CN108739390A - A kind of quick-breeding method of cymose buckwheat rhizome - Google Patents
A kind of quick-breeding method of cymose buckwheat rhizome Download PDFInfo
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- CN108739390A CN108739390A CN201810595363.2A CN201810595363A CN108739390A CN 108739390 A CN108739390 A CN 108739390A CN 201810595363 A CN201810595363 A CN 201810595363A CN 108739390 A CN108739390 A CN 108739390A
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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
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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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Abstract
The present invention relates to a kind of quick-breeding method of cymose buckwheat rhizome, main improvements are, by the composition of improved MS medium, to propose a kind of method for tissue culture suitable for cymose buckwheat rhizome, key step includes inducing clumping bud, adventitious buds proliferation and culture of rootage.The method of the present invention can rapidly breed a large amount of cymose buckwheat rhizome plant; it is calculated by 20 seeds; after sterile breeding of method establishes 40 days; every 25 days 6 generations of Multiplying culture in generation again; and 30 days culture of rootage; about 1,200,000 plants of tissue culture rooted seedlings are can get, are lasted less than 1 year, can be that large-scale planting provides a large amount of seedlings in a short time.
Description
Technical field
The present invention relates to the reproduction technique fields of cymose buckwheat rhizome, and in particular to one kind quickly breeding golden buckwheat by tissue cultures
The method of wheat.
Background technology
Cymose buckwheat rhizome Fagopyrum dibotrys (D.Don) Hara is buckwheat plant, is used as medicine with root, has heat-clearing
The effect of removing toxic substances, apocenosis dissolving stasis, is mainly used for lung carbuncle pyemesis, dyspnea and cough due to lung-heat, acute tonsillitis swelling and pain.Cymose buckwheat rhizome be distributed mainly on southwest,
South China, Central China, East China each province and Shaanxi are born in the mountain valley wetland of 250~3200 meters of height above sea level, hillside shrubbery.At present with this
Plant is mainly wild resource as the source of medicinal material, is excessively developed recently as soil and various environmental pollutions and ecology
The destruction of environment, cymose buckwheat rhizome wild resource has largely reduced, endangered, is put within 1999 II grade of country and lays special stress on protecting plant
Object.Cymose buckwheat rhizome planting technology research is carried out in various regions one after another in recent years, numerous based on seminal propagation, and rhizome is cut supplemented by bud breeding.
But that there are germination percentages is low for seed, germinates situations such as uneven, and to cut bud breeding breeding potential extremely low for stem tuber, significantly limits plantation skill
The breakthrough of art and planting scale.
Invention content
For present technology the problem of cymose buckwheat rhizome breeds, the application proposes one kind by tissue cultures come quickly
The method for breeding cymose buckwheat rhizome, includes the following steps：
1) the cymose buckwheat rhizome seed after surface sterilization is inoculated in budding culture medium, induces Multiple Buds, the budding training
The formula for supporting base is modified MS medium+1.0~4.0mg/L gibberellin；
2) Multiple Buds are cut into simple bud, be inoculated in root media, root induction obtains cymose buckwheat rhizome seedling, described
The formula of root media is+0.5~1.0g/L of improvement MS+0.5~1.0mg/L methyl α-naphthyl acetate+0.5~2.0mg/L heteroauxins
Activated carbon.
The formula of the modified MS medium is：Ammonium nitrate 2.15g/L, potassium dihydrogen phosphate 0.17g/L, magnesium sulfate 0.37g/
L, potassium nitrate 2.30g/L, calcium chloride 0.44g/L, EDTA-Na237.3mg/L, iron sulfite 27.8mg/L, inositol 100.0mg/
L, niacin 0.5mg/L, glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, potassium iodide 0.83mg/
L, boric acid 6.20mg/L, magnesium sulfate 16.90mg/L, cobalt chloride 0.025mg/L, zinc sulfate 8.6mg/L, sodium molybdate 0.25mg/L and
Copper sulphate 0.025mg/L.
The present invention be directed to cymose buckwheat rhizome tissue cultures, the formula of MS culture mediums is optimized, and propose it is suitable
The operating procedure of tissue cultures gives suitable culture medium for each operating procedure, can breed to rapid, high volume golden buckwheat
Wheat.
Preferably, after induction obtains Multiple Buds in the step 1), the Multiple Buds are cut into the stem segment with bud of about 1~2cm
Section, be inoculated in improvement MS+0.05~0.20mg/L heteroauxin+0.5~2.0mg/L 6- benzylaminopurines+0.5~
It is carried out from Multiplying culture of sprouting in the proliferated culture medium of 2.0mg/L kinetins.Cymose buckwheat rhizome seed of the present invention is taken from open country
Outside, therefore the miscellaneous bacteria of explant carrying is more, pollution is susceptible in the budding stage, if directly inducing life with the material after budding
The plant of root, final gained also easily pollutes.By carrying out Multiplying culture to development stage untainted bud, can effectively control
System pollution, is conducive to subsequently largely take root.Moreover, in the inducing clumping bud stage, the induction incidence of Multiple Buds is about 50%,
By carrying out Multiplying culture in above-mentioned culture medium, growth coefficient is 4~5, can greatly increase the quantity of Multiple Buds, Er Qiezhen
For same a collection of Multiple Buds, can Multiplying culture it is multiple, be conducive to the rapid, high volume breeding of bud.
Preferably, the formula of the budding culture medium is modified MS medium+2.8~3.2mg/L gibberellin；
And/or the formula of the proliferated culture medium is MS+0.09~0.11mg/L+1.4~1.6mg/L of heteroauxin 6-
Benzylaminopurine+1.4~1.6mg/L kinetins；
And/or the formula of the root media is improvement MS+0.9~1.0mg/L+1.4~1.6mg/L of methyl α-naphthyl acetate Yin
Indolylbutyric acid+0.9~1.0g/L activated carbons.
It is furthermore preferred that the formula of the budding culture medium is modified MS medium+3mg/L gibberellin；
And/or the formula of the proliferated culture medium be MS+0.1mg/L heteroauxin+1.5mg/L6- benzylaminopurines+
1.5mg/L kinetin.
And/or the formula of the root media be improvement MS+1.0mg/L methyl α-naphthyl acetate+1.5mg/L heteroauxins+
1.0g/L activated carbon；
Preferably, during practical operation, cymose buckwheat rhizome seed is cut off along three triangular prisms, removes covering of a seed,
After the cymose buckwheat rhizome seed for removing crust is carried out disinfection in access budding culture medium.
The condition of culture in step 1) the inducing clumping bud stage be 23~27 DEG C of temperature, intensity of illumination 3000~
4000lux, odd-numbered day light application time 12h；
The take root condition of culture in stage of the step 2) is 23~27 DEG C, 3000~4000lux of intensity of illumination of temperature, single
Day light application time 12h.
Preferably, the condition of culture in adventitious buds proliferation incubation be 23~27 DEG C of temperature, intensity of illumination 3000~
4000lux, odd-numbered day light application time 12h.
Preferably, the incubation time in the induction Multiple Buds stage is 20~25 days；In above-mentioned culture medium and condition of culture
Under, culture can bear a large amount of Multiple Buds in 20~25 days or so, can be because of the nutrition consumption in culture medium if continuing to cultivate for a long time
Lead to the withered of Multiple Buds to the greatest extent, therefore progress Multiplying culture effect is preferable under above-mentioned incubation time.
Preferably, the incubation time of the bud multiplicative stage is 20~25 days, and the growth of Multiple Buds at this time is in peak period,
If continuing to cultivate, the withered of Multiple Buds can be led to because the nutrition in culture medium exhausts.
Preferably, described to disinfect specially：
A, the explant concentrated sulfuric acid is impregnated into 4~6min, 3~4 times is rinsed to no sulfuric acid residual, then with tap water
By the explant and water by volume 1：3~4 mix, 2~3 drop Tween-80s of addition in every 500ml water, and shaking 10~
15min, 40~60min of explant described in tap water shower；
B, explant is transferred to superclean bench, with 75% 30~40s of alcohol disinfecting, sterile water rinses 3~4 times, then
25~30min is sterilized with saturation Eusol, sterile water rinses 3~4 times, finally in 0.1% mercuric chloride solutions+1 of every 500ml
10~11min is sterilized in the solution of~2 drop Tween-80s, sterile water rinses 6~7 times.Explant of the present invention usually picks up from
In field, miscellaneous bacteria is more and complicated, and disinfection way that can be excessively above-mentioned can be effectively removed surface miscellaneous bacteria.
Preferably, the cymose buckwheat rhizome seedling is subjected to hardening treatment, specially：The cymose buckwheat rhizome seedling is connected bottle to be placed in certainly
Seedling is shone in right light greenhouse 2~7 days；Young root seedling is taken out from culture product, after cleaning culture medium, with 500 times more of dilution
Bacterium spirit solution steeps complete stool 3min, is then planted in Nutrition Soil and is cultivated.
As a preferred option, the method for the present invention includes following steps：
1) the cymose buckwheat rhizome seed after surface sterilization is inoculated in budding culture medium, in 23~27 DEG C of temperature, intensity of illumination
It is cultivated 20~25 days under conditions of 3000~4000lux, odd-numbered day light application time 12h, induces Multiple Buds, the budding culture medium
Formula be modified MS medium+2.8~3.2mg/L gibberellin；
2) Multiple Buds are cut into the stem with bud of about 1~2cm, are inoculated in proliferated culture medium, in temperature 23
It~27 DEG C, 3000~4000lux of intensity of illumination, cultivates 20~25 days under conditions of odd-numbered day light application time 12h, obtains and largely grow thickly
The formula of bud, the proliferated culture medium is that MS+0.09~0.11mg/L heteroauxin+1.4~1.6mg/L 6- benzylaminos are fast
Purine+1.4~1.6mg/L kinetins；
3) Multiple Buds are cut into simple bud, be inoculated in root media, in 23~27 DEG C of temperature, intensity of illumination 3000
It is cultivated under conditions of~4000lux, odd-numbered day light application time 12h, root induction obtains cymose buckwheat rhizome seedling, the root media
Formula is improvement MS+0.9~1.0mg/L methyl α-naphthyl acetate+1.4~1.6mg/L heteroauxin+0.9~1.0g/L activated carbons；
The formula of the modified MS medium is：Ammonium nitrate 2.15g/L, potassium dihydrogen phosphate 0.17g/L, magnesium sulfate 0.37g/
L, potassium nitrate 2.30g/L, calcium chloride 0.44g/L, EDTA-Na237.3mg/L, iron sulfite 27.8mg/L, inositol 100.0mg/
L, niacin 0.5mg/L, glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, potassium iodide 0.83mg/
L, boric acid 6.20mg/L, magnesium sulfate 16.90mg/L, cobalt chloride 0.025mg/L, zinc sulfate 8.6mg/L, sodium molybdate 0.25mg/L and
Copper sulphate 0.025mg/L.
Multiplying culture carries out one or many as needed in above-mentioned preferred embodiment, and the time of Multiplying culture will be longer than
The time of bud culture.
Scheme more preferably, the method for the present invention includes following steps：
1) the cymose buckwheat rhizome seed after surface sterilization is inoculated in budding culture medium, in 23~27 DEG C of temperature, intensity of illumination
It is cultivated 20~25 days under conditions of 3000~4000lux, odd-numbered day light application time 12h, induces Multiple Buds, the budding culture medium
Formula be modified MS medium+3.0mg/L gibberellin；
2) Multiple Buds are cut into the stem with bud of about 1~2cm, are inoculated in proliferated culture medium, in temperature 23~27
DEG C, it is cultivated 20~25 days under conditions of 3000~4000lux of intensity of illumination, odd-numbered day light application time 12h, obtains a large amount of Multiple Buds, institute
The formula for stating proliferated culture medium is MS+0.1mg/L heteroauxin+1.5mg/L 6- benzylaminopurine+1.5mg/L kinetins；
3) Multiple Buds are cut into simple bud, be inoculated in root media, in 23~27 DEG C of temperature, intensity of illumination 3000
It is cultivated under conditions of~4000lux, odd-numbered day light application time 12h, root induction obtains cymose buckwheat rhizome seedling, the root media
Formula is improvement MS+1.0mg/L methyl α-naphthyl acetate+1.5mg/L heteroauxin+1.0g/L activated carbons；
The formula of the modified MS medium is：Ammonium nitrate 2.15g/L, potassium dihydrogen phosphate 0.17g/L, magnesium sulfate 0.37g/
L, potassium nitrate 2.30g/L, calcium chloride 0.44g/L, EDTA-Na237.3mg/L, iron sulfite 27.8mg/L, inositol 100.0mg/
L, niacin 0.5mg/L, glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, potassium iodide 0.83mg/
L, boric acid 6.20mg/L, magnesium sulfate 16.90mg/L, cobalt chloride 0.025mg/L, zinc sulfate 8.6mg/L, sodium molybdate 0.25mg/L and
Copper sulphate 0.025mg/L.
Multiplying culture carries out one or many as needed in above-mentioned preferred embodiment, and the time of Multiplying culture will be longer than
The time of bud culture.
Preferably, the kind of cymose buckwheat rhizome described herein is buckwheat plant cymose buckwheat rhizome Fagopyrum
dibotrys(D.Don)Hara。
The method of the present invention has the advantages that：
1) present invention obtains being suitable for cymose buckwheat rhizome by the adjustment of adjustment and culture medium integral formula to MS culture medium prescriptions
The culture medium system and method for tissue culture of tissue cultures.
2) method of the invention can rapidly breed a large amount of cymose buckwheat rhizome plant, be calculated by 20 seeds, through sterile numerous
Educate Establishing 40 days, after the completion of budding culture, based on first generation bud, if per 6 generation of Multiplying culture in 25 days generations, proliferation system
Number is 4~5 and 30 days culture of rootage, can get about 1,200,000 plants of tissue culture rooted seedlings, lasts less than 1 year, if continuing to be proliferated
Available up to ten million or even more than one hundred million plants of tissue culture rooted seedlings can be that large-scale planting provides a large amount of seedlings in a short time by culture.
Description of the drawings
Fig. 1 is the picture that the culture of cymose buckwheat rhizome Seed inducement is germinateed after 2 days；
Fig. 2 is the picture that the culture of cymose buckwheat rhizome Seed inducement grows cotyledon after 6 days；
Fig. 3 is that the culture of cymose buckwheat rhizome Seed inducement the picture of a large amount of Multiple Buds occurs after 20 days；
Fig. 4 is the picture of cymose buckwheat rhizome Multiple Buds culture of rootage, occurs a large amount of root systems after 25 days.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
The formula of involved modified MS medium is in embodiment：Ammonium nitrate 2.15g/L, potassium dihydrogen phosphate 0.17g/L,
Magnesium sulfate 0.37g/L, potassium nitrate 2.30g/L, calcium chloride 0.44g/L, EDTA-Na2 37.3mg/L, iron sulfite 27.8mg/L,
Inositol 100.0mg/L, niacin 0.5mg/L, glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L,
Potassium iodide 0.83mg/L, boric acid 6.20mg/L, magnesium sulfate 16.90mg/L, cobalt chloride 0.025mg/L, zinc sulfate 8.6mg/L, molybdenum
Sour sodium 0.25mg/L and copper sulphate 0.025mg/L.
Embodiment 1
The present embodiment is related to a kind of rapid propagation method of cymose buckwheat rhizome, includes the following steps：
1) kind of buckwheat plant cymose buckwheat rhizome Fagopyrum dibotrys (D.Don) Hara medicinal material cymose buckwheat rhizomes is selected
Son is used as explant；
2) surface sterilization processing is carried out to the explant, the explant concentrated sulfuric acid is impregnated into 4~6min, with originally
Water rinses 3~4 times and remains to no sulfuric acid, then by the explant and water by volume 1：1 mixes, and 2 are added in every 500ml water
~3 drop Tween-80s, shake 10~15min, 40~60min of explant described in tap water shower；
Explant is transferred to superclean bench, with 75% 30~40s of alcohol disinfecting, sterile water rinses 3~4 times, then uses
It is saturated Eusol and sterilizes 25~30min, sterile water rinses 3~4 times, finally in mercuric chloride solution+1~2 of 500ml 0.1%
It drips and sterilizes 10~11min in the solution of Tween-80, sterile water rinses 6~7 times.
3) explant after surface sterilization is inoculated in budding culture medium, in 23~27 DEG C of temperature, intensity of illumination
It is cultivated 15 days under conditions of 3000~4000lux, odd-numbered day light application time 12h, induces Multiple Buds, the budding culture medium is matched
Side is modified MS medium+1.0mg/L gibberellin；Bud ratio is 45%, inductivity 36%；
4) Multiple Buds are cut into the stem with bud of about 1~2cm, be inoculated in improvement MS+0.05mg/L heteroauxins+
In the proliferated culture medium of 0.5mg/L 6- benzylaminopurine+0.5mg/L kinetins, in 23~27 DEG C of temperature, intensity of illumination
3000~4000lux carries out under the condition of culture of odd-numbered day light application time 12h, from Multiplying culture of sprouting, cultivating 20 days or so, obtaining greatly
Measure Multiple Buds；The proliferation rate of Multiple Buds is 86%, and growth coefficient is 2~3.
5) Multiple Buds are cut into simple bud, be inoculated in root media, in 23~27 DEG C of temperature, intensity of illumination 3000
It is cultivated under conditions of~4000lux, odd-numbered day light application time 12h, root induction obtains cymose buckwheat rhizome seedling, the culture of rootage
The formula of base is improvement MS+0.5mg/L methyl α-naphthyl acetate+0.5mg/L heteroauxin+0.5g/L activated carbons；Rooting rate is 83%.
6) hardening culture is carried out to the seedling, the cymose buckwheat rhizome seedling is specially connected into bottle and is placed in natural light greenhouse
Middle solarization seedling 2~7 days；Young root seedling is taken out from culture product, after cleaning culture medium, is steeped with 500 times of carbendazim solution of dilution complete
Strain 3min, is then planted in Nutrition Soil and is cultivated, obtain cymose buckwheat rhizome plant, survival rate 77%.
Embodiment 2
The present embodiment is related to a kind of rapid propagation method of cymose buckwheat rhizome, includes the following steps：
1) kind of buckwheat plant cymose buckwheat rhizome Fagopyrum dibotrys (D.Don) Hara medicinal material cymose buckwheat rhizomes is selected
Son is used as explant；
2) surface sterilization processing is carried out to the explant, the explant concentrated sulfuric acid is impregnated into 4~6min, with originally
Water rinses 3~4 times and remains to no sulfuric acid, then by the implant and water by volume 1：3 mixing, per 300ml water in addition 2~
3 drop Tween-80s, shake 10~15min, 40~60min of explant described in tap water shower；
Explant is transferred to superclean bench, with 75% 30~40s of alcohol disinfecting, sterile water rinses 3~4 times, then uses
It is saturated Eusol and sterilizes 25~30min, sterile water is rinsed 3~4 times, finally dripped in 0.1% mercuric chloride solutions+1~2 of 500ml
10~11min is sterilized in the solution of Tween-80, sterile water rinses 6~7 times.
3) explant after surface sterilization is inoculated in budding culture medium, in 23~27 DEG C of temperature, intensity of illumination
Culture 20 days is carried out under conditions of 3000~4000lux, odd-numbered day light application time 12h, induces Multiple Buds, the budding culture medium
Formula be modified MS medium+3.0mg/L gibberellin, bud ratio 60%, inductivity 50%
4) Multiple Buds are cut into the stem with bud of about 1~2cm, be inoculated in improvement MS+0.1mg/L heteroauxins+
In the proliferated culture medium of 1.5mg/L6- benzylaminopurine+1.5mg/L kinetins, in 23~27 DEG C of temperature, intensity of illumination 3000
~4000lux carries out under the condition of culture of odd-numbered day light application time 12h, from Multiplying culture of sprouting, cultivating 22 days or so, obtaining largely clump
It sprouts, the proliferation rate of Multiple Buds is 95%, and growth coefficient is 4~5.
5) Multiple Buds are cut into simple bud, be inoculated in root media, in 23~27 DEG C of temperature, intensity of illumination 3000
It is cultivated under conditions of~4000lux, odd-numbered day light application time 12h, root induction obtains cymose buckwheat rhizome seedling, the culture of rootage
The formula of base is improvement MS+1.0mg/L methyl α-naphthyl acetate+1.5mg/L heteroauxin+1.0g/L activated carbons, rooting rate 96%；
6) hardening culture is carried out to the seedling, the cymose buckwheat rhizome seedling is specially connected into bottle and is placed in natural light greenhouse
Middle solarization seedling 2~7 days；Young root seedling is taken out from culture product, after cleaning culture medium, is steeped with 500 times of carbendazim solution of dilution complete
Strain 3min, is then planted in Nutrition Soil and is cultivated, obtain cymose buckwheat rhizome plant, survival rate 86%, the present embodiment different phase
Plant growth situation is shown in Fig. 1~4.
Embodiment 3
1) select the seed of buckwheat plant cymose buckwheat rhizome Fagopyrum dibotrys (D.Don) Hara as explant
Body；
2) surface sterilization processing is carried out to the explant, the explant concentrated sulfuric acid is impregnated into 4~6min, with originally
Water rinses 3~4 times and remains to no sulfuric acid, then by the implant and water by volume 1：4 mixing, per 500ml water in addition 2~
3 drop Tween-80s, shake 10~15min, 40~60min of explant described in tap water shower；
Explant is transferred to superclean bench, with 75% 30~40s of alcohol disinfecting, sterile water rinses 3~4 times, then uses
It is saturated Eusol and sterilizes 25~30min, sterile water is rinsed 3~4 times, finally dripped in 0.1% mercuric chloride solutions+1~2 of 500ml
10~11min is sterilized in the solution of Tween-80, sterile water rinses 6~7 times.
3) explant after surface sterilization is inoculated in budding culture medium, in 23~27 DEG C of temperature, intensity of illumination
Culture 25 days is carried out under conditions of 3000~4000lux, odd-numbered day light application time 12h, induces Multiple Buds, the budding culture medium
Formula be modified MS medium+4.0mg/L gibberellin, bud ratio 49%, inductivity 51%；
4) Multiple Buds are cut into the stem with bud of about 1~2cm, be inoculated in improvement MS+0.2mg/L heteroauxins+
In the proliferated culture medium of 2.0mg/L6- benzylaminopurine+2.0mg/L kinetins, in 23~27 DEG C of temperature, intensity of illumination 3000
~4000lux carries out under the condition of culture of odd-numbered day light application time 12h, from Multiplying culture of sprouting, cultivating 30 days or so, obtaining largely clump
It sprouts, the proliferation rate of Multiple Buds is 94%, and growth coefficient is 4~5.
5) Multiple Buds are cut into simple bud, be inoculated in root media, in 23~27 DEG C of temperature, intensity of illumination 3000
It is cultivated under conditions of~4000lux, odd-numbered day light application time 12h, root induction obtains cymose buckwheat rhizome seedling, the culture of rootage
The formula of base is improvement MS+1.0mg/L methyl α-naphthyl acetate+2.0mg/L heteroauxin+1.0g/L activated carbons, rooting rate 95%；
6) hardening culture is carried out to the seedling, the cymose buckwheat rhizome seedling is specially connected into bottle and is placed in natural light greenhouse
Middle solarization seedling 2~7 days；Young root seedling is taken out from culture product, after cleaning culture medium, is steeped with 500 times of carbendazim solution of dilution complete
Strain 3min, is then planted in Nutrition Soil and is cultivated, obtain cymose buckwheat rhizome plant, survival rate 88%.
Comparative example 1
Compared with Example 2, difference lies in do not use the MS culture mediums of improvement, using traditional MS culture mediums, as a result
Identical in the addition situation of hormone, under identical cultivated days, bud ratio 45%, inductivity is about 33%, the proliferation of Multiple Buds
Rate is 78%, and growth coefficient is 1~2, and culture of rootage rooting rate is 56%, and hardening culture survival rate is 85%.
Comparative example 2
Compared with Example 2, difference lies in the formula of the budding culture medium is that 1/2MS culture mediums+2.0mg/L is red
Mycin+25g/L white sugar+PH6.2, as a result bud ratio 45%, inductivity are about 33%.
Comparative example 3
Compared with Example 2, difference lies in the formula of, the proliferated culture medium be MS+0.1mg/L methyl α-naphthyl acetates+
2.0mg/L6- benzylaminopurine+1.0mg/L activated carbons, as a result the proliferation rate of Multiple Buds is 78%, and growth coefficient is 1~2
Comparative example 4
Compared with Example 2, difference lies in the formula of, the root media be MS+0.1mg/L methyl α-naphthyl acetates+
1.0mg/L heteroauxin+0.5g/L activated carbons, as a result rooting rate is 56%.
Although above having used general explanation, specific implementation mode and experiment, the present invention is made to retouch in detail
It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Range.
Claims (10)
1. a kind of quick-breeding method of cymose buckwheat rhizome, which is characterized in that include the following steps：
1) the cymose buckwheat rhizome seed after surface sterilization is inoculated in budding culture medium, induces Multiple Buds, the budding culture medium
Formula be modified MS medium+1.0~4.0mg/L gibberellin；
2) Multiple Buds are cut into simple bud, be inoculated in root media, root induction obtains cymose buckwheat rhizome seedling, described to take root
The formula of culture medium is improvement MS+0.5~1.0mg/L methyl α-naphthyl acetate+0.5~2.0mg/L heteroauxins+0.5~1.0g/L activity
Charcoal；
The formula of the modified MS medium is：Ammonium nitrate 2.15g/L, potassium dihydrogen phosphate 0.17g/L, magnesium sulfate 0.37g/L, nitre
Sour potassium 2.30g/L, calcium chloride 0.44g/L, EDTA-Na2 37.3mg/L, iron sulfite 27.8mg/L, inositol 100.0mg/L, cigarette
Sour 0.5mg/L, glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, potassium iodide 0.83mg/L, boron
Sour 6.20mg/L, magnesium sulfate 16.90mg/L, cobalt chloride 0.025mg/L, zinc sulfate 8.6mg/L, sodium molybdate 0.25mg/L and sulfuric acid
Copper 0.025mg/L.
2., will be described according to the method described in claim 1, it is characterized in that, after induction obtains Multiple Buds in the step 1)
Multiple Buds are cut into the stem with bud of about 1~2cm, are inoculated in+0.5~2.0mg/L of improvement MS+0.05~0.20mg/L heteroauxins
It is carried out from Multiplying culture of sprouting in the proliferated culture medium of 6- benzylaminopurine+0.5~2.0mg/L kinetins.
3. method according to claim 1 or 2, which is characterized in that the formula of the budding culture medium is improvement MS cultures
Base+2.8~3.2mg/L gibberellin；
And/or the formula of the proliferated culture medium is MS+0.09~0.11mg/L heteroauxin+1.4~1.6mg/L 6- benzyls
Adenine phosphate+1.4~1.6mg/L kinetins；
And/or the formula of the root media is improvement MS+0.9~1.0mg/L methyl α-naphthyl acetate+1.4~1.6mg/L indoles second
Acid+0.9~1.0g/L activated carbons.
4. according to claims 1 to 3 any one of them method, which is characterized in that step 1) the inducing clumping bud stage
Condition of culture is 23~27 DEG C, 3000~4000lux of intensity of illumination of temperature, odd-numbered day light application time 12h；
The step 2) take root the stage culture condition be 23~27 DEG C, 3000~4000lux of intensity of illumination of temperature, odd-numbered day light
According to time 12h.
5. according to claim 2~4 any one of them method, which is characterized in that the culture in adventitious buds proliferation incubation
Condition is 23~27 DEG C, 3000~4000lux of intensity of illumination of temperature, odd-numbered day light application time 12h.
6. according to the method described in claim 4, it is characterized in that, the time of inducing clumping bud culture is 20 in the step 1)
~25 days.
7. according to the method described in claim 5, it is characterized in that, the incubation time during the Multiplying culture is 20~25
It.
8. according to claim 1~7 any one of them method, which is characterized in that described to disinfect specially：
A, the explant concentrated sulfuric acid is impregnated into 4~6min, 3~4 times is rinsed to no sulfuric acid residual, then by institute with tap water
State explant and water by volume 1：3~4 mix, and 2~3 drop Tween-80s of addition, shake 10~15min, use in every 500ml water
40~60min of explant described in tap water shower；
B, explant is transferred to superclean bench, with 75% 30~40s of alcohol disinfecting, sterile water rinses 3~4 times, then with full
25~30min is sterilized with Eusol, sterile water rinses 3~4 times, and finally the 500ml mercuric chloride solutions+1~2 0.1% drip
10~11min is sterilized in the solution of Tween-80, sterile water rinses 6~7 times.
9. according to claim 1~8 any one of them method, which is characterized in that include the following steps：
1) the cymose buckwheat rhizome seed after surface sterilization is inoculated in budding culture medium, in 23~27 DEG C of temperature, intensity of illumination 3000
It is cultivated 20~25 days under conditions of~4000lux, odd-numbered day light application time 12h, induces Multiple Buds, the budding culture medium is matched
Side is modified MS medium+2.8~3.2mg/L gibberellin；
2) Multiple Buds are cut into the stem with bud of about 1~2cm, are inoculated in proliferated culture medium, in 23~27 DEG C of temperature, light
According to 3000~4000lux of intensity, is cultivated 20~25 days under conditions of odd-numbered day light application time 12h, obtain a large amount of Multiple Buds, the proliferation
The formula of culture medium be MS+0.09~0.11mg/L heteroauxin+1.4~1.6mg/L 6- benzylaminopurines+1.4~
1.6mg/L kinetin；
3) Multiple Buds are cut into simple bud, be inoculated in root media, in 23~27 DEG C of temperature, intensity of illumination 3000~
It is cultivated under conditions of 4000lux, odd-numbered day light application time 12h, root induction obtains cymose buckwheat rhizome seedling, and the root media is matched
Side is improvement MS+0.9~1.0mg/L methyl α-naphthyl acetate+1.4~1.6mg/L heteroauxin+0.9~1.0g/L activated carbons；
Multiplying culture progress is one or many, and the time of single Multiplying culture will be longer than the time of budding culture；
The formula of the modified MS medium is：Ammonium nitrate 2.15g/L, potassium dihydrogen phosphate 0.17g/L, magnesium sulfate 0.37g/L, nitre
Sour potassium 2.30g/L, calcium chloride 0.44g/L, EDTA-Na2 37.3mg/L, iron sulfite 27.8mg/L, inositol 100.0mg/L, cigarette
Sour 0.5mg/L, glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, potassium iodide 0.83mg/L, boron
Sour 6.20mg/L, magnesium sulfate 16.90mg/L, cobalt chloride 0.025mg/L, zinc sulfate 8.6mg/L, sodium molybdate 0.25mg/L and sulfuric acid
Copper 0.025mg/L.
10. according to claim 1~9 any one of them method, which is characterized in that the cymose buckwheat rhizome is buckwheat plant
Cymose buckwheat rhizome Fagopyrum dibotrys (D.Don) Hara.
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| CN111134123A (en) * | 2019-12-19 | 2020-05-12 | 东莞市东阳光冬虫夏草中药有限公司 | Seed germination treatment agent and application |
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| CN111134123A (en) * | 2019-12-19 | 2020-05-12 | 东莞市东阳光冬虫夏草中药有限公司 | Seed germination treatment agent and application |
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