CN103718969B - A kind of logical cultured in vitro regeneration plant method with a smile of poem beautiful jade - Google Patents
A kind of logical cultured in vitro regeneration plant method with a smile of poem beautiful jade Download PDFInfo
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- CN103718969B CN103718969B CN201410041803.1A CN201410041803A CN103718969B CN 103718969 B CN103718969 B CN 103718969B CN 201410041803 A CN201410041803 A CN 201410041803A CN 103718969 B CN103718969 B CN 103718969B
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- 238000000034 method Methods 0.000 title claims abstract description 18
- 238000000338 in vitro Methods 0.000 title claims abstract description 15
- 230000008929 regeneration Effects 0.000 title claims abstract description 15
- 238000011069 regeneration method Methods 0.000 title claims abstract description 15
- 241000196324 Embryophyta Species 0.000 claims abstract description 35
- 239000007921 spray Substances 0.000 claims abstract description 13
- 238000009395 breeding Methods 0.000 claims abstract description 9
- 230000001488 breeding effect Effects 0.000 claims abstract description 9
- 239000000463 material Substances 0.000 claims abstract description 9
- 238000004321 preservation Methods 0.000 claims abstract description 7
- 238000005286 illumination Methods 0.000 claims description 35
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 30
- 239000011159 matrix material Substances 0.000 claims description 25
- 230000012010 growth Effects 0.000 claims description 21
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- 238000005507 spraying Methods 0.000 claims description 10
- 230000001954 sterilising effect Effects 0.000 claims description 10
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- 239000003814 drug Substances 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 6
- 230000000050 nutritive effect Effects 0.000 claims description 6
- 235000013162 Cocos nucifera Nutrition 0.000 claims description 5
- 241000238631 Hexapoda Species 0.000 claims description 5
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
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- 230000035755 proliferation Effects 0.000 claims description 5
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- 238000005406 washing Methods 0.000 claims description 5
- 239000005972 6-Benzyladenine Substances 0.000 claims description 4
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- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 claims description 4
- YGGXZTQSGNFKPJ-UHFFFAOYSA-N methyl 2-naphthalen-1-ylacetate Chemical compound C1=CC=C2C(CC(=O)OC)=CC=CC2=C1 YGGXZTQSGNFKPJ-UHFFFAOYSA-N 0.000 claims description 4
- 230000006698 induction Effects 0.000 claims description 2
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- 241000218377 Magnoliaceae Species 0.000 abstract description 7
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- 241000218394 Magnolia liliiflora Species 0.000 description 3
- 241000894007 species Species 0.000 description 2
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Abstract
The invention discloses a kind of poem beautiful jade and lead to cultured in vitro regeneration plant method with a smile, comprise that (1) takes explant and material processed thereof, (2) set up sterile propagation body, (3) Multiplying culture, (4) culture of rootage, (5) are transplanted, (6) seedling culture six steps.The logical cultured in vitro regeneration plant method inductivity with a smile of poem beautiful jade of the present invention and survival rate high, step is simple and feasible, very urgent and significance is had with the critically endangered seeds of breeding to preservation and rescue world Precious, Rare, Endangered Magnoliaceae kind, for the large tree spray of research Magnoliacea plant carries out tissue cultures, overcome the tissue culture propagation technology such as pollution rate is high, brownization is serious, inductivity is low, transplanting success is low and there is extremely valuable research value.
Description
Technical field
The present invention relates to a kind of cultured in vitro regeneration plant method of plant, be specifically related to a kind of poem beautiful jade and lead to cultured in vitro regeneration plant method with a smile, belong to biological excised cotyledon seedling growing process field.
Background technology
Poem beautiful jade logical (formal name used at school: Micheliasirindhorniae (Noot.etChal.) N.H.XiaetX.H.Zhangj) is with a smile current, also generally poem beautiful jade wooden supporter orchid (MagnoliasirindhorniaeNoot. & Chalermglin) is used to be one of world's Precious, Rare, Endangered Magnoliaceae kind in the world, critically endangered seeds, original producton location Thailand, belong to aiphyllium, leaf, Huadu are very fragrant, can refine famous and precious plant essence.
Poem beautiful jade is logical is the evergreen megaphanerophyte of Magnoliaceae with a smile, and trunk is straight and upright, Gao Keda 30-35m, and flower is ivory white, and Dan Duohua can open 2-3 days, and the florescence is in the 6-7 month.Poem beautiful jade leads to a smile happiness to be born in the glam woods of high humidity, and be the seeds that uniquely can grow in water in Magnoliaceae extended familys, and growth fast, it is wooden excellent famous and precious, and the imperial palace noble of Thailand locality leads to the top-grade furniture of making or article for honor to have poem beautiful jade.Poem beautiful jade leads to that to be Thailand king with a smile lead to princess because compare the daughter's poem beautiful jade making a pet of oneself, and with its naming, the precious Ochnaceae in imminent danger of this Thailand treasures seeds.Therefore poem beautiful jade is logical is also called as " treasured of Thailand " with a smile, and rareness is classified as protected trees by World Conservation Union because it is precious.Within 2012, be chosen as the best arbor gold medal of form by China (middle mountain) south greening fair.It is excellent use material and flower garden ornamental tree species with a smile that poem beautiful jade leads to.
At present, there is the difficult problem that pollution rate is high, brownization is serious, inductivity is low, transplanting success is low in the tissue culture propagation of Magnoliacea plant, and lead to these bottlenecks hindering its propagation technique to develop especially of breeding of this kind of endangered species with a smile for poem beautiful jade.
Summary of the invention
The object of the invention is to provide a kind of poem beautiful jade to lead to cultured in vitro regeneration plant method with a smile, to preserve and to rescue world's Precious, Rare, Endangered Magnoliaceae kind, realize the breeding of critically endangered Magnoliaceae seeds, for the large tree spray of research Magnoliacea plant carries out tissue cultures, overcome the academic research experience that the tissue culture propagation technology such as pollution rate is high, brownization is serious, inductivity is low, transplanting success is low provide valuable.
Object of the present invention is achieved by the following technical programs: a kind of logical cultured in vitro regeneration plant method with a smile of poem beautiful jade, comprises the following steps:
(1) take explant and material processed thereof: take robust growth, the poem beautiful jade without damage by disease and insect leads to large the tree on mobile jib of life in 7 years with a smile sprouts the shoot, and after Preservation Treatment, low temperature takes back laboratory; Described material processed is the blade cut off on shoot, with the rear running water of bromogeramine cleaning, is cut into the stem section that 2cm is with 1-2 axillalry bud;
(2) sterile propagation body is set up: the stem section of having pruned under lab is used the mercury chloride sterilization 3-10 minute of 0.1%, again with aseptic washing 3-4 time, one bottle one section is inoculated on Initial culture base MS1, be 20-30 DEG C in room temperature, illumination every day 8-12 hour, 15-30 days is cultivated under the condition of intensity of illumination 800-2000LX, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, induce axillalry bud and indefinite bud and can growth and breeding continuously, complete the foundation of sterile propagation body;
(3) Multiplying culture: sterile propagation body is inoculated in proliferated culture medium MS2, being placed on manual control room temperature is 20-30 DEG C, illumination every day 8-12 hour, cultivates 15-30 days under the condition of intensity of illumination 800-2000LX, sprouts aseptic green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, bud clump breeds, and cultivates after 15-30 days, and bud clump proliferation times 2-4 doubly, aseptically split switching again, through squamous subculture expanding propagation repeatedly;
(4) culture of rootage: the Multiple Buds obtained after Multiplying culture is inoculated in root media and cultivates, namely, when Multiple Buds high at more than 15mm time, bud is cut from base portion, individual plant is divided into be inoculated in root media MS3, be placed on room temperature 20-30 DEG C, illumination every day 8-12 hour, intensity of illumination is 15-20 days under the condition of 1000-10000LX, seedling starts root, then to be placed under 75% shading condition short root in greenhouse and hardening 20-30 days, and bottle seedling leaf of taking root launches, elongation is healthy and strong, forms whole plant;
(5) transplant: the tissue-culture container seedling of taking root is transplanted in seedling-raising cup, complete bottle seedling from Artificial Control condition of culture provisions to the process of seedling autotrophy, concrete grammar is: carefully taken out by the seedling in bottle, cleans and sticks the medium on seedling, be transplanted to and fill up in the seedling-raising cup of matrix, a young plant planted by each cup, transplant thickness of earth-fill cover and just cover root system, be generally no more than 1 centimetre, compress matrix, make shoot root and matrix close contact, final singling water of drenching after having transplanted;
(6) seedling culture: bottle seedling keeps matrix moistening after the transfer, and relative air humidity, more than 95%, prevents leaves water loss, progressively control water supply in media afterwards, relative moisture controls at 80%-60%, control cultivation temperature 25-28 DEG C, cool canopy with 70% shading net shading; Within 3rd day, adopt 0.1% tpn spraying disinfection sterilizing after the transfer, can effectively prevent germ from growing, afterwards every spraying in 10-15 days once; Sprout when there being sprouting, new root is when growing, spray 1500 times of nutrient solutions, according to seedling growth, spray once every two weeks; After transplanting, growth stabilizes, and after growing sprouting and Xin Gen, progressively increases intensity of illumination, until carry out full exposure Routine Management, when seedling culture to 15 is centimetre high, moves to bigger flowerpot cultivating large seedling;
Wherein,
A. described MS1 medium is Initial culture base, and contain the drug variety of following nutritive element and the ratio of quality in every 1 liter, be dissolved in the water and be mixed with, pH is adjusted to 5.8:
Add water to 1000ml, additional in addition: 6-benzyladenine 0.5-5.0mg/l, methyl α-naphthyl acetate 0.01-0.5mg/l;
B. described MS2 medium is proliferated culture medium, and contain the drug variety of following nutritive element and the ratio of quality in every 1 liter, be dissolved in the water and be mixed with, pH is adjusted to 5.8:
Add water to 1000ml, additional in addition: 6-benzyladenine 0.2-2.0mg/l, 3-indolebutyric acid 0.01-0.5mg/l;
C. described MS3 medium is root induction medium, and contain the drug variety of following nutritive element and the ratio of quality in every 1 liter, be dissolved in the water and be mixed with, pH is adjusted to 5.8:
Add water to 1000ml, additional in addition: 3-indolebutyric acid 0.2-2.0mg/l, methyl α-naphthyl acetate 0.05-0.8mg/l.
Described matrix is fully mixed to get in the ratio of 2:1 by peat soil and coconut palm chaff.
Described transplanting carries out after bottle seedling good for root growth is moved into greenhouse domestication again, the bottle seedling starting root is directly moved into greenhouse domestication, bottle seedling is tamed in greenhouse, continue to grow root in the process of hardening, can shift to an earlier date 8-12 days and be transplanted by the bottle seedling of having tamed.
The logical cultured in vitro regeneration plant method inductivity with a smile of poem beautiful jade of the present invention and survival rate high, step is simple and feasible, very urgent and significance is had with the critically endangered seeds of breeding to preservation and rescue world Precious, Rare, Endangered Magnoliaceae kind, for the large tree spray of research Magnoliacea plant carries out tissue cultures, overcome the tissue culture propagation technology such as pollution rate is high, brownization is serious, inductivity is low, transplanting success is low and there is extremely valuable research value.
Embodiment
Be described in further details the present invention below by embodiment, these embodiments are only used for the present invention is described, do not limit the scope of the invention.
Embodiment 1 adopts following steps to realize poem beautiful jade to lead to cultured in vitro regeneration plant with a smile:
(1) take explant and material processed thereof: in lily magnolia garden, Xuwen County Divine Land, Guangdong Province, take robust growth, the poem beautiful jade without damage by disease and insect leads to large the tree on mobile jib of life in 7 years with a smile sprouts the shoot, and after on-the-spot Preservation Treatment, low temperature takes back laboratory; Cut off the blade on shoot in laboratory, with the rear running water of bromogeramine cleaning, be cut into the stem section of a band 1-2 axillalry bud of 2cm;
(2) sterile propagation body is set up: the stem section of having pruned under lab is used the mercury chloride sterilization 3 minutes of 0.1%, again with aseptic washing 4 times, one bottle one section is inoculated on Initial culture base MS1, be 30 DEG C in room temperature, illumination every day 8 hours, cultivates 15 days under the condition of intensity of illumination 2000LX, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, induces axillalry bud and indefinite bud and can growth and breeding continuously, completes the foundation of sterile propagation body;
(3) Multiplying culture: sterile propagation body is inoculated in proliferated culture medium MS2, being placed on manual control room temperature is 30 DEG C, illumination every day 8 hours, cultivates 28 days under the condition of intensity of illumination 1000LX, sprouts aseptic green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, bud clump breeds, and cultivates after 30 days, bud clump proliferation times 4 times, aseptically split switching again, through squamous subculture expanding propagation repeatedly;
(4) culture of rootage: when Multiple Buds high at more than 15mm time, bud is cut from base portion, be divided into individual plant to be inoculated in root media MS3, be placed on room temperature 28 DEG C, illumination every day 10 hours, intensity of illumination under the condition of 1000LX 15 days, seedling starts root, then to be placed under 75% shading condition short root in greenhouse and hardening 30 days, and bottle seedling leaf of taking root launches, elongation is healthy and strong, forms whole plant;
(5) transplant:
A prepares seedling medium: fully mixed with the ratio of coconut palm chaff in 2:1 by peat soil, be filled into by mixed-matrix in the seedling-raising cup in cool canopy for subsequent use;
The bottle seedling starting root is directly moved into greenhouse domestication by b, after domestication, the tissue-culture container seedling of taking root carefully is taken out, clean and stick the medium on seedling, be transplanted to and fill up in the seedling-raising cup of matrix, a young plant planted by each cup, transplants thickness of earth-fill cover and just covers root system, generally be no more than 1 centimetre, compress matrix, make shoot root and matrix close contact, final singling water of drenching after having transplanted;
(6) seedling culture: after transplanting, carries out the management of moisture, fertilizer and anti-bacteria:
A water management: bottle seedling keeps matrix moistening after the transfer, relative air humidity is 95%, and relative moisture controls 60%, control cultivation temperature 28 DEG C, cool canopy with 70% shading net shading;
The management of b fertilizer and anti-bacteria: adopt 0.1% tpn spraying disinfection sterilizing on the 3rd day after the transfer, afterwards every spraying in 15 days once; Sprout when there being sprouting, new root is when growing, spray 1500 times of nutrient solutions, according to seedling growth, spray once every two weeks;
C treats that growth stabilizes, and after growing sprouting and Xin Gen, progressively increases intensity of illumination, until carry out full exposure Routine Management, when seedling culture to 15 is centimetre high, moves to bigger flowerpot cultivating large seedling.
Embodiment 2 adopts following steps to realize poem beautiful jade to lead to cultured in vitro regeneration plant with a smile:
(1) take explant and material processed thereof: in lily magnolia garden, Xuwen County Divine Land, Guangdong Province, take robust growth, the poem beautiful jade without damage by disease and insect leads to large the tree on mobile jib of life in 7 years with a smile sprouts the shoot, and after on-the-spot Preservation Treatment, low temperature takes back laboratory; Cut off the blade on shoot in laboratory, with the rear running water of bromogeramine cleaning, be cut into the stem section of a band 1-2 axillalry bud of 2cm;
(2) sterile propagation body is set up: the stem section of having pruned under lab is used the mercury chloride sterilization 10 minutes of 0.1%, again with aseptic washing 3 times, one bottle one section is inoculated on Initial culture base MS1, be 20 DEG C in room temperature, illumination every day 12 hours, cultivates 20 days under the condition of intensity of illumination 1000LX, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, induces axillalry bud and indefinite bud and can growth and breeding continuously, completes the foundation of sterile propagation body;
(3) Multiplying culture: sterile propagation body is inoculated in proliferated culture medium MS2, being placed on manual control room temperature is 24 DEG C, illumination every day 10 hours, cultivates 30 days under the condition of intensity of illumination 800LX, sprouts aseptic green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, bud clump breeds, and cultivates after 30 days, bud clump proliferation times 3 times, aseptically split switching again, through squamous subculture expanding propagation repeatedly;
(4) culture of rootage: when Multiple Buds high at more than 15mm time, bud is cut from base portion, be divided into individual plant to be inoculated in root media MS3, be placed on room temperature 26 DEG C, illumination every day 8 hours, intensity of illumination under the condition of 20000LX 17 days, seedling starts root, then to be placed under 75% shading condition short root in greenhouse and hardening 28 days, and bottle seedling leaf of taking root launches, elongation is healthy and strong, forms whole plant;
(5) transplant:
A prepares seedling medium: fully mixed with the ratio of coconut palm chaff in 2:1 by peat soil, be filled into by mixed-matrix in the seedling-raising cup in cool canopy for subsequent use;
The bottle seedling starting root is directly moved into greenhouse domestication by b, after domestication, the tissue-culture container seedling of taking root carefully is taken out, clean and stick the medium on seedling, be transplanted to and fill up in the seedling-raising cup of matrix, a young plant planted by each cup, transplants thickness of earth-fill cover and just covers root system, generally be no more than 1 centimetre, compress matrix, make shoot root and matrix close contact, final singling water of drenching after having transplanted;
(6) seedling culture: after transplanting, carries out the management of moisture, fertilizer and anti-bacteria:
A water management: bottle seedling keeps matrix moistening after the transfer, relative air humidity is 95%, and relative moisture controls 88%, control cultivation temperature 25 DEG C, cool canopy with 70% shading net shading;
The management of b fertilizer and anti-bacteria: adopt 0.1% tpn spraying disinfection sterilizing on the 3rd day after the transfer, afterwards every spraying in 15 days once; Sprout when there being sprouting, new root is when growing, spray 1500 times of nutrient solutions, according to seedling growth, spray once every two weeks;
C treats that growth stabilizes, and after growing sprouting and Xin Gen, progressively increases intensity of illumination, until carry out full exposure Routine Management, when seedling culture to 15 is centimetre high, moves to bigger flowerpot cultivating large seedling.
Embodiment 3 adopts following steps to realize poem beautiful jade to lead to cultured in vitro regeneration plant with a smile:
(1) take explant and material processed thereof: in lily magnolia garden, Xuwen County Divine Land, Guangdong Province, take robust growth, the poem beautiful jade without damage by disease and insect leads to large the tree on mobile jib of life in 7 years with a smile sprouts the shoot, and after on-the-spot Preservation Treatment, low temperature takes back laboratory; Cut off the blade on shoot in laboratory, with the rear running water of bromogeramine cleaning, be cut into the stem section of a band 1-2 axillalry bud of 2cm;
(2) sterile propagation body is set up: the stem section of having pruned under lab is used the mercury chloride sterilization 5 minutes of 0.1%, again with aseptic washing 3 times, one bottle one section is inoculated on Initial culture base MS1, be 26 DEG C in room temperature, illumination every day 10 hours, cultivates 30 days under the condition of intensity of illumination 800LX, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, induces axillalry bud and indefinite bud and can growth and breeding continuously, completes the foundation of sterile propagation body;
(3) Multiplying culture: sterile propagation body is inoculated in proliferated culture medium MS2, being placed on manual control room temperature is 30 DEG C, illumination every day 12 hours, cultivates 15 days under the condition of intensity of illumination 1200LX, sprouts aseptic green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, bud clump breeds, and cultivates after 22 days, bud clump proliferation times 4 times, aseptically split switching again, through squamous subculture expanding propagation repeatedly;
(4) culture of rootage: when Multiple Buds high at more than 15mm time, bud is cut from base portion, be divided into individual plant to be inoculated in root media MS3, be placed on room temperature 26 DEG C, illumination every day 8 hours, intensity of illumination under the condition of 4000LX 20 days, seedling starts root, then to be placed under 75% shading condition short root in greenhouse and hardening 20 days, and bottle seedling leaf of taking root launches, elongation is healthy and strong, forms whole plant;
(5) transplant:
A prepares seedling medium: fully mixed with the ratio of coconut palm chaff in 2:1 by peat soil, be filled into by mixed-matrix in the seedling-raising cup in cool canopy for subsequent use;
The bottle seedling starting root is directly moved into greenhouse domestication by b, after domestication, the tissue-culture container seedling of taking root carefully is taken out, clean and stick the medium on seedling, be transplanted to and fill up in the seedling-raising cup of matrix, a young plant planted by each cup, transplants thickness of earth-fill cover and just covers root system, generally be no more than 1 centimetre, compress matrix, make shoot root and matrix close contact, final singling water of drenching after having transplanted;
(6) seedling culture: after transplanting, carries out the management of moisture, fertilizer and anti-bacteria:
A water management: bottle seedling keeps matrix moistening after the transfer, relative air humidity is 98%, and relative moisture controls 80%, control cultivation temperature 28 DEG C, cool canopy with 70% shading net shading;
The management of b fertilizer and anti-bacteria: adopt 0.1% tpn spraying disinfection sterilizing on the 3rd day after the transfer, afterwards every spraying in 15 days once; Sprout when there being sprouting, new root is when growing, spray 1500 times of nutrient solutions, according to seedling growth, spray once every two weeks;
C treats that growth stabilizes, and after growing sprouting and Xin Gen, progressively increases intensity of illumination, until carry out full exposure Routine Management, when seedling culture to 15 is centimetre high, moves to bigger flowerpot cultivating large seedling.
Claims (3)
1. the logical cultured in vitro regeneration plant method with a smile of poem beautiful jade, is characterized in that: comprise the following steps:
(1) take explant and material processed thereof: take robust growth, the poem beautiful jade without damage by disease and insect leads to large the tree on mobile jib of life in 7 years with a smile sprouts the shoot, and after Preservation Treatment, low temperature takes back laboratory; Described material processed is the blade cut off on shoot, with the rear running water of bromogeramine cleaning, is cut into the stem section that 2cm is with 1-2 axillalry bud;
(2) sterile propagation body is set up: the stem section of having pruned under lab is used the mercury chloride sterilization 3-10 minute of 0.1%, again with aseptic washing 3-4 time, one bottle one section is inoculated on Initial culture base MS1, be 20-30 DEG C in room temperature, illumination every day 8-12 hour, 15-30 days is cultivated under the condition of intensity of illumination 800-2000LX, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, induce axillalry bud and indefinite bud and can growth and breeding continuously, complete the foundation of sterile propagation body;
(3) Multiplying culture: sterile propagation body is inoculated in proliferated culture medium MS2, being placed on manual control room temperature is 20-30 DEG C, illumination every day 8-12 hour, cultivates 15-30 days under the condition of intensity of illumination 800-2000LX, sprouts aseptic green bud from axil, the green bud of clip is inoculated on proliferated culture medium MS2, bud clump breeds, and cultivates after 15-30 days, and bud clump proliferation times 2-4 doubly, aseptically split switching again, through squamous subculture expanding propagation repeatedly;
(4) culture of rootage: the Multiple Buds obtained after Multiplying culture is inoculated in root media and cultivates, namely, when Multiple Buds high at more than 15mm time, bud is cut from base portion, individual plant is divided into be inoculated in root media MS3, be placed on room temperature 20-30 DEG C, illumination every day 8-12 hour, intensity of illumination is 15-20 days under the condition of 1000-10000LX, seedling starts root, then to be placed under 75% shading condition short root in greenhouse and hardening 20-30 days, and bottle seedling leaf of taking root launches, elongation is healthy and strong, forms whole plant;
(5) transplant: the tissue-culture container seedling of taking root is transplanted in seedling-raising cup, complete bottle seedling from Artificial Control condition of culture provisions to the process of seedling autotrophy, concrete grammar is: carefully taken out by the seedling in bottle, cleans and sticks the medium on seedling, be transplanted to and fill up in the seedling-raising cup of matrix, a young plant planted by each cup, transplant thickness of earth-fill cover and just cover root system, be no more than 1 centimetre, compress matrix, make shoot root and matrix close contact, final singling water of drenching after having transplanted;
(6) seedling culture: bottle seedling keeps matrix moistening after the transfer, and relative air humidity, more than 95%, prevents leaves water loss, progressively control water supply in media afterwards, relative moisture controls at 80%-60%, control cultivation temperature 25-28 DEG C, cool canopy with 70% shading net shading; Within 3rd day, adopt 0.1% tpn spraying disinfection sterilizing after the transfer, can effectively prevent germ from growing, afterwards every spraying in 10-15 days once; Sprout when there being sprouting, new root is when growing, spray 1500 times of nutrient solutions, according to seedling growth, spray once every two weeks; After transplanting, growth stabilizes, and after growing sprouting and Xin Gen, progressively increases intensity of illumination, until carry out full exposure Routine Management, when seedling culture to 15 is centimetre high, moves to bigger flowerpot cultivating large seedling;
Wherein,
A. described MS1 medium is Initial culture base, and contain the drug variety of following nutritive element and the ratio of quality in every 1 liter, be dissolved in the water and be mixed with, pH is adjusted to 5.8:
Add water to 1000ml, additional in addition: 6-benzyladenine 0.5-5.0mg/l, methyl α-naphthyl acetate 0.01-0.5mg/l;
B. described MS2 medium is proliferated culture medium, and contain the drug variety of following nutritive element and the ratio of quality in every 1 liter, be dissolved in the water and be mixed with, pH is adjusted to 5.8:
Add water to 1000ml, additional in addition: 6-benzyladenine 0.2-2.0mg/l, 3-indolebutyric acid 0.01-0.5mg/l;
C. described MS3 medium is root induction medium, and contain the drug variety of following nutritive element and the ratio of quality in every 1 liter, be dissolved in the water and be mixed with, pH is adjusted to 5.8:
Add water to 1000ml, additional in addition: 3-indolebutyric acid 0.2-2.0mg/l, methyl α-naphthyl acetate 0.05-0.8mg/l.
2. the logical cultured in vitro regeneration plant method with a smile of poem beautiful jade according to claim 1, is characterized in that: described matrix is fully mixed to get in the ratio of 2:1 by peat soil and coconut palm chaff.
3. the logical cultured in vitro regeneration plant method with a smile of poem beautiful jade according to claim 1, it is characterized in that: described transplanting carries out after bottle seedling good for root growth is moved into greenhouse domestication again, the bottle seedling starting root is directly moved into greenhouse domestication, bottle seedling is tamed in greenhouse, continue to grow root in the process of hardening, within 8-12 days, is transplanted by the bottle seedling of having tamed in advance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410041803.1A CN103718969B (en) | 2014-01-28 | 2014-01-28 | A kind of logical cultured in vitro regeneration plant method with a smile of poem beautiful jade |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410041803.1A CN103718969B (en) | 2014-01-28 | 2014-01-28 | A kind of logical cultured in vitro regeneration plant method with a smile of poem beautiful jade |
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| CN104082139A (en) * | 2014-06-28 | 2014-10-08 | 玉林师范学院 | Tissue-culture rapid propagation method of Micheliamacclurei Dandy |
| CN105027741B (en) * | 2015-09-16 | 2018-08-17 | 广东神州木兰园林有限公司 | The logical seed germination with a smile of poem beautiful jade and the method for nursery |
| CN105900842B (en) * | 2016-05-09 | 2017-12-08 | 中国林业科学研究院热带林业研究所 | A kind of Michelia macclurei tissue-culturing quick-propagation method for culturing seedlings |
| CN107173155A (en) * | 2017-06-21 | 2017-09-19 | 潍坊市园林管理处 | A kind of North America yulan test tube seedling acclimation method |
| CN117598204B (en) * | 2024-01-24 | 2024-03-22 | 西南林业大学 | Tissue culture and rapid propagation method and application of michelia yunnanensis |
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