CN109479728A - A kind of micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling - Google Patents
A kind of micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling Download PDFInfo
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- CN109479728A CN109479728A CN201910077563.3A CN201910077563A CN109479728A CN 109479728 A CN109479728 A CN 109479728A CN 201910077563 A CN201910077563 A CN 201910077563A CN 109479728 A CN109479728 A CN 109479728A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of micro- Rooting ex vitro methods of Populus hopeiensis tissue-cultured seedling；Specifically comprise the following steps: that a, explant are established, b, Multiplying culture, c, strong seedling culture, d, topping rush stem hardening, f, e, the transplanting of Promoting bacteria root dipping spray collecting carbonic anhydride agent culture；Wherein the tissue-cultured seedling for breaking apical dominance is continued to cultivate after covering bottle cap in step d, until new terminal bud is formed, stem is sturdy, thickening vanes, color is dark green, corkage lid when nursery stock is healthy and strong neat, the sturdy micro- branch of clip simultaneously makes tissue-cultured seedling base portion stay 1 ~ 2 bud, in this way during transplanting, without cleaning culture medium, and micro- branch of clip only forms terminal bud, do not open up spire, there is no tender stem, tender point, acclimatization and transplants damage ratio is small, the operation is conducive to improve transplanting efficiency, and step e and f can promote plant and take root, conducive to promotion root survival, the method of the present invention transplanting efficiency improves 45% or more than original, rooting rate can reach 95% or more.
Description
Technical field
The invention belongs to technical field of plant propagation, are related to tissue cultivating and seedling technology, and in particular to a kind of Populus hopeiensis group
Train Miao Wei Rooting ex vitro methods.
Background technique
Populus hopeiensis (scientific name:Populus hopeiensis Hu et Chow) it is Salicaceae poplar, arbor, up to
30 meters.Bark yellow green is smooth to canescence；Tree crown circle is big.Leaf is oval or subcircular, 3-8 centimetres long, 2-7 centimetres wide.Male flower
Sequence is about 5 centimetres, and rachis is by close hair；Female inflorescence is 3-5 centimetres long.Capsule long avette, there is short handle.April at florescence, the fruiting period 5-6 month.
It is distributed in North China, each provinces and regions in northwest, is one of common poplar in Hebei Mountain Regions, there are cultivation in various regions, are grown on sea more
It pulls out on 700-1600 meters of river two sides, cheuch Schattenseite and alluvial terrace.
It is more particularly suitable to do food steamer material for the use such as building, farm implements, boxboard for timber.For North China, northwest loess hill loess hills top,
Liang Po, the water and soil conservation of cheuch and sandy beach ground or Timber stands reproducting tree species.It also is garden, trade fine tree species.
Populus hopeiensis tissue-cultured seedling is in conventional Rooting ex vitro incubation, and there are the low skills low with rooting rate of transplanting efficiency
Art problem, routine culture operation are only able to maintain 78% or so rooting rate.
Summary of the invention
The purpose of the present invention is being directed to the deficiencies in the prior art, providing one kind can be improved transplanting efficiency and rooting rate
Micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling.
To achieve the above object the technical solution adopted by the present invention is that:
A kind of micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling, comprising the following steps:
A, explant is established: preferred logical straight, robust growth, no disease and pests harm the annual resting shoot of elite plant strain of dry type, processing
After be inoculated into bud inducement cultivation base and cultivated, to form tender stem；
B, Multiplying culture；It the tender stem of induction is cut into section transfers and carry out the differentiation of induced bundle branch in culture medium, Multiplying culture generates micro-
Branch；
C, strong seedling culture；Tender stem is cut into 1.5 ~ 2 cm segments, accesses in culture medium and is cultivated, most test tube seedlings are micro- after 30 d
Branch grows to 3 ~ 5 roots or more；
D, topping promotees stem hardening；When required output is arrived in Populus hopeiensis tissue-cultured seedling micro- breeding, on the super-clean bench, by tissue-cultured seedling top
The tissue-cultured seedling for breaking apical dominance is continued to cover to be transferred in hardening environment after bottle cap and be cultivated in hardening by bud clip, hardening
Time controls at 10 days or so, and until new terminal bud is formed, stem is sturdy, and thickening vanes, color is dark green, and nursery stock opens when healthy and strong neat
Bottle cap, the sturdy micro- branch of clip, clip micro- when, make tissue-cultured seedling base portion stay 1-2 bud；
E, Promoting bacteria root dipping is transplanted；By the sturdy micro- branch of the Populus hopeiensis of high 5 cm or so, gently clamped with tweezers, 1-2 centimetres of base portion
The wide precious stoste of quickly dipped plant growth-promoting bacteria battalion, then cuttage enter and are transplanted in 63# seedling culture hole plate dress Denmark Pin Shi seedling medium,
Under the conditions of intelligent greenhouse, relative humidity control is 85% or so, and for light control system in 1000Lx, temperature control is raw at 25 DEG C or so
Root culture；
F, collecting carbonic anhydride agent culture is sprayed；The moisturizing of Populus hopeiensis tissue-cultured seedling has been transplanted to relative humidity 85%, has sprayed 1000 times of liquid
Collecting carbonic anhydride agent-light carbon core fertilizer, forms Populus hopeiensis tissue-cultured seedling good gas environment, sprays within every 15 days later once, daily
Morning ventilation 20 minutes；Control illumination 8000Lx is stepped up to 15000Lx.
Further, in the step d, the terminal bud of clip is seeded in needs and takes root culture medium 1/2MS+IBA0.5+ sugarcane of emerging
It is cultivated in sugared 15g/L+ agar 5%g/L.
Further, in the step d, there are add appropriate sterile liquid culture in the tissue-cultured seedling test tube of 1-2 bud for base portion
Base 1/2MS+IAA 0.5 continues lid bottle cap culture.
During micro- acclimatization and transplants of Populus hopeiensis tissue-cultured seedling are taken root, tissue-cultured seedling top children is tender, micro- branch is in cleaning base portion adhesion culture
It is easily damaged when base fracture, easily wilt in transplanting culture, Yi Zisheng mushroom causes hardening to fail, and the present invention is cut in step d
It takes tissue-cultured seedling terminal bud and the tissue-cultured seedling for losing apical dominance is cultivated, be conducive to obtain neat nursery stock in this way, after culture
Tissue-cultured seedling base portion is set to stay 1 ~ 2 bud at tissue-cultured seedling clip micro-, micro- branch of such clip is not deposited without root system compared to routine operation
In cleaning base portion culture medium operation, and micro- branch only forms terminal bud before transplantation rooting, does not open up spire, and without tender stem, tender point, hardening is moved
It is small to plant damage ratio, conducive to the raising of transplanting efficiency；It is wide precious former using quickly dipped plant growth-promoting bacteria battalion to tissue-cultured seedling terminal bud in step e
Liquid operation, be conducive to the micro- branch of tissue-cultured seedling take root rapidly growth and prevent and treat other mushrooms breed infection, formed protection advantage field planting
Bacterium and Promoting bacteria；Tissue-cultured seedling is conducive to inhibit dark respiration, be promoted using collecting carbonic anhydride agent is sprayed by operation in step f
Entering light breathing, and higher concentration carbonless base paper carbon dioxide gas environment is formed, it is unfavorable for mushroom and insect pest is bred, makes plant
The micro- branch of tissue-cultured seedling has sufficient photosynthetic environment, promotes being sufficiently formed for plant photosynthesis product, is transported to base portion conducive to by nutrient, promotees
Into the further development growth taken root with plant, achieve the purpose that with leaf promoting root growth, the raising of hestening rooting rate.
It the terminal bud of clip is seeded in needs in step d of the present invention takes root for emerging and cultivate in culture medium, it in this way can benefit
With the good feature of terminal bud detoxification efficiency, high quality tissue-cultured seedling is obtained under realization in generation culture.
By base portion, there are add appropriate aseptic liquid nutrient medium 1/ in the tissue-cultured seedling test tube of 1-2 bud in step d of the present invention
2MS+IAA 0.5 continues lid bottle cap culture, in this way can by there are bud continuously form micro- branch, and it is fast under the operation of step d
Speed, which obtains, can transplant micro- branch, which, without carrying out inoculation operation again, can be conducive to save when needing to obtain tissue-cultured seedling again
Obtain the tissue-cultured seedling operating time.
Specific embodiment
Below in conjunction with concrete operations, the present invention will be further described:
A kind of micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling, comprising the following steps:
A, explant is established: preferred dry type is logical straight, robust growth, the annual resting shoot of the elite plant strain of no disease and pests harm, originally
Water rinses 20 min, cuts 1 bud of section band.On superclean bench, 20 s are impregnated with 75% alcohol, then to be placed in 0.1% mercuric chloride molten
15 min are sterilized in liquid, are then used aseptic water washing 5 times, are blotted stem section surface moisture with sterilized filter paper；It is inoculated into bud induction training
Support 0.5 mg.L of base MS+6-BA-1After illumination cultivation 30d, there can be tender stem to be formed.
B, Multiplying culture；After 1 month, the tender stem of induction is cut into section and is transferred into culture medium MS+6-BA 0.3+NAA 0. 05
In, Multiple Buds generate after about 20 d, after 40 d, have 5~6 sprouts long to 5~6 cm, at this time can rolling bottle cut section in culture medium MS
Continue to expand in+6-BA 0.3+NAA 0. 05 numerous.
C, strong seedling culture；Tender stem is cut into 1.5 ~ 2 cm segments, accesses in culture medium 1/2MS+IAA 0.5 and is cultivated,
After about 30 d more than 3 ~ 5 roots of most test tube seedlings.
D, topping promotees stem hardening；When required output is arrived in Populus hopeiensis tissue-cultured seedling micro- breeding, on the super-clean bench, by terminal bud
The tissue-cultured seedling for breaking apical dominance is continued to cover to be transferred in hardening environment after bottle cap and be cultivated in hardening, preferably changed by clip
With ventilating cover culture, 1~5d is placed in set the 5000Lx illumination cultivation there are two fluorescent tube under the conditions of, 6~10d of culture is placed in equipped with three
It is cultivated under the 8000Lx illumination condition of a fluorescent tube, the hardening time controls at 10 days or so, and until new terminal bud is formed, stem is thick
Strong, thickening vanes, color is dark green, corkage lid when nursery stock is healthy and strong neat, the sturdy micro- branch of clip, and clip micro- when makes to be located at culture medium
In tissue-cultured seedling base portion stay 1-2 bud, micro- branch of clip can be used for transplanting；Terminal bud after clip be seeded in needs take root emergence training
Support base 1/2MS+IBA0.5+ sucrose 15gL-1+ agar 5%gL-1In, medium pH 6.0,23~25 DEG C of cultivation temperature, when illumination
Between 12 h.d-1, 40 umol.m of intensity of illumination-2.s-1, terminal bud detoxification efficiency is good, it is conducive to obtain the tissue-cultured seedling of high quality after culture,
Tissue-cultured seedling obtains the micro- branch of transplanting after can continuing terminal bud operation again；Test tube after the micro- branch of clip adds appropriate aseptic liquid nutrient medium
1/2MS+IAA 0.5 continues lid bottle cap culture, base portion there are 1-2 bud can continue culture and form new micro- branch, needing to obtain
New micro- branch when being transplanted, which can be quickly obtained the micro- branch of transplanting, be conducive to save the time without carrying out inoculation operation.
E, Promoting bacteria root dipping is transplanted；The sturdy micro- branch of Populus hopeiensis of high 5 cm by clip or so, is gently clamped, base with tweezers
The wide precious stoste of quickly dipped plant growth-promoting bacteria battalion, 1-2 centimetres of portion, then cuttage enter in 63# seedling culture hole plate dress Denmark Pin Shi seedling medium into
Row transplanting, under the conditions of intelligent greenhouse in, 85% or so, light control system exists in 1000Lx, temperature control for relative humidity control
25 DEG C or so culture of rootage, the quickly dipped wide precious stoste of battalion, which is conducive to the micro- branch of test tube seedling, takes root rapidly and growth and prevents and treats other mushrooms and breed sense
Dye forms field planting bacterium and the Promoting bacteria of protection advantage.
F, collecting carbonic anhydride agent culture is sprayed；The moisturizing of Populus hopeiensis tissue-cultured seedling has been transplanted to relative humidity 85%, has sprayed 1000
Times liquid collecting carbonic anhydride agent-light carbon core fertilizer, forms Populus hopeiensis tissue-cultured seedling good gas environment, spray within every 15 days later it is primary,
Ventilation every morning 20 minutes.Control illumination 8000Lx is stepped up to 15000Lx, accelerates raising Populus hopeiensis tissue-cultured seedling micro-
Branch photoautotrophy ability, hestening rooting form intact plant；Collecting carbonic anhydride agent is sprayed after transplanting, is conducive to blade and is carried out light
Cooperation is used, and micro- surrounding air environment of high concentration carbon dioxide is created, and is inhibited aerobic respiration mushroom and insect pest, is tried using Populus hopeiensis
The advantage of Guan Miaowei stomata mis-tie misclosures, conducive to allowing the chlorella of trapping carbon dioxide to promote carbon dioxide by stomata
Trapping reaches or approaches saturation, promotes micro- leaf photosynthesis of Populus hopeiensis, generates photosynthate, nutrient is conveyed, in this way from root
Portion, stem, blade have all ensured that other mushrooms do not infect test tube seedling by the external world, while creating good photosynthetic environment, are conducive to
Plant takes root rapidly growth.The probability of the Populus hopeiensis tissue-cultured seedling samping off and root knot cultivated in this way substantially reduces, and promotes life
The raising of root rate.
Using the method for the present invention, 520,000 plants of Populus hopeiensis tissue-cultured seedling are produced in practice, 49.6 ten thousand plants of survival or more of actually taking root,
Average rooting rate can be improved to 95% or more；Everyone average treatment tissue-cultured seedling is at 800 plants or so in conventional transplanting operating process,
It can be promoted using this method to 1700 plants or more, transplanting efficiency improves 45% or more than original in production, and nursery stock is neatly good for
Strong, commodity is good；The method of the present invention whole process does not use sterilizing insecticide and plant growth regulator, green and pollution-free, can
It preserves the ecological environment.
Claims (3)
1. a kind of micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling, which comprises the following steps:
A, explant is established: preferred logical straight, robust growth, no disease and pests harm the annual resting shoot of elite plant strain of dry type, processing
After be inoculated into bud inducement cultivation base and cultivated, to form tender stem；
B, Multiplying culture；It the tender stem of induction is cut into section transfers and carry out the differentiation of induced bundle branch in culture medium, Multiplying culture generates micro-
Branch；
C, strong seedling culture；Tender stem is cut into 1.5 ~ 2 cm segments, accesses in culture medium and is cultivated, most test tube seedlings are micro- after 30 d
Branch grows to 3 ~ 5 roots or more；
D, topping promotees stem hardening；When required output is arrived in Populus hopeiensis tissue-cultured seedling micro- breeding, on the super-clean bench, by tissue-cultured seedling top
The tissue-cultured seedling for breaking apical dominance is continued to cover to be transferred in hardening environment after bottle cap and be cultivated in hardening by bud clip, hardening
Time controls at 10 days or so, and until new terminal bud is formed, stem is sturdy, and thickening vanes, color is dark green, and nursery stock opens when healthy and strong neat
Bottle cap, the sturdy micro- branch of clip, clip micro- when, make tissue-cultured seedling base portion stay 1-2 bud；
E, Promoting bacteria root dipping is transplanted；By the sturdy micro- branch of the Populus hopeiensis of high 5 cm or so, gently clamped with tweezers, 1-2 centimetres of base portion
The wide precious stoste of quickly dipped plant growth-promoting bacteria battalion, then cuttage enter and are transplanted in 63# seedling culture hole plate dress Denmark Pin Shi seedling medium,
Under the conditions of intelligent greenhouse, relative humidity control is 85% or so, and for light control system in 1000Lx, temperature control is raw at 25 DEG C or so
Root culture；
F, collecting carbonic anhydride agent culture is sprayed；The moisturizing of Populus hopeiensis tissue-cultured seedling has been transplanted to relative humidity 85%, has sprayed 1000 times of liquid
Collecting carbonic anhydride agent-light carbon core fertilizer, forms Populus hopeiensis tissue-cultured seedling good gas environment, sprays within every 15 days later once, daily
Morning ventilation 20 minutes；Control illumination 8000Lx is stepped up to 15000Lx.
2. a kind of micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling as described in claim 1, which is characterized in that the step
In d, the terminal bud of clip, which is seeded in needs, takes root for emerging and carries out in culture medium 1/2MS+IBA0.5+ sucrose 15g/L+ agar 5%g/L
Culture.
3. a kind of micro- Rooting ex vitro method of Populus hopeiensis tissue-cultured seedling as described in claim 1, which is characterized in that the step
In d, base portion continues lid bottle there are appropriate aseptic liquid nutrient medium 1/2MS+IAA 0.5 is added in the tissue-cultured seedling test tube of 1-2 bud
Lid culture.
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