Intermediate propagation method for Diqing wild Lores comiculatus Linn.

Abstract

The invention relates to an intermediate propagation method for Diqing wild Lores comiculatus Linn., comprising the following steps: preparation and sterilization of an MS culture medium: performing pressure maintaining sterilization for 30min by using a high pressure sterilization pot, wherein each litre of the culture medium is prepared from 50ml of major elements, 5ml of microelements, 5ml of organic elements, 5ml of ferric salt, 30g of saccharose, 0.1g of inositol and 8g of agar powder, and has the pH of 5.8; disinfection of Lores comiculatus Linn. seeds and sterile inoculation: when two seed leaves are unfolded after seed germination on the 14d, taking a hypocotyledonary axis, cutting into sections, inoculating on the MS culture medium containing 2,4-D and KT and having the pH of 5.8, and grafting a callus subjected to subculture for 2 to 4 times into an MS culture medium containing NAA and 6-BA; when induced buds grow to 5 to 8cm, cutting adventitious buds into small stem sections of 1.0cm, and inserting the lower ends of the small stem sections into a 1 / 2 MS culture medium containing NAA and carbon powder for rooting, wherein each small stem section has at least one internode. The intermediate propagation method for the Diqing wild Lores comiculatus Linn. is short in propagation expanding time and large in propagation coefficient, and the propagation coefficient of the Diqing wild Lores comiculatus Linn. is increased, and a propagation cycle is shortened.

Description

technical field

[0001] The invention relates to the technical field of plant propagation, in particular to a method for rapid propagation of Diqing wild-type japonicus japonicus. Background technique

[0002] Lorescomiculatus Linn., also known as five-leaf clover (four-leaf clover), is distributed in Yunnan, Sichuan, Guizhou, Gansu, Hubei and other places in my country. The development of animal husbandry in Yunnan requires a large amount of leguminous forage. Since 1983, a large-scale introduction of leguminous forage has been introduced, and 21 varieties have been screened out. Although they grow well in various places, they have not been widely used in production. Diqing wild-type japonicus japonicus is widely distributed in mountains, forest gaps, and resting areas at an altitude of 1400-3400m in western Yunnan, and is an important leguminous herbage in natural grasslands. Diqing wild type japonicus is heat-resistant, drought-resistant, and aluminum-resistant. Its branc...

Examples

Embodiment 1

[0017] 1) preparation of medium: select MS medium for use (include in every liter of medium: macronutrient 50ml, trace element 5ml, organic element 5ml, iron salt 5ml, sugar 30g, inositol 0.1g, agar powder 8g) pH is 5.8, the details are as follows: Take about 400ml of tap water and pour it into a small aluminum pot to heat. After the water boils, add 8g of agar powder and stir while heating. Be careful to prevent the bottom from sticking. It is better to melt completely and see the bottom. After 25 minutes, turn off the heat, add 30g of sucrose, 0.1g of inositol and the measured mother liquor medicine, and stir for 5 minutes. Pour into a 1000ml constant volume beaker to constant volume. Use a pH meter to measure the pH of the medium, and adjust it to about 5.8 pH with 0.1% NaOH or HCl solution when necessary. After adjusting the pH, be sure to stir evenly, and be careful not to use acid or alkali solutions that have been left for too long.

[0018] Described mother liquor me...

Embodiment 2

[0026] Step 1), step 2) are the same as embodiment 1,

[0027] 3) Induction of callus: when the seeds germinated on 14 days, two cotyledons were unfolded, and the hypocotyls were taken, cut into 0.5 cm long sections, and inoculated in a seed containing 2,4-D (2,4-dichlorobenzene). Oxyacetic acid) 2mg / L, KT (kinetin) 1mg / L, on the MS medium of pH=5.8, each treatment combination is repeated three times;

[0028] 4) Induced differentiation of buds: the hormone concentration combination medium for inducing bud differentiation is: MS medium containing 0.5 mg / L of NAA (naphthalene acetic acid) and 0.5 mg / L of 6-BA (6-benzylaminoadenine), followed by 18-32 days after the calli of passage 2-4 were inserted into the culture medium, the shoot length was observed;

[0029] 5) Root induction: When the induced buds grow to 5-8cm, cut the adventitious buds into 1.0cm small stem segments, each of which has at least one internode, and insert the lower end of the small stem segments containin...

Embodiment 3

[0031] Step 1), step 2) are the same as embodiment 1,

[0032] 3) Induction of callus: when the seeds germinated on 14 days, two cotyledons were unfolded, and the hypocotyls were taken, cut into 0.5 cm long sections, and inoculated in a seed containing 2,4-D (2,4-dichlorobenzene). Oxyacetic acid) 4mg / L, KT (kinetin) 5mg / L, on the MS medium of pH=5.8, each treatment combination is repeated three times;

[0033] 4) Induced differentiation of buds: The hormone concentration combination medium for inducing bud differentiation is: MS medium containing 1.5 mg / L of NAA (naphthalene acetic acid) and 0.5 mg / L of 6-BA (6-benzylaminoadenine), followed by 18-32 days after the calli of passage 2-4 were inserted into the culture medium, the shoot length was observed;

[0034] 5) Root induction: When the induced buds grow to 5-8cm, cut the adventitious buds into 1.0cm small stem segments, each of which has at least one internode, and insert the lower end of the small stem segments containin...

technical field

[0001] The invention relates to the technical field of plant propagation, in particular to a method for rapid propagation of Diqing wild-type japonicus japonicus. Background technique

[0002] Lorescomiculatus Linn., also known as five-leaf clover (four-leaf clover), is distributed in Yunnan, Sichuan, Guizhou, Gansu, Hubei and other places in my country. The development of animal husbandry in Yunnan requires a large amount of leguminous forage. Since 1983, a large-scale introduction of leguminous forage has been introduced, and 21 varieties have been screened out. Although they grow well in various places, they have not been widely used in production. Diqing wild-type japonicus japonicus is widely distributed in mountains, forest gaps, and resting areas at an altitude of 1400-3400m in western Yunnan, and is an important leguminous herbage in natural grasslands. Diqing wild type japonicus is heat-resistant, drought-resistant, and aluminum-resistant. Its branc...

Examples

Embodiment 1

[0017] 1) preparation of medium: select MS medium for use (include in every liter of medium: macronutrient 50ml, trace element 5ml, organic element 5ml, iron salt 5ml, sugar 30g, inositol 0.1g, agar powder 8g) pH is 5.8, the details are as follows: Take about 400ml of tap water and pour it into a small aluminum pot to heat. After the water boils, add 8g of agar powder and stir while heating. Be careful to prevent the bottom from sticking. It is better to melt completely and see the bottom. After 25 minutes, turn off the heat, add 30g of sucrose, 0.1g of inositol and the measured mother liquor medicine, and stir for 5 minutes. Pour into a 1000ml constant volume beaker to constant volume. Use a pH meter to measure the pH of the medium, and adjust it to about 5.8 pH with 0.1% NaOH or HCl solution when necessary. After adjusting the pH, be sure to stir evenly, and be careful not to use acid or alkali solutions that have been left for too long.

[0018] Described mother liquor me...

Embodiment 2

[0026] Step 1), step 2) are the same as embodiment 1,

[0027] 3) Induction of callus: when the seeds germinated on 14 days, two cotyledons were unfolded, and the hypocotyls were taken, cut into 0.5 cm long sections, and inoculated in a seed containing 2,4-D (2,4-dichlorobenzene). Oxyacetic acid) 2mg / L, KT (kinetin) 1mg / L, on the MS medium of pH=5.8, each treatment combination is repeated three times;

[0028] 4) Induced differentiation of buds: the hormone concentration combination medium for inducing bud differentiation is: MS medium containing 0.5 mg / L of NAA (naphthalene acetic acid) and 0.5 mg / L of 6-BA (6-benzylaminoadenine), followed by 18-32 days after the calli of passage 2-4 were inserted into the culture medium, the shoot length was observed;

[0029] 5) Root induction: When the induced buds grow to 5-8cm, cut the adventitious buds into 1.0cm small stem segments, each of which has at least one internode, and insert the lower end of the small stem segments containin...

Embodiment 3

[0031] Step 1), step 2) are the same as embodiment 1,

[0032] 3) Induction of callus: when the seeds germinated on 14 days, two cotyledons were unfolded, and the hypocotyls were taken, cut into 0.5 cm long sections, and inoculated in a seed containing 2,4-D (2,4-dichlorobenzene). Oxyacetic acid) 4mg / L, KT (kinetin) 5mg / L, on the MS medium of pH=5.8, each treatment combination is repeated three times;

[0033] 4) Induced differentiation of buds: The hormone concentration combination medium for inducing bud differentiation is: MS medium containing 1.5 mg / L of NAA (naphthalene acetic acid) and 0.5 mg / L of 6-BA (6-benzylaminoadenine), followed by 18-32 days after the calli of passage 2-4 were inserted into the culture medium, the shoot length was observed;

[0034] 5) Root induction: When the induced buds grow to 5-8cm, cut the adventitious buds into 1.0cm small stem segments, each of which has at least one internode, and insert the lower end of the small stem segments containin...

Images