing the rooting seedlings for 2-3 days and acclimatizing the rooting seedlings with a film for 10 days and then transplanting the rooting seedlings. The tissue culture and rapid propagation method disclosed by the invention improves the propagation coefficient of ilex purpurea, shortens the rooting culture period greatly and improves the propagation speed. Compared with a conventional ilex purpurea tissue culture method, the method provided by the invention has higher survival rate and higher adaptability.

Description

Technical field

[0001] The invention relates to the technical field of plant tissue culture, in particular to a tissue culture and rapid propagation method for superior plants of Ilex rubra. technical background

[0002] Ilex purpurea Hassk, an evergreen tree of Ilex in Aquifoliaceae. Red holly has both medicinal value and ornamental value. Chinese Pharmacopoeia clearly records that red holly Hollyleaf can be used as medicine, which has great medicinal value, and is also the main component of commonly used Chinese patent medicine Jizhi syrup formula. Ilex rubra is dioecious, and the female plant is full of red fruits in autumn, which is bright and beautiful, and has a long fruiting period, which can last from October of that year to April to May of the following year. It is a landscape tree species with high ornamental value.

[0003] Ilex pedunculata is a chemical substance with great value in Ilex rubra, which has the functions of clearing away heat and toxic materials, reducin...

Examples

Embodiment 1

[0034] Example 1: Tissue Culture and Rapid Propagation Method 1:

[0035] A. obta4m8mining explants: selecting 200 buds of high-quality female Ilex rubra plants in the same year, cutting off redundant leaves, leaving bud tips, washing them with clean water, washing them with detergent for 4 minutes, then sterilizing them with 75wt％% alcohol for 23s, washing them with sterile water, filtering them off, then sterilizing them with 0.1wt％% mercuric chloride solution for 8 minutes by shaking, and washing them with sterile water.

[0036] B, induction culture: transferring sterile explants into bud induction medium, the induction medium is made of MS medium +NAA0.2mg / L+6BA 1.5mg / L+ agar 6.5g / L+ sucrose 30g / L, the pH value is 5.8, and the culture temperature is 25℃, the light intensity is 1800lx, and the light time is 12h / d for 40 days.

[0037] C. proliferation culture: transferring the induced new buds into a proliferation culture medium, which is made of MS culture medium +NAA0.2mg / L+...

Embodiment 2

[0041] Example 2: Tissue Culture and Rapid Propagation Method 2:

[0042] A. Obtaining explants: selecting 200 buds of the same high-quality female plant of Ilex rubra as in Example 1, cutting off redundant leaves, leaving bud tips, washing them with clean water, washing them with detergent for 3min, then sterilizing them with 75wt％% alcohol for 20s, cleaning them with sterile water, filtering them out, sterilizing them with 0.1wt％% mercuric chloride solution for 8min, and sterilizing them with sterile water.

[0043] B, induction culture: transferring sterile explants into bud induction medium, the induction medium is made of MS medium +NAA0.2mg / L+6BA 1.5mg / L+ agar 6.5g / L+ sucrose 30g / L, pH = 5.8, and culturing for 30 days under the conditions of culture temperature of 25℃, light intensity of 1800lx, and light time of 12h / d, to obtain

[0044] C. proliferation culture: transferring the induced new buds into a proliferation culture medium, which is made of MS culture medium +NAA0....

Technical field

[0001] The invention relates to the technical field of plant tissue culture, in particular to a tissue culture and rapid propagation method for superior plants of Ilex rubra. technical background

[0002] Ilex purpurea Hassk, an evergreen tree of Ilex in Aquifoliaceae. Red holly has both medicinal value and ornamental value. Chinese Pharmacopoeia clearly records that red holly Hollyleaf can be used as medicine, which has great medicinal value, and is also the main component of commonly used Chinese patent medicine Jizhi syrup formula. Ilex rubra is dioecious, and the female plant is full of red fruits in autumn, which is bright and beautiful, and has a long fruiting period, which can last from October of that year to April to May of the following year. It is a landscape tree species with high ornamental value.

[0003] Ilex pedunculata is a chemical substance with great value in Ilex rubra, which has the functions of clearing away heat and toxic materials, reducin...

Examples

Embodiment 1

[0034] Example 1: Tissue Culture and Rapid Propagation Method 1:

[0035] A. obta4m8mining explants: selecting 200 buds of high-quality female Ilex rubra plants in the same year, cutting off redundant leaves, leaving bud tips, washing them with clean water, washing them with detergent for 4 minutes, then sterilizing them with 75wt％% alcohol for 23s, washing them with sterile water, filtering them off, then sterilizing them with 0.1wt％% mercuric chloride solution for 8 minutes by shaking, and washing them with sterile water.

[0036] B, induction culture: transferring sterile explants into bud induction medium, the induction medium is made of MS medium +NAA0.2mg / L+6BA 1.5mg / L+ agar 6.5g / L+ sucrose 30g / L, the pH value is 5.8, and the culture temperature is 25℃, the light intensity is 1800lx, and the light time is 12h / d for 40 days.

[0037] C. proliferation culture: transferring the induced new buds into a proliferation culture medium, which is made of MS culture medium +NAA0.2mg / L+...

Embodiment 2

[0041] Example 2: Tissue Culture and Rapid Propagation Method 2:

[0042] A. Obtaining explants: selecting 200 buds of the same high-quality female plant of Ilex rubra as in Example 1, cutting off redundant leaves, leaving bud tips, washing them with clean water, washing them with detergent for 3min, then sterilizing them with 75wt％% alcohol for 20s, cleaning them with sterile water, filtering them out, sterilizing them with 0.1wt％% mercuric chloride solution for 8min, and sterilizing them with sterile water.

[0043] B, induction culture: transferring sterile explants into bud induction medium, the induction medium is made of MS medium +NAA0.2mg / L+6BA 1.5mg / L+ agar 6.5g / L+ sucrose 30g / L, pH = 5.8, and culturing for 30 days under the conditions of culture temperature of 25℃, light intensity of 1800lx, and light time of 12h / d, to obtain

[0044] C. proliferation culture: transferring the induced new buds into a proliferation culture medium, which is made of MS culture medium +NAA0....