CN104285814B - A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind - Google Patents

A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind Download PDF

Info

Publication number
CN104285814B
CN104285814B CN201410590375.8A CN201410590375A CN104285814B CN 104285814 B CN104285814 B CN 104285814B CN 201410590375 A CN201410590375 A CN 201410590375A CN 104285814 B CN104285814 B CN 104285814B
Authority
CN
China
Prior art keywords
culture
days
explant
alpine ash
plantlet
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201410590375.8A
Other languages
Chinese (zh)
Other versions
CN104285814A (en
Inventor
沈云
莫继有
兰俊
陈德文
苏勇
庞贞武
梁秀莉
黄全东
吴兵
陈允椿
梁庭辉
唐再生
韦炳俭
吴满芬
俸荣娣
邓冬丽
付淼
吕华丽
石前
陈东林
李丽芳
熊涛
王建忠
张磊
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
GUANGXI ZHUANG AUTONOMOUS REGION STATE-OWNED DONGMEN FOREST FARM
Original Assignee
GUANGXI ZHUANG AUTONOMOUS REGION STATE-OWNED DONGMEN FOREST FARM
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by GUANGXI ZHUANG AUTONOMOUS REGION STATE-OWNED DONGMEN FOREST FARM filed Critical GUANGXI ZHUANG AUTONOMOUS REGION STATE-OWNED DONGMEN FOREST FARM
Priority to CN201410590375.8A priority Critical patent/CN104285814B/en
Publication of CN104285814A publication Critical patent/CN104285814A/en
Application granted granted Critical
Publication of CN104285814B publication Critical patent/CN104285814B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/22—Improving land use; Improving water use or availability; Controlling erosion
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/40—Afforestation or reforestation

Abstract

The invention discloses a kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind, realized by the foundation of explant, explant process, Initial culture, squamous subculture, culture of rootage, hardening and transplanting and other steps, special medium composition is have selected for Initial culture, subculture and culture of rootage, after rational hardening measure makes tail alpine ash transplant, growth is fast, survival rate is high, reach reproduction coefficient high, the object that reproductive effect is good.Simultaneously the inventive method low production cost, the plantlet in vitro obtained heredity shape is consistent, is the numerous soon effective way of Eucalyptus Tissue Cultured, and applied widely.

Description

A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind
Technical field
The present invention relates to Eucalyptus Planting technical field, particularly a kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind.
Background technology
Eucalyptus (Eucalyptus) is the general designation that Myrtaceae (Myrtaceae) eucalyptus belongs to (Eucalyptus) plant, originate in Australia, through the introduction and acclimatization in country variant for many years and area, develop into more than 600 kind, wherein Important Economic commerical tree species about more than 100 are planted, and are had kind more than 40 by what extensively cultivate.In recent years, eucalyptus plantation is all being greatly developed in China various places, and in Guangdong and Guangxi Provinces, Hainan, Sichuan, Yunnan etc. economizes and take on a certain scale, and creates significantly ecological and economic benefit.
Tail alpine ash DH32-43 kind ( e.urophylla × E.Grandis), be forest farm, east gate, Guangxi, with Indonesia's provenance east gate Eucalyptus urophylla select tree U16( eucalyptusurophylla) as maternal, with Australian provenance east gate alpine ash select tree G46( eucalyptusgrandis) as paternal hybrid sociales, by the excellent eucalypt species that vegetative propagation obtains, this varietal characteristic is as follows: nursery stock leaf base in seedling stage is slightly roomy, blade is long by point, and edge is wavy, terminal bud kermesinus, vein is clear, not protruding, and comparatively other product are long in Seedling Stage seedling footpath.Simultaneously, there is kind of batch general performance excellent, form is logical straight, branch is little, forest form in good order, without damage by disease and insect, average annual accumulation is large, economic benefit is obvious, and the advantage of plantation suitable for large-scale promotion, all can plant in each department, Guangxi, inventor in Bobai forest farm, Guangxi, forest farm, state-run peak, Guangxi, Guangxi state-ownedly sends forest farm, Yangshan, extend trial is carried out for many years in state-run seven forest farms, slope, Guangxi, result shows: 6.5 years raw time accumulation reach 280.5m 3/ hm 2, year accumulation reaches 43.2m 3/ hm 2.In regional test, 3 years raw time average annual Stock growth reach 31 ~ 41m 3/ hm 2, fast growing, rotation short (industrial fiber material); Form is logical directly, branch is little, forest form is neat; Material is good, and texture is straight, barren-resistant, is applicable to large area region plantation.Economy and ecological benefits are very remarkable, and by excellent kind amount reproduction at short notice, acquired character performance is consistent, genetic stability, the offspring that trait segregation does not occur is significant, eucalyptus is the woody plant of perennial cross pollination, the phenomenon that between kind, natural hybrization hybridizes is frequent, offspring is seriously separated, therefore sexual propagation means are adopted to be difficult to keep select tree characteristic, and asexual reproduction method such as cuttage, press strip, grafting etc. are unfavorable for amount reproduction, it is comparatively effective method that tissue culture technique cultivates Fast-propagation eucalyptus.
Tissue cultures (TissueCulture) is aseptically, and be separated also the technology of culture of ex vivo plant tissue in the medium, it has following significance: (1) promotes the Fast-propagation of excellent genetic stocks; (2) cultivation of detoxification breeding seedling and the amount reproduction of virus-free seedling; (3) Fast-propagation of special breeding material; (4) Fast-propagation of gene engineering plant; (5) Fast-propagation of Plantlet in vitro kind matter.According to statistics, current existing more than 60 plant eucalyptus tissue cultures succeeds, first reported eucalyptus camaldulensis seedling explant obtains callus from the Susses sixties in last century, both at home and abroad successively in eucalyptus citriodora (1969), alpine ash (1974), beautiful flower eucalyptus (1976), Bai An (1975), hybrid eucalyptus (1980), grey eucalyptus, Wang An (1980), Lei Lin mono-eucalyptus (1981), willow gallery F1 hybridizes generation (1981), eucalyptus camaldulensis (1992) has carried out linked groups's culture technique research.
Meanwhile, the research for the tissue cultures aspect of eucalyptus also has many:
1, title: the stereoscopic culture of tail alpine ash and Fast-propagation; Author: Liu Yiqing, Wang Daping, Xiong Yunhai; Gardening journal the 32nd volume; Summary: the tissue culture and rapid propagation method discussing No. T13, tail alpine ash.Gather the semi-lignified spray of No. T13, tail alpine ash in the season of growth, remove blade, be cut into stem-segment with single bud, use tap water 15min, 0.5% washing powder solution soaks 15min, cleaner with tap water.Material is put into respectively the empty cup of aseptic plastic, put 15-20,75% alcohol-pickled 30s for every glass, 0.1% mercuric chloride sterilization 10-12min, finally uses aseptic water washing 3-5 time, is inoculated in inducing culture.The minimal medium of induction axillary bud sprouting and proliferation and subculture is kept to 1/3 for improvement MS(ammonium nitrate, is called for short EU), the minimal medium of taking root is 1/2MS, BA, the IBA of additional variable concentrations, NAA, active carbon (AC), sucrose 3%, agar 5.0g./L, pH6.0.Fiber differentiation utilizes Indoor Natural scattered light (not turning on light), and proliferation and subculture, strengthening seedling and rooting are cultivated, illumination 12h/d, light intensity 1500-2000lu, temperature 26 ± 3 DEG C, ambient humidity 405-70%.Results and analysis: EU+BA0.5+NAA0.2 evoked callus is best, and cultivation effect is best.This research uses traditional sucrose as carbon source, and culture medium cost is relatively high, and the nutrient component of sucrose is single, more comprehensively for cell division provides nutriment, can not affect culture effect to a certain extent.In addition, do not add the composition improving resistance and suppress living contaminants in medium, the survival rate of plantlet in vitro can be affected.
2, title: eucalyptus tissue culture rapid propagating technology; Author: Lin Yingyan; Guangxi Vocational & Technical College's thesis (design), summary: more detailed discuss the formula (MS+BA0.5+IBA0.2) of eucalyptus from the sterilizing of the sampling of Initial culture, material, inoculation, cultivation and medium; Inoculation method in subculture, medium (MS+BA1.0+KT0.5+IBA0.1-0.5) and condition of culture; The formula (1/2MS+ABT1.5+IBA0.1+ active carbon 0.25%) of the medium in culture of rootage, inoculation method and cultivation and condition; Various control measures etc. in hardening in nursery, the preparation of matrix, transplanting and seedling raising process.Equally, do not add the composition improving resistance and suppress living contaminants in the medium of this research, the survival rate of plantlet in vitro can be affected.
3, Authorization Notice No. is the patent of CN103155872B: a kind of tail alpine ash embryoid induction seedling-cultivating method, and it for explant with tail alpine ash aseptic seedling stem sections, is sprouted through embryonic callus induction, embryoid induction and embryoid and can obtain complete regeneration plant; Adopt method of the present invention can obtain through embryoid way regeneration plant, result is stablized, and repeatability is good, can apply and produce upper large-scale commercial nursery, for the breeding of tail alpine ash is laid a good foundation.Above-mentioned patent adopts tail alpine ash aseptic seedling stem sections to be that explant carrys out induced embryonic callus, regeneration plant is obtained by somatic embryo generation Regeneration Ways, operation is comparatively complicated, enforcement difficulty is large, use tail alpine ash aseptic seedling stem sections to be explant simultaneously, maternal good characteristic needs to be investigated, and simple this technology commercialization nursery of employing can not effective hereditary parent's good characteristic.
Visible, although more for the research of eucalyptus tissue culture technique at present, but it is numerous that the purposive merit for select tree carries out expansion, the tissue culture and rapid propagation method simultaneously can effectively applied also has a lot of need of work to grope, because DH32-43 is perennial tail alpine ash kind, market plantlet in vitro is less, resource is well sold and in short supply, and for this kind tissue cultures work and not yet in effectly to carry out, Study on tissue culture is carried out to tail alpine ash DH32-43 new varieties, for the Fast-propagation of these improved seeds, meet market supply, the ecological benefits and the economic benefit that improve these improved seeds are significant.
Summary of the invention
The object of the invention is for above deficiency, provide that a kind of operation easier is low, reproduction coefficient is high, the control cycle is short, can genetic stability and the tissue culture and rapid propagation method of the high tail alpine ash DH32-43 kind of survival rate.
Technical scheme of the present invention is as follows:
The tissue culture and rapid propagation method of tail alpine ash DH32-43 kind, concrete steps are as follows:
1. the foundation of explant: select the 4-5 age of tree, robust growth, trunk be thick, without the select tree of damage by disease and insect, cuts down from overhead 20-25cm, grows to 10cm, semi-lignified, eye be not when also sprouting, adopt back budling as explant material until stub rudiment.For reducing material contamination source, when taking budling, should select within continuous more than three days, carry out after fair weather.After the budling adopted back is removed most of blade, use tap water 15min, 0.5% washing powder solution soaks 15min, cleaner with tap water.
2. explant sterilization: the explant that 1. step obtains with 75% alcohol-pickled 30S, aseptic water washing 4-5 time, drains the water, then the mercuric chloride solution being placed in 0.1% is sterilized 9min, aseptic water washing 4-5 time, cut rudiment bar terminal bud get off second section rise 1-2 joint stem with bud, long 1-2cm.
3. Initial culture: stem section step 2. processed aseptically is inoculated into Initial culture base, and Initial culture based component is: MS+IAA2mg/L+6-BA1.6mg/L+3% molasses, medium pH 5.9; Intensity of illumination 1000Lux, cultivation temperature 25 ± 2 DEG C, incubation time 35 ± 2 days.
4. squamous subculture: Initial culture goes out clump bud to joint director, and get clump bud and carry out squamous subculture, squamous subculture based component is: MS+6-BA1.5mg/L+IAA0.15mg/L+0.08% amino-oligosaccharide+5% molasses; Cultivation temperature 25 ± 2 DEG C, light application time 16h/d, medium pH 5.8-6.0; Cultivate 20 days, form the budling with 3 stipes that 2cm is long, be cut into the segment of band stipes, each bud is divided into 2 sections; Every 20 days subcultures like this once, expand numerous test-tube plantlet.
5. culture of rootage: treat that 4. step expands numerous test-tube plantlet and take out, proceed to culture of rootage, culture of rootage based component is: 1/5MS+IBA1.5mg/L+ABT0.2mg/L+1.0% amino-oligosaccharide+15% molasses; Intensity of illumination is 3000Lux, cultivation temperature 25 ± 2 DEG C, and be cultured to 25 days, root grows to 1-3cm, seedling grow tall 3cm time bottle outlet.
In subculture medium and root media, the Main Function of amino-oligosaccharide is: (1) excites clump bud tissue to produce phenolic compound, lignin and pathogenesis-related proteins, improves clump bud anti-adversity ability; (2) plant cell is activated, Promoting plant growth, the expression of regulating plant resistant gene, use amino-oligosaccharide as subculture medium and root media, inhibitory action can not be produced to the cultivation of explant under described dosage, in incubation, not be vulnerable to living contaminants, through multiple authentication, inventor proves that amino-oligosaccharide plays an important role in subculture medium of the present invention and root media, be its important component part.
And in three kinds of medium, add molasses (it is non crystallized containing sugar liquors that sugar refinery sugar industry is separated) also play an important role: (1), as the carbon source of tissue rapid propagation medium, substitutes sucrose, reduces production cost; (2) be easier to the various monose, thick protein and the mineral matter that utilize containing a large amount of plant, comparatively sucrose nutrition is more comprehensive.
Other medium compound can the textbooks of chemically dictionary or arviculture to its title and effect, the present invention adopts these compounds to carry out combining tail alpine ash DH32-43 kind the most applicable, particularly add the molasses being rich in nutrient component, be through that a lot of screening just finally determines.
6. hardening: select test-tube plantlet bottle outlet that is healthy and strong, semi-lignified to transplant at peat soil: coconut palm chaff proportioning is in the nutrient matrix of 1:3, keeps air humidity 85-90%, illumination 10000lux.
7. transplant; Hardening moved on to outdoor cultivation after 15 days, when seedling height in seedling-cultivating tray is 15-20cm, transplanted afforestation.
The tissue culture and rapid propagation method of tail alpine ash DH32-43 kind, also comprises the management after plantlet in vitro transplanting, is specially: before transplanting, plantlet in vitro is soaked 1-2h in normal root water; Use sufficient base manure, topdress in time, and prevention and elimination of disease and pests in time.
Described normal root water consists of: 3% metalaxyl+3% dislikes mould spirit+0.05% indolebutyric acid+0.05% Nafusaku+0.5% Sodium Polyacrylate+pure water surplus.
Described prevention and elimination of disease and pests is: prevent and treat damping off, fusarium wilt in seedling stage; Gray mold, leaf spot, defoliation is prevented and treated in vegetative period; Use bait formulation control termites, grub, cutworm; Thermal fog is used to prevent and treat eucalyptus caterpillar, looper in vegetative period.
Beneficial effect of the present invention is;
One, the merit of DH32-43 is maintained: fast growing, rotation short (industrial fiber material); Form is logical directly, branch is little, forest form is neat; Material is good, and texture is straight, barren-resistant, is applicable to large area region plantation.
Two, Eucalyptus Tissue Cultured quick-breeding method operation easier of the present invention is low, production cost is low, is easy to promote the use of on a large scale.In extensive tissue culture procedures, medium uses molasses as important component, instead of molasses, directly reduces the cost of medium; Meanwhile, in medium, amino-oligosaccharide appropriate use in the medium effectively inhibits miscellaneous bacteria to infect, and minimizing group training risk is the tissue culture and rapid propagation method of comparatively mature and reliable safety.
Three, the plantlet in vitro survival rate utilizing Eucalyptus Tissue Cultured quick-breeding method of the present invention to produce is high, and DH32-43 plantlet in vitro survival rate, more than 95%, is adapted at Guangxi most area plantation simultaneously and promotes, directly enhance economic benefit and social benefit.
Embodiment
Be described in detail preferred embodiment of the present invention below, consciousness advantages and features of the invention can be easier to be readily appreciated by one skilled in the art, thus make more explicit defining to protection scope of the present invention.
embodiment 1
The tissue culture and rapid propagation method of tail alpine ash DH32-43 kind, concrete steps are as follows:
1. the foundation of explant: select the 4-5 age of tree, robust growth, trunk be thick, without the select tree of damage by disease and insect, cuts down from overhead 20-25cm, grows to 10cm, semi-lignified, eye be not when also sprouting, adopt back budling as explant material until stub rudiment.For reducing material contamination source, when taking budling, should select within continuous more than three days, carry out after fair weather.After the budling adopted back is removed most of blade, use tap water 15min, 0.5% washing powder solution soaks 15min, cleaner with tap water.
2. explant sterilization: the explant that 1. step obtains with 75% alcohol-pickled 30S, aseptic water washing 5 times, drains the water, then the mercuric chloride solution being placed in 0.1% is sterilized 9min, aseptic water washing 5 times, cut rudiment bar terminal bud get off second section rise 1-2 joint stem with bud, long 1-2cm.
3. Initial culture: stem section step 2. processed aseptically is inoculated into Initial culture base, and Initial culture based component is: MS+IAA2mg/L+6-BA1.6mg/L+3% molasses, medium pH 5.9; Intensity of illumination 1000Lux, cultivation temperature 25 ± 2 DEG C, incubation time 35 ± 2 days.
4. squamous subculture: Initial culture goes out clump bud to joint director, and get clump bud and carry out squamous subculture, squamous subculture based component is: MS+6-BA1.5mg/L+IAA0.15mg/L+0.08% amino-oligosaccharide+5% molasses; Cultivation temperature 25 ± 2 DEG C, light application time 16h/d, medium pH 5.8-6.0; Cultivate 20 days, form the budling with 3 stipes that 2cm is long, be cut into the segment of band stipes, each bud is divided into 2 sections; Every 20 days subcultures like this once, expand numerous test-tube plantlet.
5. culture of rootage: treat that 4. step expands numerous test-tube plantlet and take out, proceed to culture of rootage, culture of rootage based component is: 1/5MS+IBA1.5mg/L+ABT0.2mg/L+1.0% amino-oligosaccharide+15% molasses; Intensity of illumination is 3000Lux, cultivation temperature 25 ± 2 DEG C, and be cultured to 25 days, root grows to 1-3cm, seedling grow tall 3cm time bottle outlet.
6. hardening: select test-tube plantlet bottle outlet that is healthy and strong, semi-lignified to transplant at peat soil: coconut palm chaff proportioning is in the nutrient matrix of 1:3, keeps air humidity 85-90%, illumination 10000lux.
7. transplant; Hardening moved on to outdoor cultivation after 15 days, when seedling height in seedling-cultivating tray is 15-20cm, transplanted afforestation.
embodiment 2
The tissue culture and rapid propagation method of tail alpine ash DH32-43 kind, concrete steps are as follows:
1. the foundation of explant: select the 4-5 age of tree, robust growth, trunk be thick, without the select tree of damage by disease and insect, cuts down from overhead 20-25cm, grows to 10cm, semi-lignified, eye be not when also sprouting, adopt back budling as explant material until stub rudiment.For reducing material contamination source, when taking budling, should select within continuous more than three days, carry out after fair weather.After the budling adopted back is removed most of blade, use tap water 15min, 0.5% washing powder solution soaks 15min, cleaner with tap water.
2. explant sterilization: the explant that 1. step obtains with 75% alcohol-pickled 30S, aseptic water washing 4 times, drains the water, then the mercuric chloride solution being placed in 0.1% is sterilized 9min, aseptic water washing 4 times, cut rudiment bar terminal bud get off second section rise 1-2 joint stem with bud, long 1-2cm.
3. Initial culture: stem section step 2. processed aseptically is inoculated into Initial culture base, and Initial culture based component is: MS+IAA2mg/L+6-BA1.6mg/L+3% molasses, medium pH 5.9; Intensity of illumination 1000Lux, cultivation temperature 25 ± 2 DEG C, incubation time 35 ± 2 days.
4. squamous subculture: Initial culture goes out clump bud to joint director, and get clump bud and carry out squamous subculture, squamous subculture based component is: MS+6-BA1.5mg/L+IAA0.15mg/L+0.08% amino-oligosaccharide+5% molasses; Cultivation temperature 25 ± 2 DEG C, light application time 16h/d, medium pH 5.8-6.0; Cultivate 20 days, form the budling with 3 stipes that 2cm is long, be cut into the segment of band stipes, each bud is divided into 2 sections; Every 20 days subcultures like this once, expand numerous test-tube plantlet.
5. culture of rootage: treat that 4. step expands numerous test-tube plantlet and take out, proceed to culture of rootage, culture of rootage based component is: 1/5MS+IBA1.5mg/L+ABT0.2mg/L+1.0% amino-oligosaccharide+15% molasses; Intensity of illumination is 3000Lux, cultivation temperature 25 ± 2 DEG C, and be cultured to 25 days, root grows to 1-3cm, seedling grow tall 3cm time bottle outlet.
6. hardening: select test-tube plantlet bottle outlet that is healthy and strong, semi-lignified to transplant at peat soil: coconut palm chaff proportioning is in the nutrient matrix of 1:3, keeps air humidity 85-90%, illumination 10000lux.
7. transplant; Hardening moved on to outdoor cultivation after 15 days, when seedling height in seedling-cultivating tray is 15-20cm, transplanted afforestation.
8. the management after transplanting: before transplanting, plantlet in vitro is soaked 1-2h in normal root water, normal root water consists of: 3% metalaxyl+3% dislikes mould spirit+0.05% indolebutyric acid+0.05% Nafusaku+0.5% Sodium Polyacrylate+pure water surplus; Every plantlet in vitro uses 10kg high-quality well-rotted farmyard manure, additional 2kgNPK15:15:15 composite fertilizer; Within latter 3 months, 6 months, respectively topdress 1 time in transplanting, topdress at every turn and use urea 1kg/, topdress every year once later, each use 2kg urea/, and prevention and elimination of disease and pests in time.Prevention and elimination of disease and pests and make with medicament as follows in DH32-43 process of growth: seedling blight and fusarium wilt only have fragmentary generation, after using normal root water to dilute 800 times to the plantlet in vitro that disease occurs, every plantlet in vitro perfusion water 500-800 gram; Gray mold uses the 5000 times of sprayings of BASF medicament 50% Boscalid water-dispersible grain dilution agent; Leaf spot and defoliation use 25% pyraclostrobin missible oil of BASF, dilute 8000 times of even spraying, and the effectively pre-disease prevention of decapacitation meets accident, and also has and obviously promotes Eucalyptus effect; Use 0.5% fipronil bait control soil insect grub cutworm and termite, using method is for be evenly sprinkling upon bait formulation around plantlet in vitro, and every uses bait formulation 20 grams-30 grams; When 3-5 age period eucalyptus outburst looper and caterpillar, the 1.8% A Wei chlorine cyanogen thermal fog control using Guangxi Tianyuan Biochemical Co., Ltd. to produce, saves man-hour, improves work efficiency, ensure that the high yield of eucalyptus.
Be below tail alpine ash DH32-43 plantlet in vitro breeding results that the inventive method obtains:
Can find out, tail alpine ash DH32-43 kind tissue culture and rapid propagation method of the present invention, reproduction coefficient is high, rooting rate reaches more than 99%, and survival rate is more than 95%, and plantlet in vitro can adapt to open-air atmosphere rapidly after transplanting, growth rapidly, anti-extraneous poor environment ability is strong, has planted popularization 5000 mu, has created larger economic benefit, ecological benefits and social benefit.

Claims (2)

1. a tissue culture and rapid propagation method for tail alpine ash DH32-43 kind, is characterized in that: step is as follows:
1. the foundation of explant: select the 4-5 age of tree, robust growth, trunk be thick, without the select tree of damage by disease and insect, cuts down from overhead 20-25cm, grows to 10cm, semi-lignified, eye be not when also sprouting, adopted back by budling as explant material until stub rudiment; For reducing material contamination source, when taking budling, should select within continuous more than three days, carry out after fair weather; After the budling adopted back is removed most of blade, use tap water 15min, 0.5% washing powder solution soaks 15min, cleaner with tap water;
2. explant sterilization: the explant that 1. step obtains with 75% alcohol-pickled 30s, aseptic water washing 4-5 time, drains the water, then the mercuric chloride solution being placed in 0.1% is sterilized 9min, aseptic water washing 4-5 time, cut rudiment bar terminal bud get off second section rise 1-2 joint stem with bud, long 1-2cm;
3. Initial culture: stem section step 2. processed aseptically is inoculated into Initial culture base, and Initial culture based component is: MS+IAA2mg/L+6-BA1.6mg/L+3% molasses, medium pH 5.9; Intensity of illumination 1000Lux, cultivation temperature 25 ± 2 DEG C, incubation time 35 ± 2 days;
4. squamous subculture: Initial culture goes out clump bud to joint director, and get clump bud and carry out squamous subculture, squamous subculture based component is: MS+6-BA1.5mg/L+IAA0.15mg/L+0.08% amino-oligosaccharide+5% molasses; Cultivation temperature 25 ± 2 DEG C, light application time 16h/d, medium pH 5.8-6.0; Cultivate 20 days, form the budling with 3 stipes that 2cm is long, be cut into the segment of band stipes, each bud is divided into 2 sections; Every 20 days subcultures like this once, expand numerous test-tube plantlet;
5. culture of rootage: treat that 4. step expands numerous test-tube plantlet and take out, proceed to culture of rootage, culture of rootage based component is: 1/5MS+IBA1.5mg/L+ABT0.2mg/L+1.0% amino-oligosaccharide+15% molasses; Intensity of illumination is 3000Lux, cultivation temperature 25 ± 2 DEG C, and be cultured to 25 days, root grows to 1-3cm, seedling grow tall 3cm time bottle outlet;
6. hardening: select test-tube plantlet bottle outlet that is healthy and strong, semi-lignified to transplant at peat soil: coconut palm chaff proportioning is in the nutrient matrix of 1:3, keeps air humidity 85-90%, illumination 10000lux;
7. transplant: hardening moved on to outdoor cultivation after 15 days, when seedling height in seedling-cultivating tray is 15-20cm, transplant afforestation;
The tissue culture and rapid propagation method of described tail alpine ash DH32-43 kind, also comprises the management after plantlet in vitro transplanting, is specially: before transplanting, plantlet in vitro is soaked 1-2h in normal root water; Use sufficient base manure, topdress in time, and prevention and elimination of disease and pests in time; Described normal root water consists of: 3% metalaxyl+3% dislikes mould spirit+0.05% indolebutyric acid+0.05% Nafusaku+0.5% Sodium Polyacrylate+pure water surplus.
2. the tissue culture and rapid propagation method of tail alpine ash DH32-43 kind according to claim 1, is characterized in that: described prevention and elimination of disease and pests is: prevent and treat damping off, fusarium wilt in seedling stage; Gray mold, leaf spot, defoliation is prevented and treated in vegetative period; Use bait formulation control termites, grub, cutworm; Thermal fog is used to prevent and treat eucalyptus caterpillar, looper in vegetative period.
CN201410590375.8A 2014-10-29 2014-10-29 A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind Active CN104285814B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201410590375.8A CN104285814B (en) 2014-10-29 2014-10-29 A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201410590375.8A CN104285814B (en) 2014-10-29 2014-10-29 A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind

Publications (2)

Publication Number Publication Date
CN104285814A CN104285814A (en) 2015-01-21
CN104285814B true CN104285814B (en) 2016-04-06

Family

ID=52305988

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410590375.8A Active CN104285814B (en) 2014-10-29 2014-10-29 A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind

Country Status (1)

Country Link
CN (1) CN104285814B (en)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105230489A (en) * 2015-11-02 2016-01-13 大新县生产力促进中心 Tissue culture fast propagation method for dendrobium candidum
CN105379624B (en) * 2015-12-10 2017-09-12 广西壮族自治区国有东门林场 A kind of tissue culture and rapid propagation method of Eucalyptus pellita
CN105454047B (en) * 2015-12-23 2017-09-26 广西壮族自治区国有东门林场 A kind of tissue culture and rapid propagation method of Eucalyptus cloeziana
CN105815223A (en) * 2016-04-21 2016-08-03 中国林业科学研究院热带林业研究所 Building method of eucapyptus urophylla leaf-induced plantlet regeneration system
CN105993766A (en) * 2016-05-24 2016-10-12 象州县科学技术局 Cultivation method for skiranui tangerine
CN108651283A (en) * 2018-05-17 2018-10-16 邵武市闽澳农业发展有限公司 A kind of narrow leaved tea tree tissue-cultured seedling culture of rootage method

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101884301A (en) * 2010-04-08 2010-11-17 浙江省农业科学院 Method for preventing pollution in plant tissue culture

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101884301A (en) * 2010-04-08 2010-11-17 浙江省农业科学院 Method for preventing pollution in plant tissue culture

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
‘T13’号桉树再生体系的建立;刘奕清;《中国种业》;20061231(第4期);第31-32页 *
尾巨桉提纯复壮初步研究;吴建宇等;《现代农业科技》;20101231(第4期);第234-235页 *
桉树组织培养与再生系统建立研究;王笑春等;《河北建筑科技学院学报》;20060630;第23卷(第2期);第71-74页 *

Also Published As

Publication number Publication date
CN104285814A (en) 2015-01-21

Similar Documents

Publication Publication Date Title
CN104285814B (en) A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind
CN104273037B (en) A kind of tissue culture and rapid propagation method of tail alpine ash DH33-27 kind
CN101401524B (en) Cutting propagation method for beach plum epicormic branch
CN104365479B (en) A kind of tissue culture and rapid propagation method of tail alpine ash DH32-28 kind
CN104585033B (en) Blood aspidistra quick breeding method for tissue culture
CN107018896B (en) A kind of method of facility cuttage tilia miqueliana
CN104285815B (en) A kind of tissue culture and rapid propagation method of tail alpine ash DH32-13 kind
CN104686338A (en) In-vitro culture technique of anther of angelica dahurica
CN106613718A (en) Cultivation method for grape cutting seedlings with high cold resistance
CN103461121A (en) Ex-vitro rooting method for tissue culture seedlings of pinus massoniana
CN104304028B (en) A kind of tissue culture and rapid propagation method of tail alpine ash DH32-26 kind
CN104094848B (en) The method of the induction of tung oil tree hypocotyledonery axis callus and highly efficient regeneration plant
CN103299909A (en) Method for breeding cherry seedlings in large scales through tissue culturing
CN105941155A (en) Method for rapidly propagating fraxinus mandshurica by utilizing suspension culture technology
CN103975855A (en) Haploid breeding method of dendrobium candidum
CN104823790B (en) A kind of bulla walnut cuttage fast seedling-cultivating method
CN104304027B (en) A kind of tissue culture and rapid propagation method of tail alpine ash DH32-29 kind
CN104304026B (en) A kind of tissue culture and rapid propagation method of tail alpine ash DH30-1 kind
CN102668991B (en) Application of penicillin to simple test-tube breeding of grapes and novel technology for test-tube breeding of grapes
CN105638482B (en) The method of walnut and Juglans mandshurica interspecific hybridization IMMATURE EMBRYOS CULTURE
CN109362524A (en) A kind of cultural method of African Chrysanthemum new varieties
CN104054579B (en) A kind of method of tung oil tree petiole directly regenerated plant
CN103004600A (en) Regeneration method for rosa wichuriana plant taking leaf as explant
CN102119664B (en) Method for culturing muscari botryoides mill with plant tissues
CN104920215A (en) Simplified sugarcane test-tube plantlet rooting and seedling method

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant