Rapid propagation method for cyclobalanopsis glauca tissue culture

A technology of tissue culture and qinggang tree, which is applied in the field of plant propagation, can solve the problems of slow growth and low bud multiplication coefficient, and achieve the effects of large yield of subculture buds, simple operation process and reduced production cost

Inactive Publication Date: 2017-06-06

唐春艳 +1

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AI-Extracted Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a method for tissue culture and propagation of twigs with good growth effect a...

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Abstract

The invention discloses a rapid propagation method for cyclobalanopsis glauca tissue culture. The rapid propagation method comprises the work procedures of selecting explant, processing the explant, inducing and culturing initial buds, multiplication culturing, culturing strong buds and rooting culturing and comprises the steps of collecting underyearling twigs of cyclobalanopsis glauca which is in half lignifications and grows robustly as the explant, pruning and sterilizing the explant, inoculating the explant into an initial bud inducing and culturing medium to obtain the initial buds, then inoculating the initial buds into a multiplication culturing medium, forming cluster buds after 3 to 4 periods of multiplication culture, inoculating the cluster buds into a strong bud culture medium to be cultured and finally inserting single buds with the height larger than or equal to 2cm into a rooting culture medium to be induced and rooted. The rapid propagation method disclosed by the invention shortens a seedling culture period of the cyclobalanopsis glauca, saves seedling culture materials, overcomes the problems that a traditional seedling culture method needs a lot of seedling culture materials and has a long period, greatly reduces production cost, improves production efficiency and has better economical benefit, social benefit and ecological benefit.

Application Domain

Horticulture methodsPlant tissue culture

Technology Topic

BudCulture mediums +5

Examples

Experimental program(3)

Example Embodiment

[0024] Example 1:

[0025] (1) Explant selection: collect semi-lignified and robust growing green shoots of the year as explants;

[0026] (2) Explant treatment: Rinse the explants under tap water for 10 minutes, then remove the leaves of the explants, and then soak the explants in a 0.1% mercury solution, and control the soaking time for 15 minutes. Finally, wash the medicine with pure water, cut the explants into 1.5-3cm long stems with at least one axillary bud, place them in containers according to the degree of lignification of the stems, and reserve them;

[0027] (3) Initial bud induction culture: inoculate the explants obtained in step (2) in the initial bud induction medium and cultivate 30-35 in an environment of light culture, 500 lx light, and culture temperature of 20±1℃ d, initial bud germination and growth; the formula of the initial bud induction medium is: 1/2 modified WPM medium+IBA 2.0 mg/L+6-BA 1.0 mg/L+ZT 1.5 mg/L+ VC 15 mg /L+Folic acid 10mg/L+VB 2 15 mg/+agar 3.0 g/L+carrageenan 25 g/L;

[0028] (4) Proliferation culture: insert the initial buds with height ≥ 1 cm into the proliferation medium for proliferation and culture, and place them in light culture, 2500 lx light, 16 h daily light, and culture temperature 25-28 ℃, 35- It is transferred once in 40 d, cyclically, after 3-4 cycles of cultivation, clumping buds are formed; the formula of the proliferation medium is: modified WPM medium +6-BA 6.0 mg/L+IBA 2.0 mg/L +ZT 0.5 mg/L+VC 15 mg/L+VB 2 15 mg+L-cysteine 20 mg/L+agar 3.6 g/L+carrageenan 30 g/L;

[0029] (5) Cultivation of strong buds: cut the clump buds obtained in step (4), insert 4-5 buds/cluster, into the strong bud culture medium, place in light culture, 2500 lx light, 16 h daily light, and cultivate Cultivate 35-40 d in an environment with a temperature of 25-28℃ to form strong buds; the formula of the strong bud medium is: modified WPM medium +6-BA 3.0 mg/L+IBA 1.0 mg/L+VC 15mg /L+VB 2 15 mg+L-cysteine 20 mg/L+agar 3.5 g/L+carrageenan 25 g/L;

[0030] (6) Rooting culture: insert the single buds with height ≥ 2cm obtained in step (5) into the rooting medium, and inoculate the remaining small bud clusters in the proliferation medium of step (4) to continue to multiply, and then repeat step (5) ), cyclically, to obtain a large number of strong buds; rooting culture is placed in light culture, light 2500 lx, daily light for 16 h, culture temperature 25-28 ℃ to induce rooting, the rooting rate is 95%; said rooting The medium formula is: 1/2 modified WPM medium+6-BA 1.5 mg/L+IBA 1.0mg/L+agar 3.5 g/L+carrageenan 30 g/L.

[0031] The composition and volume-weight ratio of the above-mentioned modified WPM medium are: NH 4 NO 3 230mg/L, CaCl 2 ·2H 2 O76 mg/L, MgSO 4 ·7H 2 0 340 mg/L, KH 2 PO 4 170 mg/L, Ca(NO 3 ) 2 ·4H 2 O 180 mg/L, KI 0.83mg/L, MnSO 4 ·H 2 O 22.4 mg/L, ZnSO 4 ·7H 2 O 8.6 mg/L, Na 2 MoO 4 ·2H 2 O 0.25 mg/L, CuSO 4 ·5H 2 O 0.025 mg/L, Na 2 ·EDTA 37.4 mg/L, FeSO 4 ·7H 2 O 25.2 mg/L, inositol 200 mg/L, niacin 0.5 mg/L, pyridoxine hydrochloride 0.5 mg/L, thiamine hydrochloride 1.0 mg/L, and glycine 2.0 mg/L.

Example Embodiment

[0032] Example 2:

[0033] (1) Explant selection: collect semi-lignified and robust growing green shoots of the year as explants;

[0034] (2) Explant treatment: Rinse the explants under running water for 15 minutes, then remove the leaves of the explants, and then soak the explants in a mercury solution with a volume concentration of 0.1%, and control the soaking time for 20 minutes. Finally, wash the medicine with pure water, cut the explants into 1.5-3cm long stems with at least one axillary bud, place them in containers according to the degree of lignification of the stems, and reserve them;

[0035] (3) Initial bud induction culture: inoculate the explants obtained in step (2) in the initial bud induction medium and cultivate 30-35 in an environment of light culture, 1000 lx light, and culture temperature of 20±1℃ d, initial bud germination and growth; the formula of the initial bud induction medium is: 1/2 modified WPM medium+IBA 3.0 mg/L+6-BA 1.5 mg/L+ZT 1.5mg/L+ VC 15 mg /L+Folic acid 10mg/L+VB 2 15 mg/+agar 3.0 g/L+carrageenan 25 g/L;

[0036] (4) Proliferation culture: Put the initial buds with height ≥1cm into the proliferation medium for proliferation and culture, and place them in light culture, 3000 lx light, 16 hours light per day, and culture temperature 25-28℃, 35- Transfer once in 40 days, cyclically, after 3-4 cycles of culture, clumping buds are formed; the formula of the proliferation medium is: modified WPM medium +6-BA 7.0 mg/L+IBA 2.5 mg/L +ZT 1.0 mg/L+VC 15 mg/L+VB 2 15 mg+L-cysteine 20 mg/L+agar 3.6 g/L+carrageenan 30 g/L;

[0037] (5) Cultivation of strong buds: cut the clump buds obtained in step (4), insert 4-5 buds per clump, and insert them into the strong bud culture medium, and place them in light culture, 3000 lx light, 16 h daily light, and cultivate Cultivate 35-40 d in an environment with a temperature of 25-28℃ to form strong buds; the formula of the strong bud medium is: modified WPM medium+6-BA 3.5 mg/L+IBA 1.0 mg/L+VC 15mg /L+VB 2 15 mg+L-cysteine 20 mg/L+agar 3.5 g/L+carrageenan 25 g/L;

[0038] (6) Rooting culture: insert the single buds with height ≥ 2cm obtained in step (5) into the rooting medium, and inoculate the remaining small bud clusters in the proliferation medium of step (4) to continue to multiply, and then repeat step (5) ), cyclically, to obtain a large number of strong buds; rooting culture is placed in light culture, 3000 lx light, 16 h daily light, and a culture temperature of 25-28℃ to induce rooting, and the rooting rate is 91%; The medium formula is: 1/2 modified WPM medium+6-BA 2.0 mg/L+IBA 1.5mg/L+agar 3.5 g/L+carrageenan 30 g/L.

[0039] The composition and volume-weight ratio of the above-mentioned modified WPM medium are: NH 4 NO 3 230mg/L, CaCl 2 ·2H 2 O76 mg/L, MgSO 4 ·7H 2 0 340 mg/L, KH 2 PO 4 170 mg/L, Ca(NO 3 ) 2 ·4H 2 O 180 mg/L, KI 0.83mg/L, MnSO 4 ·H 2 O 22.4 mg/L, ZnSO 4 ·7H 2 O 8.6 mg/L, Na 2 MoO 4 ·2H 2 O 0.25 mg/L, CuSO 4 ·5H 2 O 0.025 mg/L, Na 2 ·EDTA 37.4 mg/L, FeSO 4 ·7H 2 O 25.2 mg/L, inositol 200 mg/L, niacin 0.5 mg/L, pyridoxine hydrochloride 0.5 mg/L, thiamine hydrochloride 1.0 mg/L, and glycine 2.0 mg/L.

Example Embodiment

[0040] Example 3:

[0041] (1) Explant selection: collect semi-lignified and robust growing green shoots of the year as explants;

[0042] (2) Explant treatment: Rinse the explants under running water for 15 minutes, then remove the leaves of the explants, and then soak the explants in a mercury solution with a volume concentration of 0.1%, and control the soaking time for 20 minutes. Finally, wash the medicine with pure water, cut the explants into 1.5-3cm long stems with at least one axillary bud, place them in containers according to the degree of lignification of the stems, and reserve them;

[0043] (3) Initial bud induction culture: inoculate the explants obtained in step (2) in the initial bud induction medium and cultivate 30-35 in an environment of light culture, 1000 lx light, and culture temperature of 20±1℃ d, initial bud germination and growth; the formula of the initial bud induction medium is: 1/2 modified WPM medium+IBA 2.5 mg/L+6-BA 2.0 mg/L+ZT 2.0mg/L+ VC 15 mg /L+Folic acid 10mg/L+VB 2 15 mg/+agar 3.0 g/L+carrageenan 25 g/L;

[0044] (4) Proliferation culture: insert the initial buds with a height of ≥1cm into a proliferation medium for proliferation and culture, and place them in a light culture, 4000 lx light, 16 hours of light per day, and a culture temperature of 25-28℃, 35- It is transferred once in 40 d, cyclically, after 3-4 cycles of culture, clumping buds are formed; the formula of the proliferation medium is: modified WPM medium +6-BA 8.0 mg/L+IBA 3.0 mg/L +ZT 1.0 mg/L+VC 15 mg/L+VB 2 15 mg+L-cysteine 20 mg/L+agar 3.6 g/L+carrageenan 30 g/L;

[0045] (5) Cultivation of strong buds: cut the clump buds obtained in step (4), insert 4-5 buds/cluster into the strong bud culture medium, and place them in light culture with 4000 lx light, 16 hours of light daily, and cultivate Cultivate 35-40 d in an environment with a temperature of 25-28℃ to form strong buds; the formula of the strong bud medium is: modified WPM medium +6-BA 4.0 mg/L+IBA 1.5 mg/L+VC 15mg /L+VB 2 15 mg+L-cysteine 20 mg/L+agar 3.5 g/L+carrageenan 25 g/L;

[0046] (6) Rooting culture: insert the single buds with height ≥ 2cm obtained in step (5) into the rooting medium, and inoculate the remaining small bud clusters in the proliferation medium of step (4) to continue to multiply, and then repeat step (5) ), cyclically, to obtain a large number of strong buds; rooting culture is placed in light culture, 4000 lx light, 16 hours of light per day, culture temperature 25-28 ℃ to induce rooting, the rooting rate is 96%;; The formula of rooting medium is: 1/2 modified WPM medium+6-BA 2.5 mg/L+IBA 1.5mg/L+agar 3.5 g/L+carrageenan 30 g/L.

[0047] The composition and volume-weight ratio of the above-mentioned modified WPM medium are: NH 4 NO 3 230mg/L, CaCl 2 ·2H 2 O76 mg/L, MgSO 4 ·7H 2 0 340 mg/L, KH 2 PO 4 170 mg/L, Ca(NO 3 ) 2 ·4H 2 O 180 mg/L, KI 0.83mg/L, MnSO 4 ·H 2 O 22.4 mg/L, ZnSO 4 ·7H 2 O 8.6 mg/L, Na 2 MoO 4 ·2H 2 O 0.25 mg/L, CuSO 4 ·5H 2 O 0.025 mg/L, Na 2 ·EDTA 37.4 mg/L, FeSO 4 ·7H 2 O 25.2 mg/L, inositol 200 mg/L, niacin 0.5 mg/L, pyridoxine hydrochloride 0.5 mg/L, thiamine hydrochloride 1.0 mg/L, and glycine 2.0 mg/L.

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Classification and recommendation of technical efficacy words

Shorten the breeding cycle

Rapid propagation method for cyclobalanopsis glauca tissue culture

A technology of tissue culture and qinggang tree, which is applied in the field of plant propagation, can solve the problems of slow growth and low bud multiplication coefficient, and achieve the effects of large yield of subculture buds, simple operation process and reduced production cost

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

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Description & Claims & Application Information

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