CN107155875A - A kind of allopolyploid induction production method of hybrid Chinese pennisetum - Google Patents
A kind of allopolyploid induction production method of hybrid Chinese pennisetum Download PDFInfo
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- CN107155875A CN107155875A CN201710534194.7A CN201710534194A CN107155875A CN 107155875 A CN107155875 A CN 107155875A CN 201710534194 A CN201710534194 A CN 201710534194A CN 107155875 A CN107155875 A CN 107155875A
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- 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
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- 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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- 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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Abstract
The present invention relates to a kind of multiploid induction growing method of hybrid Chinese pennisetum, methods described includes allotriploid hybrid Chinese pennisetum generation seed carrying out immersion treatment in the mutagens containing colchicin and dimethyl sulfoxide (DMSO), then carries out Initial culture, polyploid identification, Multiplying culture, expanding production and transplants.The method of the present invention can obtain a large amount of mutagenesis seedlings in a short time, greatly improve culture efficiency, saved production cost.
Description
Technical field
The present invention relates to plant breeding and tissue culture technology field, and in particular to a kind of allopolyploid of hybrid Chinese pennisetum is lured
Lead production method.
Background technology
Hybrid Chinese pennisetum (Pennisetum americanum × P.purpureum) category grass family Pennisetum is perennial
Herbage, is tetraploid napier grass (Pennisetumpurpureum Schum) and diploid American pennisetum alopecuroides
Generally with asexual in the inter-species allotriploid hybrid that (Pennisetumalopecuroides (L.) Spreng) is produced, production
Breeding or first-filial generation seminal propagation.The merit that hybrid Chinese pennisetum combines parents has obvious hybrid vigour, fresh grass
Middle crude fat, crude protein, crude fibre, NFE and ash content content it is high, it is nutritious, be that a kind of good quality and high output is adapted to feeding
Feed the high-quality greenfeed of a variety of plant-eating animals such as ox, sheep, fish.In addition, hybrid Chinese pennisetum is alternatively arranged as high-quality paper pulp and wood-based plate
Raw material, or important biomass energy crop.
Based on above advantage, researcher proceeds the work of kind genetic improvement to it again, is desirably to obtain biological yield
Higher, adaptable and best in quality using energy source new varieties.Allopolyploid breeding is one kind that plant innovates breeding
Means, by chromosome doubling technology, artificial creation's polyploid is the effective way for obtaining high yield of biomass new variety of plant
One of.However, for hybrid Chinese pennisetum, the report on multiploid induction breeding method is there is no at present.
The content of the invention
The method that the present inventor is combined by distant hybridization and chromosome doubling, can obtain a large amount of in the most short time
Mutagenesis test tube seedling, i.e. allohexaploid hybrid Chinese pennisetum.By planting experimental verification, the allohexaploid hybridization that the present invention is obtained
Chinese pennisetum has higher yield and quality than allotriploid hybrid Chinese pennisetum, so as to complete the present invention.
Therefore, the invention provides a kind of multiploid induction growing method of hybrid Chinese pennisetum, methods described includes：
(1) immersion treatment：After allotriploid hybrid Chinese pennisetum generation seed disinfection, mutagens are soaked at 4 DEG C
In, immersion treatment 30-90min, preferably 60min, the mutagens are sterilized containing colchicin and dimethyl sulfoxide (DMSO)
Mixed liquor；
(2) Initial culture：Seed after immersion treatment in step (1) is taken out, is placed on aseptic filter paper and dries, then
Directly it is inoculated into comprising on 0.5-3.0mg/L colchicins, the preferably Initial culture base of 1.5mg/L colchicins, in 20-30
DEG C, preferably 25 DEG C, under 1000lx intensities of illumination, daily illumination 10h, illumination cultivation 25-35 days, preferably 30 days obtains plant；
(3) polyploid is identified：The obvious plant of phenotypic variation in the plant of step (2) culture acquisition is selected, in training of taking root
Support to cultivate in base and take root, then take the tip of a root to include the part at meristematic zone position, carry out karyotype identification；Chromosome number
What is be multiplied in units of genome is polyploid；
(4) Multiplying culture：The plant that polyploid is accredited as in step (3) is inoculated into Multiplying culture 6 on proliferated culture medium
Individual month, obtain bottle seedling；
(5) produce：The bottle seedling that step (4) is obtained, is inoculated into one-step-seedling formation culture medium and is cultivated, big obtaining
While measuring clump bud, root also forms the root system of a large amount of stalwartnesses, obtains propagation rooted seedling；
(6) transplant：The propagation rooted seedling that step (5) is obtained, is placed in natural light lower refining seedling 3-5 days；Then take out, clean
Culture medium on root system, selection cloudy day or directly transplant to crop field the dusk time-division.
Preferably, in step (1), colchicin and 1.5-2.0mg/L comprising 1.0-3.0mg/L in the mutagens
The dimethyl sulfoxide (DMSO) of dimethyl sulfoxide (DMSO), the more preferably colchicin comprising 1.5mg/L and 1.5mg/L；
Preferably, in step (1), the seed disinfection is concretely comprised the following steps：The seed is placed on sterile working first
Washed 5-7 times, then infiltrated -1 minute 30 seconds with 70% alcohol with sterile on platform, is then placed in 0.1% liquor hydrargyri perchloridi
Sterilizing 12-18 minutes, then with aseptic water washing 5-6 times.
Preferably, in step (2), the Initial culture base is to include N6+6-BA3.0-5.0mg/L+ cycocels 3.0-
5.0mg/L Solid agar culture, preferably comprises N6+6-BA5.0mg/L+ cycocels 5.0mg/L or N6+6-BA4.0mg/L+
Cycocel 5.0mg/L Solid agar culture.
Preferably, in step (3), the root media is solid for the agar comprising 1/2MS+ paclobutrazols 0.5-3.0mg/L
Body culture medium；Preferably comprise 1/2MS+ paclobutrazol 1.0mg/L or 1/2MS+ paclobutrazols 1.5mg/L Solid agar culture.
Preferably, in step (4), the proliferated culture medium is to include 1/2MS+6-BA1.0-2.5mg/L+ paclobutrazols 3.0-
5.0mg/L Solid agar culture, preferably comprises 1/2MS+6-BA1.0mg/L+ paclobutrazols 5.0mg/L or 1/2MS+6-
BA2.0mg/L+ paclobutrazols 5.0mg/L Solid agar culture.
Preferably, in step (5), the one-step-seedling formation culture medium is to include 1/2MS+6-BA1.0-2.5mg/L+ multiple-effect
Azoles 3.0-5.0mg/L Solid agar culture.
Technical scheme has the following advantages and effect：
1st, present invention selection allotriploid hybrid Chinese pennisetum generation seed directly uses mutagens as explant after sterilization
Processing, easy to operate, materials are easy；
2nd, the present invention only needs training using mutagenesis agent solution infusion method with adding the processing mode that medium culture method is combined
Support 20 days or so, just can obtain a large amount of variation plants；Efficiency of inducing mutation is substantially increased, mutagenesis is saved and explores the time；
3rd, the plant of the present invention only selection phenotypic variation obvious (as shown in Figure 2) carries out karyotype identification；Will identification
Be transferred in proliferated culture medium and cultivate 30 days or so for the plant of allohexaploid, obtain a large amount of Multiple Buds, proliferation times reach 7 with
On；More than 30 plants of the effective seedling of every bottle of acquisition, substantially increases culture efficiency, has saved production cost；
4th, mass production stage synchronous induced bud and root, step completion is reduced to by propagation with taking root, i.e., " step into
Seedling " method, forming intact plant is only needed 20 days or so, and allohexaploid hybrid Chinese pennisetum is applied to production by short time interior energy
In；
5th, present invention selection allotriploid hybrid Chinese pennisetum plant is control (as shown in Fig. 5-Fig. 9)；Both transplant bar
Part is consistent, and allohexaploid plant is high compared to adjoining tree and stem color is dark green, stem is thick, internode is short and axillary bud is more, leaf short-wide,
Tiller number is more, and aerial root is more, and especially nutritional ingredient is significantly improved.
Therefore, can be with most fast speed by achievement in research the invention provides the cultivating system of a set of efficient low-consume amount
It is converted into actual productivity.
Brief description of the drawings
Fig. 1 shows the growing state of allotriploid hybrid Chinese pennisetum plant；
Fig. 2 shows the growing state of allohexaploid hybrid Chinese pennisetum plant；
Fig. 3 shows allotriploid hybrid Chinese pennisetum (3 ×) idiogram (1000 ×)；
Fig. 4 shows allohexaploid hybrid Chinese pennisetum (6 ×) idiogram (1000 ×)；
Fig. 5 shows the growing state of allotriploid hybrid Chinese pennisetum field plant.
Fig. 6 shows the growing state of allohexaploid hybrid Chinese pennisetum field plant.
Fig. 7 is allotriploid hybrid Chinese pennisetum crop field seedling leaf (left side) and allohexaploid hybrid Chinese pennisetum field plant leaf
The photo of piece (right side).
Fig. 8 is allotriploid hybrid Chinese pennisetum field plant bar (left side) and allohexaploid hybrid Chinese pennisetum field plant bar
The photo on (right side).
Fig. 9 is allotriploid hybrid Chinese pennisetum field plant fringe (left side) and allohexaploid hybrid Chinese pennisetum field plant fringe
The photo on (right side).
Embodiment
The present invention is intended to further illustrate with reference to embodiments, rather than technical scheme is limited.
Embodiment 1
Multiploid induction production is carried out to hybrid Chinese pennisetum using following methods：
First, culture medium is prepared and sterilized
Initial culture base：
1. the agar of N6+6-BA3.0mg/L+ cycocels 3.0mg/L+3% sucrose+0.75%；
2. the agar of N6+6-BA4.0mg/L+ cycocels 5.0mg/L+3% sucrose+0.75%；
3. the agar of N6+6-BA5.0mg/L+ cycocels 5.0mg/L+3% sucrose+0.75%.
Proliferated culture medium：
1. the agar of 1/2MS+6-BA1.0mg/L+ paclobutrazols 5.0mg/L+3% sucrose+0.75%；
2. the agar of 1/2MS+6-BA2.0mg/L+ paclobutrazols 5.0mg/L+3% sucrose+0.75%；
3. the agar of 1/2MS+6-BA2.5mg/L+ paclobutrazols 3.0mg/L+3% sucrose+0.75%.
At 121 DEG C of above culture medium, sterilize 20 minutes.
Root media：
1. the agar of 1/2MS+ paclobutrazols 0.5mg/L+2% sucrose+0.75%；
2. the agar of 1/2MS+ paclobutrazols 1.0mg/L+2% sucrose+0.75%；
3. the agar of 1/2MS+ paclobutrazols 1.5mg/L+2% sucrose+0.75%；
4. the agar of 1/2MS+ paclobutrazols 2.0mg/L+2% sucrose+0.75%；
5. the agar of 1/2MS+ paclobutrazols 3.0mg/L+2% sucrose+0.75%.
One-step-seedling formation culture medium：
1. 1/2MS+6-BA1.0mg/L+ paclobutrazols 3.0mg/L；
2. 1/2MS+6-BA2.0mg/L+ paclobutrazols 5.0mg/L；
3. 1/2MS+6-BA2.5mg/L+ paclobutrazols 5.0mg/L.
2nd, immersion treatment
Hybrid Chinese pennisetum (Juncus effusus L.) allotriploid first-filial generation seed is taken, on aseptic operating platform
Washed 6 times, infiltrated 1 minute with 70% alcohol with sterile, placed into 0.1% liquor hydrargyri perchloridi and sterilize 15 minutes, it is sterile
After water is rinsed 6 times, the mutagens of colchicin 1.5mg/L and the 1.5mg/L dimethyl sulfoxide (DMSO) containing filtration sterilization are soaked in
Middle immersion 60min, treatment temperature be 4 DEG C or so, then take out clean up it is rear standby.
3rd, Initial culture
The filter paper suck dry moisture of the seed after immersion treatment is respectively connected to equipped with first comprising 1.5mg/L colchicins
For in the plastic culture bottle (60 milliliters of capacity) of culture medium 1., 2. and 3., each blake bottle explant number is 10, inoculation 30
Bottle, in triplicate.
Cultivated under following condition of culture：25 DEG C or so, daily illumination 10 hours, intensity of illumination is 1000-
2000lx or so；Seed culture starts rudiment for 7 days or so, but the bud sprouted is weaker, grows tall rapidly, phenotypic variation is not obvious；Training
Support the bud phenotypic variation sprouted for 30 days or so substantially, bud is thick and base portion expands；But the easy browning of bud sprouted for more than 35 days is cultivated, by
It is gradually dead.
4th, polyploid is identified
The plant of phenotypic variation obvious (as described in Figure 2, bar stem is more and big, darker compared with thick, blade) is selected, by it
Root media is inoculated into respectively 1., 2., 3., 4. and 5. above to be cultivated, and is cut the plant tip of a root and is included meristematic zone position
Part, carries out karyotype identification, and what chromosome number was multiplied in units of genome is polyploid.
5th, Multiplying culture
The plant of polyploid will be accredited as, proliferated culture medium 1., 2. and 3. upper Multiplying culture 6 months are inoculated into respectively, are made
It breeds sprouting, and proliferation times reach more than 7, obtain bottle seedling；
6th, produce
Bottle seedling is inoculated into 1/2MS+6-BA2.0mg/L+ paclobutrazols 5.0mg/L one-step-seedling formation culture medium and cultivated,
While a large amount of clump buds are obtained, root also forms the root system of a large amount of stalwartnesses, obtains propagation rooted seedling；As a result show, bud propagation
Multiple is more than 5, and sprout is healthy and strong, more than 30 plants of every bottle of effective seedling, synchronous rooting rate up to 100%,；
7th, transplant
Propagation rooted seedling is put bottle cap is opened under natural light, hardening 3-5 days；Then the training on cleaning root system is taken out with tweezers
Base is supported, selection cloudy day or directly transplant to crop field the dusk time-division.Transplant 1 month or so, survive and reach more than 90%.
To the plant in each step, bottle seedling, breed rooted seedling growing state observe, find Initial culture base 2. with
3. it is better than Initial culture base 1.；1., 4., 5. 2. and 3. root media be better than root media；1. and 2. proliferated culture medium
Better than proliferated culture medium 3.；1. 2. and 3. one-step-seedling formation culture medium be better than one-step-seedling formation culture medium.
EXPERIMENTAL EXAMPLE 1
In this EXPERIMENTAL EXAMPLE, using mutagenesis agent solution infusion method, add medium therapy, solution immersion and add and cultivate
Three kinds of methods of base combined method carry out multiploid induction experiment, and every kind of method is provided with some groups of processing times and concentration gradient,
It has studied the influence of different temperatures, different disposal method, various concentrations mutagens and different disposal time to multiploid induction rate.
A, mutagenesis agent solution infusion method：Take hybrid Chinese pennisetum (Juncus effususL.) allotriploid first-filial generation kind
Son, after being sterilized on aseptic operating platform, be soaked in respectively containing filtration sterilization colchicin (1.0mg/L, 2.0mg/L,
3.0mg/L) and in the mutagens of dimethyl sulfoxide (DMSO) (1.5mg/L, 2.0mg/L) soak 30min, 60min, 90min, treatment temperature
For 4 DEG C, then take out and clean up, be seeded in Initial culture base 2. on, each processing inoculation explant number is 10,30
Bottle, respectively in triplicate.
B, addition medium therapy：Hybrid Chinese pennisetum (Juncus effususL.) allotriploid first-filial generation seed is taken,
On aseptic operating platform sterilize after, use filter paper suck dry moisture, be respectively connected to be equipped with comprising 0.5mg/L, 1.0mg/L, 1.5mg/L,
In the plastic culture bottle (60 milliliters of capacity) of the Initial culture base of 2.0mg/L, 2.5mg/L, 3.0mg/L colchicin 2., often
Individual blake bottle explant number is 10, is inoculated with 30 bottles, in triplicate.
C, solution immersion and addition culture medium mixing method：Take hybrid Chinese pennisetum (Juncus effususL.) allotriploid
First-filial generation seed, after being sterilized on aseptic operating platform, is soaked in the colchicin containing filtration sterilization respectively at 4 DEG C
In the mutagens of (1.0mg/L, 2.0mg/L, 3.0mg/L) and dimethyl sulfoxide (DMSO) (1.5mg/L, 2.0mg/L) soak 30min,
60min, 90min, then take out and clean up, and use filter paper suck dry moisture, be respectively connected to be equipped with comprising 0.5mg/L, 1.0mg/L,
The plastic culture bottle (60 milliliters of capacity) of the Initial culture base of 1.5mg/L, 2.0mg/L, 2.5mg/L, 3.0mg/L colchicin 2.
In, each blake bottle explant number is 10, is inoculated with 30 bottles, in triplicate.
As a result with analysis
(1) using mutagenesis agent solution infusion method, addition medium therapy, solution immersion and addition culture in this EXPERIMENTAL EXAMPLE
These three processing methods of base combined method can obtain the mutagenesis seedling of certain ratio, but long-time solution immersion treatment fatal rate is high
Up to more than 90%, addition medium therapy fatal rate is low but induced mutation rate is also low.The solution immersion and addition culture medium combination of the present invention
Method inductivity highest, best of breed parameter is：In the mutagens containing 1.5mg/L colchicines and 1.5mg/L dimethyl sulfoxide (DMSO)s
In, then the immersion treatment 60min in 4 DEG C of refrigerators is inoculated into comprising the 2. upper training of 1.5mg/L colchicins Initial culture base
Support.The fatal rate that aseptic seedling is poisoned in the combination in the presence of mutagens is 27.07%, therefore, solution immersion and addition culture
Base combined method effect is substantially better than solution infusion method, or adds medium therapy；Allotriploid hybrid Chinese pennisetum with heterologous six times
The growing state difference of body hybrid Chinese pennisetum plant is as shown in Figure 1 and Figure 2.
(2) what chromosome number was multiplied in units of genome is polyploid.The present invention is by largely identifying
As shown by data, the Chinese pennisetum allotriploid chromosome number is 3n=3x=21；Hexaploid is 6n=6x=42；The present invention is most
Excellent combination allohexaploid inductivity reaches 66.67%, and chimera inductivity reaches 26.67%.Allotriploid hybridizes wolf tail
Grass and allohexaploid hybrid Chinese pennisetum plant idiogram difference are as shown in Figure 3, Figure 4.
(3) present invention selection allotriploid hybrid Chinese pennisetum plant is control；Both transplanting conditions are consistent, and seedling is different
The plant of source hexaploid hybrid Chinese pennisetum is high compared to adjoining tree and stem color is dark green, stem is thick, internode is short and axillary bud is more, leaf is short and
Wide, tiller number is more, and fibrous root is sturdy, and inflorescence becomes big, and especially nutritional ingredient is significantly improved.Allotriploid hybrid Chinese pennisetum with it is different
The main morphological features of source hexaploid hybrid Chinese pennisetum are more as shown in table 1, and both main nutrient compositions compare such as the institute of table 2
Show, the growing way of allotriploid hybrid Chinese pennisetum and allohexaploid hybrid Chinese pennisetum plant is as shown in figures 5-9.
The allotriploid hybrid Chinese pennisetum of table 1 is compared with the main morphological features of allohexaploid hybrid Chinese pennisetum
The allotriploid hybrid Chinese pennisetum of table 2 and the comparison of the main nutrient composition of allohexaploid hybrid Chinese pennisetum
| Sequence number | Type | Dry matter content | Crude protein content | Crude fat content | Crude fiber content |
| 1 | Allotriploid | 15.82±0.06 | 13.39±0.21 | 4.69±0.17 | 33.49±0.08 |
| 2 | Allohexaploid | 16.74±0.07 | 15.91±0.85 | 6.37±0.12 | 33.58±0.46 |
Above only describes the better embodiment of patent of the present invention, but patent of the present invention is not limited to above-described embodiment.
It will be appreciated by persons skilled in the art that any same or similar means of patented technology effect of the present invention can be realized,
In the protection domain that patent of the present invention should be fallen into.
Claims (7)
1. a kind of multiploid induction growing method of hybrid Chinese pennisetum, methods described includes：
(1) immersion treatment：After allotriploid hybrid Chinese pennisetum generation seed disinfection, it is soaked in mutagens, soaks at 4 DEG C
Bubble processing 30-90min, preferably 60min, the mutagens are the sterilized mixing containing colchicin and dimethyl sulfoxide (DMSO)
Liquid；
(2) Initial culture：Seed after immersion treatment in step (1) is taken out, is placed on aseptic filter paper and dries, then directly
Be inoculated into comprising 0.5-3.0mg/L colchicins, preferably in the Initial culture base of 1.5mg/L colchicins, 20-30 DEG C, it is excellent
Select under 25 DEG C, 1000lx intensities of illumination, daily illumination 10h, illumination cultivation 25-35 days, preferably 30 days, acquisition plant；
(3) polyploid is identified：The obvious plant of phenotypic variation in the plant of step (2) culture acquisition is selected, in root media
Middle culture is taken root, and then takes the tip of a root to include the part at meristematic zone position, carries out karyotype identification；Chromosome number is to contaminate
Colour solid group is the as polyploid that unit is multiplied；
(4) Multiplying culture：The plant that polyploid is accredited as in step (3) is inoculated into Multiplying culture 6 months on proliferated culture medium,
Obtain bottle seedling；
(5) produce：The bottle seedling that step (4) is obtained, is inoculated into one-step-seedling formation culture medium and is cultivated, and is obtaining a large amount of clumps
While bud, root also forms the root system of a large amount of stalwartnesses, obtains propagation rooted seedling；
(6) transplant：The propagation rooted seedling that step (5) is obtained, is placed in natural light lower refining seedling 3-5 days；Then take out, clean root system
On culture medium, selection cloudy day or the dusk time-division directly transplant to crop field.
2. 1.0-3.0mg/ according to the method described in claim 1, it is characterised in that in step (1), is included in the mutagens
The two of L colchicin and 1.5-2.0mg/L dimethyl sulfoxide (DMSO), the more preferably colchicin comprising 1.5mg/L and 1.5mg/L
Methyl sulfoxide.
3. according to the method described in claim 1, it is characterised in that in step (1), the seed disinfection is concretely comprised the following steps：
First by the seed be placed on aseptic operating platform with it is sterile washing 5-7 time, then with 70% alcohol infiltration -1 minute 30 seconds, then
It is put into 0.1% liquor hydrargyri perchloridi and sterilizes 12-18 minutes, then with aseptic water washing 5-6 times.
4. according to the method described in claim 1, it is characterised in that in step (2), the Initial culture base is to include N6+6-
BA3.0-5.0mg/L+ cycocels 3.0-5.0mg/L Solid agar culture, preferably comprises N6+6-BA5.0mg/L+ cycocels
5.0mg/L or N6+6-BA4.0mg/L+ cycocels 5.0mg/L Solid agar culture.
5. according to the method described in claim 1, it is characterised in that in step (3), the root media is to include 1/2MS+
Paclobutrazol 0.5-3.0mg/L Solid agar culture；Preferably comprise 1/2MS+ paclobutrazol 1.0mg/L or 1/2MS+ paclobutrazols
1.5mg/L Solid agar culture.
6. according to the method described in claim 1, it is characterised in that in step (4), the proliferated culture medium is to include 1/2MS+
6-BA1.0-2.5mg/L+ paclobutrazol 3.0-5.0mg/L Solid agar culture, preferably comprises 1/2MS+6-BA1.0mg/L+
Paclobutrazol 5.0mg/L or 1/2MS+6-BA2.0mg/L+ paclobutrazol 5.0mg/L Solid agar culture.
7. according to the method described in claim 1, it is characterised in that in step (5), the one-step-seedling formation culture medium be comprising
1/2MS+6-BA1.0-2.5mg/L+ paclobutrazols 3.0-5.0mg/L Solid agar culture.
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| CN108935067A (en) * | 2018-05-17 | 2018-12-07 | 北海市绿邦生物创景有限公司 | The preparation method of Pennisetum inter-species triploid hydrid kind under natural conditions |
| CN110301354A (en) * | 2019-07-23 | 2019-10-08 | 北京市农林科学院 | A kind of breeding method of ' purple leaf ' Chinese pennisetum polyploid |
| CN110338057A (en) * | 2019-07-23 | 2019-10-18 | 北京市农林科学院 | A kind of Chinese pennisetum method for mutation breeding |
| CN114747478A (en) * | 2021-01-15 | 2022-07-15 | 冯永德 | Compound breeding method of pasture |
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| CN101953304A (en) * | 2010-10-19 | 2011-01-26 | 赵国平 | Pennisetum americanum rapid propagation method and pennisetum americanum rapid virus-free propagation method |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101953304A (en) * | 2010-10-19 | 2011-01-26 | 赵国平 | Pennisetum americanum rapid propagation method and pennisetum americanum rapid virus-free propagation method |
| CN106332781A (en) * | 2016-09-29 | 2017-01-18 | 遵义市龙驰生物科技有限公司 | High-efficiency expanding propagation method for crossbred Chinese pennisetum |
| CN106818482A (en) * | 2017-01-25 | 2017-06-13 | 厦门市葳迎生物科技有限公司 | A kind of method of hybrid Chinese pennisetum micropropagation |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108935067A (en) * | 2018-05-17 | 2018-12-07 | 北海市绿邦生物创景有限公司 | The preparation method of Pennisetum inter-species triploid hydrid kind under natural conditions |
| CN110301354A (en) * | 2019-07-23 | 2019-10-08 | 北京市农林科学院 | A kind of breeding method of ' purple leaf ' Chinese pennisetum polyploid |
| CN110338057A (en) * | 2019-07-23 | 2019-10-18 | 北京市农林科学院 | A kind of Chinese pennisetum method for mutation breeding |
| CN114747478A (en) * | 2021-01-15 | 2022-07-15 | 冯永德 | Compound breeding method of pasture |
| CN114747478B (en) * | 2021-01-15 | 2023-10-13 | 冯永德 | Composite breeding method for pasture |
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