CN102138529A - Quick rooting method for 'Heyun' blueberry seedlings formed by tissue culture - Google Patents
Quick rooting method for 'Heyun' blueberry seedlings formed by tissue culture Download PDFInfo
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Abstract
The invention discloses a quick rooting method for 'Heyun' blueberry seedlings formed by tissue culture, which comprises the following steps: dipping the 'Heyun' blueberry seedlings formed by tissue culture into root stimulate, planting and rooting to obtain rooted seedlings. Compared with the conventional seedling culture method, the new seedling survival rate of the 'Heyun No.1 and No.2' blueberry seedlings is improved to 90 percent from 50 percent, the seedling raising period is reduced to 6 months from 12 months, and the quality of the new seedlings is improved obviously.
Description
Technical field
The present invention relates to the method that blueberry seedling breeding is taken root fast and bred, belong to the vegetative propagation technique field of plant.
Background technology
Blueberry formal name used at school cowberry, belong to Ericaceae (Ericaceae) Vaccinium (Vaccinium spp.) plant, the sweet acid of fruit is fragrant refreshing, unique flavor, have abundant nutritive value and multiple health care, being described as " gold berry ", is one of the mankind's five big healthy food of FAO (Food and Agriculture Organization of the United Nation) (FAO) keypoint recommendation.Blueberry not only is used to eat raw, and develops multiple product, makes its nutritive value bring bigger effectiveness to the consumer.
The artificial cultivation blueberry starts from the U.S. the earliest, so far T﹠B.Last century, the eighties rose, and China has introduced a large amount of cultivars from external, and had begun relevant popularization.But because of developing rapidly, the blueberry nurseries put upon the full stretch becomes the technical bottleneck of Jilin Province and even whole nation development blueberry industry.Blueberry has acid resistance soil environment, low temperature resistant, barren-resistant, stronger drought-resistant ability and diseases and insect pests resistance; Biological properties such as not anti-chemical fertilizer (dislike calcium, dislike chlorine, dislike sodium), weed killer herbicide possess the exclusive advantage of first developing.Simultaneously, along with the raising of the life and the level of consumption, people are more and more higher to the safety and the quality requirements of food, and the market prospects of blueberry nursery stock are more wide.No. 1, standing grain rhythm " (Lan Feng) " blueberry seedling of Jilin lucky Kanggong department authentication belong to Gao Cong, large fruit, in late-maturing cowberry kind; No. 2, standing grain rhythm " (Elliot) " blueberry seedling belongs to half Gao Cong, middle fruit type, extremely late-maturing cowberry kind, and the two resistance is all very strong, is adapted at the suitable zone of northern China and carries out commerial growing and promote.
Current, utilize method for plant tissue culture can breed various plants in a short time fast, reproduction rate height not only, and, can keep the merit of former stock because it is vegetative propagation, on producing, use more and more wider in recent years.Simultaneously, cuttage also earns widespread respect because of economical, convenient as the important means of seedling-wood breeding.Green wood cutting is because the existence cutting is drawn materials and the result should not promote in commodity fruit plantation with a contradiction of vying each other.
Summary of the invention
The object of the present invention is to provide the method for a kind of blueberry group training quick reproduction technique, for the blueberry seedling provide a kind of simple to operate, save time, the fast breeding method of with low cost, stability and high efficiency.
The method of blueberry group provided by the invention training quick reproduction technique is characterized in that, comprises the steps: that tissue cultivating seedling with blueberry dips in to carry out cuttage root-taking behind the root-growing agent and cultivate, and obtains the seedling of taking root.
Above-mentioned root-growing agent be following a) or b):
A) solution of methyl and heteroauxin composition: the compound method of the described root-growing agent of 1L is as follows: 1000mg methyl and 1000mg heteroauxin is soluble in water, be settled to 1L;
B) solution of root-inducing powder ABT preparation: the compound method of the described root-growing agent of 1L is as follows: after root-inducing powder ABT 0.05g added the dissolving of 5ml alcohol, it was fixed molten to 1L to add water again.This alcohol can be the ethanol water of 70% (percent by volume).
The time that above-mentioned tissue cultivating seedling dips in root-growing agent can be 3-10 second, preferably 5 seconds.
The spacing in the rows of tissue cultivating seedling was 2-5cm, preferably 3cm when above-mentioned cuttage root-taking was cultivated; Line-spacing is 3-8cm, is preferably 5cm; The cuttage degree of depth is 1-1.5cm.
Above-mentioned cuttage root-taking culture condition is: temperature 25-35 ℃, relative moisture is 50-60%, and the light transmittance that shelters from heat or light is 45%-65%.
Further, said method also comprises the preliminary hardening of described tissue cultivating seedling before dipping in root-growing agent;
The time of described preliminary hardening is 7-8 days; Temperature is 25-35 ℃; Relative moisture is 50%-60%; Intensity of illumination is 2500-3500LX, preferably 3000LX.
A nearlyer step says that said method also comprises described seedling greenhouse hardening and the land for growing field crops hardening afterwards that obtain taking root; Described greenhouse hardening and land for growing field crops hardening comprise the steps: the described seedling of taking root green house hardening 40-60 days, and between 25～35 ℃ of the condition degree of being of green house hardening, relative moisture is advisable with 50%-60%, and the light transmittance that shelters from heat or light is 45%～65%; The time of carrying out the land for growing field crops hardening then is 150-180 days.
The acquisition step of the tissue cultivating seedling of above-mentioned blueberry is as follows: the explant of blueberry is placed on the initial culture base induce differentiation culture, obtain just for tissue cultivating seedling; Described explant is preferably with the stem section of axillalry bud;
Described initial culture base is to add the solid culture medium that following material obtains in basic culture solution: zeatin, NAA, carbon source and gel; The final concentration of zeatin is 2-5mg/L in the described initial culture base, and the final concentration of NAA is 0.01-0.12mg/L; The solvent of described basic culture solution is that water, solute are as shown in table 1; Intensity of illumination is controlled between 4000～4500LX; Illumination cultivation time every day is 15h, and the dark culturing time is 9h; Temperature is controlled at 20-24 ℃ during illumination cultivation, and temperature is controlled at 16-20 ℃ during dark culturing; Humidity is controlled between the 60%-75%.
The final concentration of zeatin and NAA is following 1 in the above-mentioned initial culture base)-3) in arbitrary described: 1) final concentration of zeatin is 3-4mg/L, and the final concentration of NAA is 0.05-0.1mg/L; 2) final concentration of zeatin is 3mg/L or 4mg/L, and the final concentration of NAA is 0.05-0.1mg/L; 3) final concentration of zeatin is 3-4mg/L, and the final concentration of NAA is 0.05mg/L or 0.1mg/L.
The acquisition step of the tissue cultivating seedling of above-mentioned blueberry also can comprise successive transfer culture, and described successive transfer culture comprises the steps: just to place subculture medium to carry out successive transfer culture for tissue cultivating seedling with above-mentioned, obtains the tissue cultivating seedling of described blueberry;
Described subculture medium is to add the solid culture medium that following material obtains in basic culture solution: zeatin, NAA, IBA, carbon source and gel; The final concentration of zeatin is 1.5-4mg/L in the described subculture medium, and the final concentration of NAA is 0.04-0.1mg/L, and the final concentration of IBA is 0.1-0.5mg/L; The solvent of described basic culture solution is that water, solute are as shown in table 1.The successive transfer culture condition: intensity of illumination is about 2500-3500LX; Illumination cultivation time every day is 15h, and the dark culturing time is 9h; Temperature is controlled at 20-24 ℃ during the subculture illumination cultivation, and temperature is controlled at 18-22 ℃ during dark culturing; Humidity is controlled between the 60%-75%.
The final concentration of zeatin, NAA and IBA is as 1 in the described subculture medium)-3) in arbitrary as described in: 1) final concentration of zeatin is 2-3mg/L, and the final concentration of NAA is 0.06-0.08mg/L, and the final concentration of IBA is 0.2-0.3mg/L; 2) final concentration of zeatin is 2mg/L, and the final concentration of NAA is 0.08mg/L, and the final concentration of IBA is 0.2mg/L; 3) final concentration of zeatin is 3mg/L, and the final concentration of NAA is 0.06mg/L, and the final concentration of IBA is 0.3mg/L.
Gel in above-mentioned initial culture base and the described subculture medium all is agar, carragheen or Gelrite, preferably all is agar; Described carbon source all is glucose, maltose or sucrose, preferably is sucrose; Described agar all is 1300mg/cm in the intensity of described initial culture base and described subculture medium
2The final concentration of described agar in described initial culture base and described subculture medium all is 4-5g/L, is preferably 4.5g/L; The final concentration of described sucrose in described initial culture base and described subculture medium all is 25-35g/L, preferably is 30g/L.
The present invention has set up a kind of blueberry seedling " hardening is integrated in group training---cuttage---" method of taking root fast and breed, comprise method steps such as explantation tissue's cultivation, tissue cultivating seedling greenhouse cuttage hardening, new transplantation of seedlings land for growing field crops, greenhouse hardening, cultivating seedling-growing method with the routine group compares, the new shoot survival percent of blueberry seedling " standing grain rhythm No. 1 and No. 2 " is risen to more than 90% by 50%, growing-seedling period shortened to 6 months by 12 months, and simultaneously the new talent quality has also had and significantly improves.Be convenient to the large-scale industrialized blueberry seedling of breeding.The advantage of the technical program is: " integrated " the method cultivation of not taking root in group training process of group training---cuttage---hardening, directly tissue cultivating seedling being dipped in the root-growing agent cuttage buries and carries out hardening, on average like this save 20～30 day time, shortened growing-seedling period, in addition, the tissue cultivating seedling rooting rate can reach more than 85%, and the new shoot survival percent of taking root can reach more than 90%, and the new talent quality has also had and significantly improves.This method has fast, and economy is succinct, and advantage has efficiently increased substantially the reproduction speed and the quality of blueberry seedling, provides safeguard thereby breed the blueberry seedling for large-scale commercial.The tissue cultivating seedling cuttage that the present invention proposes not only can be saved seedling raise period, and avoided the deficiency of green wood cutting, has more outstanding advantage, is convenient to the large-scale industrialized blueberry seedling of breeding.
Description of drawings
Fig. 1 is the tissue culture bottle hardening photo of uncapping.
Fig. 2 lays photo for the seedbed, greenhouse.
Fig. 3 is a greenhouse cuttage tissue cultivating seedling photo.
Fig. 4 is a green house hardening initial stage photo.
Fig. 5 is a green house hardening later stage photo.
Fig. 6 is a land for growing field crops hardening photo.
Embodiment
The invention will be further described below in conjunction with specific embodiment, but the present invention is not limited to following examples.
Among the following embodiment, if no special instructions, be conventional method.
Embodiment 1,
One, the training of the group of standing grain rhythm blueberry seedling is bred
At " standing grain rhythm No. 1 and No. 2 " blueberry seedling, adopt method for tissue culture to carry out fast breeding, this process is only carried out breeding of explant, has saved rooting process.The blueberry tissue culture is taked the shoot proliferation mode.Pickup kind operation just adopts test tube to cultivate, the stem section of 1 band of test tube inoculation axillalry bud.The subinoculation operation adopts tissue culture bottle to cultivate, 15 of each tissue culture bottle inoculations.Concrete group training medium mixture and method of operating are referring to patent: standing grain rhythm blueberry seedling group training medium and tissue culture method thereof.Concrete steps are as follows:
(1), " standing grain rhythm No. 1 and No. 2 " first preparation and statistical observation for tissue cultivating seedling
1, the acquisition of explant and preliminary treatment thereof
1) explant selection
The blueberry tissue culture is taked the mode of shoot proliferation in the present embodiment.Annual 6 to September, in " No. 1, standing grain rhythm ", " No. 2, standing grain rhythm " blueberry (available from the lucky Kanggong in Jilin department) seed resource garden, selects no damage by disease and insect, and tree-like good, the setting proterties is good, and the blueberry nursery stock of getting bumper crops is got explant.Explant is chosen the spray of New Development then, and 6-10 bud arranged on the branch.
2) preliminary treatment of explant
The explant of adopting 2 kinds that shear off in the above-mentioned steps 1 is put in the beaker that fills with clear water, splash into 4-6 and drip abluent, softly scrub axillalry bud, the stem bar of branch with banister brush, blade with explant gets rid of and puts into another beaker down then, use flushing with clean water 15min, the explant rinsed well and other inoculation outfits are put into the superclean bench of prior ultraviolet-sterilization, with tweezers explant is sandwiched and carry out sterilization treatment in the sterilized wide-mouth bottle.
The concrete steps of sterilization treatment are for pouring the alcohol of 75% (percent by volume) rapidly in the wide-mouth bottle of the above-mentioned explant of packing into, rolling was washed 30 seconds, wash 4 times with the sterile water rolling, every all over 2min, the explant that sterilization is intact sandwiches in the lunch box with aseptic nipper again, carry out shearing manipulation, explant bottom blackout stem section is wiped out, and by one section of a bud stem section of axillalry bud (promptly with), and bud is longer apart from lower cut, lack (deciding) apart from upper cut on the explant blade gap distance, but a terminal bud 2-3 bud.Promptly obtain the good explant of sterilization treatment.
2, first acquisition and statistical observation for tissue cultivating seedling
1) obtains just for tissue cultivating seedling
By the inoculation operational procedure, the explant that obtains the good explant of sterilization treatment in the step 2 with above-mentioned steps one inserts and is equipped with in the sterile test tube of initial culture base, the stem section of a band of test tube inoculation axillalry bud is sealed membrane closure with bilayer, induces differentiation culture.Induce the condition of culture of differentiation culture to be: intensity of illumination is controlled between 4000～4500LX; Illumination cultivation time every day is 15h, and the dark culturing time is 9h; Temperature is controlled at 20-24 ℃ during illumination cultivation, and temperature is controlled at 16-20 ℃ during dark culturing; Humidity is controlled between the 60%-75%.Simultaneously, can a week open uviol lamp once, each 30min, the calm rainy day opens 30min with culturing room's window and ventilates.The cultivation cycle of initial culture is 60 days-70 days, promptly cultivates just can obtain in 60 days-70 days just for tissue cultivating seedling, and what obtain just can carry out next step amplification switching according to the quantity of required tissue cultivating seedling for tissue cultivating seedling.
Above-mentioned initial culture base is to have added the solid culture medium that zeatin (ZT), NAA, sucrose and agar obtain in basic culture solution.Wherein the final concentration of zeatin in the initial culture base is 3mg/L, and the final concentration of NAA is 0.05mg/L, and the final concentration of sucrose is 30g/L, and it is 1300mg/cm that agar adopts intensity
2Agar, the final concentration of its agar is 4.5g/L; The solvent of basic culture solution is that water, solute are as shown in table 1, and the pH of medium is 5.2.
The solute of table 1. basic culture solution
| Macroelement | Concentration (mgL in the medium -1) |
| NH 4NO 3 | 400 |
| K 2SO 4 | 990 |
| MgSO 4·7H 2O | 370 |
| KH 2PO 4 | 170 |
| Ca(NO 3) 2·4H 2O | 556 |
| CaCl 2 | 92 |
| Molysite | Concentration (mgL in the medium -1) |
| FeSO 4·7H 2O | 27.8 |
| Na 2EDTA | 37.3 |
| Trace element | Concentration (mgL in the medium -1) |
| MnSO 4·4H 2O | 22.4 |
| ZnSO 4·7H 2O | 8.6 |
| H 3BO 3 | 6.2 |
| Na 2MoO 4·2H 2O | 0.25 |
| CuSO 4·5H 2O | 0.25 |
| Organic principle | Concentration (mgL in the medium -1) |
| Thiamine hydrochloride | 1.0 |
| Puridoxine hydrochloride | 0.5 |
| Nicotinic acid | 0.5 |
| Glycine | 2.0 |
| Inositol | 100 |
Be convenient preparation medium, macroelement in the table 1, trace element, molysite, organic matter and required hormone (being zeatin, NAA) can be mixed with the mother liquor of high concentration, as concentrating the mother liquor of 100 times or 50 times, get mother liquor again during actual preparation and prepare by above-mentioned final concentration.
The compound method concrete steps of above-mentioned initial culture base are:
(1) measures mother liquor by the usefulness of the final concentration of each material in the above-mentioned initial culture base, add the water of required solvent,, prepared culture medium is carried out acidity adjustment, make its pH value in the 5.2-5.3 scope by pH meter with 0.1mol/L NaOH or 0.1mol/L HCl.
(2) by above-mentioned initial culture base sucrose the consumption of final concentration take by weighing sucrose and add to boil and begin heating in the pot, to medium when at a simmer, the agar powder that takes by weighing institute's expense adds in the pot, and constantly stir with glass bar rapidly, boil after the 1-2min, stop heating, polishing is to 1000ml when treating that the medium temperature is reduced to 45 ℃ of left and right sides, reheat and stir, to about 50 ℃, stop, when the medium ot-yet-hardened, divide while hot to install in the selected culture vessel, the culture test tube of available 18mm * 180mm, every pipe packing medium 12ml seals, 121 ℃ of sterilization 15min, medium is at room temperature cooled off, promptly obtain the initial culture base for preparing.
2) statistical observation
The stem section of the blueberry band axillalry bud of inoculating in the step 1) of above-mentioned steps 2 was cultivated 55-65 days, can observe and induce the blueberry tender shoots that differentiates, experiment repeats 3 times, carry out statistical observation above-mentioned steps 1 when cultivating 60 days) in axillalry bud induce situation, the inductivity of " No. 1, standing grain rhythm ", " No. 2, standing grain rhythm " bud all can reach more than 90%.
(2), the preparation of successive transfer culture seedling and statistical observation
1, the acquisition of successive transfer culture seedling
Choose pollution-free, what obtain in the good above-mentioned steps that can be used for transferring () of growing way is first for tissue cultivating seedling or subculture seedling (promptly passing through the successive transfer culture seedling of successive transfer culture), press the subinoculation operational procedure, folder adds aqua sterilisa and soaks into filter paper in case dehydration is cut into 1 section of 1 bud with seedling to the sterilization cassette that contains filter paper, be transferred on the shoot proliferation medium, 15 of every tissue culture bottle inoculations move to group training chamber and cultivate after sealing, pollution rate is controlled in 1%.It is about 3000LX that the tissue cultivating seedling that is transferred to the shoot proliferation medium places intensity of illumination; Illumination cultivation time every day is 15h, and the dark culturing time is 9h; Temperature is controlled at 20-24 ℃ during the subculture illumination cultivation, and temperature is controlled at 18-22 ℃ during dark culturing; Humidity is controlled between the 60%-75%.Simultaneously, a week is opened uviol lamp once, each 30min, and the calm rainy day opens 30min with culturing room's window and ventilates.The successive transfer culture cycle is just switchable about 60 days.The number of times of subculture is decided according to the numerous amount of required expansion.
Above-mentioned subculture medium is to have added the solid culture medium that zeatin, NAA, IBA, sucrose and agar obtain in basic culture solution.Wherein the final concentration of zeatin in subculture medium is 2mg/L, and the final concentration of NAA is 0.08mg/L, and the final concentration of IBA is 0.2mg/L, and the final concentration of sucrose is 30g/L, and it is 1300mg/cm that agar adopts intensity
2Agar, the final concentration of its agar is 4.5g/L; The solvent of basic culture solution is that water, solute are as shown in table 1, and the pH of medium is 5.2.The solvent of above-mentioned basic culture solution is that water, solute are as shown in table 1, and the pH of medium is 5.2.
2, statistical observation
The first of inoculation cultivated 55-65 days for tissue cultivating seedling or subculture seedling in the step 1 of above-mentioned steps two, can observe and induce the blueberry tender shoots that differentiates, experiment repeats 3 times, carry out the first situation of inducing for tissue cultivating seedling or subculture seedling in the statistical observation above-mentioned steps 1 when cultivating 60 days, the growth coefficient of " No. 1, standing grain rhythm ", " No. 2, standing grain rhythm " successive transfer culture all can reach 4.
Two, standing grain rhythm blueberry tissue cultivating seedling greenhouse cuttage hardening
Concrete steps are as follows:
(1) after the group training new talent undercarriage of above-mentioned 2 kinds, move to and open tissue culture bottle lid hardening 7 days (Fig. 1) in the green house, keep 25～35 ℃ of temperature, relative moisture 50%～60% is about intensity of illumination 3000LX.
(2) lay the seedbed, bottom is the mashed bark in shop or pine needle 5cm (1.1m wide * 50m is long) earlier, repaves one deck liver moss (being divided into the ridge), and about 10cm is thick in whole seedbed, and 2cm is wide, interval 5cm, height of bed 4cm (Fig. 2);
(3) prepare the cuttage tissue cultivating seedling, press from both sides out, clean subsidiary medium with clear water, the black callus bottom removing with scissors with the blueberry seedling of tweezers after with the hardening in (1);
(4) with cuttage seeding in that a) root-growing agent is (soluble in water with 1000mg methyl and 1000mg heteroauxin, be settled to the solution that 1L obtains) or b) root-growing agent (root-inducing powder ABT 0.05g is dissolved in earlier in the 5ml alcohol, add water again and be settled to the solution that 1L obtains) in dip in 5s, be inserted on the seedbed according to spacing in the rows 3cm * line-spacing 5cm, the degree of depth is inserted 1～1.5cm, about 34 strains of every ridge cuttage tissue cultivating seedling;
(5) with Polypropylence Sheet canopy is built, generally striden whole seedbed, high 0.6m, and add lid layer black sunshade net (Fig. 3).Between 25～35 ℃ of the maintenance temperature of shed, relative moisture is advisable with 50%-60%, and the light transmittance that shelters from heat or light is 45%～65%;
(6) observe in good time and carry out the statistics of table 2, choose and remove dead seedling, weak seedling, the stalwartness of selecting and remain, the seedling that insect pest is few are shown in Fig. 4,5, consistent in the green house hardening 40～60 days (in good time raising sunshade net and Polypropylence Sheet), the temperature of green house hardening, humidity and (5).
Table 2. is used a) and is added up the shoot survival percent of taking root behind the root-growing agent
Use b) statistical data of root-growing agent and a) the statistical data there was no significant difference of root-growing agent.
(7) survive the winter, after the 2nd year beginning of spring, carry out land for growing field crops hardening process with kind transplantation of seedlings land for growing field crops, greenhouse the first tenday period of a month in May.The greenhouse hardening is generally carried out in annual 5～September, and stop October substantially.
Three, standing grain rhythm blueberry new talent field-transplanting hardening (photo as shown in Figure 6)
Concrete steps are as follows:
(1) changes the cave
Change the cave annual autumn, generally dig the soil pit of 35cm * 35cm * 35cm, seedbed 70cm, operation road 80cm, 2/3 of soil pit volume is filled composite soil earlier, and (three skin soil: the peat composed of rotten mosses=2: 1), other 1/3 volume is filled fertilizer (peat composed of rotten mosses: chicken manure or pig manure 2: 1), and mixing forms the cave and improves the soil, adjust soil pH value 4.5～5.5, the content of organic matter 8～10%.Adopting black Polypropylence Sheet to carry out plastic mulching (membrane interaction: insulation, weeding, water conservation) survives the winter.
(2) plant transplantation of seedlings
The first tenday period of a month in late April, second to May, choose that growing way is good, the greenhouse cuttage seeding of no damage by disease and insect is carried out planting seedlings, the field planting degree of depth is according to different cultivars and different, general 7～8cm, dark 10～the 15cm that reaches, water water (general 7 days water one time water) after the field planting, mid or late May chases after fertilizer one time, and the breast that imposes the soya-bean cake fermentation at the beginning of 7 months then is fertile.In the hardening of land for growing field crops, note water and fertilizer management, reject sick and weak seedling at any time, and the control weed growth.
(3) emerge
About annual October, the blueberry seedling of land for growing field crops hardening can be transplanted to small flower, carries out commercialization and sells.
Claims (10)
1. the method for blueberry group training quick reproduction technique is characterized in that, comprises the steps: that tissue cultivating seedling with blueberry dips in to carry out cuttage root-taking behind the root-growing agent and cultivate, and obtains the seedling of taking root.
2. the method for claim 1 is characterized in that: described root-growing agent be following a) or b):
A) solution of methyl and heteroauxin composition: the compound method of the described root-growing agent of 1L is as follows: 1000mg methyl and 1000mg heteroauxin is soluble in water, be settled to 1L;
B) solution of root-inducing powder ABT preparation: the compound method of the described root-growing agent of 1L is as follows: after root-inducing powder ABT 0.05g added the dissolving of 5ml alcohol, it was fixed molten to 1L to add water again.
3. method as claimed in claim 1 or 2 is characterized in that: the time that described tissue cultivating seedling dips in root-growing agent is 3-10 second, preferably 5 seconds.
4. as arbitrary described method among the claim 1-3, it is characterized in that: the spacing in the rows of tissue cultivating seedling was 2-5cm, preferably 3cm when described cuttage root-taking was cultivated; Line-spacing is 3-8cm, is preferably 5cm; The cuttage degree of depth is 1-1.5cm; Described cuttage root-taking culture condition is: temperature 25-35 ℃, relative moisture is 50-60%, and the light transmittance that shelters from heat or light is 45%-65%.
5. as arbitrary described method among the claim 1-4, it is characterized in that: described method also comprises the preliminary hardening of described tissue cultivating seedling before dipping in root-growing agent; The time of described preliminary hardening is 7-8 days; Temperature is 25-35 ℃; Relative moisture is 50%-60%; Intensity of illumination is 2500-3500, preferably 3000LX.
6. as arbitrary described method among the claim 1-5, it is characterized in that: described method also comprises described seedling greenhouse hardening and the land for growing field crops hardening afterwards that obtain taking root; Described greenhouse hardening and land for growing field crops hardening comprise the steps: the described seedling of taking root green house hardening 40-60 days, and between 25～35 ℃ of the condition degree of being of green house hardening, relative moisture is 50%-60%, and the light transmittance that shelters from heat or light is 45%～65%; The time of carrying out the land for growing field crops hardening then is 150-180 days.
7. as arbitrary described method among the claim 1-6, it is characterized in that: the acquisition step of the tissue cultivating seedling of described blueberry is as follows: the explant of blueberry is placed on the initial culture base induce differentiation culture, obtain just for tissue cultivating seedling; Described explant is preferably with the stem section of axillalry bud;
Described initial culture base is to add the solid culture medium that following material obtains in basic culture solution: zeatin, NAA, carbon source and gel; The final concentration of zeatin is 2-5mg/L in the described initial culture base, and the final concentration of NAA is 0.01-0.12mg/L; The solvent of described basic culture solution is that water, solute are as shown in table 1.
8. the method described in claim 7, it is characterized in that: the acquisition step of the tissue cultivating seedling of described blueberry also comprises successive transfer culture, described successive transfer culture comprises the steps: just to place subculture medium to carry out successive transfer culture for tissue cultivating seedling with described, obtains the tissue cultivating seedling of described blueberry;
Described subculture medium is to add the solid culture medium that following material obtains in basic culture solution: zeatin, NAA, IBA, carbon source and gel; The final concentration of zeatin is 1.5-4mg/L in the described subculture medium, and the final concentration of NAA is 0.04-0.1mg/L, and the final concentration of IBA is 0.1-0.5mg/L; The solvent of described basic culture solution is that water, solute are as shown in table 1.
9. method as claimed in claim 8 is characterized in that: the final concentration of zeatin and NAA is following 1 in the described initial culture base)-3) in arbitrary described: 1) final concentration of zeatin is 3-4mg/L, and the final concentration of NAA is 0.05-0.1mg/L; 2) final concentration of zeatin is 3mg/L or 4mg/L, and the final concentration of NAA is 0.05-0.1mg/L; 3) final concentration of zeatin is 3-4mg/L, and the final concentration of NAA is 0.05mg/L or 0.1mg/L;
The final concentration of zeatin, NAA and IBA is as 1 in the described subculture medium)-3) in arbitrary as described in: 1) final concentration of zeatin is 2-3mg/L, and the final concentration of NAA is 0.06-0.08mg/L, and the final concentration of IBA is 0.2-0.3mg/L; 2) final concentration of zeatin is 2mg/L, and the final concentration of NAA is 0.08mg/L, and the final concentration of IBA is 0.2mg/L; 3) final concentration of zeatin is 3mg/L, and the final concentration of NAA is 0.06mg/L, and the final concentration of IBA is 0.3mg/L.
10. method as claimed in claim 8 or 9, it is characterized in that: the gel in described initial culture base and the described subculture medium all is agar, carragheen or Gelrite, preferably all is agar; Described carbon source all is glucose, maltose or sucrose, preferably is sucrose;
Described agar all is 1300mg/cm in the intensity of described initial culture base and described subculture medium
2The final concentration of described agar in described initial culture base and described subculture medium all is 4-5g/L, is preferably 4.5g/L;
The final concentration of described sucrose in described initial culture base and described subculture medium all is 25-35g/L, preferably is 30g/L.
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102524061A (en) * | 2011-12-03 | 2012-07-04 | 韦庆和 | Rapid breeding technology for blueberry tissue culture seedling micro cuttings at high and cold area |
| CN103875515A (en) * | 2014-02-18 | 2014-06-25 | 杭州佑国生物科技有限公司 | Ex-vitro rooting method for tissue culture blueberry seedlings |
| CN103891495A (en) * | 2014-01-16 | 2014-07-02 | 安徽和美蓝莓生物科技有限公司 | Multivariate space ecological blueberry planting pattern |
| CN104350912A (en) * | 2014-10-23 | 2015-02-18 | 湖南省核农学与航天育种研究所 | Cutting propagation method for southern blueberries |
| CN106171980A (en) * | 2016-07-12 | 2016-12-07 | 安徽师范大学 | Blueberry tissue cultural method |
| CN106550750A (en) * | 2016-11-09 | 2017-04-05 | 广东融和生态农业集团有限公司 | It is a kind of to improve the method that blue berry seedling cultivates quality |
| CN108651276A (en) * | 2017-03-29 | 2018-10-16 | 北京林业大学 | Rapid Rooting cultural method in blueberry tissue bag |
| CN109661960A (en) * | 2018-08-24 | 2019-04-23 | 伊春林业科学院 | A kind of wild bog bilberry spray micro cuttage seedling-raising technique |
| CN110192525A (en) * | 2019-06-28 | 2019-09-03 | 大连大学 | A kind of cultural method of blueberry polyploid |
| CN110741931A (en) * | 2019-10-15 | 2020-02-04 | 贵州省生物研究所 | Preparation method of culture medium for promoting tissue culture and rooting of blueberries |
| CN111011216A (en) * | 2019-12-31 | 2020-04-17 | 青海省农林科学院 | Efficient regeneration culture medium and culture method for Atlantic potatoes |
| CN112021181A (en) * | 2020-10-11 | 2020-12-04 | 江苏东郁植物科技有限公司 | Rooting medium for blueberry tissue culture seedlings and rooting method thereof |
| CN112167034A (en) * | 2020-08-07 | 2021-01-05 | 兰婷 | Method for quickly breeding blueberries and natural light microcirculation cultivation box |
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| CN102524061A (en) * | 2011-12-03 | 2012-07-04 | 韦庆和 | Rapid breeding technology for blueberry tissue culture seedling micro cuttings at high and cold area |
| CN103891495A (en) * | 2014-01-16 | 2014-07-02 | 安徽和美蓝莓生物科技有限公司 | Multivariate space ecological blueberry planting pattern |
| CN103875515A (en) * | 2014-02-18 | 2014-06-25 | 杭州佑国生物科技有限公司 | Ex-vitro rooting method for tissue culture blueberry seedlings |
| CN103875515B (en) * | 2014-02-18 | 2017-01-04 | 杭州佑国生物科技有限公司 | A kind of blueberry tissue culture outside sprout-cultivating-bottle radication method |
| CN104350912A (en) * | 2014-10-23 | 2015-02-18 | 湖南省核农学与航天育种研究所 | Cutting propagation method for southern blueberries |
| CN104350912B (en) * | 2014-10-23 | 2016-09-21 | 湖南省核农学与航天育种研究所 | The cuttage breeding method of south blue berry |
| CN106171980A (en) * | 2016-07-12 | 2016-12-07 | 安徽师范大学 | Blueberry tissue cultural method |
| CN106550750A (en) * | 2016-11-09 | 2017-04-05 | 广东融和生态农业集团有限公司 | It is a kind of to improve the method that blue berry seedling cultivates quality |
| CN108651276A (en) * | 2017-03-29 | 2018-10-16 | 北京林业大学 | Rapid Rooting cultural method in blueberry tissue bag |
| CN109661960A (en) * | 2018-08-24 | 2019-04-23 | 伊春林业科学院 | A kind of wild bog bilberry spray micro cuttage seedling-raising technique |
| CN110192525A (en) * | 2019-06-28 | 2019-09-03 | 大连大学 | A kind of cultural method of blueberry polyploid |
| CN110192525B (en) * | 2019-06-28 | 2022-05-17 | 大连大学 | Culture method of blueberry polyploids |
| CN110741931A (en) * | 2019-10-15 | 2020-02-04 | 贵州省生物研究所 | Preparation method of culture medium for promoting tissue culture and rooting of blueberries |
| CN111011216A (en) * | 2019-12-31 | 2020-04-17 | 青海省农林科学院 | Efficient regeneration culture medium and culture method for Atlantic potatoes |
| CN112167034A (en) * | 2020-08-07 | 2021-01-05 | 兰婷 | Method for quickly breeding blueberries and natural light microcirculation cultivation box |
| CN112021181A (en) * | 2020-10-11 | 2020-12-04 | 江苏东郁植物科技有限公司 | Rooting medium for blueberry tissue culture seedlings and rooting method thereof |
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