CN111587788A - Induction medium for carnation anther culture - Google Patents
Induction medium for carnation anther culture Download PDFInfo
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- CN111587788A CN111587788A CN202010558849.6A CN202010558849A CN111587788A CN 111587788 A CN111587788 A CN 111587788A CN 202010558849 A CN202010558849 A CN 202010558849A CN 111587788 A CN111587788 A CN 111587788A
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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Abstract
The invention provides an induction culture medium for carnation anther culture, which comprises KNO3、NH4NO3、KH2PO4、CaCl2·2H2O、Mg2SO4·7H2O、KCl、FeSO4·7H2O、Na2‑EDTA、KI、H3BO3、MnSO4·4H2O、ZnSO4·7H2O、Na2MoO4·2H2O、CuSO4·5H2O、CoCl2·6H2O, vitamin B1, vitamin B6, biotin, riboflavin, D-calcium pantothenate, aspartic acid, glycine, sodium pyruvate, fumaric acid, cysteine hydrochloride, hydrolyzed casein, inositol, 5-azacytosine nucleoside, phytohemagglutinin, maleic hydrazide, 2,4-D, 6-BA, carnation bleeding liquid, sodium bisulfite, trehalose, sucrose and plant gel. The culture medium is suitable for callus induction of carnation, and has highest induction rate20.15 percent, and lays a foundation for establishing a high-efficiency and stable dianthus chinensis anther culture system.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly relates to an induction culture medium for carnation anther culture.
Background
Dianthus caryophyllus (C)Dianthus caryophyllusL.) The common names of carnation, Shizhu, Dianthus chinensis, Musk Shizhu and the like are perennial root herbaceous plants in the genus Dianthus of the family Dianthus. Dianthus caryophyllus originated in southern Europe and Mediterranean region, and is famous for its delicate color and fragrant smell. Carnation, one of the four major cut flowers in the world, has quite high ornamental and application values and is mainly applied to the cut flowers. Due to rich colors, the flower-arranging material can be used as a material for art flower arrangement or used for manufacturing flower baskets, flower bundles, brooches and the like. Beginning in 1907, pink carnation was regarded as the symbol of love of mother, and was known as the flower of mother's festival, representing the mother's love of indifference, warm, true and full, and was widely loved by people.
The flower industry is one of the most well-recognized industries with development potential in the world and is called the "gold industry". In recent years, with the development of the flower industry, the cultivation area of carnation in China is continuously enlarged. However, the varieties of carnation cultivated in China at present are mainly introduced from foreign countries, and countries such as the Netherlands, Italy, Columbia, Israel and the like are the major countries for producing the carnation seedlings. Although the mode can keep up with the popular trend of the carnation varieties in the world, the production cost is high and is generally restricted by people, so that the research on the breeding of the carnation is enhanced, and the new variety of the carnation which is suitable for the climate conditions of China, has high disease resistance, stress resistance and high ornamental value and has the independent intellectual property right of China is cultivated. The conventional breeding mode of the carnation is crossbreeding, the crossbreeding is an important source of biological genetic variation, and through the combination of different types of genotypes, people can breed hybrids meeting the production and life requirements. However, the limitations of complex operation, long breeding cycle, poor predictability, low selection efficiency and the like exist in cross breeding, so that the modern biotechnology means needs to be fully utilized to assist breeding on the basis of conventional breeding, and the breeding process of new varieties is accelerated.
Anther culture is a part of plant tissue culture technology, and is one of the main ways of producing haploids by inoculating the whole anther on a culture medium and inducing microspores of the anther to form a haploid plant under the condition of isolated culture. Anther culture has important significance: firstly, obtaining pure line breeding material and carrying out haploid breeding to utilize heterosis, shorten the breeding period and improve the breeding efficiency; secondly, the obtained homozygous diploid material is used for researching the genetic variation rule of the fruit tree, so that the plant breeding has a solid genetic theory basis, and the breeding blindness is reduced; thirdly, overcoming the sterility of the distant hybrid and obtaining the fertile distant hybrid with the excellent characteristics of parents. Haploid breeding has been combined with conventional crossbreeding, distant crossbreeding, mutation breeding and transgenic technology, and now becomes one of the most widely and effectively applied methods of biotechnology in crop breeding.
Anther culture is a special isolated culture of sexual organs, so the requirements are higher and the difficulty is higher than that of general tissue culture techniques, and anther culture research of many plants is still in a blank state so far. The genotype of the donor plant is a key factor affecting anther culture. Anthers of plants with different genotypes react differently to the culture, which is reflected in the difference in anther culture power such as callus induction rate, differentiation rate, embryoid induction rate, plant induction rate, etc., and some materials do not react to some culture media. The pollen development stage is the critical stage for inducing microspore division. Generally, pollen in the metaphase to late stage of the mononuclear is better cultured. Because neither the young anthers, which are in the pollen meiosis stage, nor the older anthers, which are pollen filled with starch grains, can produce embryoid bodies. The selection of a proper culture medium is also one of the important links of the anther culture technology. Generally, the selected cultured gene is different from plant species to plant species, and until now, culture media such as MS, B5, N6, White, etc. have been widely used in plant anther culture. In addition, growth regulating substances, additional components and the like in the culture medium have important influence on the induction and differentiation of the callus.
At present, few research reports about the dianthus caryophyllus anther culture at home and abroad are reported, a dianthus caryophyllus anther culture system is not established, and a dianthus caryophyllus haploid or DH plant is not obtained, so that deep research needs to be carried out by people.
Disclosure of Invention
The invention aims to provide an induction culture medium for carnation anther culture, which lays a foundation for establishment of a carnation anther culture system and future carnation haploid breeding work.
The purpose of the invention is realized by the following technical scheme:
an induction culture medium for carnation anther culture is characterized in that the components and the using concentration of the culture medium are as follows: KNO31700-2000mg/L，NH4NO3600-800mg/L，KH2PO4140-180mg/L，CaCl2·2H2O 490-530mg/L，Mg2SO4·7H2O 280-320mg/L，KCl 95-115mg/L，FeSO4·7H2O 26.4-30.2mg/L，Na2-EDTA 35.7-39.3mg/L，KI 0.7-0.8mg/L，H3BO35.5-6.5mg/L，MnSO4·4H2O 12.6-14.0mg/L，ZnSO4·7H2O 7.7-8.5mg/L，Na2MoO4·2H2O 0.2-0.3mg/L，CuSO4·5H2O 0.04-0.06mg/L，CoCl2·6H20.02-0.03mg/L of O, 10.4-0.6 mg/L of vitamin B, 60.4-0.6 mg/L of vitamin B, 0.01-0.03mg/L of biotin, 0.2-0.4mg/L of riboflavin, 0.5-1.0mg/L of D-calcium pantothenate, 1.0-2.0mg/L of aspartic acid, 1.0-2.0mg/L of glycine, 15-25mg/L of sodium pyruvate, 35-45mg/L of fumaric acid, 20-30mg/L of cysteine hydrochloride, 150mg/L of hydrolyzed casein 110, 140mg/L of inositol 100, 2.5-3.5mg/L of 5-azacytosine nucleoside, 4.6-5.4mg/L of phytohemagglutinin, 1.8-2.2mg/L of maleic hydrazide, 2, 4-0.6mg/L of D, 0.4-0.6mg/L of 6-BA, 20-30ml/L of carnation bleeding sap, 0.07-0.09mg/L of sodium bisulfite, 2.0-4.0g/L of trehalose, 40-50g/L of sucrose and 6-8g/L of plant gel.
The pH of the medium is adjusted to 5.6-6.0.
The preparation method of the dianthus caryophyllus bleeding wound liquid in the culture medium comprises the following steps: cutting off the carnation stalk at a distance of 2cm from the root by using a clean scissors, discarding the 1 st dripping liquid to prevent pollution, sleeving a sealed plastic bag filled with absorbent cotton, enabling the absorbent cotton to be tightly attached to the cut, fastening the bag opening by using a rubber band, taking down the absorbent cotton and the plastic bag after 5h, and collecting the bleeding liquid into a container by a suction filtration method to be frozen and stored at the temperature of-20 ℃.
The culture medium is used for culturing and inducing callus of carnation anther.
The invention has the following beneficial effects:
the induction culture medium is optimized on the basis of a conventional culture medium, the use concentrations of major elements and trace elements are adjusted, additional components such as sodium pyruvate, fumaric acid, cysteine hydrochloride, hydrolyzed casein, 5-azacytosine nucleoside, phytohemagglutinin, carnation bleeding sap, sodium bisulfite, trehalose and the like are added, maleic hydrazide and 2,4-D and 6-BA are used as plant growth regulators, the induction rate of carnation anther callus can be obviously improved, the culture efficiency is improved by 2-3 times compared with other culture media, and the foundation is laid for establishing a high-efficiency and stable carnation anther culture system.
Detailed Description
The following examples are provided to further illustrate the beneficial effects of the present invention, and are not intended to limit the invention.
Example 1
Test materials and methods
1.1 test materials
'Master' and 'liberty' 2 varieties of carnation were from the Shaoxing International flower City of Living Yunnan harbor, Jiangsu province.
1.2 pretreatment
Picking up flower buds of carnation when no dew exists in 8: 00-9: 00 am in sunny weather, adopting carmine acetate for dyeing, checking the development period of pollen, selecting out the flower buds with the development period of pollen in the mononuclear border period, wrapping with moist gauze, and then placing in a refrigerator at 4 ℃ for low-temperature pretreatment for 3 d.
1.2 Sterilization and inoculation of materials
Washing the treated bud with tap water, placing on a clean bench, soaking in 75% alcohol for 30s, sterilizing the bud surface, and adding 0.1% HgCl2Sterilizing for 10min, and washing with sterile water for 4 times; placing the flower bud in a sterile culture dish, peeling off the flower bud with sterilized forceps, taking out anther, cutting off filament, inoculating on induction culture medium, and culturing in dark at 25 deg.C in a constant temperature incubator.
The composition and concentration of the induction medium were as follows: KNO31850mg/L，NH4NO3700mg/L，KH2PO4160mg/L，CaCl2·2H2O 510mg/L，Mg2SO4·7H2O 300mg/L，KCl 105mg/L，FeSO4·7H2O28.3mg/L，Na2-EDTA 37.5mg/L，KI 0.75mg/L，H3BO36.0mg/L，MnSO4·4H2O 13.3mg/L，ZnSO4·7H2O 8.1mg/L，Na2MoO4·2H2O 0.25mg/L，CuSO4·5H2O 0.05mg/L，CoCl2·6H2O0.025mg/L, vitamin B10.5mg/L, vitamin B60.5mg/L, biotin 0.02mg/L, riboflavin 0.3mg/L, D-calcium pantothenate 0.75mg/L, aspartic acid 1.5mg/L, glycine 1.5mg/L, sodium pyruvate 20mg/L, fumaric acid 40mg/L, cysteine hydrochloride 25mg/L, hydrolyzed casein 130mg/L, inositol 120mg/L, 5-azacytosine nucleoside 150. mu. mol/L, phytohemagglutinin 5.0mg/L, maleic hydrazide 2.0mg/L, 2, 4-D0.5 mg/L, 6-BA 0.5mg/L, carnation bleeding sap 25ml/L, sodium bisulfite 0.08mg/L, trehalose 3.0g/L, sucrose 45g/L, vegetable gel 7g/L, the pH was adjusted to 5.8.
The preparation method of the carnation bleeding sap comprises the following steps: cutting off the carnation stalk at a distance of 2cm from the root by using a clean scissors, discarding the 1 st dripping liquid to prevent pollution, sleeving a sealed plastic bag filled with absorbent cotton, enabling the absorbent cotton to be tightly attached to the cut, fastening the bag opening by using a rubber band, taking down the absorbent cotton and the plastic bag after 5h, and collecting the bleeding liquid into a container by a suction filtration method to be frozen and stored at the temperature of-20 ℃.
2 design of the experiment
2.1 Effect of different concentrations of 5-azacytosine nucleosides on callus induction
The anther of carnation was inoculated on induction media of different 5-azacytidine concentrations (50. mu. mol/L, 100. mu. mol/L, 150. mu. mol/L, 200. mu. mol/L, 250. mu. mol/L), the other components of the media were unchanged with the control of the induction media containing no 5-azacytidine, and the callus induction rate was counted after dark culture in a 25 ℃ incubator for 40-50 days, with the results shown in Table 1.
Callus induction rate (%) = (number of anthers from which callus was induced/number of anthers inoculated) × 100.
As can be seen from Table 1, the callus induction rate of the carnation anther shows a tendency of rising first and then falling with the increase of the concentration of 5-azacytidine in the induction medium, and when the concentration of the 5-azacytidine is 150 μmol/L, the callus induction rates of 'malaster' and 'free' reach the highest values, respectively 19.90% and 15.67%, which are remarkably improved compared with the control group; when the concentration of 5-azacytidine exceeds 150. mu. mol/L, the callus induction rate is rather decreased, so that the suitable concentration of 5-azacytidine is 150. mu. mol/L.
2.2 Effect of different concentrations of phytohemagglutinin on callus induction
Anthers were inoculated on induction media of different phytohemagglutinin concentrations (0.5 mg/L, 1.0mg/L, 2.0mg/L, 5.0mg/L, 10.0mg/L), the other components of the media were unchanged with the induction media without phytohemagglutinin as a control, and the callus induction rates were counted after dark culture in a 25 ℃ incubator for 40-50 days, with the results shown in Table 2.
As can be seen from Table 2, the callus induction rates of carnation anther were improved to different degrees on the respective induction media to which different concentrations of phytohemagglutinin were added, wherein the phytohemagglutinin concentration of 5.0mg/L had the best effect on the callus induction of ` Masster ` and ` free ` anther, and the induction rates reached 20.22% and 14.25%, respectively, which were significantly improved compared to the control group. Therefore, a suitable concentration of phytohemagglutinin in the induction medium is 5.0 mg/L.
2.3 Effect of different Induction Medium on anther callus Induction
The induction culture medium of the invention and MS +3% sucrose +2, 4-D2.0 mg/L + BAP 1.0mg/L (research on preliminary breeding and anther culture of Caryophyllum self-bred line) are used as a contrast culture medium, anther culture is carried out on 'Master' and 'free' under the same conditions, and the induction rate of callus is counted after 40-50 days, and the results are shown in Table 3.
As can be seen from table 3, the induction rate of the induction medium of the present invention on the callus of the carnation variety 'master' is 20.15% which is 2.45 times that of the control medium, and the induction rate of the induction medium of the carnation variety 'free' is 13.48% which is 3.15 times that of the control medium, so that the induction medium provided by the present invention has a significant improvement effect on the induction capability of the carnation anther callus. The induction culture medium is optimized on the basis of a conventional culture medium, the use concentrations of major elements and trace elements are adjusted, additional components such as sodium pyruvate, fumaric acid, cysteine hydrochloride, hydrolyzed casein, 5-azacytosine nucleoside, phytohemagglutinin, carnation bleeding sap, sodium bisulfite, trehalose and the like are added, and maleic hydrazide, 2,4-D and 6-BA are combined to be used as a plant growth regulator, so that the aim of remarkably improving the callus induction rate of the carnation anther is fulfilled.
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (4)
1. An induction culture medium for carnation anther culture is characterized in that the components and the using concentration of the culture medium are as follows: KNO31700-2000mg/L，NH4NO3600-800mg/L，KH2PO4140-180mg/L，CaCl2·2H2O 490-530mg/L，Mg2SO4·7H2O 280-320mg/L，KCl 95-115mg/L，FeSO4·7H2O 26.4-30.2mg/L，Na2-EDTA 35.7-39.3mg/L，KI 0.7-0.8mg/L，H3BO35.5-6.5mg/L，MnSO4·4H2O 12.6-14.0mg/L，ZnSO4·7H2O 7.7-8.5mg/L，Na2MoO4·2H2O 0.2-0.3mg/L，CuSO4·5H2O 0.04-0.06mg/L，CoCl2·6H20.02-0.03mg/L of O, 10.4-0.6 mg/L of vitamin B, 60.4-0.6 mg/L of vitamin B, 0.01-0.03mg/L of biotin and riboflavin0.2-0.4mg/L, 0.5-1.0mg/L of D-calcium pantothenate, 1.0-2.0mg/L of aspartic acid, 1.0-2.0mg/L of glycine, 15-25mg/L of sodium pyruvate, 35-45mg/L of fumaric acid, 20-30mg/L of cysteine hydrochloride, 150mg/L of hydrolyzed casein 110, 140mg/L of inositol 100, 2.5-3.5mg/L of 5-azacytosine nucleoside, 4.6-5.4mg/L of phytohemagglutinin, 1.8-2.2mg/L of maleic hydrazide, 0.4-0.6mg/L of 2,4-D, 0.4-0.6mg/L of 6-BA, 20-30ml/L of carnation bleeding sap, 0.07-0.09mg/L of sodium bisulfite, 2.0-4.0g/L of trehalose, 40-50g/L of sucrose and 6-8g/L of plant gel.
2. The induction medium for the cultivation of dianthus caryophyllus anther according to claim 1, wherein the pH of the medium is adjusted to 5.6-6.0.
3. The induction medium for carnation anther culture as claimed in claim 1, wherein the preparation method of the carnation bleeding sap in the medium is as follows: cutting off the carnation stalk at a distance of 2cm from the root by using a clean scissors, discarding the 1 st dripping liquid to prevent pollution, sleeving a sealed plastic bag filled with absorbent cotton, enabling the absorbent cotton to be tightly attached to the cut, fastening the bag opening by using a rubber band, taking down the absorbent cotton and the plastic bag after 5h, and collecting the bleeding liquid into a container by a suction filtration method to be frozen and stored at the temperature of-20 ℃.
4. The induction medium for carnation anther culture as claimed in claim 1, wherein the medium is used for carnation anther culture to induce callus.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111972293A (en) * | 2020-10-19 | 2020-11-24 | 江苏春之雨生物科技发展有限公司 | Callus induction method for in vitro culture of sesame unpolarized ovaries |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992017056A1 (en) * | 1991-04-01 | 1992-10-15 | Florigene B.V. | Carnation plants and methods for their transformation and propagation |
| CN1951174A (en) * | 2005-10-20 | 2007-04-25 | 上海师范大学 | A culture medium composition adaptable for carnation stalk induced bud and application thereof |
| CN101874471A (en) * | 2009-11-10 | 2010-11-03 | 华中农业大学 | Plant regeneration method of dianthus caryophyllus direct somatic embryo generating path and special culture medium |
| CN108112475A (en) * | 2017-12-13 | 2018-06-05 | 云南中烟工业有限责任公司 | A kind of method that tobacco Brown is prevented with vitamin C, L-cysteine and sodium hydrogensulfite |
| CN108522270A (en) * | 2018-03-06 | 2018-09-14 | 山东寿光蔬菜种业集团有限公司 | A method of based on cucumber anther culture and improvement virus elimination rate and hardening survival rate |
| CN108559733A (en) * | 2017-12-31 | 2018-09-21 | 青岛袁策生物科技有限公司 | A method of promoting the fusion of rice many cells |
| CN110338062A (en) * | 2019-08-16 | 2019-10-18 | 杨迪 | A kind of method of Radix Glycyrrhizae Anther Culture evoked callus |
-
2020
- 2020-06-18 CN CN202010558849.6A patent/CN111587788A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992017056A1 (en) * | 1991-04-01 | 1992-10-15 | Florigene B.V. | Carnation plants and methods for their transformation and propagation |
| CN1951174A (en) * | 2005-10-20 | 2007-04-25 | 上海师范大学 | A culture medium composition adaptable for carnation stalk induced bud and application thereof |
| CN101874471A (en) * | 2009-11-10 | 2010-11-03 | 华中农业大学 | Plant regeneration method of dianthus caryophyllus direct somatic embryo generating path and special culture medium |
| CN108112475A (en) * | 2017-12-13 | 2018-06-05 | 云南中烟工业有限责任公司 | A kind of method that tobacco Brown is prevented with vitamin C, L-cysteine and sodium hydrogensulfite |
| CN108559733A (en) * | 2017-12-31 | 2018-09-21 | 青岛袁策生物科技有限公司 | A method of promoting the fusion of rice many cells |
| CN108522270A (en) * | 2018-03-06 | 2018-09-14 | 山东寿光蔬菜种业集团有限公司 | A method of based on cucumber anther culture and improvement virus elimination rate and hardening survival rate |
| CN110338062A (en) * | 2019-08-16 | 2019-10-18 | 杨迪 | A kind of method of Radix Glycyrrhizae Anther Culture evoked callus |
Non-Patent Citations (1)
| Title |
|---|
| 程晓岩: "香石竹愈伤组织的诱导及遗传转化的研究", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111972293A (en) * | 2020-10-19 | 2020-11-24 | 江苏春之雨生物科技发展有限公司 | Callus induction method for in vitro culture of sesame unpolarized ovaries |
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Application publication date: 20200828 |