Efficient one-step regeneration method taking root system of broussonetia papyrifera as explant

Abstract

The invention discloses an efficient one-step regeneration method taking a root system of broussonetia papyrifera as an explant. The method comprises the following steps: S1, adventitious bud induction: selecting a root system of a broussonetia papyrifera aseptic seedling as the explant, cutting out a wound, inoculating the explant to an adventitious bud induction medium, and conducting inducing to form adventitious buds; and S2, rooting culture: cutting the adventitious buds in the step S1, inoculating the adventitious buds to a rooting culture medium for rooting induction, and culturing the adventitious buds until the adventitious buds root to form regenerated seedlings. According to the method, the broussonetia papyrifera in-vitro regeneration plant is successfully obtained through the steps of adventitious bud induction, adventitious bud rooting, domestication and transplanting and the like by taking the root system of the broussonetia papyrifera as the explant, and compared with a culture method taking other broussonetia papyrifera tissues as explants, the method is simpler and more convenient to operate, wider in material taking range and more efficient in propagation.

Description

technical field

[0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a high-efficiency one-step regeneration method using the root system of a mulberry tree as an explant. Background technique

[0002] Broussonetia Papyrifera (L.) Vent., also known as hair peach, mulberry peach, paper mulberry, valley tree, etc., is a perennial deciduous tree belonging to the family Moraceae. The adult tree is 10-20m high; the bark is dark gray; the whole plant contains latex; the branchlets are densely pilose; the leaves are broadly ovate to oblong-ovate and spirally arranged, the apex is acuminate, the base is heart-shaped, and the edge is coarsely serrated. The leaves of small trees often have obvious 3-5 lobes, and the back is densely covered with fluff; the petiole is 2.5-8cm long and densely covered with strigose; flowers are dioecious; the male inflorescence is a catkin, and the female inflorescence is spherical-headed; the fleshy c...

Examples

Embodiment 1

[0036] (1) Inducing adventitious buds: select the root system of the aseptic seedlings of S. mulberry as explants, cut 2 to 3 cuts to increase the wound; inoculate it on the adventitious bud induction medium in a horizontal manner, at a temperature of 25°C and light intensity 2500lx, and the light time is 12h per day for cultivating until adventitious buds are induced. The adventitious bud induction medium is: 1 / 2MS+1.0mg / L 6-BA+0.1mg / L NAA+0.05mg / L IBA+30g / L sucrose+6.0g / L agar, with a pH of 5.8.

[0037] (2) Rooting culture: Cut the adventitious buds up to 2 cm in step (1) and inoculate them on the rooting medium to induce rooting. After inoculation, first cultivate them in total darkness at 25°C for 3 days, and then place them under light for 12 hours a day , the light intensity is 1500lx, and the cultivation temperature is cultivated until rooting under the condition of 25°C. The rooting medium is: 1 / 2MS+0.1mg / L NAA+20g / L sucrose+4.0g / L agar, pH is 5.8.

[0038] (3) Hard...

Embodiment 2

[0040] (1) Inducing adventitious buds: select the root system of the aseptic seedlings of S. mulberry as explants, cut 2 to 3 cuts to increase the wound; inoculate it on the adventitious bud induction medium in a horizontal manner, at a temperature of 25°C and light intensity 2500lx, and the light time is 15h per day for cultivating until adventitious buds are induced. The adventitious bud induction medium is: 1 / 2MS+1.5mg / L 6-BA+0.05mg / L NAA+0.01mg / L IBA+20g / L sucrose+6.0g / L agar, with a pH of 6.0.

[0041] (2) Rooting culture: Cut the adventitious buds up to 4 cm in step (1) and inoculate them on the rooting medium to induce rooting. After inoculation, first culture them in total darkness at 25°C for 3 days, and then place them under light for 15 hours a day , the light intensity is 2500lx, and the cultivation temperature is cultivated until rooting under the condition of 28° C.; the rooting rate reaches 85%. The rooting medium is: 1 / 2MS+0.1mg / LNAA+15g / L sucrose+4.0g / L agar,...

Embodiment 3

[0044] (1) Inducing adventitious buds: select the root system of the aseptic seedlings of S. mulberry as explants, cut 2 to 3 cuts to increase the wound; inoculate it on the adventitious bud induction medium in a horizontal manner, at a temperature of 28°C and light intensity 1500lx, light time is 15h under the condition of cultivation, until induction to form adventitious buds. The adventitious bud induction medium is: 1 / 2MS+1.5mg / L 6-BA+0.2mg / LNAA+0.1mg / L IBA+25g / L sucrose+6.0g / L agar, with a pH of 6.2.

[0045] (2) Rooting culture: Cut the adventitious buds up to 3 cm in step (1) and inoculate them on the rooting medium to induce rooting. After the inoculation, first cultivate them in the dark at 28°C for 5 days, and then place them in the light for 12 hours every day. , under the condition that the light intensity is 2500lx and the culture temperature is 25°C, it is cultivated until rooting. Described rooting medium is: 1 / 2MS+0.4mg / L NAA+30g / L sucrose+6.0g / L agar, pH is 5...

technical field

[0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a high-efficiency one-step regeneration method using the root system of a mulberry tree as an explant. Background technique

[0002] Broussonetia Papyrifera (L.) Vent., also known as hair peach, mulberry peach, paper mulberry, valley tree, etc., is a perennial deciduous tree belonging to the family Moraceae. The adult tree is 10-20m high; the bark is dark gray; the whole plant contains latex; the branchlets are densely pilose; the leaves are broadly ovate to oblong-ovate and spirally arranged, the apex is acuminate, the base is heart-shaped, and the edge is coarsely serrated. The leaves of small trees often have obvious 3-5 lobes, and the back is densely covered with fluff; the petiole is 2.5-8cm long and densely covered with strigose; flowers are dioecious; the male inflorescence is a catkin, and the female inflorescence is spherical-headed; the fleshy c...

Examples

Embodiment 1

[0036] (1) Inducing adventitious buds: select the root system of the aseptic seedlings of S. mulberry as explants, cut 2 to 3 cuts to increase the wound; inoculate it on the adventitious bud induction medium in a horizontal manner, at a temperature of 25°C and light intensity 2500lx, and the light time is 12h per day for cultivating until adventitious buds are induced. The adventitious bud induction medium is: 1 / 2MS+1.0mg / L 6-BA+0.1mg / L NAA+0.05mg / L IBA+30g / L sucrose+6.0g / L agar, with a pH of 5.8.

[0037] (2) Rooting culture: Cut the adventitious buds up to 2 cm in step (1) and inoculate them on the rooting medium to induce rooting. After inoculation, first cultivate them in total darkness at 25°C for 3 days, and then place them under light for 12 hours a day , the light intensity is 1500lx, and the cultivation temperature is cultivated until rooting under the condition of 25°C. The rooting medium is: 1 / 2MS+0.1mg / L NAA+20g / L sucrose+4.0g / L agar, pH is 5.8.

[0038] (3) Hard...

Embodiment 2

[0040] (1) Inducing adventitious buds: select the root system of the aseptic seedlings of S. mulberry as explants, cut 2 to 3 cuts to increase the wound; inoculate it on the adventitious bud induction medium in a horizontal manner, at a temperature of 25°C and light intensity 2500lx, and the light time is 15h per day for cultivating until adventitious buds are induced. The adventitious bud induction medium is: 1 / 2MS+1.5mg / L 6-BA+0.05mg / L NAA+0.01mg / L IBA+20g / L sucrose+6.0g / L agar, with a pH of 6.0.

[0041] (2) Rooting culture: Cut the adventitious buds up to 4 cm in step (1) and inoculate them on the rooting medium to induce rooting. After inoculation, first culture them in total darkness at 25°C for 3 days, and then place them under light for 15 hours a day , the light intensity is 2500lx, and the cultivation temperature is cultivated until rooting under the condition of 28° C.; the rooting rate reaches 85%. The rooting medium is: 1 / 2MS+0.1mg / LNAA+15g / L sucrose+4.0g / L agar,...

Embodiment 3

[0044] (1) Inducing adventitious buds: select the root system of the aseptic seedlings of S. mulberry as explants, cut 2 to 3 cuts to increase the wound; inoculate it on the adventitious bud induction medium in a horizontal manner, at a temperature of 28°C and light intensity 1500lx, light time is 15h under the condition of cultivation, until induction to form adventitious buds. The adventitious bud induction medium is: 1 / 2MS+1.5mg / L 6-BA+0.2mg / LNAA+0.1mg / L IBA+25g / L sucrose+6.0g / L agar, with a pH of 6.2.

[0045] (2) Rooting culture: Cut the adventitious buds up to 3 cm in step (1) and inoculate them on the rooting medium to induce rooting. After the inoculation, first cultivate them in the dark at 28°C for 5 days, and then place them in the light for 12 hours every day. , under the condition that the light intensity is 2500lx and the culture temperature is 25°C, it is cultivated until rooting. Described rooting medium is: 1 / 2MS+0.4mg / L NAA+30g / L sucrose+6.0g / L agar, pH is 5...

Images