CN103798143A - Out-bottle cutting and rooting method for sequoia sempervirens tissue culture seedling - Google Patents
Out-bottle cutting and rooting method for sequoia sempervirens tissue culture seedling Download PDFInfo
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Abstract
The invention provides an out-bottle cutting and rooting method for a sequoia sempervirens tissue culture seedling. The out-bottle cutting and rooting method comprises the following steps: taking and inserting a sterile sequoia sempervirens tissue culture multiplied seedling taken as a cutting slip into a base material composed of perlite, turf and red soil in a ratio of 1 to 1 to 2; putting the cutting slip into a greenhouse with the light shielding rate being 50-70%; keeping the air humidity to be more than 90% in one to two weeks after cottage is implemented at the temperature of 23+/-2 DEG C; keeping the humidity to be 60-80% within 30-40 days after two weeks to obtain a rooting seedling; transplanting the rooting seedling into a mixed base material of the perlite, humus and the red soil in a ratio of 1 to 1 to 2; putting the rooting seedling into the greenhouse with the light shielding rate being 50-70%; and growing until young leaves grow to obtain a sequoia sempervirens plant with a developed root system, a healthy and strong plant and the transplanting survival rate being more than 90%. The problems that the production period is long, the transplanting survival rate of the tissue culture seedling is low and the industrial production cannot be realized in the tissue culture process of sequoia sempervirens are solved.
Description
Technical field
The present invention relates to a kind of plant cultivation method, especially the outer cuttage root-taking cultural method of sequoia sempervirens group training seedling bottle, belongs to technical field of plant culture.
Background technology
Sequoia sempervirens (
sequoia sempervirens(Larnb.) Endl.), originate in U.S. pacific rim, have another name called Chinese larch, world grandfather, American Red Chinese firs etc., for Taxodiaceae sequoia sempervirens belongs to, are the rare few survivors seeds in the world.Sequoia sempervirens has very high ornamental value, is the fast-growing commerical tree species of world's preciousness and fabulous green tree species.Why belonging to rare seeds is because its seed lacks, and exists germination rate low in seminal propagation process, is only 4.5%～11.9%, and seedling genetic variation is large.Face cottage propagation is taken root unstable, and because adopting, fringe maternal plant difference is minimum reaches 16% to rooting rate, reaches as high as 100%, and generally lower than 50%, and cuttage seeding plant type is not good, thus though cause sequoia sempervirens successful introduction already, its provenance is less, always quite concerned.Plant Tissue Breeding is undoubtedly numerous plants and solves the effective way of planting source problem.
The tissue culture technique of sequoia sempervirens just started to have studied from the eighties in 20th century, but study more or after 2000, after Hu Qing etc. are sterile-processed with sequoia sempervirens seed, be inoculated into the aseptic explant stem section obtaining in MS medium, for experiment material is set up tissue culture propagation technical system, rootage duration is about 50d left and right, rooting rate 65%, after practicing seedling 30～40d, transplant, survival rate is more than 65%.The sequoia sempervirens tissue culture technique that Chen Fang etc. set up, rooting rate is 72.16%, after transplanting, under the suitable condition of temp. and humidity, survival rate can reach more than 80%.The first superfine experiment of hormon on the impact of sequoia sempervirens in-vitro propagate of having carried out of week, has set up corresponding tissue culture technology system, and rooting rate is 74%.The refined bright grade of jade pendant is by the asexual quick propagating technology research of cold resistance sequoia sempervirens is found, the root media rooting rate screening is 72%.Dong Linna etc., by studying the impact of different hormones on sequoia sempervirens in-vitro propagate, have set up corresponding tissue culture technology system, and in root media, cultivating its rooting rate after 45 days is 86.7%.
In summary it can be seen that researcher adopts spray, seed, stem section etc. for explant, all set up the tissue culture technique system of sequoia sempervirens, but rooting rate is lower, transplanting survival rate is also low, and growth cycle is long, therefore, be necessary to research and develop new culture technique and solve the key issue such as rooting rate, survival rate.
Summary of the invention
The object of the present invention is to provide a kind of method that can improve sequoia sempervirens group training seedling rooting rate and transplanting survival rate, shorten growth cycle simultaneously, promote the tissue-culturing rapid propagation of sequoia sempervirens, meet the market demand of sequoia sempervirens tree.
The present invention completes by following technical proposal: a kind of outer cuttage root-taking method of sequoia sempervirens group training seedling bottle, using aseptic sequoia sempervirens group training propagation seedling as cuttings, is characterized in that comprising the following steps:
A, by aseptic cuttings cuttage in the sterilization matrix of following volume ratio: perlite: the peat composed of rotten mosses: laterite=1:1:2, be placed in shading rate and be in 50～70% booth, and in after cuttage 1～2 week, keep air humidity more than 90%, temperature is at 23 ± 2 ℃, after 2 weeks, between 30～40 days, keeps humidity 60～80%, to keep nursery stock blade not wilt, the seedling of must taking root;
B, the steps A gained seedling of taking root is moved in the mixed-matrix of following volume ratio: perlite: humus soil: laterite=1:1:2, be placed in shading rate and be 50～70% booth, keep when high temperature daytime ventilating, and routinely with broad-spectrum germicide spray disinfection and water, and after noting while watering doing, rewater, grow to extraction young leaves, the group that obtains sequoia sempervirens is cultivated seedling, after stem lignification, can transplant to land for growing field crops.
Described aseptic sequoia sempervirens group training propagation seedling obtains through the following steps:
1), take the spray of sequoia sempervirens as explant, after induction is sprouted routinely, proceed in following medium:
MS medium
6-BA 0.1—0.3mg/L
NAA 0.1—0.3mg/L
KT 0.1—0.3mg/L
Sugar 20-30g/L
Agar 6g/L
Be 21～25 ℃ in temperature, intensity of illumination is 2000～2500Lx, and light application time is 12～16h/ days, under the condition that medium pH is 5.6～5.8, cultivates 20～30 days;
2) grow to 3-4cm when high when the group training propagation seedling seedling strain of step 1), be placed in shading rate and be 70% booth, hardening 5～8d, must organize training and breed seedling;
3) by step 2) group training propagation seedling take out, clean the medium on it with clear water, then use 1000 times of liquid of tpn wetting powder of 75% mass ratio, disinfect after 1～2min, obtain aseptic sequoia sempervirens group training propagation seedling.
The cuttage of described steps A is: first group training propagation seedling is cut into individual plant seedling, and each strain seedling base portion is immersed in the following water of taking root 30 seconds:
NAA 500—700mg/L
IBA 100—300mg/L
Cuttage afterwards, in matrix, is sprayed normal root water, the 1000 times of liquid of the thiophanate methyl wetting powder that sprays 70% appropriate mass ratio for second day after cuttage.
The sterilization matrix of described steps A is: every cubic metre of matrix, the carbendazol wettable powder sterilization that carries out disinfection of 20 gram of 50% mass ratio of spray, is sub-packed in the standard cave dish in 105 holes or 128 holes afterwards, and waters permeablely, obtains sterilization matrix.
The present invention has following advantages and effect: adopts such scheme, can cultivate heritability stable, and well developed root system, robust plant, transplanting survival rate reaches more than 90% plant.Simultaneously by the present invention perfect sequoia sempervirens tissue-culturing rapid propagation system, do not need through the program of taking root in tissue culture bottle, saving group training space, chamber reaches more than 50%, saving group training seedling cost is more than 30%, greatly shorten the group training cycle, improve the efficiency of sequoia sempervirens culture of rootage, solved that production cycle in sequoia sempervirens tissue culture procedures is long, group training transplantation of seedlings survival rate is low and cannot realize the problem that industrialization is produced.
Embodiment
Below in conjunction with embodiment, the present invention is described further.
Embodiment 1
A kind of outer cuttage root-taking method of sequoia sempervirens group training seedling bottle, comprises the following steps:
1), take the spray of sequoia sempervirens as explant, after induction is sprouted routinely, proceed in following medium:
MS medium
6-BA 0.1mg/L
NAA 0.3mg/L
KT 0.1mg/L
Sugar 20g/L
Agar 6g/L
Be 21 ℃ in temperature, intensity of illumination is 2500Lx, and light application time is 12h/ days, under the condition that medium pH is 5.6, cultivates 30 days;
2) grow to 3-4cm when high when the group training propagation seedling seedling strain of step 1), be placed in shading rate and be 70% booth, hardening 5d, must organize training and breed seedling;
3) by step 2) group training propagation seedling take out, clean the medium on it with clear water, then use 1000 times of liquid of tpn wetting powder of 75% mass ratio, disinfect after 1min, obtain aseptic sequoia sempervirens group training propagation seedling;
4) step 3) gained group training propagation seedling is cut into individual plant seedling, and each strain seedling base portion is immersed in the following water of taking root 30 seconds:
NAA 500mg/L
IBA 300mg/L；
5) by the seedling cuttage of step 4) in the sterilization matrix in the standard cave dish in 105 holes or 128 holes, spray normal root water, 1000 times of liquid of the thiophanate methyl wetting powder that sprays 70% appropriate mass ratio for second day after cuttage, described sterilization matrix is: every cubic metre of matrix, the carbendazol wettable powder sterilization that carries out disinfection of 20 gram of 50% mass ratio of spray; Described sterilization matrix is following volume ratio: perlite: the peat composed of rotten mosses: laterite=1:1:2;
6), the cuttage seeding of step 5) is placed in to shading rate is in 50% booth, and in after cuttage 1 week, keeps air humidity more than 90%, temperature, at 23 ± 2 ℃, between 40d, keeps humidity 80% after 2 weeks, to keep nursery stock blade not wilt, the seedling of must taking root;
7), step 6) gained is taken root seedling moves in the mixed-matrix of following volume ratio: perlite: humus soil: laterite=1:1:2, be placed in shading rate and be 50% booth, keep when high temperature daytime ventilating, and routinely with broad-spectrum germicide spray disinfection and water, and after noting while watering doing, rewater, grow to extraction young leaves, the group that obtains sequoia sempervirens is cultivated seedling, after stem lignification, can transplant to land for growing field crops.
Embodiment 2
A kind of outer cuttage root-taking method of sequoia sempervirens group training seedling bottle, comprises the following steps:
1), take the spray of sequoia sempervirens as explant, after induction is sprouted routinely, proceed in following medium:
MS medium
6-BA 0.3mg/L
NAA 0.1mg/L
KT 0.3mg/L
Sugar 30g/L
Agar 6g/L
Be 25 ℃ in temperature, intensity of illumination is 2000Lx, and light application time is 16h/ days, under the condition that medium pH is 5.8, cultivates 30 days;
2) grow to 3-4cm when high when the group training propagation seedling seedling strain of step 1), be placed in shading rate and be 70% booth, hardening 8d, must organize training and breed seedling;
3) by step 2) group training propagation seedling take out, clean the medium on it with clear water, then use 1000 times of liquid of tpn wetting powder of 75% mass ratio, disinfect after 2min, obtain aseptic sequoia sempervirens group training propagation seedling;
4) step 3) gained group training propagation seedling is cut into individual plant seedling, and each strain seedling base portion is immersed in the following water of taking root 30 seconds:
NAA 700mg/L
IBA 100mg/L；
5) by the seedling cuttage of step 4) in the sterilization matrix in the standard cave dish in 105 holes or 128 holes, spray normal root water, 1000 times of liquid of the thiophanate methyl wetting powder that sprays 70% appropriate mass ratio for second day after cuttage, described sterilization matrix is: every cubic metre of matrix, the carbendazol wettable powder sterilization that carries out disinfection of 20 gram of 50% mass ratio of spray; Described sterilization matrix is following volume ratio: perlite: the peat composed of rotten mosses: laterite=1:1:2;
6), the cuttage seeding of step 5) is placed in to shading rate is in 70% booth, and in after cuttage 2 weeks, keeps air humidity more than 90%, temperature, at 23 ± 2 ℃, between 40d, keeps humidity 60% after 2 weeks, to keep nursery stock blade not wilt, the seedling of must taking root;
7), step 6) gained is taken root seedling moves in the mixed-matrix of following volume ratio: perlite: humus soil: laterite=1:1:2, be placed in shading rate and be 70% booth, keep when high temperature daytime ventilating, and routinely with broad-spectrum germicide spray disinfection and water, and after noting while watering doing, rewater, grow to extraction young leaves, the group that obtains sequoia sempervirens is cultivated seedling, after stem lignification, can transplant to land for growing field crops.
For showing technique effect of the present invention, through experimental study, its result is as follows:
1, material: material therefor is the spray of sequoia sempervirens;
2, control group adopts rooting method in conventional organization culturing room, and experimental group is embodiment 1, and it the results are shown in Table 1.
The present invention is compared with cellar culture, and its advantage applies, in perfect sequoia sempervirens tissue-culturing rapid propagation system, does not need through the program of taking root in tissue culture bottle, saving group training space, chamber reaches more than 50%, saving group training seedling cost more than 30%, has shortened the group training cycle greatly, and specific experiment report the test is as follows:
Table 1 conventional organization is cultivated to take root and is trained the comparison of the outer cuttage root-taking of seedling bottle with the group of embodiment 1
As can be seen from the table, the sequoia sempervirens group training seedling (experimental group) that the present invention obtains, its average rootage duration is identical with the time that conventional organization is cultivated take root (control group) substantially, but the average rooting rate of the outer cuttage of the group of the embodiment of the present invention 1 training seedling bottle is higher, can reach 90%, and more conventional group of the seedling training seedling stalwartness of taking root, transplant and easily survive, transplanting survival rate reaches 90%, transplant and can become commodity seedling after 40 days, and conventional organization cultivation rooting rate is only just to become commodity seedling after 78%, 60 day, and transplanting survival rate only has 75%.From saving group training space, chamber, conventional production group is that breeding seedling does together with the seedling of taking root substantially while cultivating seedling, and the present invention is without through taking root program, therefore for Zu Pei culturing room has saved a large amount of spaces, approximately can save the space of 50% left and right.From being produced into originally, due to the raising of saving, rooting rate and the transplanting survival rate in Zu Pei culturing room space, the shortening of production cycle, make every strain sequoia sempervirens seedling can save the production cost of 0.10 yuan, approximately save the cost of 30% left and right.
Table 2 is the impact of water on the outer cuttage root-taking of sequoia sempervirens group training seedling bottle of taking root of different formulations.
Table 2
Can find out from table 2, adopt the water of taking root of different formulations all can make the outer cuttage root-taking of sequoia sempervirens group training seedling bottle, and rooting rate be more than 80%, root is white, and root system is sturdy, the seedling growing way of taking root stalwartness, but the optimum water formula of taking root is NAA500mg/L+IBA100mg/L, and rooting rate reaches 90%.
Claims (4)
1. the outer cuttage root-taking method of sequoia sempervirens group training seedling bottle, using aseptic sequoia sempervirens group training propagation seedling as cuttings, is characterized in that comprising the following steps:
A, by aseptic cuttings cuttage in the sterilization matrix of following volume ratio: perlite: the peat composed of rotten mosses: laterite=1:1:2, be placed in shading rate and be in 50～70% booth, and in after cuttage 1～2 week, keep air humidity more than 90%, temperature is at 23 ± 2 ℃, after 2 weeks, between 30～40 days, keeps humidity 60～80%, to keep nursery stock blade not wilt, the seedling of must taking root;
B, the steps A gained seedling of taking root is moved in the mixed-matrix of following volume ratio: perlite: humus soil: laterite=1:1:2, be placed in shading rate and be 50～70% booth, keep when high temperature daytime ventilating, and routinely with broad-spectrum germicide spray disinfection and water, and after noting while watering doing, rewater, grow to extraction young leaves, the group that obtains sequoia sempervirens is cultivated seedling, after stem lignification, can transplant to land for growing field crops.
2. the outer cuttage root-taking method of sequoia sempervirens group training seedling bottle as claimed in claim 1, is characterized in that described aseptic sequoia sempervirens group training propagation seedling obtains through the following steps:
1), take the spray of sequoia sempervirens as explant, after induction is sprouted routinely, proceed in following medium:
MS medium
6-BA 0.1—0.3mg/L
NAA 0.1—0.3mg/L
KT 0.1—0.3mg/L
Sugar 20-30g/L
Agar 6g/L
Be 21～25 ℃ in temperature, intensity of illumination is 2000～2500Lx, and light application time is 12～16h/ days, under the condition that medium pH is 5.6～5.8, cultivates 20～30 days;
2) grow to 3-4cm when high when the group training propagation seedling seedling strain of step 1), be placed in shading rate and be 70% booth, hardening 5～8d, must organize training and breed seedling;
3) by step 2) group training propagation seedling take out, clean the medium on it with clear water, then use 1000 times of liquid of tpn wetting powder of 75% mass ratio, disinfect after 1～2min, obtain aseptic sequoia sempervirens group training propagation seedling.
3. the outer cuttage root-taking method of sequoia sempervirens as claimed in claim 1 group training seedling bottle, is characterized in that the cuttage of described steps A is: first group training propagation seedling is cut into individual plant seedling, and by the following water of taking root of each strain seedling base portion immersion 30 seconds:
NAA 500—700mg/L
IBA 100—300mg/L
Cuttage afterwards, in matrix, is sprayed normal root water, the 1000 times of liquid of the thiophanate methyl wetting powder that sprays 70% appropriate mass ratio for second day after cuttage.
4. the outer cuttage root-taking method of sequoia sempervirens group training seedling bottle as claimed in claim 1, the sterilization matrix that it is characterized in that described steps A is: every cubic metre of matrix, the carbendazol wettable powder sterilization that carries out disinfection of 20 gram of 50% mass ratio of spray, be sub-packed in afterwards in the standard cave dish in 105 holes or 128 holes, and water permeablely, obtain sterilization matrix.
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| CN105009908A (en) * | 2015-07-29 | 2015-11-04 | 广西壮族自治区林业科学研究院 | Out-bottle one-step plantlet developing method for rootless test-tube cunninghamia lanceolata seedlings |
| CN105613238A (en) * | 2015-12-29 | 2016-06-01 | 江苏美尚生态景观股份有限公司 | Seedling hardening method for sequoia sempervirens |
| CN109041850A (en) * | 2018-06-11 | 2018-12-21 | 南京林业大学 | A kind of cryptomeria cuttage breeding method |
| CN109997537A (en) * | 2019-05-21 | 2019-07-12 | 黑龙江伊蓝生物科技有限公司 | The micro- plug bottle ex vitro rooting technique of indigo fruit tissue culture subculture unrooted test tube seedling |
| CN110249818A (en) * | 2019-07-31 | 2019-09-20 | 云南省林业科学院 | A kind of rapid propagation method of the Water culture sequoia sempervirens based on wet flower mud matrix |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105009908A (en) * | 2015-07-29 | 2015-11-04 | 广西壮族自治区林业科学研究院 | Out-bottle one-step plantlet developing method for rootless test-tube cunninghamia lanceolata seedlings |
| CN105009908B (en) * | 2015-07-29 | 2017-11-24 | 广西壮族自治区林业科学研究院 | The method of the outer forming seedling through one step culture of China fir unrooted test tube seedling bottle |
| CN105613238A (en) * | 2015-12-29 | 2016-06-01 | 江苏美尚生态景观股份有限公司 | Seedling hardening method for sequoia sempervirens |
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| CN110249818A (en) * | 2019-07-31 | 2019-09-20 | 云南省林业科学院 | A kind of rapid propagation method of the Water culture sequoia sempervirens based on wet flower mud matrix |
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Application publication date: 20140521 |