CN105123750A - Mold bacteriostatic agent for open culture of potato tissue culture seedlings and using method thereof - Google Patents
Mold bacteriostatic agent for open culture of potato tissue culture seedlings and using method thereof Download PDFInfo
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- CN105123750A CN105123750A CN201510663986.5A CN201510663986A CN105123750A CN 105123750 A CN105123750 A CN 105123750A CN 201510663986 A CN201510663986 A CN 201510663986A CN 105123750 A CN105123750 A CN 105123750A
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Abstract
The invention discloses a mold bacteriostatic agent for open culture of potato tissue culture seedlings and a using method thereof. Sodium hypochlorite of the certain concentration is adopted as the bacteriostatic agent for open tissue culture of potatoes. The mold bacteriostatic agent has the ideal mold inhibiting effect, has the small effect on potato rooting and growth, can be used for open tissue culture of potatoes, can lower mold pollution of potato test-tube seedlings, and provides the feasible scheme for lowering the contamination rate and production cost in industrial potato production of test-tube seedlings. The mold bacteriostatic agent is simple and easy to use, low in cost and good in using effect.
Description
Technical field
The present invention relates to biological technical field, especially a kind of mould bacteriostatic agent of open cultivation potato tissue culture seedling.
Background technology
The Fast-propagation of Virus-free Tube Potato Plantlets is the basic link that potato primary stock is produced, and ensures and improve test-tube plantlet quality numerous soon to produce significant to seed potato within the relatively short time.But often because the pollution of different mushroom appears in a variety of causes in Vitro Rapid Propagation of Virus-free Potato process, as bacterium, mould, saccharomycete, actinomycetes etc., these growth of microorganism speed are fast.Bacterium, saccharomycete, actinomycetes pollute and can remove pollution after 2 ~ 3 switchings.And mould contamination directly can invade test-tube plantlet organization internal, be difficult to process with general sterilization method, the diffusion of spore can cause large-area pollution, and expansion rate is very fast, and after diffusion, Regenerated plant still all shows as pollution.Test-tube plantlet growth after mould contamination is slow, and growing way is weak, takes root and takes root less or not, pollutes transplantation of seedlings survival rate low, has a strong impact on the growth of test-tube plantlet, bring loss to production.In order to solve the problem polluted in the fast numerous process of potato, for the research work that Control pollution in Vitro Rapid Propagation of Virus-free Potato occurs, there has been some reports.
Chen Yalan (2012) with the medical antibiotic of variable concentrations, preservative, chemical pesticide for bacteriostatic agent, medium direct syringe after high-temperature sterilization injects antibacterial substance, the fungistatic effect in research potato tissue culture seedling and propagating and the impact growing plantlet in vitro and take root.Result of the test shows, 70% thiophanate-methyl wetting powder and 5O% carbendazol wettable powder potato tissue culture seedling is produced in modal Penicillium notatum, bacterium and saccharomycete 3 kinds of bacterium all there is better inhibitory action, and the negatively influencing produced the growth of potato test-tube plantlet and taking root is minimum.
Peng Guanglin etc. (2012) add 5 respectively in MS medium, l0,2O, 3O and 40mg/L clorox, with potato tissue culture seedling for test material, analyze the control efficiency polluted plantlet in vitro hay bacillus (bacterium).Result shows, after 2 ~ 3 switchings, 5 and 10mg/L clorox better to the fungistatic effect of potato tissue culture seedling hay bacillus, polluting does not appear in the plantlet in vitro of this concentration processed group, and seedling growth is better, but obviously decline along with the rising control efficiency of concentration.
(2012) employing plate growth inhibition assay such as Su Fei flies, use
no. mono-, kuh-seng, Chinese herbal medicine and Hei Nong3 Plants source bactericides,
penicillium notatum fungistatic effect in Plant Tissue Breeding is studied.being mixed with the liquid of variable concentrations gradient respectively, contrasting the medium for not adding botanical fungicide, after autoclave sterilization, accurately measure a certain amount of liquid and join in the medium of heat.Result shows, add black agriculture bacteriostatic agent in the medium and suppress the effect of Penicillium notatum best, concentration is 2.0% time, and bacteriostasis rate reaches 100.0%.
Su Fei to fly etc. (2010) add in the medium kanamycin, penicillin, streptomycin and penicillin and streptomycin with the use of antibiotic, research
bacteriostatic agentthe effect of the anti-bacteria in the fast numerous process of potato.Result shows: the mixture adding kanamycin, novocillin, penicillin and streptomycin in the medium can the pollution of anti-bacteria, but penicillin respectively processes and can not only pollute by anti-bacteria, and have obvious facilitation to the growth of test-tube plantlet, wherein the effect of penicillin 60mgL-is best, and the bacteriostasis of bacteriostatic agent has ageing simultaneously.
Wang Chun (2004) tests 4 kinds of antibiotic fungistatic effects in potato tissue is cultivated.Result shows, when medium adds 20mg/L ampicillin sodium or streptomycin sulphate, effectively can suppress the bacillary dip-dye of kind Atlantic Ocean test-tube plantlet
.
The Sodium Benzoate of Yang Chen (2012) experimental observation variable concentrations and potassium sorbate pressing down in Vitro Rapid Propagation of Virus-free Potato
bacterium effect.Result shows, in MS medium, add potassium sorbate has significantly
fungistatic effect, and have obvious facilitation to the growth of test-tube plantlet, wherein best with the consumption effect of 20mg/L.
Summary of the invention
The object of the invention is: the mould bacteriostatic agent and the using method thereof that provide a kind of open cultivation potato tissue culture seedling, it can reduce the pollution of mould in potato test-tube plantlet, reduces pollution rate and production cost.
Technical solution problem of the present invention adopts following technical scheme: the mould bacteriostatic agent of open cultivation potato tissue culture seedling, using clorox as sole component.
The using method of the mould bacteriostatic agent of open cultivation potato tissue culture seedling, adds clorox in the medium of open cultivation potato tissue culture seedling, makes it account for 0.5% of medium gross mass.
Before carrying out open cultivation, first adopt the utensil of liquor natrii hypochloritis to open cultivation to clean, then use
cold boiling waterscrubber, then carry out open cultivation by flow process.
In order to verify technique effect of the present invention, inventors performed following experiment:
1 test material
1.1 mould
The mold species of test is separated to obtain from the plantlet in vitro polluted.
1.2 antibacterial class reagent
Allicin: Solarbio, 1g import packing, the boiled thawing of medium or sterilizing slightly cold time add, because valency is high, only for testing 2.
fresh garlic: market is bought.Take 50g fresh garlic, fruit agitator is smashed, and dissolves, 3 layers of filtered through gauze, then use the sterile filter of import with 50ml sterile water.Allicin is at high temperature easily destroyed, and loses bactericidal action, therefore the boiled thawing of medium or sterilizing slightly cold time add, for testing 3.About calculate (data source: net news), containing 0.5g(and 500mg in 50g garlic containing the allicin of 1% by garlic) allicin.The liquor capacity obtained by filtration is containing allicin 7.8mg in the fresh garlic liquid of 64ml, 1ml, and the fresh garlic liquid consumption of test 3 is 5ml, namely containing the about 40mg/L of allicin.
Clorox: Tianjin Fu Yu Fine Chemical Co., Ltd produces, effective chlorine composition is >=10%, 500ml dress.
Plant training spirit: Yu Han bio tech ltd, Shanghai produces, and batch number is 141218,
100ml.
Benefit training is grand: Yu Han bio tech ltd, Shanghai, batch number 141015,
10g.
Carbendazim a:Aladdin, C
9h
9n
3o
2, MW=191.19, analyze pure (%) 1g, valency is high, only for testing 1.
Carbendazim b: biochemical science (group) Co., Ltd of Canadian triumphant nurse Cole produces, and import 400g fills, 80%WP, for testing 2 and test 3.
1.3 medium
Mushroom and Potato Plantlets in vitro medium are the MS medium (permanent in company's purchase) of purchase.
1.4 potato tissue culture seedlings
This test adopts " Favorita " mutant breeding 5-6 to be test material for stable strain 37A and at potato " western red " plantlet in vitro of the red Beijing opera meat of Tibet introduction,
with the former strain with bacteriostatic agent for contrast.
test method and result
2.1 tests 1
2.1.1 date of test and place: on February 10th, 2015, Guizhou Bio-technology Inst. laboratory.
2.1.2 test process is arranged
1) clorox (C): concentration gradient is 0.0% (CK), 0.005%, 0.01%, 0.02%, 0.03%; The strain of every bottle graft kind potato tissue culture seedling 8, each process 5 bottles (following process all with);
2) carbendazim a (DJ): 0% (CK), 15mg/L, 30mg/L, 45mg/L, 60mg/L, 75mg/L;
3) benefit training grand (Y): 0.0% (CK), 0.3%(manufacturer's recommended concentration);
4) training spirit (Z): 0.0% (CK) is planted, 2.5ml/L(manufacturer's recommended concentration).
2.1.3 medium treatment and data statistics: all medium autoclave sterilization treatment.Randomly draw 3 bottle of 20 strain and observe plant height and situation of taking root.
2.1.4 result of the test
1) plant height (5.99cm) of C process is apparently higher than contrast
(5.19cm), DJ
(3.87cm), Y (3.58cm), Z (3.65cm) process.The plant height (3.87cm) of DJ process compared with contrast (2.51cm) a little more than contrasting (table 1).
2) C process increases gradually with process increasing concentrations plant height; The plant height of DJ process increases with the increasing of concentration in 45mg/L concentration range, and exceed this concentration, plant height reduces (table 1) with increasing concentrations.
3) according to observations, compared with CK, the obvious undergrowth of root of DJ, Y, Z3 process, the root of more than 95% plant is 0-2 bar, and shorter (0.3-0.5cm).And the root of C process is 3-8 bar, length is 3-5cm, close with CK.
2.1.5 the 1st result of the test is summed up
1) concentration that adopts of test on plant height between process to affect difference not obvious;
2) root growth of DJ, Y, Z3 process is suppressed;
3) total Test does not pollute.
According to the 1st result of the test, Adjustment Tests designs, and strengthens the concentration gradient between process, under open condition, inoculates mould test.
test 2
2.2.1 date of test and place: on February 11st, 2015, Guizhou Bio-technology Inst. laboratory.
2.2.2 setting is tested
1) clorox (C): concentration is treated to 0.0% (CK), 0.01% (C
1), 0.05% (C
2), 0.1% (C
3), 0.5% (C
4), 1.0% (C
5); Each process disinfection inoculation plantlet in vitro 5 bottles, every bottle of 15 strains; Open inoculation mould, each 3, concentration inoculated and cultured ware, each inoculation mould bacterium block 1 piece (2-3mm is shown in Fig. 1), contrast is not with bacteriostatic agent and opens the process of same time under the same conditions;
2) carbendazim b (DJ): concentration is treated to 0mg/L (CK), 20mg/L (DJ
1), 40mg/L (DJ
2), 60mg/L (DJ
3), 80mg/L (DJ
4), 100mg/L (DJ
5), every bottle of 10 strains, each process disinfection inoculation plantlet in vitro 5 bottles, every bottle of 15 strains; Open inoculation mould, each 3, concentration inoculated and cultured ware, each inoculation mould bacterium block 1 piece (2-3mm is shown in Fig. 1), contrast is not with bacteriostatic agent and opens the process of same time under the same conditions;
3) benefit training grand (Y): concentration is treated to 0.0% (CK), 0.1% (Y
1), 0.2% (Y
2), 0.3%(Y
3, manufacturer's recommended concentration), 0.4% (Y
4), 0.5% (Y
5); Every bottle of 10 strains, each process disinfection inoculation plantlet in vitro 5 bottles, every bottle of 15 strains; Open inoculation mould, each 3, concentration inoculated and cultured ware, each inoculation mould bacterium block 1 piece (2-3mm is shown in Fig. 1), contrast is not with bacteriostatic agent and opens the process of same time under the same conditions;
4) training spirit (Z) is planted: concentration is treated to 0.0ml/L (CK), 1.5ml/L (Z
1), 2.0ml/L (Z
2), 2.5ml/L(manufacturer's recommended concentration) (Z
3), 3.0ml/L (Z
4), 3.5ml/L (Z
5), each process disinfection inoculation plantlet in vitro 5 bottles, every bottle of 15 strains; Open inoculation mould, each 3, concentration inoculated and cultured ware, each inoculation mould bacterium block 1 piece (2-3mm is shown in Fig. 1), contrast is not with bacteriostatic agent and opens the process of same time under the same conditions;
5) allicin (DS): concentration is treated to
0% (cK), 1mg/L (DS
1), 2mg/L (DS
2), 3mg/L (DS
3), 4mg/L (DS
4), 5mg/L (DS
5), each process disinfection inoculation plantlet in vitro 5 bottles, every bottle of 15 strains; Open inoculation mould, each 3, concentration inoculated and cultured ware, each inoculation mould bacterium block 1 piece (2-3mm is shown in Fig. 1), contrast is not with bacteriostatic agent and opens the process of same time under the same conditions;
2.2.3 medium and data statistics
1) inoculate the whole autoclave sterilizing of medium of plantlet in vitro process, the culture dish of inoculation mould is only boiled by medium, (doorway, laboratory) inoculation under open condition.
2)
after cultivating 15d, investigate potato tissue culture seedling pollution rate, randomly draw 20 strain plantlet in vitro and investigate plant heights and situations of taking root, and the growing state of mushroom under open condition.
2.2.4 result of the test
1) mould test is inoculated under open condition
A. from result of the test, the Y process of variable concentrations and DS process all pollute (Fig. 2), and these 2 bacteriostatic agents do not have depression effect to mould;
B.C
4and C
5whole 3 culture dishes 100% of process inoculation do not have mould contamination (Fig. 2), show that the C process of high concentration has good fungistatic effect to mould;
C.Z process only has Z
3process 100% does not have mould contamination, all seriously infects (Fig. 2) by mycelia higher or lower than this concentration.This is an interesting result, seems to think that the mould of this concentration to inoculation has the effect invading and kill, and just as the alcohol energy sterilization of 75%, and the alcohol of 95% does not just have bactericidal effect the same.
The DJ of d.DJ process
2, DJ
4and DJ
5there is certain fungistatic effect, but can not the growth of 100% complete mould fungus inhibition.At DJ
2inoculation 3 culture dishes in, have the mycelia of 2 culture dishes slightly to grow; At DJ
4and DJ
5process in, have the mycelia of 1 culture dish slightly to grow, DJ
5bacterial plaque be greater than DJ
4bacterial plaque (Fig. 2).Therefore, comparatively speaking, relatively better with the effect of DJ4.
E. from contrast, certain bacterium and mould contamination is all had,
withgerm contamination is main, but the mushroom expansion rate polluted relatively slow (Fig. 3).
2) plantlet in vitro test is inoculated under different disposal sterilising conditions
Be more or less the same from CK and the C1 of Fig. 4, C process, C2, C3 plant height, about 10-12cm, radical is at 3-7 root, and root is long is 5-10cm, and leaf look green, and the number of blade is many.Only C4 slightly difference, plant height is 5-8cm, and root is long substantially identical with the process of other C concentration with radical.
Plant height and the C process of DJ, DS, Z, Y process are basically identical, but the growth of root is subject to suppression in various degree, and with the increase of concentration, depression effect is further obvious.Whole process, comprising contrast does not have pollution.
2.2.5 the 2nd result of the test is summed up
1) with regard to the concentration that test adopts, between process, plant height difference when low concentration is not obvious, shows certain depression effect when high concentration;
2) under a high concentration condition, except C process, the growth of root suppresses by serious, and this is consistent with the result of test 1;
3) under the condition of medium sterilization, all the test of inoculation plantlet in vitro is not polluted;
4) under open condition, the inoculation test result of mould shows, the bacteriostatic agent selected by this test has certain inhibitory action to mould, and wherein, the best process of fungistatic effect has C
4, C
5, Z
3, process preferably has DJ
4.
The data display of afore-mentioned test 1, test 2, certain density bacteriostatic agent C, Z, DJ have certain inhibitory action to group training mould contamination, and C process is further remarkable with the increasing fungistatic effect of concentration.But result of the test shows, the growth of bacteriostatic agent to plantlet in vitro of higher concentration has certain inhibitory action, also can seriously suppress the formation of root even to cause the death of plantlet in vitro.Therefore, testing 3 selects certain concentration and the concentration that filtered out to test.
2.3 tests 3
2.3.1 date of test and place: on July 30th, 2015, Guizhou Bio-technology Inst. laboratory.
2.3.2 experimental scheme and method
According to afore-mentioned test result, the concentration that test 3 selects test 2 to screen, namely C process only adopts C
4concentration, Z process only adopts Z
3concentration, DJ process only adopts DJ
4concentration.Due to a large amount of reporting, the fungistatic effect of allicin (DS) is fine, test 3
do againthe bacteriostatic test of a DS.DS adopt 5ml/L fresh garlic liquid (
be about 40mg/L allicin).Aforementioned 4 kinds of bacteriostatic agents are mixed by afore mentioned concentration and is called mixed culture medium (mixing).Namely test 3 and have 5 process (DS, DJ, C, Z, mix), 1 concentration is selected in each process.
The cultivation basigamy 400ml of each process, 150ml are used for autoclaving,
in addition 250ml medium boiled after under open condition packing inoculation.autoclaving group often processes inoculation 4 bottles, every bottle graft kind 5 strain plantlet in vitro.Open inoculation group often processes inoculation 4 bottles, every bottle graft kind 5 strain plantlet in vitro;
meanwhile, under open condition, every component fills 4 culture dishes, and inoculates 3 kinds of moulds under open condition(No.-1, mould, grey mold-No. 2,
no.-3, white mould).The bottle of open inoculation group, bottle cap, instrument and culture dish all soak about 1min with the liquor natrii hypochloritis of 10%, use after then cleaning 2 times with the boiling water of cooling.
2.3.3 result of the test
1) plantlet in vitro of sterilization method inoculation is adopted not pollute.Adopt the plantlet in vitro of Open-Architecture Controller, DS, DJ all pollute; C
4, Z
3, mixed 3 process plantlet in vitro do not have pollution.Can be used for the open group of training of potato.
2) different disposal inoculation mould test result
DJ process: medium overgrows with white hypha (No. 3 bacterium), No. 1 and No. 2 mycelia also grow to some extent.
DS material: DJ process is slightly better than to the inhibitory action of No. 3 mycelia, but the mycelia of No. 1 and No. 2 mould grows to some extent.
Z process: close with DS to the inhibitory action of 1, No. 2 mould, is slightly better than DS process to the inhibitory action of No. 3 moulds.
Mixed culture medium: have very strong depression effect to No. 2 bacterium is close with Z to the inhibitory action of No. 3 bacterium.
C process: have very strong depression effect to 1, No. 2 mould.50% inhibitory action is shown as to No. 3 bacterium.In all process
resultthe process behaved oneself best.
sum up
1) benefit training grand (Y) is in the tissue-culturing quick-propagation of potato, does not have depression effect to mould, and to the growth of potato tissue culture seedling with take root and have obvious inhibitory action.
2) allicin (DS): in the tissue-culturing quick-propagation of potato, depression effect is not had to mould, to the growth of potato tissue culture seedling and Rooting effect little.
3) carbendazim (DJ): not obvious to the depression effect of the fast numerous middle mould of potato tissue culture, and to the growth of potato tissue culture seedling with take root and have obvious inhibitory action.
4) plant training spirit (Z): concentration be 2.5ml/L plant training spirit numerous soon to potato tissue culture in mould have good depression effect, to the growth of potato tissue culture seedling with take root and have certain inhibitory action, can be used for the open group of training of potato.
5) clorox (C): certain density clorox process (>0.5%) has the very strong depression effect (white mould of test 2 to mould in potato tissue culture is numerous soon, and the suppression of the equal energy 100% of test 3 No. 1 and No. 2 moulds), to the growth of potato tissue culture seedling and rooting inhibition of cuttings effect not obvious.Under finite concentration, can be used for the open group of training of potato.
6) theoretically, the fungistatic effect of mixed culture medium should be better than single factor test medium, but in mould inoculation test, its fungistatic effect inferior to C process, may be the mixing of various chemistry or biological bacteriostatic agent make its have certain mutual work and
the effect of mutual suppression.Our result of the test shows, mixed culture medium also can be used for the open group of training of potato.
7) bacteriostatic agent cultivated for potato tissue is reported few.In our data-searching, have no report with clorox as the bacteriostatic agent of mould in potato tissue culture.Chen Yalan's (2012) is that thiophanate-methyl and carbendazim are as bacteriostatic agent, the control efficiency that Peng Guanglin etc. (2012) pollute potato tissue culture seedling hay bacillus (bacterium) with clorox research, Su Fei (2010), the Wang Chun (2004) such as to fly and utilizes the antibiosis usually bacterium of inhibition of potato plantlet in vitro and the pollution of fungi, and Su Fei to fly etc. (2012) and uses
3 kindspenicillium notatum fungistatic effect in the cultivation of botanical fungicide research organization, what this test adopted is sterilization method, is inoculated on plating medium and cultivates, do not test with potato tissue culture seedling.Yang Chen (2012) adopts in Sodium Benzoate and potassium sorbate 2 kinds of preservative experimental observation Vitro Rapid Propagation of Virus-free Potatos
antibacterialeffect, potassium sorbate has significantly
fungistatic effect.Above-mentioned test is the result of the test obtained under sterilising conditions mostly.
conclusion
1) plant that the clever concentration of training is 2.5ml/L, open tissue-culturing rapid propagation that clorox process (0.5%) and mixed culture medium can be considered for potato.
2) consider production cost, and on the impact that plantlet in vitro grows and takes root, recommend the bacteriostatic agent using 0.5% clorox as potato tissue culture, produce for sterilizing or open group of training in conjunction with the operation sequence described in this test.
3)
this test period is in spring, summer, and in Guiyang Region, Guizhou Province, this period is the season that in 1 year, humidity is comparatively large, temperature is higher, is also one of fungus growth the most vigorous period.Therefore, this result of the test for group training in other production effects in season more obvious with have reference.
Owing to have employed above technical scheme, compared with prior art, the bacteriostatic agent that the present invention adopts certain density clorox to train as open group of potato, it is ideal to the inhibition of mould, and taking root and growth effect to potato
comparativelylittle, can be used for the open group of training of potato, the pollution of potato test-tube plantlet mould can be reduced, for reducing pollution rate in potato factorial praluction test-tube plantlet and production cost provides feasible program.The present invention is simple, with low cost, and result of use is good.
Accompanying drawing explanation
Accompanying drawing 1
for testing 2open inoculation mould; Each culture dish inoculation white mould bacterium 1 (inoculation on March 11st, 2015)
Accompanying drawing 2 is
test 2mould inoculation test result under different disposal open condition, each culture dish inoculation white mould bacterium 1 (investigation date: on March 26th, 2015);
Accompanying drawing 3 is
test 2the pollution result of CK under open condition (investigation date: on March 26th, 2015);
Accompanying drawing 4 is
test 2the plantlet in vitro growing state (investigation date: on March 11st, 2015) of different disposal;
Accompanying drawing 5
for testing 3 kinds of separating moulds of 3 inoculations, be from left to right respectively mould No. 1, No. 2, No. 3;
Accompanying drawing 6 is
test 3the result (observing time: on August 11st, 2015) of different disposal Open-Architecture Controller mould.
Embodiment
Embodiments of the invention 1: the mould bacteriostatic agent of open cultivation potato tissue culture seedling, using clorox as sole component.
Embodiments of the invention 2: the use of the mould bacteriostatic agent of open cultivation potato tissue culture seedling: before carrying out open cultivation, first adopt the utensil of liquor natrii hypochloritis to open cultivation to clean, then use
cold boiling waterscrubber; Get out the open medium of potato tissue culture seedling, add in this medium account for medium gross mass 0.5% clorox, carry out open group of training by flow process.
Claims (3)
1. a mould bacteriostatic agent for open cultivation potato tissue culture seedling, is characterized in that: using clorox as sole component.
2. a using method for bacteriostatic agent as claimed in claim 1, is characterized in that: added by clorox in the medium of open cultivation potato tissue culture seedling, make it account for 0.5% of medium gross mass.
3. using method according to claim 2, is characterized in that: before carrying out open cultivation, first adopts the utensil of liquor natrii hypochloritis to open cultivation to clean, then uses
cold boiling waterscrubber, then carry out open cultivation by flow process.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110999790A (en) * | 2019-12-26 | 2020-04-14 | 云南师范大学 | Method for simultaneously inhibiting bacteria and prolonging subculture time of potato tissue culture seedlings and subculture method |
| CN115589945A (en) * | 2022-10-10 | 2023-01-13 | 长江大学（Cn） | Method for openly culturing ginger test-tube plantlets |
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| CN102498793A (en) * | 2011-10-22 | 2012-06-20 | 中国科学院合肥物质科学研究院 | Method for removing endophytes from pistacia chinensis bunge seeds and quickly germinating pistacia chinensis bunge seeds |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110999790A (en) * | 2019-12-26 | 2020-04-14 | 云南师范大学 | Method for simultaneously inhibiting bacteria and prolonging subculture time of potato tissue culture seedlings and subculture method |
| CN110999790B (en) * | 2019-12-26 | 2021-08-31 | 云南师范大学 | Method for simultaneously inhibiting bacteria and prolonging subculture time of potato tissue culture seedlings and subculture method |
| CN115589945A (en) * | 2022-10-10 | 2023-01-13 | 长江大学（Cn） | Method for openly culturing ginger test-tube plantlets |
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