CN107295970A - A kind of method for inducing castor-oil plant flower pesticide explant formation callus - Google Patents
A kind of method for inducing castor-oil plant flower pesticide explant formation callus Download PDFInfo
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- CN107295970A CN107295970A CN201710571480.0A CN201710571480A CN107295970A CN 107295970 A CN107295970 A CN 107295970A CN 201710571480 A CN201710571480 A CN 201710571480A CN 107295970 A CN107295970 A CN 107295970A
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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 invention discloses a kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprise the following steps：S1. castor-oil plant flower pesticide explant is placed in 10~20min of immersion in 15~30 mg/L BA solution；S2. it will be inoculated on callus inducing medium and cultivated by the S1 castor-oil plant flower pesticide explants handled, obtain callus；The MS culture mediums that the callus inducing medium is the SNP that with the addition of 4~16mg/L, or the MS culture mediums of 1~2mg/L copper ions are with the addition of, or with the addition of the MS culture mediums of 3~6mg/L glutamine.The present invention can promote flower pesticide explant to form more callus with higher efficiency dedifferentiation, be that the work of castor-oil plant associated biomolecule technology Breeding is had laid a good foundation while improving the quality of obtained callus.
Description
Technical field
The present invention relates to field of plant cell engineering technology, in particular it relates to which a kind of induction castor-oil plant flower pesticide explant is formed
The method of callus.
Background technology
Since Guhas reports that mao Thorn-Apple Anther Culture obtains haplobiont first, on grinding for Anther Culture
Study carefully and develop rapidly, many countries including China carry out the research work of this respect in succession, plant more make about 200 so far
Thing is cultivated by flower pesticide and obtains haplobiont.
In breeding field, Breeding by anther culture and conventional cross-breeding, distant hybridization breeding, mutation breeding and transgenosis
Technology is combined, and has become biotechnology one of relatively broad and effective method in crop breeding, be crop improvement and
Science of heredity and physiology, the important method of Biochemical Research.Up to the present, existing 40 various plants of China are obtained with anther culture method
Obtained pollen plant, mainly have wheat, corn, paddy rice, grape, citrus, apple, longan, rubber, willow and ginseng etc. some
Medicinal plant.More than 80 paddy pollen new lines and new varieties, more than 20 Wheat Pollen new varieties and new lines are turned out.
Castor-oil plant is that Euphorbiaceae Ricinus is annual or herbaceos perennial, is one of ten big oil crops, tool in the world
Have widely used, the features such as economic value is high, be with special industry purposes essential industry oil, its seed, blade, cane,
Shell comprehensive utilization value is high.At present, competitive improved seeds are also lacked.
For a long time because castor-oil plant is main traditional mode such as to hybridize, select, introduce a fine variety carry out breeding, largely
Limit its development.Haplobiont can be obtained using Anther Culture, then it is carried out after chromosome doubling, can quickly be obtained
Wide variety of pure line cultivar in castor-oil plant breeding；And trait segregation will not occur for the offspring for the excellent homozygous line therefrom selected,
Neat and consistent is showed, can significantly shorten the breeding time limit.But at present, castor-oil plant Anther Culture is also in the starting stage, relevant castor-oil plant
The report of Anther Culture is also seldom, also there is that the inductivity of castor-oil plant anther callus is low and callus quality is poor etc. asks
Topic, seriously constrains the progress of castor-oil plant vitro anther culture.Therefore, efficient induction castor-oil plant flower pesticide explant is set up to be formed
The tissue culture system of callus, all has important to the haploid breeding for accelerating castor-oil plant and the castor-oil plant industry for further developing China
Meaning.
The content of the invention
The invention aims to overcome the deficiencies in the prior art to be cured there is provided one kind induction castor-oil plant flower pesticide explant formation
The method of injured tissue.
To achieve these goals, the present invention is achieved by the following technical programs：
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
S1. castor-oil plant flower pesticide explant is placed in 10~20min of immersion in 15~30 mg/L BA solution；
S2. it will be inoculated on callus inducing medium and cultivated by the S1 castor-oil plant flower pesticide explants handled, is cured
Injured tissue；The MS culture mediums that the callus inducing medium is the SNP that with the addition of 4~16mg/L, or with the addition of 1~2mg/
The MS culture mediums of L copper ions, or with the addition of the MS culture mediums of 3~6mg/L glutamine.
The method of tradition induction castor-oil plant flower pesticide explant formation callus is all to select flower pesticide explant being seeded in contain
Directly induced on the culture medium for there are various hormones, creativeness of the invention is first to carry out an early stage to flower pesticide explant
Immersion treatment, explant is then seeded on the culture medium of copper ions, SNP or glutamine progress callus induction group again
Knit.Early stage immersion treatment is carried out to flower pesticide explant and is advantageous in that the BA of high concentration can promote flower pesticide explant with higher
Efficiency dedifferentiation forms more callus, while improving the quality of obtained callus.
Preferably, castor-oil plant flower pesticide explant is placed in 15mg/L BA solution and soaks 10min.
Although identical explant generating process of evoked callus on different culture mediums be substantially it is consistent,
But its callus quality and inductivity are then different and different because of culture medium prescription；In addition, different explant types is in phase
The quality and inductivity of the callus induced on same culture medium are also different.
Preferably, the MS culture mediums that the callus inducing medium is the SNP that with the addition of 8mg/L.
Preferably, the callus inducing medium is the MS culture mediums that with the addition of 2mg/L copper ions.
Preferably, the callus inducing medium is the MS culture mediums that with the addition of 6mg/L glutamine.
Preferably, the acquisition methods of the castor-oil plant flower pesticide explant are：Obtain without insect food take, full and perianth not yet
The male flower of opening, surface sterilizing is carried out to castor-oil plant male flower using 2% sodium hypochlorite, and peels off the perianth on male flower surface, careful to obtain
Castor-oil plant flower pesticide explant therein.
The collocation method of the BA solution of various concentrations of the present invention is：Accurately weigh a certain amount of 6-benzyl aminopurine（6-BA,
Abbreviation BA）（Sigma, USA）Powder, is fully dissolved with 1 mol/L NaOH solution, then uses deionized water constant volume, is configured to dense
Degree is respectively 0,7.5,15,30,60,120 mg/L BA processing solutions.Then BA processing is adjusted with 1 mol/L HCl solution
The pH of solution to 5.8~6.0, uses the preceding water system filter membrane with 0.22 micron（Millipore, USA）To at the BA that has prepared
Manage solution and carry out filtration sterilization.
The MS culture mediums：The inorganic constituents of MS culture medium prescriptions（16.5 g/L ammonium nitrate, 19 g/L potassium nitrate, 1.7
G/L potassium dihydrogen phosphates, 3.7 g/L epsom salts, 4.4 g/L calcium chloride dihydrates, 16.9 mg/L manganese sulfate monohydrates, 8.6 mg/
L white vitriols, 6.2 mg/L boric acid, 0.83 mg/L KIs, 0.25 mg/L sulfate dihydrate molybdenums disodium, 0.025 mg/L five
Brochanite, 0.025 mg/L CoCL2 6H2Os, the water sodium ethylene diamine tetracetates of 37.3 mg/L bis-, the water sulfuric acid of 27.8 mg/L seven are sub-
Iron）The organic principle of+MS culture medium prescriptions（2 mg/L glycine, 0.1 mg/L thiamine hydrochlorides, 0.5 mg/L hydrochloric acid pyrroles are trembled
Alcohol, 0.5 mg/L nicotinic acid）The g/L agar of+30 g/L sucrose of+100 mg/L inositols+7, pH is adjusted extremely with 1 mg/L NaOH solutions
5.8~6.0.Culture medium sterilizes 15 minutes under conditions of 121 DEG C, 0.1 MPa.
Condition of culture described in step S2 is：Room temperature is 25 DEG C, intensity of illumination is that 2000 lx, light application time are daily 12
Hour.
Compared with prior art, the present invention has the advantages that：
The novelty of the present invention is the method for changing tradition induction castor-oil plant flower pesticide explant formation callus, that is, utilizes height
Concentration BA solution carries out short-term immersion treatment to flower pesticide explant, thus promotes flower pesticide explant with higher efficiency dedifferentiation shape
Into more callus.The quality of obtained callus is improved simultaneously, is that the work of castor-oil plant associated biomolecule technology Breeding is established
Good basis is determined.
Embodiment
The present invention is made with reference to specific embodiment and further being elaborated, the embodiment is served only for explaining this hair
It is bright, it is not intended to limit the scope of the present invention.Test method used in following embodiments is routine unless otherwise specified
Method；Used material, reagent etc., are the reagent and material commercially obtained unless otherwise specified.
Embodiment 1
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
(1) flower pesticide explant is obtained：Choose without insect food is taken, full and perianth is not yet opened in spherical castor-oil plant male flower.Take
It should be selected in that temperature is low the time, during the weak fine day morning 10 of illumination or so, now the bud on castor-oil plant can reduce pollution, light without fog
According to weaker and the forfeiture of flower pesticide moisture can be prevented.The male flower bud of collection is lived with tissue-wrapped, first soaked with distilled water, then gently
Unnecessary moisture is gently squeezed out, is then put into vial, is handled 3 days in 4 DEG C of refrigerator.First with 75% on superclean bench
Ethanol solution first sterilizes 1 min to castor-oil plant male flower, then with 2% NaClO solution disinfection 20min, rinsed with sterile water castor-oil plant male flower 5
It is secondary.Removed afterwards with the careful perianth by male flower outer layer of sterilized dissecting needle, then by flower pesticide careful separation therein, you can obtain
Obtain castor-oil plant flower pesticide explant.
(2) castor-oil plant flower pesticide explant is seeded on addition various concentrations BA MS minimal mediums, room temperature be 25 DEG C,
Intensity of illumination is that 2000 lx, light application time are evoked callus under conditions of daily 12 hours.As a result as shown in table 1, addition
Formation of the appropriate BA to castor-oil plant anther callus has facilitation, and when BA concentration is 1 mg/L, castor-oil plant flower pesticide is cured
The inductivity of injured tissue reaches maximum, is 10.67%；And the continuation increase of the concentration with BA, castor-oil plant anther callus lures
Conductance is gradually reduced.Meanwhile, experimental result is also shown that induces castor-oil plant flower pesticide explant formation callus using conventional method
During, the induced efficiency for obtaining callus is relatively low and second-rate, small volume.
Table 1 is influences of the BA to castor-oil plant induction of anther callus efficiency
Note：Data carry out variance analysis and Duncan Multiple range test using the statistical analysis softwares of SPSS Statistics 17.0（P
≦ 0.05）, significant difference between the different expressions processing of letter after data.
Embodiment 2
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
(1) flower pesticide explant is obtained：Be the same as Example 1.
(2) castor-oil plant flower pesticide explant is placed in 15 mg/L BA solution and soaks different time（0~80 min）；
(3) and then castor-oil plant flower pesticide explant is placed on sterile blotting paper and sucked after surplus liquid, careful is seeded to without sharp
Cultivated 30 days on plain MS culture mediums.
Test result indicates that, when the time of immersion treatment explant is 10 min, explant callus induction efficiency effect
Fruit is optimal, up to 41.25%（Table 2）.Continue to extend processing time, castor-oil plant flower pesticide explant callus induction rate will be reduced gradually.
Table 2 is influence of the processing time to castor-oil plant induction of anther callus efficiency
Note：Data carry out variance analysis and Duncan Multiple range test using the statistical analysis softwares of SPSS Statistics 17.0（P
≦ 0.05）, significant difference between the different expressions processing of letter after data.
Embodiment 3
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
(1) flower pesticide explant is obtained：Be the same as Example 1.
(2) castor-oil plant flower pesticide explant is placed in various concentrations (0~120 mg/L) BA solution and soaks 10min.
(3) and then castor-oil plant flower pesticide explant is placed on sterile blotting paper and sucked after surplus liquid, careful is seeded to
Without culture 30 days on hormone MS culture mediums.
Test result indicates that, with the increase of BA solution concentrations, castor-oil plant flower pesticide explant callus induction efficiency will be by
Step is improved, when the concentration of immersion treatment explant is 15 mg/L, explant callus induction best results, callus
Inductivity reaches maximum, is 41.25%（Table 3）.And continue to improve BA treatment fluid concentration, castor-oil plant flower pesticide explant callus is lured
Leading efficiency will gradually reduce.Meanwhile, induce castor-oil plant flower pesticide explant in the method using short-term high concentration BA solution immersion treatments
During body formation callus, it was found that obtain callus quality preferably, volume is larger.
Table 3 is influence of the BA solution concentrations to castor-oil plant induction of anther callus efficiency
Note：Data carry out variance analysis and Duncan Multiple range test using the statistical analysis softwares of SPSS Statistics 17.0（P
≦ 0.05）, significant difference between the different expressions processing of letter after data.
From above-mentioned experimental result, compared with using conventional method induction castor-oil plant flower pesticide explant formation callus,
Significantly improved using the callus induction rate of high concentration BA solution immersion treatment castor-oil plant flower pesticide explants institute induced synthesis（Callus
Tissue inductivity improves 30.58%）, and the better quality of the callus obtained, volume is bigger.Therefore, using 15
Mg/L BA solution carries out 10 min immersion treatment to castor-oil plant flower pesticide explant, can obtain more, quality for reaching
The purpose of more preferable callus.
Embodiment 4
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
(1) flower pesticide explant is obtained：Be the same as Example 1.
(2) castor-oil plant flower pesticide explant is placed in 15mg/L BA solution and soaks 10min.
(3) and then castor-oil plant flower pesticide explant is placed on sterile blotting paper and sucked after surplus liquid, careful is seeded to
It with the addition of the sodium nitroprussiate of various concentrations（SNP）（0th, 2,4,8,16 and 32mg/L）MS culture mediums on cultivate 30 days.
Test result indicates that, with the raising of SNP concentration, castor-oil plant induction of anther callus rate works as SNP also with raising
When concentration is 8 mg/L, callus induction rate reaches maximum, is 60.37%, 49.70% is improved than conventional method, than passing through
It is seeded to after the processing of high concentration BA solution on no hormone MS culture mediums and improves 19.12%；But when being to continue with improving SNP concentration,
Callus induction will be suppressed, and induced efficiency will be gradually reduced.
Table 4 is influences of the SNP to castor-oil plant induction of anther callus efficiency
Note：Data carry out variance analysis and Duncan Multiple range test using the statistical analysis softwares of SPSS Statistics 17.0（P
≦ 0.05）, significant difference between the different expressions processing of letter after data.
Embodiment 5
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
(1) flower pesticide explant is obtained：Be the same as Example 1.
(2) castor-oil plant flower pesticide explant is placed in 15mg/L BA solution and soaks 10min.
(3) and then castor-oil plant flower pesticide explant is placed on sterile blotting paper and sucked after surplus liquid, careful is seeded to
It with the addition of the copper ion of various concentrations（Anhydrous cupric sulfate）（0th, 0.5,1,2,4 and 8 mg/L）MS culture mediums on cultivate 30 days.
Test result indicates that, with the raising of copper ion concentration, castor-oil plant induction of anther callus rate also with raising, when
When copper ion concentration is 2 mg/L, callus induction rate reaches maximum, is 68.36%, 57.69% is improved than conventional method,
27.11% is improved than being seeded to after being handled through excessive concentrations BA solution on no hormone MS culture mediums；；But be to continue with improve copper from
During the concentration of son, callus induction will be suppressed, and induced efficiency will be gradually reduced.
Table 5 is influence of the copper ion to castor-oil plant induction of anther callus efficiency
Note：Data carry out variance analysis and Duncan Multiple range test using the statistical analysis softwares of SPSS Statistics 17.0（P
≦ 0.05）, significant difference between the different expressions processing of letter after data.
Embodiment 6
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
(1) flower pesticide explant is obtained：Be the same as Example 1.
(2) castor-oil plant flower pesticide explant is placed in 15mg/L BA solution and soaks 10min.
(3) and then castor-oil plant flower pesticide explant is placed on sterile blotting paper and sucked after surplus liquid, careful is seeded to
It with the addition of the glutamine of various concentrations（0th, 1.5,3,6,12 and 24 mg/L）MS culture mediums on cultivate 30 days.
Test result indicates that, with the raising of Gln concentration, castor-oil plant induction of anther callus rate works as paddy also with raising
When glutamine concentration is 6 mg/L, callus induction rate reaches maximum, is 54.63%, 43.96% is improved than conventional method,
13.38% is improved than being seeded to after being handled through excessive concentrations BA solution on no hormone MS culture mediums；But it is to continue with improving glutamy
During the concentration of amine, callus induction will be suppressed, and induced efficiency will be gradually reduced.
Table 6 is influence of the glutamine to castor-oil plant induction of anther callus efficiency
Note：Data carry out variance analysis and Duncan Multiple range test using the statistical analysis softwares of SPSS Statistics 17.0（P
≦ 0.05）, significant difference between the different expressions processing of letter after data.
Embodiment 7
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, step (1) and (3) be the same as Example 2, with embodiment 2
Unique different place is that step (2) is soaked using different hormones, and the present embodiment is different arranged side by side including following 4
Processing：Processing 1 is the TDZ solution immersion flower pesticide explant 10min using 15mg/L；Processing 2 is molten using 15mg/L NAA
Immersion flower bulb medicine explant 10min；Processing 3 is the IBA solution immersion flower pesticide explant 10min using 15mg/L；Handling 4 is
Use 15mg/L IAA solution immersion flower pesticide explant 10min.Callus induction after four kinds of different disposals the results are shown in Table
7。
Table 7 is influence of the hormon immersion to castor-oil plant induction of anther callus efficiency
Embodiment 8
A kind of method for inducing castor-oil plant flower pesticide explant formation callus, comprises the following steps：
(1) flower pesticide explant is obtained：Be the same as Example 1.
(2) castor-oil plant flower pesticide explant is seeded on the MS minimal mediums of addition hormon, is 25 DEG C, light in room temperature
It is that 2000 lx, light application time are evoked callus under conditions of daily 12 hours according to intensity.The present embodiment includes following 2
Different and column processing：Processing 1 uses addition 3mg/L BA+0.2mg/L NAA MS culture mediums as callus induction
Culture medium；Processing 2 uses addition 2mg/L BA+2mg/L 2,4-D MS culture mediums as callus inducing medium；Knot
Fruit is shown in Table 8.
Table 8 is influence of the different callus inducing mediums to castor-oil plant induction of anther callus efficiency
Embodiment 9
(1) flower pesticide explant is obtained：Be the same as Example 1.
(2) castor-oil plant flower pesticide explant is placed in 15mg/L BA solution and soaks 10min.
(3) and then castor-oil plant flower pesticide explant is placed on sterile blotting paper and sucked after surplus liquid, careful is seeded to
On the MS minimal mediums for adding hormon, room temperature be 25 DEG C, intensity of illumination be that 2000 lx, light application time are daily 12
Evoked callus under conditions of hour.The present embodiment includes following 4 different and column processing：Processing 1 is using addition
The MS culture mediums of 8mg/L SNP+2mg/L copper ions are used as callus inducing medium；Processing 2 is using addition 2mg/L
The MS culture mediums of copper ion+6mg/L glutamine are used as callus inducing medium；Processing 3 is using addition 8mg/L
The MS culture mediums of SNP+6mg/L glutamine are used as callus inducing medium；Processing 4 is the SNP using addition 8mg/L
+ add the MS culture mediums of 2mg/L copper ion+6mg/L glutamine to be used as callus inducing medium；It the results are shown in Table 9.
Table 9 is influence of the different callus inducing mediums to castor-oil plant induction of anther callus efficiency
Above example is served only for explaining the present invention, is not intended to limit the scope of the present invention.
SEQUENCE LISTING
<110>Guangdong Jie Shi agricultural science and technologys Co., Ltd
<120>One plant of plant azotobacteria DJG211 and its application in soil and plant vigor is improved
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1345
<212> DNA
<213>16S rDNA sequences
<400> 1
ggtagcacag ggagcttgct ctcgggtgac gagtggggga cgggtgagta atgtctggga 60
aactgcccga tggaggggga taactactgg aaacggtagc taataccgca taatgtggca 120
agaccaaaga gggggacctt cgggcctctt gccatcggat gtgcccagat gggattagct 180
tgttggtgag gtaatggctc accaaggcga caatccctag ctggtctgag aggatgacca 240
gccacactgg aactgagaca cggtccagac tcctacggga ggcagcagtg gggaatattg 300
cacaatgggc gcaagcctga tgcagccatg ccgcgtgtgt gaagaaggcc ttcgggttgt 360
aaagcacttt cagcggggag gaaggtgtta aggttaataa ccttgtcgat tgacgttacc 420
cgcagaagaa gcaccggcta actccgtgcc agcagccgcg gtaatacgga gggtgcaagc 480
gttaatcgga attactgggc gtaaagcgca cgcaggcggt ctgtcaagtc ggatgtgaaa 540
tccccgggct caacctggga actgcattcg aaactggcag gcttgagtct tgtagagggg 600
ggtagaattc caggtgtagc ggtgaaatgc gtaaagatct ggaggaatac cggtggcgaa 660
ggcggccccc tggacaaaga ctgacgctca ggtgcgaaag cgtggggagc aaacaggatt 720
agataccctg gtagtccacg ccgtaaacga tgtcgacttg gaggttgtgc ccttgaggcg 780
tggcttccgg agctaacgcg ttaagtcgac cgcctgggga gtacggccgc aaggttaaaa 840
ctcaaatgaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgatgcaa 900
cgcgaagaac cttacctggt cttgacatcc acggaatttg gcagagatgc cttagtgcct 960
tcgggaaccg tgagacaggt gctgcatggc tgtcgtcagc tcgtgttgtg aaatgttggg 1020
ttaagtcccg caacgagcgc aacccttatc ctttgttgcc agcggtccgg ccgggaactc 1080
aaaggasact gccagtgata aactggagga aggtggggat gacgtcaagt catcatggcc 1140
cttacgacta gggctacaca cgtgctacaa tggcatatac aaagagaagc gacctcgcga 1200
gagcaagcgg acctcataaa gtatgtcgta gtccggattg gagtctgcaa ctcgactcca 1260
tgaagtcgga atcgctagta atcgtggatc agaatgccac ggtgaatacg ttcccgggcc 1320
ttgtacacac cgcccgtcac accat 1345
Claims (5)
1. a kind of method for inducing castor-oil plant flower pesticide explant formation callus, it is characterised in that comprise the following steps：
S1. castor-oil plant flower pesticide explant is placed in 10~20min of immersion in 15~30 mg/L BA solution；
S2. it will be inoculated on callus inducing medium and cultivated by the S1 castor-oil plant flower pesticide explants handled, is cured
Injured tissue；The MS culture mediums that the callus inducing medium is the SNP that with the addition of 4~16mg/L, or with the addition of 1~2mg/
The MS culture mediums of L copper ions, or with the addition of the MS culture mediums of 3~6mg/L glutamine.
2. according to the method described in claim 1, it is characterised in that castor-oil plant flower pesticide explant is placed in 15mg/L BA solution
Soak 10min.
3. according to the method described in claim 1, it is characterised in that the callus inducing medium is that with the addition of 8mg/L
SNP MS culture mediums.
4. according to the method described in claim 1, it is characterised in that the callus inducing medium is that with the addition of 2mg/L
The MS culture mediums of copper ion.
5. according to the method described in claim 1, it is characterised in that the callus inducing medium is that with the addition of 6mg/L
The MS culture mediums of glutamine.
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| CN108633741A (en) * | 2018-07-16 | 2018-10-12 | 河南农业大学 | The method for tissue culture of four seasons begonia |
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| CN101385442A (en) * | 2008-09-27 | 2009-03-18 | 云南大学 | Method for obtaining haploid plant of calla by anther culture |
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| 陈永胜等: "蓖麻花药愈伤组织诱导及防褐化研究", 《江苏农业科学》 * |
| 黄凤兰等: "蓖麻花药愈伤组织诱导的研究", 《作物杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108633741A (en) * | 2018-07-16 | 2018-10-12 | 河南农业大学 | The method for tissue culture of four seasons begonia |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107295970B (en) | 2019-02-05 |
| CN107295970A9 (en) | 2018-05-04 |
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