CN103493736B - The method of scale Fast-propagation Tillandsia-Si Chuikete test-tube plantlet - Google Patents
The method of scale Fast-propagation Tillandsia-Si Chuikete test-tube plantlet Download PDFInfo
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- CN103493736B CN103493736B CN201310456770.2A CN201310456770A CN103493736B CN 103493736 B CN103493736 B CN 103493736B CN 201310456770 A CN201310456770 A CN 201310456770A CN 103493736 B CN103493736 B CN 103493736B
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Abstract
The invention discloses the method for the efficient Fast-propagation Tillandsia of a kind of scale-Si Chuikete test-tube plantlet: sterilized by the capsule of Tillandsia Si Chuikete, rinsing, cut, get seed; Cultivated in solid culture medium by planting seed, seed germination forms the small and weak seedling of children of tool three leaves; Be placed in by seedling and proliferation and subculture medium is housed continues cultivation 3 ~ 4 months, seedling to be tillered propagation from base portion, and growth coefficient reaches 3 ~ 4; Seedling of growing thickly on propagation seedling is separated, is transferred to and is equipped with in the film bag of strong seedling culture base, move on outer plastic greenhouse seedbed of shading, utilize light source to cultivate and obtain seedling in 3 ~ 4 months; Seedling cleaned, dislocation coils without the flat cave of culture matrix and puts in green house to be cultivated 24 ~ 36 months.The advantages such as the present invention can be directly used in the seeling industry of industrial massive, and have and pollute less, seed germination rate is high, and aberration rate is low, and seedling is healthy and strong, and production cost is low, convenient transportation, and transplanting survival rate is high.
Description
Technical field
The invention belongs to cultivation of plants technical field, be specifically related to a kind of method of breeding Tillandsia-Si Chuikete test-tube plantlet.
Background technology
Tillandsia (Tillandsia) belongs to CAM class plant, and night, absorbing carbon dioxide, added the oxygen concentration of local environment relatively, but also can absorb and occupy the poisonous and harmful substance such as indoor formaldehyde, benzene homologues, and purification indoor air, is beneficial to physical and mental health.Meanwhile, Tillandsia has higher ornamental value.It is of a great variety, comes in every shape, and growth is in atmosphere, without the need to earth, clean, light, and the bright-coloured flower that can burst forth, can leaf be appreciated, again considerable flower, there is the feature such as good decorating effect, strong adaptability.Can put arbitrarily in indoor, hang from above, can be bonded on ancient tree stake, rockwork, wall, be placed in bamboo basket, on shell, also can be hung up, make curtain etc., intersperse the places such as room, parlor, balcony, fashion is pure and fresh, is imbued with nature rustic charm, both can be used as interior decoration to beautify the environment, can purify the air of a room again, DEVELOPMENT PROSPECT is wide, and Si Chuikete is the kind of wherein most Development volue.
Summary of the invention
Technical problem to be solved by this invention, is to provide the method for the efficient Fast-propagation Tillandsia of a kind of scale-Si Chuikete test-tube plantlet.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
A method for the efficient Fast-propagation Tillandsia of scale-Si Chuikete test-tube plantlet, the method comprises the following steps:
(1) by Si Chuikete capsule first with alcohol-pickled 45 ~ 60s, then be positioned in mercuric chloride solution and soak 15-20min;
(2) the extraordinary son of Si Chuike step (1) obtained is seeded in solid culture medium, 22 ~ 25 DEG C, illuminance 500 ~ 1000 lux, light application time 11 ~ 13h/d condition under, cultivate 40 ~ 50d, seed germination forms 2 ~ 4mm size and has the weak seedling of three leaf childrens, gets seedling for subsequent use; Wherein, described solid culture medium consists of: heteroauxin (IAA) 0.5 ~ 1.0mg/L+ murphy juice 60 ~ 80g/L+ sucrose 25 ~ 30g/L+1/2MS medium+pH5.4 ~ 5.6;
(3) seedling that step (2) obtains is placed in proliferation and subculture medium, 25 ~ 28 DEG C, illuminance 1000 ~ 2000 lux, light application time 11 ~ 13h/d condition under cultivate 3 ~ 4 months, Si Chuikete to be tillered propagation from base portion, and growth coefficient reaches 3-4; Wherein, described subculture multiplication medium consists of: heteroauxin 0.5 ~ 1.0mg/L+6-benayl aminopurine 0.2 ~ 0.3mg/L+ murphy juice 60 ~ 80g/L+ sucrose 25 ~ 30g/L+1/2MS medium+agar 4.5 ~ 6g/L+pH 5.6 ~ 5.8;
(4) seedling of growing thickly obtained in step (3) is separated into individual plant on superclean bench, transfer in the strong seedling culture base be packaged in film bag, 25 ~ 28 DEG C, illuminance 1000 ~ 2000 lux, light application time 11 ~ 13h/d condition under cultivate 3 ~ 4 months, move on outer plastic greenhouse seedbed of shading, temperature controls at 25 ~ 30 DEG C, utilize sunshine or natural daylight to carry out domestication as light source to cultivate, cultivate and obtain seedling in 20 ~ 30 days; Wherein, described strong seedling culture base consists of: heteroauxin 1.0 ~ 1.5mg/L+ sucrose 25 ~ 30g/L+1/4MS medium+agar 4.5 ~ 6g/L+pH5.6 ~ 5.8;
(5) seedling step (4) obtained is rinsed from taking-up clear water in bottle and is removed after medium, and dislocation, without in the cave dish of matrix, moves in green house and cultivates 24 ~ 30 months, namely can be used for various indoor and decorate more.
In step (1), described alcohol, concentration expressed in percentage by volume is 70% ~ 75%.
In step (1), in described mercuric chloride solution, the mass percentage of mercury chloride is 0.1% ~ 0.15%.
In step (2), described solid culture medium consists of: heteroauxin 1.0mg/L+ murphy juice 80g/L+ sucrose 30g/L+1/2MS medium+agar 6g/L+pH5.6.
In step (3), described subculture multiplication medium consists of: heteroauxin 1.0mg/L+6-benayl aminopurine 0.2mg/L+ murphy juice 80g/L+ sucrose 30g/L+1/2MS medium+agar 4.5 ~ 6g/L+pH5.8.
In step (4), described strong seedling culture base consists of: heteroauxin 1.0mg/L+ sucrose 30g/L+1/4MS medium+agar 6g/L+pH5.8.
In step (4), the film bag of described encapsulation medium is polypropylene screen bag.
Beneficial effect: compared with prior art, tool has the following advantages in the present invention:
1, after the sterilization of Tillandsia-Si Chuikete capsule, seed is sent out in the solid culture medium in step (2), adopt 1/2MS and be equipped with suitable hormone and natural additives, compared with existing seed germination medium, the method seed germination time shortens 20 ~ 25d greatly, and germination rate is up to more than 90%, aberration rate is less than 1%, sprouts neat.
2, in the subculture multiplication medium described in step (3), adopt 1/2MS and be equipped with suitable hormone, compared with traditional subculture multiplication medium, the method can promote tillering of seedling better faster, and growth coefficient can reach 3-4.
3, seedling is inoculated in film bag by step (4), moves in green house, utilize sunshine or natural daylight to cultivate after inoculation, the growth that the advantages such as sunshine spectrum is wide, spectrum segment Energy distribution is even promote seedling can be made full use of, compared with the method for routine, the method can improve growth rate, the robustness of seedling, reduces energy consumption, reduce and pollute, be convenient to transport, after seedling, transplanting adaptability is good, survival rate is high by more than 95%, is a kind of training mode of low-carbon high-efficiency.
Embodiment
According to following embodiment, the present invention may be better understood.But those skilled in the art will readily understand, the concrete content described by embodiment only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.
Embodiment 1: the method for the efficient Fast-propagation Tillandsia of a kind of scale-Si Chuikete test-tube plantlet, the method comprises the following steps:
(1) on transfer room superclean bench, by the capsule of uncracked Tillandsia-Si Chuikete, be positioned in wide-mouth bottle, in bottle, pour 70% (v/v) alcohol into, soak about 50s, alcohol is leached, then uses the mercuric chloride solution of 0.1% (w/w) to soak about 12 ~ 15min, horizontal oscillations repeatedly in immersion process, make capsule surface sterilization thorough, again with sterile water rinsing 3 ~ 4 times repeatedly, the capsule after sterilization is cut, gets seed for subsequent use.
(2) extraordinary for Tillandsia-Si Chuike is seeded in the solid culture medium of heteroauxin 1.0mg/L+ murphy juice 80g/L+ sucrose 30g/L+1/2MS medium+agar 6g/L, pH5.6; Condition of culture is set to temperature 25 DEG C, illuminance 500 ~ 1000 lux, light application time 12h/d, cultivates 40 ~ 50d, and seed germination forms 2-4mm and has the seedling of 3 leaves.
(3) budlet obtained is transferred in the subculture multiplication medium of heteroauxin 1.0mg/L+6-benayl aminopurine 0.2mg/L+ murphy juice 80g/L+ sucrose 30g/L+1/2MS medium+agar 6g/L, pH5.8; Culturing room's ambient As is under the condition of temperature 25 ~ 28 DEG C, illuminance 950 ~ 1200 lux, light application time 12h/d, and cultivate 3 ~ 4 months, the growth coefficient of seedling reaches 3-4.
(4) seedling obtained is inoculated into is equipped with in the polypropylene screen bag of strong seedling culture base, culture medium prescription is heteroauxin 1.0mg/L+ sucrose 30g/L+1/4MS medium+agar 6g/L, pH5.8, after inoculation, with sealing machine, film bag is sealed, outer plastic greenhouse seedbed of shading is moved to after sealing, temperature controls at 25 ~ 30 DEG C, sunshine or natural daylight is utilized to cultivate as light source, cultivation is continued 3 ~ 4 months in greenhouse, because film bag seals with sealing machine, good sealing effect, the problem that plantlet in vitro pollutes under green house external environment can be effectively reduced.
(5) seedling obtained is taken out in film bag, after rinsing removal medium with clear water, directly be positioned in green house and coil upper cultivation 24 ~ 36 months without the flat cave of culture matrix, Dai Qichui section speciality to 10 ~ 20cm is large, just can be used for the decoration of various indoor and outdoor.
Embodiment 2:
Substantially the same manner as Example 1, difference is: step (5) green house natural daylight and sunshine are cultivated and changed into outside culturing room's fluorescent lamp cultivation; Seed germination in step (2), step (3), step (4), shoot proliferation is cultivated, strong seedling culture three phases is combined into row filter with medium listed by table 1, respectively with preferred optimum medium combination formula.Table 1 is described as follows:
1. culture medium prescription optimization mainly designs different basal mediums, hormone concentration, natural additive, concentration of activated carbon etc. and is optimized design to medium, all solid culture medium is designed in this example, in often kind of formula, agar concentration is 5g/L, sucrose is 25g/L, and culture environment is in temperature 25 DEG C, illuminance 1000 ~ 2000 lux, light application time 12h/d culturing room.2. for production, different cultivation stages devises three groups of culture medium prescriptions, and the index such as speed, growing way, color (emerald green is good) of planting seed stage main detection seed germination judges whether formula is reasonable; Shoot proliferation stage main detection seedling tiller the indexs such as propagation (The more the better), vitrifying degree judge formula whether reasonable; In the strong seedling culture stage, the index such as growth potential, vitrifying degree (more low better), color of main clearance observation seedling judges the quality of filling a prescription.By observing and the mensuration of indices, and represent the quality of filling a prescription with the how much of " ★ " and " ☆ ", ★ more multilist show better, a ☆ grade lower than ★.
Table 1 screening of medium formula
Test shows, in seed germination stage (step 2) 18 kinds of formula combination, basal medium, hormone and natural additive combination range are within the scope of 1/2MS+ heteroauxin 0.5 ~ 1.0mg/L+ murphy juice 60 ~ 80g/L medium, seed germination rate is high, growth is fast, be the combination formula combination be comparatively suitable for, after planting seed, about 45d can be divided into seedling; In shoot proliferation stage (step 3) 18 kinds of formula combination, basal medium, hormone and natural additive combination range are within the scope of 1/2MS+6-benayl aminopurine 0.2 ~ 0.3mg/L+ murphy juice 60 ~ 80g/L medium, the growth potential of seedling is good, growth rate fast, vitrifying degree is low, 3 ~ 4 months, the growth coefficient of seedling can reach 3 ~ 4; Stage in strong sprout (step 4), result shows, 1/4MS+ heteroauxin 1.0 ~ 1.5mg/L is preferred culture medium formula combination.Also find that in test the propagation of potato to seed germination and seedling has sent out good facilitation.
Embodiment 3:
Substantially the same manner as Example 1, difference is: take following 4 kinds of different culture environment respectively by after step (4) switching seedling: 1. transfer in tissue culture bottle by seedling, move on outer plastic greenhouse seedbed of shading, by wet curtain and blower fan, temperature is controlled at 25 ~ 30 DEG C, utilize sunshine or natural daylight to cultivate as light source; 2. seedling is transferred in film bag, move on outer plastic greenhouse seedbed of shading, by wet curtain and blower fan, temperature is controlled at 25 ~ 30 DEG C, utilize sunshine and natural daylight to cultivate as light source; 3. seedling is transferred in tissue culture bottle, is positioned in culturing room, culturing room's ambient As is temperature 25 ~ 28 DEG C, illuminance 1000 ~ 1200 lux, light application time 12h/d condition under cultivate; 4. seedling is transferred in film bag, is positioned in culturing room, culturing room's ambient As is temperature 25 ~ 28 DEG C, illuminance 1000 ~ 1200 lux, light application time 12h/d condition under cultivate.
Four kinds of training methods are carried out simultaneously.2. and 4. result cultivates about three months in 4 kinds of training methods, and little seedling leaf is emerald green, healthy and strong, the seedling of long 2 ~ 3cm, particularly 2., because warm indoor illumination is sufficient, leaf look becomes bottle green, more healthy and stronger, 2. compared with 1., pollution rate aspect has obvious advantage; Because external condition is complicated, because of 2. direct sealing machine sealing after inoculation, good airproof performance, pollution rate only has about 5%, and pollution rate 1. reaches more than 50%, than 2. about 5% pollution rate obviously high, in four kinds of training modes, pollution rate is 4. minimum, about 1%.Because 3. this training mode energy consumption is high, production cost is large, and adopt pattern 2., pollution rate also within acceptable scope, simultaneously because of with sunshine and natural daylight for light source, after seedling, can directly transplant, do not need through practicing seedling link, survival rate can up to more than 95%, and therefore resultant effect is higher than 4..
Embodiment 4: the method for the efficient Fast-propagation Tillandsia of a kind of scale-Si Chuikete test-tube plantlet, the method comprises the following steps:
(1) on transfer room superclean bench, by the capsule of uncracked Tillandsia-Si Chuikete, be positioned in wide-mouth bottle, in bottle, pour 75% (v/v) alcohol into, soak 45 ~ 60s, alcohol is leached, then uses the mercuric chloride solution of 0.15% (w/w) to soak about 12 ~ 15min, horizontal oscillations repeatedly in immersion process, make capsule surface sterilization thorough, again with sterile water rinsing 3 ~ 4 times repeatedly, the capsule after sterilization is cut, gets seed for subsequent use.
(2) extraordinary for Tillandsia-Si Chuike is seeded in the solid culture medium of heteroauxin 0.5mg/L+ murphy juice 60g/L+ sucrose 25g/L+1/2MS medium+agar 4.5g/L, pH5.4; Condition of culture is set to temperature 25 DEG C, illuminance 500 ~ 1000 lux, light application time 12h/d, cultivates 40 ~ 50d, and seed germination forms 2-4mm and has the seedling of 3 leaves.
(3) budlet obtained is transferred in the subculture multiplication medium of heteroauxin 0.5mg/L+6-benayl aminopurine 0.3mg/L+ murphy juice 60g/L+ sucrose 25g/L+1/2MS medium+agar 4.5g/L, pH5.6; Culturing room's ambient As is under the condition of temperature 25 ~ 28 DEG C, illuminance 1000 ~ 1200 lux, light application time 11 ~ 13h/d, and cultivate 3 ~ 4 months, the growth coefficient of seedling reaches 3-4.
(4) seedling obtained is inoculated into is equipped with in the polypropylene screen bag of strong seedling culture base, culture medium prescription is heteroauxin 1.5mg/L+ sucrose 25g/L+1/4MS medium+agar 4.5g/L, pH5.6, after inoculation, with sealing machine, film bag is sealed, outer plastic greenhouse seedbed of shading is moved to after sealing, temperature controls at 25 ~ 30 DEG C, sunshine or natural daylight is utilized to cultivate as light source, cultivation is continued 3 ~ 4 months in greenhouse, because film bag seals with sealing machine, good sealing effect, the problem that plantlet in vitro pollutes under green house external environment can be effectively reduced.
(5) seedling obtained is taken out in film bag, after rinsing removal medium with clear water, directly be positioned in green house and coil upper cultivation 24 ~ 36 months without the flat cave of culture matrix, Dai Qichui section speciality to 10 ~ 20cm is large, just can be used for the decoration of various indoor and outdoor.
Claims (5)
1. a method for scale Fast-propagation Tillandsia-Si Chuikete test-tube plantlet, it is characterized in that, the method comprises the following steps:
(1) by Si Chuikete capsule first with alcohol-pickled 45 ~ 60s, then be positioned in mercuric chloride solution and soak; Described alcohol, concentration expressed in percentage by volume is 70% ~ 75%; In described mercuric chloride solution, the mass percentage of mercury chloride is 0.1% ~ 0.15%;
(2) the extraordinary son of Si Chuike step (1) obtained is seeded in solid culture medium, 22 ~ 25 DEG C, illuminance 300 ~ 1000 lux, light application time 11 ~ 13h/d condition under, cultivate 40 ~ 50d, seed germination forms the weak seedling of tool three leaf childrens, gets seedling for subsequent use; Wherein, described solid culture medium consists of: heteroauxin 0.5 ~ 1.0 mg/L+ murphy juice 60 ~ 80 g/L+ sucrose 25 ~ 30g/L+1/2MS medium+agar 4.5 ~ 6 g/L+pH 5.4 ~ 5.6;
(3) seedling that step (2) obtains is placed in subculture multiplication medium, 25 ~ 28 DEG C, illuminance 1000 ~ 2000 lux, light application time 11 ~ 13h/d condition under cultivate 3 ~ 4 months, Si Chuikete to be tillered propagation from base portion, and growth coefficient reaches 3-4; Wherein, described subculture multiplication medium consists of: heteroauxin 0.5 ~ 1.0 mg/L+6-benayl aminopurine 0.2-0.3mg/L+ murphy juice 60 ~ 80 g/L+ sucrose 25 ~ 30 g/L+1/2MS medium+agar 4.5 ~ 6 g/L+pH 5.6 ~ 5.8;
(4) seedling of the growing thickly cutting and separating on superclean bench obtained in step (3) is become individual plant, transfer in the strong seedling culture base be packaged in film bag, move on outer plastic greenhouse seedbed of shading, temperature controls at 25 ~ 30 DEG C, utilize sunshine or natural daylight to cultivate as light source, cultivate and obtain seedling in 3 ~ 4 months; Wherein, described strong seedling culture base consists of: heteroauxin 1.0 ~ 1.5 mg/L+ sucrose 25 ~ 30 g/L+1/4MS medium+agar 4.5 ~ 6 g/L+pH 5.6 ~ 5.8;
(5), after seedling step (4) obtained rinses removal medium from taking-up clear water in bottle, dislocation coils interior without the cave of matrix or sticks on cystosepiment, and in immigration green house, cultivation 24 ~ 30 months, can be used for various interior/exterior decoration.
2. the method for scale Fast-propagation Tillandsia-Si Chuikete test-tube plantlet according to claim 1; it is characterized in that; in step (2), described solid culture medium consists of: heteroauxin 1.0mg/L+ murphy juice 80 g/L+ sucrose 30g/L+1/2MS medium+agar 6 g/L+pH 5.6.
3. the method for scale Fast-propagation Tillandsia-Si Chuikete test-tube plantlet according to claim 1; it is characterized in that; in step (3), described subculture multiplication medium consists of: heteroauxin 1.0 mg/L+6-benayl aminopurine 0.2mg/L+ murphy juice 80 g/L+ sucrose 30 g/L+1/2MS medium+agar 6 g/L+pH 5.8.
4. the method for scale Fast-propagation Tillandsia-Si Chuikete test-tube plantlet according to claim 1; it is characterized in that; in step (4), described strong seedling culture base consists of: heteroauxin 1.0 mg/L+ sucrose 30 g/L+1/4MS medium+agar 6 g/L+pH5.8.
5. the method for scale Fast-propagation Tillandsia-Si Chuikete test-tube plantlet according to claim 1, is characterized in that, in step (4), the film bag of described encapsulation medium is polypropylene screen bag.
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| CN104094844A (en) * | 2014-06-26 | 2014-10-15 | 江苏农林职业技术学院 | Tissue culture and inducing culture method for Tillandsia |
| CN104255400B (en) * | 2014-08-04 | 2016-08-17 | 江苏农林职业技术学院 | A kind of cultural method of Tillandsia tissue cultured seedling |
| CN104255321A (en) * | 2014-09-17 | 2015-01-07 | 江苏农林职业技术学院 | Tillandsia stricta rigid leaf tissue culture seedling strengthening method |
| CN110547195B (en) * | 2019-09-23 | 2021-09-07 | 江苏农林职业技术学院 | Method for obtaining air pineapple seedlings through aseptic germination of seeds |
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