CN109997689A - A method of cultivating polyploid succulent - Google Patents

A method of cultivating polyploid succulent Download PDF

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CN109997689A
CN109997689A CN201910315567.0A CN201910315567A CN109997689A CN 109997689 A CN109997689 A CN 109997689A CN 201910315567 A CN201910315567 A CN 201910315567A CN 109997689 A CN109997689 A CN 109997689A
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plant
polyploid
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谢宗宜
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Guangxi Nan Bazai Science And Technology Ltd
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G31/00—Soilless cultivation, e.g. hydroponics
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general
    • A01G7/04—Electric or magnetic or acoustic treatment of plants for promoting growth
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
    • A01H1/08—Methods for producing changes in chromosome number
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05B—PHOSPHATIC FERTILISERS
    • C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
    • 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
    • Y02P60/14—Measures for saving energy, e.g. in green houses

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Health & Medical Sciences (AREA)
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  • Biodiversity & Conservation Biology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention belongs to technical field of plant cultivation, and in particular to a method of cultivate polyploid succulent, the specific steps are as follows: the fleshiness blade that selection grows adventitious root is put into the refrigerating chamber for placing into 4 DEG C in liquid MS medium, Fiber differentiation 36h；The tetraploid cell for taking gained blade root cells detects its cell membrane resonant frequency, is powered and irradiates screening and culturing with ultrasonic resonance simultaneously；Thereafter gained blade is cultivated when growing seedling to development, takes plant shoots crop leaf measuring cell membrane resonant frequency, with ultrasonic resonance irradiation and the culture of lignin culture medium；Cultivating the mature generation plant population of gained is F1, screens F1 plant, cultivates to obtain second generation plant population F2 in batches, select pure tetraploid F2 for target seedling.Both can be improved chimera screening rate by the processing method of the application, improve polyploid take root, developmental rate, while reaching that plant is higher containing the amount of supporting, and root system is more flourishing, and have and stablize genetic effect.

Description

A method of cultivating polyploid succulent
[technical field]
The invention belongs to technical field of plant cultivation, and in particular to a method of cultivate polyploid succulent.
[background technique]
Polyploid breeding refers to using induced mutations or natural variation etc., doubles to obtain polyploid by cell chromosome group Breeding material meets the excellent variety of people's needs to breeding.Polyploid refers to from development of fertilized ova and body cell In containing there are three or three or more genomic individuals.The most frequently used, most effective polyploid breeding method is to use colchicine Or low temperature induction handles seed or the seedling of sprouting.There are polyploid acquisition polyploid cultivating process is cumbersome and time-consuming for existing technology Long, mutagens act on greatly plant poisoning, and chimera phenomenon is serious, and it is too low that pure polyploid obtains rate, although polyploid plant can It educates, but slow growth, hypoevolutism.
Ultrasonic wave can make cell membrane resonate, at this time cell vibration of membrane when identical as the resonant frequency of cell membrane Amplitude is maximum, and acoustic wave energy is made to enter cell to greatest extent, the height generated due to the mechanism of ultrasonic wave, heat effect, cavitation Temperature is changed plant cell membrane molecular level, increases the Drug delivery rate of DNA, to generate a series of effect of biophysics.It is logical Cell membrane surface tension and superficial density, the optimum frequency of ultrasonic irradiation when can obtain breeding.But the shortcomings that ultrasonic wave is cultivated It is, while certain energy can be supplied to the cell other than target dna cell, and ultrasonic wave cannot be provided to plant cell and be supported Point.
Electric current has the function of that cell growth and guidance cell is promoted to tend to.Carrying out stimulation to histocyte, can change can Excitatory cells are to Na+ or Ca+ permeability, to generate action potential, Na+, K+ ion channel for keeping it internal, which change, to be drawn Play the change of internal environment, the final state for changing cell.The disadvantage is that since the resistance of plant cell wall and cell membrane is made With current strength is small can not to stimulate plant cell internal intensity to cross conference damage plant cell.And while function of current plant, Culture medium cell reaction can occur, generate contaminants plant cell membrane.
[summary of the invention]
The invention reside in shortcoming and deficiency of the existing technology are overcome, a kind of side for cultivating polyploid succulent is provided Method.Method of the invention both can be improved chimera screening rate, improve polyploid take root, developmental rate, while plant can be allowed to contain Feeding amount is higher, and root system is more flourishing, and has and stablize genetic effect.
To achieve the goals above, The technical solution adopted by the invention is as follows:
A method of cultivating polyploid succulent, the specific steps are as follows:
1) succulent blade is chosen, immersion is taken root water 2-3 days, and the blade for growing 0.5-1.2cm adventitious root is taken；
2) blade is put into the nanofiltration membrane in liquid MS medium, culture medium is put into 4 DEG C of refrigerating chamber, induction training Support 36h；
3) gained blade root cells after inducing in step 2) is taken, observation screening obtains tetraploid cell, and detection tetraploid is thin After birth resonant frequency, give liquid MS medium be powered, while with ultrasonic resonance irradiate blade, screening and culturing 7-10 days；
4) gained blade in step 3) is laid flat and is cultivated into lignin culture medium, leaf development grows the seedling of 0.5-1cm When, plant shoots crop leaf measuring cell membrane resonant frequency is taken, irradiates culture 1-2 months with ultrasonic resonance；
5) cultivating the mature generation plant population of gained is F1, and screening F1 plant leaf leaf insert method batch cultivates to obtain the second generation Plant population F2 selects pure tetraploid F2 for target seedling.
It further illustrates, detects cell membrane with He-Ne laser doppler vialog in the step 3) or step 4) Resonant frequency.
It further illustrates, the nanofiltration membrane is aromatic series and polyacids hydrogen species composite nanometer filtering film, and position is placed in liquid MS training It supports in the middle part of based containers.
It further illustrates, the electric current being powered in the step 3) is alternating current, voltage 5-10V, electric current 500- 1000 μ A, frequency 40-60Hz.
It further illustrates, ultrasonic frequency is identical as detection gained plant cell membrane frequency in the step 3), culture medium Being powered with the spacing frequency of ultrasonic irradiation and duration is to carry out 1h every 5h.
It further illustrates, the constituent concentration of the lignin culture medium are as follows: finished product lignin 0.08-0.16g/L, glucose 3-5g/L, KH2PO41-3g/L, MgSO40.22-0.28g/L, CaCl20.08-0.12g/L, MnSO4 0.005-0.008g/ L, VB 10-12g/L, ammonium tartrate 0.1-0.3g/L, trace element solution 135-155mL/L.
It further illustrates, the constituent concentration of the trace element solution are as follows: NaCl 1.0-1.5g/L, FeSO4·7H2O 100-120mg/L, CoSO4·7H2O 100-120mg/L, ZnSO4·7H2O 100-120mg/L, CuSO4·5H2O 10- 12mg/L, KAl (SO4)2100-120mg/L, H3BO310-12mg/L, Na2MoO4 10-12mg/L。
It further illustrates, in the step 4), ultrasonic frequency is identical as detection gained plant cell membrane frequency, ultrasonic wave Irradiation time and spacing frequency are every 1 day irradiation 2-3h.
It further illustrates, the range of the plant cell membrane frequency is 30-65 ± 2.33kHz, and output intensity of acoustic wave is 20- 150db。
It further illustrates, it is Crassulaceae that blade is chosen in the step 1), and the Crassulaceae is congealed fat lotus or Huang Li or rainbow Jade or the second female heart or peach beauty, cultivation temperature be 23-28 DEG C, daily light application time 3h.
In conclusion by adopting the above-described technical solution, the beneficial effects of the present invention are:
The present invention makes chromosome doubling in Crassulaceae succulent cell mitogen by the way of low temperature induction, Tetraploid cell required for selecting, detects the resonant frequency of its cell membrane, using plant after ultrasonic resonance radiation-induced, changes Become plant cell molecular level structure, opens the channel that plant cell absorbs nutrient.It is cultivated simultaneously by electric current stimulation liquid MS Nutrient ion is largely entered into the cell under the push effect of ultrasonic wave by cell membrane channels in base.It is thin to give target tetraploid Born of the same parents' ceiling capacity carry out activity breeding, makes the quickening of target cell proliferative speed and intracellular nutritional material aggregation increases.While by The energy value suffered by tetraploid cell is utilizing the super of tetraploid cell resonant frequency much larger than energy value suffered by diploid cell Under sound wave radiation, diploid cell film is sparse, and electric current, which can enter, into the cell cuts DNA molecular, to reduce two Times somatic cell division increases chimera screening efficiency.It can be by reducing the function of current culture while nutrient ion with nanofiltration membrane Infringement of the pollutant that base generates to plant cell.Lignin can generate unproductive absorption with cellulase, promote in ultrasonic wave Into under effect, a large amount of cellulases is made to lose hydrolysis ability.Later-stage utilization ultrasonic wave cooperates the firm cultivation of lignin culture medium to plant Strain Seedlings Cell film guarantees cell under the action of first time ultrasonic wave function cells film opens cell membrane and absorbs energy conduit The robustness of film continues to reinforce plant absorption ultrasonic wave bring effect of biophysics simultaneously.Reach plant volume increase, plant leaf Nutrient content is higher, the higher effect of plant breeding rate.
[specific embodiment]
In order to make the foregoing objectives, features and advantages of the present invention clearer and more comprehensible, below with reference to of the invention specific Embodiment is described in detail.In the following description, numerous specific details are set forth in order to facilitate a full understanding of the present invention.But It is that the invention can be embodied in many other ways as described herein, those skilled in the art can be without prejudice to originally Similar improvement is done in the case where invention intension, therefore the present invention is not limited to the specific embodiments disclosed below.
Following example 1-3, the Crassulaceae succulent blade of comparative example 1-5 is by taking yellow beautiful blade as an example.
Embodiment 1:
1. choosing 50, Crassulaceae succulent blade, hot-house culture, temperature is 25 DEG C during culture, daily solar radiation 3h, immersion are taken root water 2.5 days, and the blade for growing 0.5-1.2cm adventitious root is taken.
2. culture medium is put in aromatic series and polyacids hydrogen species composite nanometer filtering film that blade is put into the middle part of liquid MS medium Enter in 4 DEG C of refrigerating chamber, Fiber differentiation 36h；
3. taking gained blade root cells after inducing in step 2), observation screening obtains tetraploid cell, with He-Ne laser Doppler effect vialog detects tetraploid cell membrane resonance frequency, and gained frequency is 45 ± 2.33kHz, and output intensity of acoustic wave is 70db.It is 8V with voltage, electric current is 800 μ A, and the exchange that frequency is 50Hz supplies electricity to fluid nutrient medium energization, while total with ultrasonic wave Vibration irradiation culture 8 days, being powered with the spacing frequency of ultrasonic irradiation and duration was to carry out 1h every 5h.
4. allotment constituent concentration is finished product lignin 0.12g/L, glucose 4g/L, KH2PO42g/L, MgSO4 0.25g/ L, CaCl20.1g/L, MnSO40.007g/L, VB 11g/L, ammonium tartrate 0.2g/L, the wood of trace element solution 145mL/L Quality culture medium.Wherein trace element solution constituent concentration is NaCl 1.3g/L, FeSO4·7H2O 110mg/L, CoSO4· 7H2O 110mg/L, ZnSO4·7H2O 110mg/L, CuSO4·5H2O 11mg/L, KAl (SO4)2110mg/L, H3BO3 11mg/L, Na2MoO411mg/L.Gained blade in step 3) is laid flat and is cultivated into lignin culture medium, leaf development is grown When the seedling of 0.5-1cm, plant shoots crop leaf measuring cell membrane resonant frequency is taken, frequency is 51 ± 2.33kHz, exports sound wave Intensity is 70db, and with ultrasonic resonance irradiation culture 1.5 months, ultrasonic irradiation time and spacing frequency were to irradiate every 1 day 2.5h；
5. cultivating the mature generation plant population of gained is F1, F1 is the plant rich in chimeric body cell, after gene duplication Polyploid and former plant size and form difference are obvious, and by observing Plant state, blade shape screens F1 plant leaf.Pass through Leaf insert method batch cultivates to obtain second generation plant population F2.It is target children that pure tetraploid F2, which can be filtered out, by active somatic cell observation Seedling.
Embodiment 2:
1. choosing 50, Crassulaceae succulent blade, hot-house culture, temperature is 28 DEG C during culture, daily solar radiation 3h, immersion are taken root water 3 days, and the blade for growing 0.5-1.2cm adventitious root is taken.
2. culture medium is put in aromatic series and polyacids hydrogen species composite nanometer filtering film that blade is put into the middle part of liquid MS medium Enter in 4 DEG C of refrigerating chamber, Fiber differentiation 36h；
3. taking gained blade root cells after inducing in step 2), observation screening obtains tetraploid cell, with He-Ne laser Doppler effect vialog detects tetraploid cell membrane resonance frequency, and gained frequency is 48 ± 2.33kHz, and output intensity of acoustic wave is 150db.It is 10V with voltage, electric current is 1000 μ A, and the exchange that frequency is 60Hz supplies electricity to fluid nutrient medium energization, while with ultrasonic Wave resonance irradiation culture 10 days, being powered with the spacing frequency of ultrasonic irradiation and duration is to carry out 1h every 5h.
4. allotment constituent concentration is finished product lignin 0.16g/L, glucose 5g/L, KH2PO43g/L, MgSO4 0.28g/ L, CaCl20.12g/L, MnSO40.008g/L, VB 12g/L, ammonium tartrate 0.3g/L, trace element solution 155mL/L's Lignin culture medium.Wherein trace element solution constituent concentration is NaCl 1.5g/L, FeSO4·7H2O 120mg/L, CoSO4· 7H2O 120mg/L, ZnSO4·7H2O 120mg/L, CuSO4·5H2O 12mg/L, KAl (SO4)2120mg/L, H3BO3 12mg/L, Na2MoO412mg/L.Gained blade in step 3) is laid flat and is cultivated into lignin culture medium, leaf development is grown When the seedling of 0.5-1cm, plant shoots crop leaf measuring cell membrane resonant frequency is taken, frequency is 53 ± 2.33kHz, output sound Intensity of wave is 150db, and with ultrasonic resonance irradiation culture 2 months, ultrasonic irradiation time and spacing frequency were every 1 day photograph Penetrate 3h；
5. cultivating the mature generation plant population of gained is F1, F1 is the plant rich in chimeric body cell, after gene duplication Polyploid and former plant size and form difference are obvious, and by observing Plant state, blade shape screens F1 plant leaf.Pass through Leaf insert method batch cultivates to obtain second generation plant population F2.It is target children that pure tetraploid F2, which can be filtered out, by active somatic cell observation Seedling.
Embodiment 3:
1. choosing 50, Crassulaceae succulent blade, hot-house culture, temperature is 23 DEG C during culture, daily solar radiation 3h, immersion are taken root water 2 days, and the blade for growing 0.5-1.2cm adventitious root is taken.
2. culture medium is put in aromatic series and polyacids hydrogen species composite nanometer filtering film that blade is put into the middle part of liquid MS medium Enter in 4 DEG C of refrigerating chamber, Fiber differentiation 36h；
3. taking gained blade root cells after inducing in step 2), observation screening obtains tetraploid cell, with He-Ne laser Doppler effect vialog detects tetraploid cell membrane resonance frequency, and gained frequency is 44 ± 2.33kHz, and output intensity of acoustic wave is 20db.It is 5V with voltage, electric current is 500 μ A, and the exchange that frequency is 40Hz supplies electricity to fluid nutrient medium energization, while total with ultrasonic wave Vibration irradiation culture 7 days, being powered with the spacing frequency of ultrasonic irradiation and duration was to carry out 1h every 5h.
4. allotment constituent concentration is finished product lignin 0.08g/L, glucose 3g/L, KH2PO41g/L, MgSO4 0.22g/ L, CaCl20.08g/L, MnSO40.005g/L, VB 10g/L, ammonium tartrate 0.1g/L, trace element solution 135mL/L's Lignin culture medium.Wherein trace element solution constituent concentration is NaCl 1.0g/L, FeSO4·7H2O 100mg/L, CoSO4· 7H2O 100mg/L, ZnSO4·7H2O 100mg/L, CuSO4·5H2O 10mg/L, KAl (SO4)2100mg/L, H3BO3 10mg/L, Na2MoO410mg/L.Gained blade in step 3) is laid flat and is cultivated into lignin culture medium, leaf development is grown When the seedling of 0.5-1cm, plant shoots crop leaf measuring cell membrane resonant frequency is taken, frequency is 48 ± 2.33kHz, output sound Intensity of wave is 20db, and with ultrasonic resonance irradiation culture 1 month, ultrasonic irradiation time and spacing frequency were to irradiate every 1 day 2h；
5. cultivating the mature generation plant population of gained is F1, F1 is the plant rich in chimeric body cell, after gene duplication Polyploid and former plant size and form difference are obvious, and by observing Plant state, blade shape screens F1 plant leaf.Pass through Leaf insert method batch cultivates to obtain second generation plant population F2.It is target children that pure tetraploid F2, which can be filtered out, by active somatic cell observation Seedling.
Comparative example 1: processing scheme is substantially same as Example 1, and difference is not carry out ultrasonic resonance in step 3) The processing of irradiation is directly cultivated after vanes low temperature induction to growing new seedling.
Comparative example 2: processing scheme is substantially same as Example 1, and difference is not carry out culture medium in step 3) to be powered Processing.
Comparative example 3: processing scheme is substantially same as Example 1, and difference is not place nanofiltration membrane.
Comparative example 4: processing scheme is substantially same as Example 1, and difference is not carry out blade in step 4) to be put into wood Quality culture medium is changed to blade and is put into sterile soil treatment.
Comparative example 5: processing scheme is substantially same as Example 1, and difference is not carry out any ultrasonic resonance irradiation It is handled with culture medium energization.
Verification test one:
It will be in embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3, comparative example 4 and comparative example 5 The probability of the pure tetraploid plant of the F2 finally obtained compares, and obtains such as data in following table:
Table 1:
Classification Pure tetraploid probability of occurrence % in F2
Embodiment 1 62.5
Embodiment 2 60.8
Embodiment 3 61.6
Comparative example 1 34.8
Comparative example 2 35.2
Comparative example 3 55.4
Comparative example 4 50.6
Comparative example 5 34.7
By embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3, comparative example 4 and comparative example 5 In the pure tetraploid plant of F2 that finally obtains, after cultivating 5 months, carry out the parameter detecting such as following table:
Table 2:
Classification Survival rate % Cell average external volume μm3 Root system average length cm Blade average length cm
Embodiment 1 98 35222 16.97 9.68
Embodiment 2 94 35125 15.96 9.21
Embodiment 3 96 34879 16.32 9.18
Comparative example 1 79 27000 6.82 5.41
Comparative example 2 80 27100 7.22 5.52
Comparative example 3 69 28700 11.36 7.06
Comparative example 4 72 31050 8.06 6.76
Comparative example 5 80 26500 6.52 5.31
By embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3, comparative example 4 and comparative example 5 Middle plant nutrition state carries out the parameter detecting such as following table with TY-ZWY02 diagnosis of plant nutrition instrument:
Table 3:
By embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3, comparative example 4 and comparative example 5 Middle plant nutrition state carries out the parameter detecting such as following table with TY-4N Chlorophyll meter:
Table 4:
Using a kind of method for cultivating polyploid succulent of the application, plant root cells are first carried out more times of low temperature induction Chemical reagent injury caused by plant cell is removed in body culture from.Somatoscopy is carried out with the root cells after inducing again, is obtained Required tetraploid cell detects its cell membrane resonant frequency.Plant is placed into nanofiltration membrane, it can be same by nutrient ion When can stop be powered caused by contaminants plant cell.It is irradiated using ultrasonic resonance, changes plant cell membrane molecular water It is flat, keep the entrance of plant cell membrane absorption nutrient and energy penetrating.It is powered while ultrasonic irradiation to fluid nutrient medium, it is micro- Electric current is not injuring plant and power frequency is minimum to impact to ultrasonic resonance.In electric current stimulation liquid MS culture medium In a large amount of importing plant cells of push effect that nutrient ion passes through ultrasonic wave, the energy supply to tetraploid cell schizogamy is improved It supports, to improve tetraploid cell fertility and proliferative speed.Tetraploid cell is than diploid and three times body cell simultaneously, Nucleus volume is significantly greater, and cell membrane is thicker, and diploid and triploid are by under the ultrasonic wave effect of tetraploid resonant frequency Cell membrane can be thin penetrating, and electric current, which can enter, into the cell cuts DNA molecular, to reduce the numerous of diploid and triploid Rate is grown, and then improves the screening rate to chimera.Lignin can generate unproductive absorption with cellulase, promote in ultrasonic wave Into under effect, a large amount of cellulases is made to lose hydrolysis ability.Ultrasonic resonance and wood are persistently used in plant shoots early days of growth The effect culture of quality culture medium can consolidate plant cell toughness and absorb the ability of nutrient, sustainable to provide energy to plant cell Amount accelerates plant development rate, promotes plant establishment.
Test proves:
It can be concluded that, averagely improved using this method by upper table and occur the probability 45-51% of pure tetraploid plant in F2；It is flat F2 is improved for pure tetraploid survival rate 15-25%；Averagely increase plant volume 32-21%；Averagely improve plant cell nutrient Content 47-85%.Above data explanation, after low temperature induction, mixoplod cell proliferation irradiates plant cell in ultrasonic resonance Under conditions of the function of current, the low cell of comparison target dna content can be reinforced and screened out, target dna content cell is reinforced Breeding potential, cell absorb nutrient it is more, cell division proliferative speed is fast, vegetative propagation survival rate improve, cell average external volume increase Greatly.But energization, which pollutes object, to make a big impact to the survival rate of plant cell, with nanofiltration membrane energy effective protection pollutant Encroach on plant cell.Plant seedlings can reinforce plant nutrient absorption with ultrasonic resonance irradiation during developing but be easy to destroy cell Membrane stability, under the repair of lignin, cell membrane is not firm fragile while plant cell can be made to keep absorbing path It is broken.Higher containing the amount of supporting to reach plant, root system is more flourishing, and survival rate is higher, and has and stablize genetic effect.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously Limitation of the scope of the invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art, Without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection model of the invention It encloses.Therefore, protection scope of the present invention should be determined by the appended claims.

Claims (10)

1. a kind of method for cultivating polyploid succulent, which is characterized in that specific step is as follows:
1) succulent blade is chosen, immersion is taken root water 2-3 days, and the blade for growing 0.5-1.2cm adventitious root is taken；
2) blade is put into the nanofiltration membrane in liquid MS medium, culture medium is put into 4 DEG C of refrigerating chamber, Fiber differentiation 36h；
3) gained blade root cells after inducing in step 2) is taken, observation screening obtains tetraploid cell, detects tetraploid cell film Resonant frequency, give liquid MS medium be powered, while with ultrasonic resonance irradiate blade, screening and culturing 7-10 days；
4) gained blade in step 3) is laid flat and is cultivated into lignin culture medium, when leaf development grows the seedling of 0.5-1cm, taken Plant shoots crop leaf measuring cell membrane resonant frequency irradiates culture 1-2 months with ultrasonic resonance；
5) cultivating the mature generation plant population of gained is F1, and screening F1 plant leaf leaf insert method batch cultivates to obtain second generation plant Group F2 selects pure tetraploid F2 for target seedling.
2. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: the detection cell Membrane resonance frequency is specifically that He-Ne laser doppler vialog is used to be detected.
3. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: in step 2), The nanofiltration membrane is aromatic series and polyacids hydrogen species composite nanometer filtering film, and position is placed in the middle part of Liquid Culture based containers.
4. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: in step 3), The electric current of the energization is alternating current, and voltage 5-10V, current range is 500-1000 μ A, frequency 40-60Hz.
5. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: in step 3), Ultrasonic frequency is identical as detection gained plant cell membrane frequency, and culture medium is powered and the spacing frequency of ultrasonic irradiation and lasting Time is to carry out 1h every 5h.
6. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: in step 4), The constituent concentration of the lignin culture medium are as follows: finished product lignin 0.08-0.16g/L, glucose 3-5g/L, KH2PO4 1-3g/ L, MgSO40.22-0.28g/L, CaCl20.08-0.12g/L, MnSO40.005-0.008g/L, VB 10-12g/L, tartaric acid Ammonium 0.1-0.3g/L, trace element solution 135-155mL/L.
7. a kind of method for cultivating polyploid succulent according to claim 6, it is characterised in that: the microelement The constituent concentration of solution are as follows: NaCl 1.0-1.5g/L, FeSO4·7H2O 100-120mg/L, CoSO4·7H2O 100- 120mg/L, ZnSO4·7H2O 100-120mg/L, CuSO4·5H2O 10-12mg/L, KAl (SO4)2100-120mg/L, H3BO310-12mg/L, Na2MoO4 10-12mg/L。
8. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: in step 4), Ultrasonic frequency is identical as detection gained plant cell membrane frequency, and ultrasonic irradiation time and spacing frequency are to irradiate every 1 day 2-3h。
9. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: the plant cell The range of film frequency is 30-65 ± 2.33kHz, and output intensity of acoustic wave is 20-150db.
10. a kind of method for cultivating polyploid succulent according to claim 1, it is characterised in that: in step 1) Selection blade is Crassulaceae, and cultivation temperature is 23-28 DEG C, daily light application time 3h.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110622864A (en) * 2019-10-23 2019-12-31 国家林业和草原局泡桐研究开发中心 Chinaberry tissue culture chromosome doubling rapid propagation method
CN112753581A (en) * 2021-02-04 2021-05-07 公孙国林 Cultivation formula of polyploid plant of Dolicholepis of Africaceae

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108004148A (en) * 2017-12-01 2018-05-08 齐鲁工业大学 A kind of fluid nutrient medium and preparation method thereof
CN108130277A (en) * 2017-12-01 2018-06-08 齐鲁工业大学 A kind of white-rot fungi culture medium and its preparation method based on lignin
US20180317413A1 (en) * 2015-11-03 2018-11-08 Tianjin Nankai University Castor Engineering Scien And Technology Co., Ltd. Breeding Method for Tetraploid Ricinus Communis

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20180317413A1 (en) * 2015-11-03 2018-11-08 Tianjin Nankai University Castor Engineering Scien And Technology Co., Ltd. Breeding Method for Tetraploid Ricinus Communis
CN108004148A (en) * 2017-12-01 2018-05-08 齐鲁工业大学 A kind of fluid nutrient medium and preparation method thereof
CN108130277A (en) * 2017-12-01 2018-06-08 齐鲁工业大学 A kind of white-rot fungi culture medium and its preparation method based on lignin

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
卢继传: "《谁将在生物学世纪称雄》", 31 January 1991, 湖北少年儿童出版社 *
白均元: ""超声辐射对植物种子细胞膜渗透性的研究"", 《中国优秀博硕士学位论文全文数据库(硕士) 基础科学辑》 *
赵欢: ""多肉花卉‘特玉莲’‘丽娜莲’再生体系建立及辐射与多倍体技术研究"", 《中国优秀博硕士学位论文全文数据库(硕士) 农业科技辑》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110622864A (en) * 2019-10-23 2019-12-31 国家林业和草原局泡桐研究开发中心 Chinaberry tissue culture chromosome doubling rapid propagation method
CN112753581A (en) * 2021-02-04 2021-05-07 公孙国林 Cultivation formula of polyploid plant of Dolicholepis of Africaceae

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