Two-step seedling formation method of daphniphyllum calycinum

Abstract

The invention belongs to the technical field of plant tissue culture, and particularly relates to a two-step seedling formation method of daphniphyllum calycinum. The two-step seedling formation method of daphniphyllum calycinum includes the following steps that 1, newly germinated tender leaves of daphniphyllum calycinum are taken as explants, after sterilization in a conventional way, the explants are placed on a super-clean worktable, the edges of the leaves are cut with an anatomical knife into small pieces of 1 square centimeter, and the small pieces are placed in a modified WPM for 35 days to obtain aseptic test-tube plantlets; 2, rooting culture is conducted; 3, seedling exercising and transplanting are conducted. The multiplication coefficient of the daphniphyllum-calycinum clusterbuds obtained by the culture method reaches 30-40 times, the cluster buds are robust and take roots easily after being inoculated into a rooting medium, and the rooting rate can reach 90% or above.

Description

technical field

[0001] The invention belongs to the technical field of plant tissue culture, in particular to a two-step seedling-forming method of the eucalyptus. Background technique

[0002] Cow ear maple (Daphniphyllum calycinum Benth.) Alias: Bell pork belly, pig belly fruit, pig belly wood, pig belly, pig Luo wood, pig network wood, pig wood, Zhuluzi, goat dung, sheep dung, tiger ear , Crow Gallbladder, Big Ear Comb, Nanling Hunan, Nanling Tiger Skin Nan and Lingnan Tiger Skin Nan. It is a plant of the genus Panacaceae.

[0003] The fruit branches, leaves and roots of the Fructus japonica can be used as medicine, and the branches and leaves: expelling wind and relieving pain; detoxifying and reducing swelling. Mainly rheumatic bone pain; sore swelling and poison; bruises and fractures; poisonous snake bites. Roots: heat-clearing and toxic substances removing; promoting blood circulation and removing blood stasis; reducing swelling and relieving pain. Indicates feve...

Examples

Embodiment 1

[0022] A two-step method for growing seedlings of maple leaves, comprising the following steps:

[0023] (1) Take the young leaves of the newly sprouted maple as explants, and put them in the ultra-clean workbench, cut the edge of the leaves with a scalpel, cut them into small pieces of 1 cm square, and place them in WPM for cultivation medium, cultivated for 35 days under the condition of culture temperature of 23°C, light intensity of 1500lux, and light time of 8h / day to obtain sterile test-tube plantlets; wherein the WPM medium contained 1.0mg / L brassinolide, 0.2 mg / L indolebutyric acid, 0.2mg / L kinetin, 1.0mg / L gibberellin, 2mg / L polyvinylpyrrolidone PVP, 30g / L sucrose and 5g / L agar, the pH value of the medium is 5.8;

[0024] (2) Rooting culture: Place the robust plants obtained in step (1) in 1 / 2MS rooting medium at a culture temperature of 23°C, a light intensity of 3000 lux, and a light time of 8 hours / day for 30 days to obtain complete Rooted shoots, wherein 1.0mg / L...

Embodiment 2

[0027] A two-step method for growing seedlings of maple leaves, comprising the following steps:

[0028] (1) Take the young leaves of the newly sprouted maple as explants, and put them in the ultra-clean workbench, cut the edge of the leaves with a scalpel, cut them into small pieces of 1 cm square, and place them in WPM for cultivation culture medium, at a culture temperature of 27°C, a light intensity of 1500 lux, and a light time of 10 hours / day to cultivate sterile test-tube plantlets for 35 days; wherein the WPM medium contains 1.5 mg / L of brassinolide, 0.2mg / L indolebutyric acid, 0.2mg / L kinetin, 0.5mg / L gibberellin, 2mg / L polyvinylpyrrolidone PVP, 30g / L sucrose and 5g / L agar, the pH of the medium The value is 5.8;

[0029] (2) Rooting culture: place the robust plants obtained in step (1) in 1 / 2MS rooting medium at a culture temperature of 27°C, with a light intensity of 3000 lux and a light time of 10 hours / day for 30 days to obtain complete Rooted shoots, wherein 1 / 2...

Embodiment 3

[0032] A two-step method for growing seedlings of maple leaves, comprising the following steps:

[0033] (1) Take the young leaves of the newly sprouted maple as explants, and put them in the ultra-clean workbench, cut the edge of the leaves with a scalpel, cut them into small pieces of 1 cm square, and place them in WPM for cultivation culture medium, at a culture temperature of 25°C, a light intensity of 1500 lux, and a light time of 9 hours / day, cultured for 35 days to obtain sterile test-tube plantlets; wherein the WPM medium contained 2.0 mg / L of brassinolide, 0.2mg / L indolebutyric acid, 0.5mg / L kinetin, 0.5mg / L gibberellin, 2mg / L polyvinylpyrrolidone PVP, 30g / L sucrose and 5g / L agar, the pH of the medium The value is 5.8;

[0034] (2) Rooting culture: place the strong plants obtained in step (1) in 1 / 2MS rooting medium at a culture temperature of 25°C, light intensity of 3000 lux, and light time of 9 hours / day under the condition of cultivating for 30 days to obtain com...

technical field

[0001] The invention belongs to the technical field of plant tissue culture, in particular to a two-step seedling-forming method of the eucalyptus. Background technique

[0002] Cow ear maple (Daphniphyllum calycinum Benth.) Alias: Bell pork belly, pig belly fruit, pig belly wood, pig belly, pig Luo wood, pig network wood, pig wood, Zhuluzi, goat dung, sheep dung, tiger ear , Crow Gallbladder, Big Ear Comb, Nanling Hunan, Nanling Tiger Skin Nan and Lingnan Tiger Skin Nan. It is a plant of the genus Panacaceae.

[0003] The fruit branches, leaves and roots of the Fructus japonica can be used as medicine, and the branches and leaves: expelling wind and relieving pain; detoxifying and reducing swelling. Mainly rheumatic bone pain; sore swelling and poison; bruises and fractures; poisonous snake bites. Roots: heat-clearing and toxic substances removing; promoting blood circulation and removing blood stasis; reducing swelling and relieving pain. Indicates feve...

Examples

Embodiment 1

[0022] A two-step method for growing seedlings of maple leaves, comprising the following steps:

[0023] (1) Take the young leaves of the newly sprouted maple as explants, and put them in the ultra-clean workbench, cut the edge of the leaves with a scalpel, cut them into small pieces of 1 cm square, and place them in WPM for cultivation medium, cultivated for 35 days under the condition of culture temperature of 23°C, light intensity of 1500lux, and light time of 8h / day to obtain sterile test-tube plantlets; wherein the WPM medium contained 1.0mg / L brassinolide, 0.2 mg / L indolebutyric acid, 0.2mg / L kinetin, 1.0mg / L gibberellin, 2mg / L polyvinylpyrrolidone PVP, 30g / L sucrose and 5g / L agar, the pH value of the medium is 5.8;

[0024] (2) Rooting culture: Place the robust plants obtained in step (1) in 1 / 2MS rooting medium at a culture temperature of 23°C, a light intensity of 3000 lux, and a light time of 8 hours / day for 30 days to obtain complete Rooted shoots, wherein 1.0mg / L...

Embodiment 2

[0027] A two-step method for growing seedlings of maple leaves, comprising the following steps:

[0028] (1) Take the young leaves of the newly sprouted maple as explants, and put them in the ultra-clean workbench, cut the edge of the leaves with a scalpel, cut them into small pieces of 1 cm square, and place them in WPM for cultivation culture medium, at a culture temperature of 27°C, a light intensity of 1500 lux, and a light time of 10 hours / day to cultivate sterile test-tube plantlets for 35 days; wherein the WPM medium contains 1.5 mg / L of brassinolide, 0.2mg / L indolebutyric acid, 0.2mg / L kinetin, 0.5mg / L gibberellin, 2mg / L polyvinylpyrrolidone PVP, 30g / L sucrose and 5g / L agar, the pH of the medium The value is 5.8;

[0029] (2) Rooting culture: place the robust plants obtained in step (1) in 1 / 2MS rooting medium at a culture temperature of 27°C, with a light intensity of 3000 lux and a light time of 10 hours / day for 30 days to obtain complete Rooted shoots, wherein 1 / 2...

Embodiment 3

[0032] A two-step method for growing seedlings of maple leaves, comprising the following steps:

[0033] (1) Take the young leaves of the newly sprouted maple as explants, and put them in the ultra-clean workbench, cut the edge of the leaves with a scalpel, cut them into small pieces of 1 cm square, and place them in WPM for cultivation culture medium, at a culture temperature of 25°C, a light intensity of 1500 lux, and a light time of 9 hours / day, cultured for 35 days to obtain sterile test-tube plantlets; wherein the WPM medium contained 2.0 mg / L of brassinolide, 0.2mg / L indolebutyric acid, 0.5mg / L kinetin, 0.5mg / L gibberellin, 2mg / L polyvinylpyrrolidone PVP, 30g / L sucrose and 5g / L agar, the pH of the medium The value is 5.8;

[0034] (2) Rooting culture: place the strong plants obtained in step (1) in 1 / 2MS rooting medium at a culture temperature of 25°C, light intensity of 3000 lux, and light time of 9 hours / day under the condition of cultivating for 30 days to obtain com...