CN103947556B - The selection of the potential test tube flowering material of a kind of fragrant camphor tree Adult form and plant regeneration method - Google Patents
The selection of the potential test tube flowering material of a kind of fragrant camphor tree Adult form and plant regeneration method Download PDFInfo
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Abstract
The invention discloses selection and the plant regeneration method of the potential test tube flowering material of a kind of fragrant camphor tree Adult form, concrete grammar is: (1) is in by the end of December between January next year, choose age of tree 8-10, the diameter of a cross-section of a tree trunk 1.3 meters above the ground is that the raw then tool bulbil branch of the fragrant camphor tree of adult healthy of about 13.5cm is originated as explant; (2) be inoculated on germination medium by the Adult form Agistemus exsertus after disinfecting, induction stem section is sprouted; (3) be inoculated in by sprouting branch on expansion breeding culture medium, propagation forms single-strain clone, sets up stem section culture seedling and propagating system; (4) cutting shoots of stem section culture seedling is proceeded to inducing culture and carry out adventitious bud induction culture, set up fragrant camphor tree stem section culture seedling cutting shoots plant regeneration system.The invention provides a kind of fragrant camphor tree method for tissue culture, this cultural method is drawn materials conveniently, and material source, quantity are sufficient, easily obtain sterilizable material; The decision method of Adult form material, method is simple, and conclusion is reliable.
Description
Technical field
The present invention relates to technical field of forestry, particularly relate to selection and the plant regeneration method of the potential test tube flowering material of a kind of fragrant camphor tree Adult form.
Background technology
Fragrant camphor tree is Lauraceae cinnamon, and this kind with luxuriant foliage and spreading branches in leafy profusion, and hat is large shady dense, and tree performance is grand, is the fine tree species of urban afforestation, is widely used as shade tree, street tree, shelter forest and amenity forest.Again because it contains special fragrance and volatile oil, and moisture-proof, anti-corruption, worm of dispelling, long shelf-life, be valuable furniture, senior building, shipbuilding and engraving etc. desirable with material, in addition each part of the whole body of fragrant camphor tree can extract important industrial chemicals: camphor, camphor oil, and this is the important outlet goods and materials of of China; Therefore, fragrant camphor tree and nanmu, Chinese catalpa, paulownia are collectively referred to as the south of the River four your name wood.In recent years, the application of fragrant camphor tree in afforestation gets more and more, but still have some aspects to need to be improved: growing up fragrant camphor tree 1 year hip number for one can more than 100,000 (LandProtection, 2001), these fruits drop on pavement, or by birdseed with after pollute road surface or pedestrian's clothing everywhere with ight soil, cause much inconvenience.Therefore, high ability limits the application of fragrant camphor tree in gardens.In some country; camphor tree is even limited plantation because of its huge hip number and extremely strong fertility; as Australia once promulgated that land protection method restriction people planted camphor tree (LandProtection near farmland, in suburb or city; 2001); in Florida; camphor tree is listed in I level invasive species and is prohibited to plant (FLEPPC, 2001).In order to make fragrant camphor tree be more widely used, give full play to its ornamental value and economic worth, be mankind's services better, the fragrant camphor tree new germ plasm that cultivation is had no result or hip number reduces just has been put on agenda.
The development of plant cell engineering and plant genetic engineering theory and technology, cultivates for transgenic technology and provides theoretical foundation and realization possibility without fruital camphor tree.The fragrant camphor tree of bio-technology improvement must be established as Research foundation with efficient plant regeneration system, but at present owing to adopting fragrant camphor tree embryo callus to be transgenosis explant material, embryo callus is lower for this species regeneration frequency of fragrant camphor tree more; In addition, after transforming foreign gene, need to add certain density antibiosis in selection formula and usually screen converting material, and antibiotic interpolation turn increases its regeneration plant difficulty of induction, finally causes fragrant camphor tree difficulty to obtain transfer-gen plant; In addition, the plant obtained by somatic embryogenesis has the brephic the same with seedling, and (namely woody plant only lives through the vegetative growth phase of a period of time, just can enter the stage of blossoming and bearing fruit), be thus also not suitable as the research material carrying out Flowering mechanism or transgenosis and suppress to bloom; All factors govern the process of the fragrant camphor tree of bio-technology improvement above.Set up a set of stable, can the test tube flowering fragrant camphor tree group training system of artificial adjustment be suppress fragrant camphor tree to bloom by transgenic approach thus reduce the Research foundation of hip number; And the selection of the potential test tube flowering material of Adult form and cultured in vitro set up the important substance basis of this group training system.
Therefore how providing a kind of to have test tube flowering potential and to grow up the plant regeneration method of material selection method and this material, is technical problem urgently to be resolved hurrily to one skilled in the art.
Summary of the invention
The object of this invention is to provide a kind of fragrant camphor tree plant regeneration method being explant with Adult form material decision method and selection Adult form material.
The technical scheme that the present invention takes is:
The selection of fragrant camphor tree Adult form of the present invention potential test tube flowering material and the concrete steps of plant regeneration method as follows:
(1) the potential test tube flowering material of fragrant camphor tree Adult form judgement:
Grow up fragrant camphor tree 8-10 in age, the diameter of a cross-section of a tree trunk 1.3 meters above the ground is about 13.5cm, land occupation condition is good, the healthy and strong raw then tool squama leaf stem section without obvious outer damage of tree body, its bulbil is in next year by the end of March at the beginning of-4 months, sprout after germinateing, tool bud sprig accounts for investigation bulbil ratio and reaches more than 95%, is defined as the potential test tube flowering material of fragrant camphor tree Adult form;
(2) selection of fragrant camphor tree explant:
In by the end of December between January next year, the raw then tool bulbil branch choosing the fragrant camphor tree of adult healthy from the potential test tube flowering material of fragrant camphor tree Adult form that step (1) is determined is originated as explant;
(3) process of fragrant camphor tree explant:
From the explant source that step (2) is selected, gather the fragrant camphor tree current-year branch of diameter at 2-3mm, branch often saves and all has bulbil; Remove bulbil, blade and petiole, running water down-flow water rinses 30min; The branch retained only there being leaf scar, with operating scissors according to the fragrant camphor tree branch of the mode process of a leaf scar on an explant, obtains the fragrant camphor tree Agistemus exsertus of length at 1.5-2cm;
(4) fragrant camphor tree plant regeneration:
Fragrant camphor tree Agistemus exsertus disinfection after step (3) being processed, is then inoculated in germination medium and induces it to sprout, illumination cultivation;
Induce in the future the many Adult form material cutting and separating of clump branch number to proceed to germination medium and carry out subculture, expanding propagation sets up the stem section culture seedling clone of high proliferation coefficient, as stable Adult form cutting shoots source of supply, i.e. Adult form stem section culture seedling and propagating system;
With Adult form stem section culture seedling cutting shoots for explant, proceed to inducing culture and induce its adventitious bud formation, set up stem section culture seedling cutting shoots regenerating system, illumination cultivation; Cutting shoots regenerated adventitious bud proceeds to MS medium and carries out growth in strong sprout.
In step (4), the formula of germination medium is: MS+BA2.0mg/L+IBA0.5mg/L+30g/L sucrose+8g/L agar powder, the pH=6.0 of germination medium.The formula of inducing culture is: MS+BA3.0mg/L+IBA0.5mg/L+30g/L sucrose+8g/L agar powder, the pH=6.0 of inducing culture.
Good effect of the present invention is as follows:
The selection of the potential test tube flowering material of fragrant camphor tree Adult form of the present invention and plant regeneration method are drawn materials conveniently, and material source, quantity are sufficient, easily obtain sterilizable material; The decision method of Adult form material, method is simple, and conclusion is reliable, for selecting the potential test tube flowering material of fragrant camphor tree Adult form, set up Adult form material plant regeneration system, so for set up a set of stable, the test tube flowering fragrant camphor tree group training system of artificial adjustment can establish Research foundation.Screening obtains the fragrant camphor tree clone of Adult form and sets up fragrant camphor tree Adult form material plant regeneration system, improves fragrant camphor tree fertility and provides effective acceptor material and plant regeneration system, effectively can shorten the fragrant camphor tree genetic transformation cycle for Agrobacterium tumefaciens-mediated Transformation; Because explant material is derived from the Adult form material of adult plants, the plant regeneration system set up with this at the tissue cultures of fragrant camphor tree, genetic transformation and bio-technology improvement, especially can be bloomed to fragrant camphor tree and suppresses to reduce solid improvement breeding, can play a significant role.
Embodiment
The following examples describe in further detail of the present invention.
In the following example, method therefor is conventional method if no special instructions.Described percentage concentration is mass percentage concentration if no special instructions.
Embodiment 1
1.1 material
By the end of December at the beginning of 1 month next year, gather land occupation condition superior, the age of tree is at 8-10, healthy, without damage by disease and insect, the adult fragrant camphor tree current-year branch setting body grace, selected branch all has bulbil; Remove unnecessary blade and bulbil, collection only retains the branch of leaf scar as research material.
Land occupation condition is superior: fragrant camphor tree plantation ground, be the growing area away from building, soil is mainly arable soil.
Land occupation condition is severe: fragrant camphor tree plantation ground, and be building periphery 8-10m, soil mostly is building waste and covers shallow soil.
1.2 method
1.2.1 the process of fragrant camphor tree Adult form material
Processing mode is as follows: gather diameter fragrant camphor tree current-year branch between 2-3mm, branch often saves and all has bulbil; Remove bulbil, blade and petiole, running water down-flow water rinses 30min; The branch retained only there being leaf scar, with operating scissors according to the fragrant camphor tree branch of the mode process of a leaf scar on an explant, obtains the fragrant camphor tree Agistemus exsertus of length at about 1.5-2cm.
Adult form material: the material with potential of blooming; This patent refers in particular to from adult (age of tree 8-10) fragrant camphor tree plant, the stem section of tool bulbil.
1.2.2 the selection of the potential test tube flowering material of Adult form and judgement
At the beginning of next year by the end of March to 4 month, when fragrant camphor tree bulbil is sprouted (fragrant camphor tree shoot in spring all pumping in bulbil), start the investigation carrying out adult fragrant camphor tree bud bulbil correlation.Investigation method is as follows: select the plantation time identical (investigate fragrant camphor tree and all plant zones of different in same residential quarters, community has built up 6 years), the age of tree the adult fragrant camphor tree of 8-10 as respondent; Fragrant camphor tree land occupation condition is divided into superior and severe by the soil quality according to survey area.3 fragrant camphor trees are chosen arbitrarily in the region that land occupation condition is different, often set investigation 100 bulbils, observe tool bud sprig and account for bulbil percentage, investigation repetition 3 times, to determine the correlation of bloom potential and Adult form material and the age of tree of tool bulbil branch, measure and calculate the diameter of a cross-section of a tree trunk 1.3 meters above the ground of investigated fragrant camphor tree; Fragrant camphor tree plant is divided into normally (explant sample tree) according to physiological situation difference and sets body impaired (summer then, this tree 1/4th tree crown rolled over by storm wind windscreen wiper), often tree investigation 100 bulbils arbitrarily, observe tool bud sprig and account for bulbil percentage, investigation repetition 3 times, to determine that tool is bloomed the correlation of potential Adult form material and tree body physiological situation, measure simultaneously and calculate the diameter of a cross-section of a tree trunk 1.3 meters above the ground of investigated fragrant camphor tree.
The diameter of a cross-section of a tree trunk 1.3 meters above the ground: the diameter referring to the liftoff surperficial 1.2m place of fragrant camphor tree trunk.
1.2.3 the sterilization of explant and inoculation
Superclean bench carries out Aseptic sterilisation to explant, and sterilization method is as follows: 70% Ethanol Treatment 30s; Adopt mercuric chloride process 8min afterwards; Finally use sterile water wash 3 times, obtain sterilizable material, explant is exposed to the two ends of disinfectant by aseptic filter paper, cuts 1-2mm respectively, afterwards explant is seeded on ready medium according to morphology upper and lower side.
1.2.4 cultural method
Fragrant camphor tree Adult form Agistemus exsertus sprouts induction process, the expanding propagation process of stem section culture seedling and the process of cutting shoots adventitious bud inducing, is illumination cultivation, and periodicity of illumination is that 16h illumination/8h is dark, and intensity of illumination is 2000-3000Lx; Wherein expanding propagation is treated to quickening reproductive schedules, the frequency subculture of the medium that more can renew according to every 15d.
1.2.5 different germination medium sprouts the impact of induction on fragrant camphor tree Adult form Agistemus exsertus
Fragrant camphor tree Adult form Agistemus exsertus being placed in different inducing culture of sprouting induces it to sprout, tests different basic element of cell division kind and the impact of inducing is sprouted in the combination of variable concentrations plant hormone on fragrant camphor tree Adult form Agistemus exsertus.Select the individual plant of growth coefficient high (forming the individual plant stem section culture seedling that clump branch number is greater than 3 ~ 5), transfer and in the medium sprouting inducing effect the best, carry out expansions propagation process, form the single-strain clone of Adult form stem section culture seedling, this stem section culture seedling and propagating system can be fragrant camphor tree Adult form genetic transformation and provides transformation receptor.Each process inoculation explant number is no less than 30, adds up Agistemus exsertus germination rate after 4w.
1.2.6 the impact of different plant hormone combinations on fragrant camphor tree stem section culture seedling cutting shoots adventitious bud inducing
The cutting shoots of fragrant camphor tree stem section culture seedling is placed in different adventitious bud induction culture base (S1, S2, S11, S22, MBDC, MBIC), study the impact of different plant hormone combination on fragrant camphor tree stem section culture seedling cutting shoots adventitious bud inducing, in order to set up fragrant camphor tree Adult form cutting shoots plant regeneration system; Wherein cutting shoots obtains in accordance with the following methods: get Adult form stem section culture seedling scalpel and excise terminal bud on branch, lateral bud and blade completely; By the branch of remainder, by joint segment (namely each cutting shoots having 1-2 joint), be about 5-7mm, access in various inducing culture as explant and carry out adventitious bud induction culture; Each process inoculation explant number is no less than 75, adds up adventitious bud induction frequency and coefficient of differentiation after 4w.
Above medium all adopts MS as minimal medium without specified otherwise, and add agar powder 8g/L, the plant hormone of sucrose 30g/L and variable concentrations, after medium pH is all adjusted to 6.0, keeps 121 DEG C of sterilizing 20min.
1.2.7 data calculate and process
Sprout stem hop count/inoculation Adult form Agistemus exsertus number × 100% that inductivity=sprouting is sprouted
Explant number/inoculation explant sum × 100% (indefinite bud length is greater than 3 ~ 5mm, records an effective bud) that adventitious bud induction frequency=induction is sprouted
Coefficient of differentiation=induction is sprouted explant sum × 100% that sum/induction sprouts
2 results and analysis
2.1 fragrant camphor tree tool squama leaf stem sections of growing up are bloomed the investigation of potentiality
At the beginning of next year by the end of March to 4 month, when fragrant camphor tree bulbil is sprouted, carry out the investigation of adult fragrant camphor tree bud bulbil correlation.Known according to table 1, land occupation condition is different, soil nutrient status is different, tool bud sprig accounts for bulbil percentage slightly difference, the fragrant camphor tree of investigation can be observed bulbil and sprout the situation that pumping goes out pure vegetative bud sprig, on the fragrant camphor tree that land occupation condition is severe, the sprig of pure vegetative bud accounts for bulbil and compares a little more than the superior fragrant camphor tree of land occupation condition.The fragrant camphor tree of fragrant camphor tree simultaneously under this land occupation condition under bud quantity and bulbil sprout time are all weaker than supremacy clause.The fragrant camphor tree of the visible identical age of tree (maturity), because nutritional condition is different, the tree body diameter of a cross-section of a tree trunk 1.3 meters above the ground there are differences, and the fragrant camphor tree mean DBH increment that land occupation condition is excellent is 13.7cm; The fragrant camphor tree mean DBH increment of inclement condition only has 9.7cm; It can thus be appreciated that Adult form material is subject to the restriction of this nutrition status of the plant in the potential ability of blooming of performance, and nutritional condition is sufficient, then flower amount is large, and the florescence early; Oligotrophy, then flower amount is bloomed late less.
But in all fragrant camphor tree of investigation, the rear pumping of bulbil sprouting goes out bud branch and accounts for investigated bulbil percentage all more than 95%; Whether the potential ability of blooming of visible fragrant camphor tree stem section material is grown up with fragrant camphor tree and be closely related age, and fragrant camphor tree body maturation is that its branch tool is bloomed the guarantee of potentiality.Therefore can think, grow up fragrant camphor tree in well-fed situation, namely its tool squama leaf stem section has potentiality of blooming, and selects it to carry out cultured in vitro as explant, has the possibility of observing its test tube flowering; If floral induction condition is suitable for, by the cultured in vitro of this Adult form material is set up stable, can the test tube flowering fragrant camphor tree group training system of artificial adjustment.The phenomenon of test tube flowering has been observed in the cultured in vitro of fragrant camphor tree Adult form material, illustrate that induction Adult form material test tube flowering is feasible, but because (<5%) occurs this phenomenon low frequency, the test tube flowering system as stablized artificial adjustment then also needs to carry out system research.
Adult fragrant camphor tree (8 ~ 10 years) bud bulbil ratio under table 1 different condition area
Note: land occupation condition is superior: fragrant camphor tree plantation ground, be the growing area away from building, soil is mainly arable soil; Land occupation condition is severe: fragrant camphor tree plantation ground, and be building periphery 8 ~ 10m, soil mostly is building waste and covers shallow soil.
The adult fragrant camphor tree that physiological situation is different, bud bulbil ratio presents larger difference.The fragrant camphor tree of sample tree under normal physiological conditions, the diameter of a cross-section of a tree trunk 1.3 meters above the ground is 13.5cm, and tool bud sprig accounts for bulbil ratio and reaches 98%; One strain is planted in the severe fragrant camphor tree of land occupation condition, and the diameter of a cross-section of a tree trunk 1.3 meters above the ground is 7.5cm, and owing to being subject to storm damage then, 1/4 tree crown is scraped folding, and the bud bulbil ratio of base portion branch only has 60% (table 2).In investigation, chancing on the diameter of a cross-section of a tree trunk 1.3 meters above the ground of a strain through transplanting is the fragrant camphor tree of 14.5cm, and it was 75% (data do not show) that the tool bud sprig of the larger branch of base portion one accounts for bulbil ratio, and all the other branch bud bulbils of this tree are than close with the fragrant camphor tree of normal condition; The appearance of the special branch that vegetative bud increases, may cause damage to cause owing to transplanting plant.As can be seen here, though grow up the tool squama leaf stem section of fragrant camphor tree tool to bloom potential, if plant corpus is subject to external injury (artificial transplanting or strong wind blow folding), its stem section is shown can be subject to certain restrictions in potential ability of blooming, namely the bulbil in stem section forms bud and reduces, and vegetative bud is formed and increases.Therefore, in the research of setting up fragrant camphor tree test tube flowering group training system, reduce as far as possible the external injury of Adult form material to guarantee that it can artificial adjustment test tube flowering as early as possible.
Adult fragrant camphor tree (8 ~ 10 years) the bud bulbil ratio of the different physiological status of table 2
2.2 different germination mediums sprout the impact of induction to fragrant camphor tree Adult form Agistemus exsertus
Fragrant camphor tree Adult form Agistemus exsertus being placed in different inducing culture of sprouting induces it to sprout, and after inoculation 3-5d, can observe pumping stem section having axillalry bud or indefinite bud, after 3w-4w, can sprout and form sprig.As shown in Table 3, the impact that the medium that variable concentrations BA with 0.5mg/LIBA combines sprouts induction to stem section is different, wherein fill a prescription A2:MS+BA2.0+IBA0.5, stem section germination rate is the highest, reach 90%, stem section can be induced simultaneously to sprout pumping and go out multiple sprig, formation clump branch number can be observed reach 3-5 branch in a stem section, afterwards along with BA concentration increases, stem section germination rate declines.Formula A2 can sprout the shoot proliferation formula of inducing culture and stem section culture seedling as fragrant camphor tree Adult form Agistemus exsertus.
From the impact that table 4, table 5 more different basic element of cell division kind are sprouted fragrant camphor tree stem section.Can find, along with the increase of KT concentration, stem section germination rate increases thereupon, and when KT is 0.3mg/L, sprouting inductivity the highest, is 69.2%; After this, the increase of KT concentration, sprouts inductivity and does not increase counter falling (table 4).On the induction formula containing TDZ, the highest sprouting inductivity only has 60% (table 5).As can be seen here, KT and TDZ is when carrying out stem section and sprouting induction, and successful is not as BA.Thus, in experimental design after this, the basic element of cell division type of fragrant camphor tree stem section culture seedling cutting shoots is induced to select BA.
The material cutting and separating many from single stem section formation clump branch number is proceeded to formula A2 and carries out subculture, expanding propagation sets up the stem section culture seedling clone of high proliferation coefficient, can be used as stable Adult form cutting shoots source of supply, i.e. Adult form stem section culture seedling and propagating system.
Table 3 different B A concentration is on the impact of fragrant camphor tree Agistemus exsertus adventitious bud inducing
Table 4 different K T concentration is on the impact of fragrant camphor tree Agistemus exsertus adventitious bud inducing
Table 5 different TDZ concentration is on the impact of fragrant camphor tree Agistemus exsertus adventitious bud inducing
2.3 impacts of different plant hormone combinations on fragrant camphor tree stem section culture seedling cutting shoots adventitious bud inducing
The cutting shoots of fragrant camphor tree stem section culture seedling is placed in different adventitious bud induction culture base (table 6), study the impact of different plant hormone combination on fragrant camphor tree stem section culture seedling cutting shoots adventitious bud inducing, in order to set up fragrant camphor tree Adult form stem section culture seedling cutting shoots plant regeneration system; After cutting shoots inoculation, not only at cutting shoots axillalry bud place, adventitious bud formation can be observed in other position of explant, and therefore this plant regeneration system can guarantee the transformant also renewable plant of non-meristem zone, and the unlikely positive transformed cells that makes is because of can not regeneration plant and leak choosing.
The impact of table 6 different plant hormone combination on fragrant camphor tree stem section culture seedling cutting shoots adventitious bud inducing
In different adventitious bud inducings formula, can be observed adventitious bud induction frequency and coefficient of differentiation is had nothing in common with each other.In formula S22:MS+BA3.0+IBA0.5, adventitious bud induction frequency is 53.3, and coefficient of differentiation is 1.75; And S11 adventitious bud induction frequency of filling a prescription is 38.6%, coefficient of differentiation is 1.9.Visible, improve inductivity and coefficient of differentiation that cytokinin concentration is conducive to improving cutting shoots indefinite bud.Formula S22 can fill a prescription as cutting shoots adventitious bud inducing, and separated from explant by evoking adventive bud, proceed to continued growth in MS medium, indefinite bud can grow up to seedlings.
Therefore, on the basis of the numerous Establishing of expansion of Adult form stem section culture seedling, carry out the induction of cutting shoots indefinite bud, progressively set up cutting shoots plant regeneration system, the genetic transformation that can be Adult form material provides Research foundation and transformation receptor.
Although illustrate and describe embodiments of the invention, for the ordinary skill in the art, be appreciated that and can carry out multiple change, amendment, replacement and modification to these embodiments without departing from the principles and spirit of the present invention, scope of the present invention is by claims and equivalents thereof.
Claims (1)
1. the selection of the potential test tube flowering material of fragrant camphor tree Adult form and a plant regeneration method, is characterized in that: the concrete steps of described method are as follows:
(1) judgement of the potential test tube flowering material of fragrant camphor tree Adult form:
Grow up fragrant camphor tree 8-10 in age, the diameter of a cross-section of a tree trunk 1.3 meters above the ground is about 13.5cm, land occupation condition is good, the healthy and strong raw then tool squama leaf stem section without obvious outer damage of tree body, its bulbil is in next year by the end of March at the beginning of-4 months, sprout after germinateing, tool bud sprig accounts for investigation bulbil ratio and reaches more than 95%, is defined as the potential test tube flowering material of fragrant camphor tree Adult form;
(2) selection of fragrant camphor tree explant:
In by the end of December between January next year, the raw then tool bulbil branch choosing the fragrant camphor tree of adult healthy from the potential test tube flowering material of fragrant camphor tree Adult form that step (1) is determined is originated as explant;
(3) process of fragrant camphor tree explant:
From the explant source that step (2) is selected, gather the fragrant camphor tree current-year branch of diameter at 2-3mm, branch often saves and all has bulbil; Remove bulbil, blade and petiole, running water down-flow water rinses 30min; The branch retained only there being leaf scar, with operating scissors according to the fragrant camphor tree branch of the mode process of a leaf scar on an explant, obtains the fragrant camphor tree Agistemus exsertus of length at 1.5-2cm;
(4) fragrant camphor tree plant regeneration:
Fragrant camphor tree Agistemus exsertus disinfection after step (3) being processed, is then inoculated in germination medium and induces it to sprout, illumination cultivation;
Proceed to germination medium carry out subculture by inducing the many Adult form material cutting and separating of clump branch number, expanding propagation sets up the stem section culture seedling clone of high proliferation coefficient, as stable Adult form cutting shoots source of supply, i.e. Adult form stem section culture seedling and propagating system; The formula of germination medium is: MS+BA2.0mg/L+IBA0.5mg/L+30g/L sucrose+8g/L agar powder, the pH=6.0 of germination medium;
With Adult form stem section culture seedling cutting shoots for explant, proceed to inducing culture and induce its adventitious bud formation, set up stem section culture seedling cutting shoots regenerating system, illumination cultivation; Cutting shoots regenerated adventitious bud proceeds to MS medium and carries out growth in strong sprout, and the formula of inducing culture is: MS+BA3.0mg/L+IBA0.5mg/L+30g/L sucrose+8g/L agar powder, the pH=6.0 of inducing culture.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102144546A (en) * | 2011-01-12 | 2011-08-10 | 福建农林大学 | Method for subculturing camphorwood tissue culture seedlings |
| CN102860258A (en) * | 2012-09-19 | 2013-01-09 | 广东省林业科学研究院 | Clonal tissue culture breeding method for camphor tree |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102144546A (en) * | 2011-01-12 | 2011-08-10 | 福建农林大学 | Method for subculturing camphorwood tissue culture seedlings |
| CN102860258A (en) * | 2012-09-19 | 2013-01-09 | 广东省林业科学研究院 | Clonal tissue culture breeding method for camphor tree |
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