CN111758569A - Method for inhibiting calli of fragrant fruit trees from browning - Google Patents
Method for inhibiting calli of fragrant fruit trees from browning Download PDFInfo
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- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY 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
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Abstract
The invention relates to a method for inhibiting calli of a fragrant fruit tree from browning, which comprises the following steps: (1) and (3) disinfection of explants: taking the petiole of the myrtle tree as an explant, and disinfecting to obtain a disinfected explant; (2) callus induction: inoculating the sterilized explant on a callus induction culture medium under an aseptic condition, and culturing to form callus on the explant; (3) anti-browning subculture: transferring the callus into a subculture medium, and performing subculture to achieve the effect of inhibiting the callus of the fragrant fruit tree from browning. Compared with the prior art, the method takes the petiole of the fragrant fruit tree as an explant, adopts the special callus induction culture medium to induce the callus, and utilizes the browning-inhibiting subculture medium to perform subculture, so that the browning rate of the callus is reduced to 3.3 percent.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a method for inhibiting calli of fragrant fruit trees from browning.
Background
The myrtle tree is a tall deciduous tree of the genus Syzygium of the family Rubiaceae, originates from the middle-aged chalk era of about 1 hundred million years from today, is a quaternary glacier wiggling plant, is a unique single-species rare tree species in China, and has great value in the aspects of systematic development and morphological evolution of the Rubiaceae plant, the research of the geographical system of the Chinese plant, appreciation, medicinal use, material use and the like.
Under the natural state, the fragrant fruit tree has typical intermittent fructification characteristics, and blossoms and fruits once in 2-4 years; the seeds have the characteristics of dormancy, tiny seeds, low vitality, only 1 year of life, extremely low natural germination rate, declining population and imminent extinction, and are classified as national secondary protection plants. The method for developing the research of the seedling breeding technology of the wild fragrant fruit tree by using the plant tissue culture method has important significance for overcoming the difficulty in seed breeding of the fragrant fruit tree, saving the germplasm resources of the rare endangered wild fragrant fruit tree and realizing the sustainable utilization of the germplasm resources of the fragrant fruit tree.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a method for inhibiting the calli of the fragrant fruit trees from browning.
The purpose of the invention can be realized by the following technical scheme:
a method for inhibiting calli of a myrtle tree from browning comprises the following steps:
(1) and (3) disinfection of explants: taking the petiole of the myrtle tree as an explant, and disinfecting to obtain a disinfected explant;
(2) callus induction: inoculating the sterilized explant on a callus induction culture medium under an aseptic condition, and culturing to form callus on the explant;
(3) anti-browning subculture: transferring the callus into a subculture medium, and performing subculture to achieve the effect of inhibiting the callus of the fragrant fruit tree from browning.
Further, the disinfection method comprises the following specific steps: sterilizing with ethanol and sodium hypochlorite, and washing with sterile water.
Further, the callus induction medium comprises the following components: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 20g of cane sugar, 6g of agar and the balance of water.
Further, the temperature of the culture in the step (2) is 25 +/-2 ℃, and the time is 20 days; the temperature of the subculture in the step (3) is 25 +/-2 ℃ and the time is 20 days.
Further, the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mgMgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mgH3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mgFeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 1-2g of polyvinylpyrrolidone, 20g of cane sugar, 6g of agar and the balance of water.
Further, the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mgMgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mgH3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mgFeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D and 2g of polyethyleneVinylpyrrolidone, 20g sucrose, 6g agar and balance water.
Further, the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mgMgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mgH3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mgFeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.5-1g of citric acid, 20g of cane sugar, 6g of agar and the balance of water.
Further, the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mgMgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mgH3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mgFeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.5g of citric acid, 20g of cane sugar, 6g of agar and the balance of water.
Further, the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mgMgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mgH3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mgFeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.01-0.2g of ascorbic acid, 20g of cane sugar, 6g of agar and the balance of water.
Further, the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mgMgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mgH3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mgFeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.01-0.1g of ascorbic acid, 20g of cane sugar, 6g of agar and the balance of water.
The calli browning mechanism of the fragrant fruit trees comprises the following steps: the activity of Phenylalanine Ammonia Lyase (PAL) and polyphenol oxidase (PPO) is improved by the higher concentration of 6-BA in the subculture medium, the PAL promotes the synthesis and accumulation of phenolic substances in the callus, and the phenolic substances are oxidized under the catalysis of the high-activity PPO to generate a large amount of brown quinone substances, thereby causing the browning of the callus and leading the culture of the callus to be incapable of being continued.
Polyvinylpyrrolidone, citric acid and ascorbic acid inhibit the cause of browning: polyvinylpyrrolidone, citric acid and ascorbic acid with proper concentration are added into the subculture medium, so that PAL activity and PPO activity can be remarkably inhibited, the synthesis of phenolic substances and the formation of quinone substances are reduced, the browning rate of callus is reduced, and the subculture of the callus can be continued.
Compared with the prior art, the method takes the petiole of the fragrant fruit tree as an explant, adopts the specific callus induction culture medium to induce the callus, and utilizes the browning-inhibiting subculture medium to perform subculture, so that the browning rate of the callus can be obviously reduced, the subculture of the callus can be continued, and a usable material is provided for the subsequent bud differentiation and plant regeneration.
Detailed Description
The following examples are given for the detailed implementation and specific operation of the present invention, but the scope of the present invention is not limited to the following examples.
Example 1
A method for inhibiting calli of a fragrant fruit tree from browning comprises the following steps:
(1) and (3) disinfection of explants: taking the leaf stalks of the fragrant fruit trees as explants, sequentially disinfecting the leaf stalks by using 75% alcohol by volume percentage for 15 seconds and 0.1% sodium hypochlorite aqueous solution by volume percentage for 5 minutes, and then washing the leaf stalks by using sterile water to obtain disinfected explants;
(2) callus induction: inoculating the sterilized explant on a callus induction culture medium under an aseptic condition, controlling the temperature to be 25 +/-2 ℃ and culturing for 20 days to form callus on the sterilized explant;
callus induction medium, calculated per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B1、0.25mgVitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 20g of cane sugar, 6g of agar and the balance of water;
(3) subculturing on a browning-inhibiting medium: transferring the callus into a subculture medium, and performing proliferation subculture for 20 days at a controlled temperature of 25 +/-2 ℃;
proliferation medium, calculated per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 2g of polyvinylpyrrolidone, 20g of sucrose, 6g of agar and the balance of water.
Example 2
The difference from example 1 is that the concentration of polyvinylpyrrolidone in the callus subculture medium in step (3) was 1 g/L.
Example 3
The difference from example 1 is that the concentration of polyvinylpyrrolidone in the callus subculture medium in step (3) was 1.5 g/L.
The anti-browning effects of examples 1-3 are shown in Table 1.
TABLE 1
| Treatment of | PVP concentration (g/L) | Browning rate% |
| Control | 0 | 44.3 |
| Example 2 | 1 | 23.3 |
| Example 3 | 1.5 | 14.0 |
| Example 1 | 2 | 3.3 |
The result shows that the browning rate of the callus is only 3.3% when the addition amount of PVP is 2g/L, and the anti-browning effect is very obvious.
Example 4
A method for inhibiting calli of a fragrant fruit tree from browning comprises the following steps:
(1) and (3) disinfection of explants: taking the leaf stalks of the fragrant fruit trees as explants, sequentially disinfecting the leaf stalks by using 75% alcohol by volume percentage for 15 seconds and 0.1% sodium hypochlorite aqueous solution by volume percentage for 5 minutes, and then washing the leaf stalks by using sterile water to obtain disinfected explants;
(2) callus induction: inoculating the sterilized explant on a callus induction culture medium under an aseptic condition, controlling the temperature to be 25 +/-2 ℃ and culturing for 20 days to form callus on the sterilized explant;
callus induction medium, calculated per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 20g of cane sugar, 6g of agar and the balance of water;
(3) subculturing on a browning-inhibiting medium: transferring the callus into a subculture medium, and performing proliferation subculture for 20 days at a controlled temperature of 25 +/-2 ℃;
proliferation medium, calculated per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.5g of citric acid, 20g of cane sugar, 6g of agar and the balance of water.
Comparative example 1
The difference from example 4 was that the concentration of citric acid in the callus subculture medium in step (3) was 0.2 g/L.
Example 5
The difference from example 4 was that the concentration of citric acid in the callus subculture medium in step (3) was 1 g/L.
Comparative example 2
The difference from example 4 was that the concentration of citric acid in the callus subculture medium in step (3) was 1.5 g/L.
The anti-browning effects of examples 4 to 5 and comparative examples 1 to 2 are shown in table 2.
TABLE 2
| Treatment of | Citric acid concentration (g/L) | Browning rate% |
| Control | 0 | 44.3 |
| Comparative example 1 | 0.2 | 46.4 |
| Example 4 | 0.5 | 24.5 |
| Example 5 | 1 | 43.1 |
| Comparative example 2 | 1.5 | 83.6 |
The result shows that the browning rate of the callus is only 24.5% when the addition amount of the citric acid is 0.5g/L, and the anti-browning effect is very obvious.
Example 6
A method for inhibiting calli of a fragrant fruit tree from browning comprises the following steps:
(1) and (3) disinfection of explants: taking the leaf stalks of the fragrant fruit trees as explants, sequentially disinfecting the leaf stalks by using 75% alcohol by volume percentage for 15 seconds and 0.1% sodium hypochlorite aqueous solution by volume percentage for 5 minutes, and then washing the leaf stalks by using sterile water to obtain disinfected explants;
(2) callus induction: inoculating the sterilized explant on a callus induction culture medium under an aseptic condition, controlling the temperature to be 25 +/-2 ℃ and culturing for 20 days to form callus on the sterilized explant;
callus induction medium, calculated per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 20g of cane sugar, 6g of agar and the balance of water;
(3) subculturing on a browning-inhibiting medium: transferring the callus into a subculture medium, and performing proliferation subculture for 20 days at a controlled temperature of 25 +/-2 ℃;
proliferation medium, calculated per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.1g of ascorbic acid, 20g of sucrose, 6g of agar and the balance of water.
Example 7
The difference from example 6 was that the ascorbic acid concentration in the callus subculture medium in step (3) was 0.01 g/L.
Example 8
The difference from example 6 was that the ascorbic acid concentration in the callus subculture medium in step (3) was 0.2 g/L.
Comparative example 3
The difference from example 6 was that the ascorbic acid concentration in the callus subculture medium in step (3) was 0.5 g/L.
The anti-browning effects of examples 6 to 8 and comparative example 3 are shown in table 3.
TABLE 3
| Treatment of | Ascorbic acid concentration (g/L) | Browning rate% |
| Control | 0 | 44.3 |
| Example 7 | 0.01 | 21.9 |
| Example 6 | 0.1 | 14.8 |
| Example 8 | 0.2 | 41.9 |
| Comparative example 3 | 0.5 | 94.4 |
The results show that the browning rate of the callus is only 14.8% when the addition amount of the ascorbic acid is 0.1g/L, and the anti-browning effect is very obvious.
In conclusion, the method for inhibiting calli of the fragrant fruit trees takes petioles of the fragrant fruit trees as explants, adopts the special callus induction culture medium to induce the calli, and utilizes the browning-inhibiting subculture medium to culture, so that the browning rate is only 3.3%. The obtained callus without browning and with good growth can be used for callus bud differentiation and plant regeneration of the fragrant fruit tree, and guarantees are provided for improving the propagation efficiency of the fragrant fruit tree seedling.
The foregoing is merely an example of the embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A method for inhibiting calli of a myrtle tree from browning is characterized by comprising the following steps:
(1) and (3) disinfection of explants: taking the petiole of the myrtle tree as an explant, and disinfecting to obtain a disinfected explant;
(2) callus induction: inoculating the sterilized explant on a callus induction culture medium under an aseptic condition, and culturing to form callus on the explant;
(3) anti-browning subculture: transferring the callus into a subculture medium, and performing subculture to achieve the effect of inhibiting the callus of the fragrant fruit tree from browning.
2. The method for inhibiting the calli of the myrtle tree from browning according to claim 1, wherein the disinfection is carried out by the following specific methods: sterilizing with ethanol and sodium hypochlorite, and washing with sterile water.
3. The method of claim 1, wherein the callus induction medium comprises: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mgKH2PO4、200mg NH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mgCuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 20g of cane sugar, 6g of agar and the balance of water.
4. The method for inhibiting the calli of the myrtle tree from browning according to claim 1, wherein the temperature of the culture in the step (2) is 25 ± 2 ℃ for 20 days; the temperature of the subculture in the step (3) is 25 +/-2 ℃ and the time is 20 days.
5. The method of claim 1, wherein the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 1-2g of polyvinylpyrrolidone, 20g of cane sugar, 6g of agar and the balance of water.
6. The method of claim 5, wherein the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 2g of polyvinylpyrrolidone, 20g of sucrose, 6g of agar and the balance of water.
7. The method of claim 1, wherein the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.5-1g of citric acid, 20g of cane sugar, 6g of agar and the balance of water.
8. The method of claim 7, wherein the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.5g of citric acid, 20g of cane sugar, 6g of agar and the balance of water.
9. The method of claim 1, wherein the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.01-0.2g of ascorbic acid, 20g of cane sugar, 6g of agar and the balance of water.
10. The method of claim 9, wherein the composition of the subculture medium is as follows: calculated as per liter, from 495mg K2SO4、185mg MgSO4·7H2O、85mg KH2PO4、200mgNH4NO3、11.25mg MnSO4·4H2O、4.3mg ZnSO4·7H2O、3.1mg H3BO3、0.125mg CuSO4·5H2O、0.125mg Na2MoO4·2H2O、278mg Ca(NO3)2·4H2O、13.9mg FeSO4·7H2O、18.65mg Na2-EDTA, 50mg inositol, 1mg glycine, 0.5mg vitamin B10.25mg vitamin B50.25mg vitamin B62mg of 6-BA, 0.5mg of 2,4-D, 0.01-0.1g of ascorbic acid, 20g of cane sugar, 6g of agar and the balance of water.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101027974A (en) * | 2007-04-10 | 2007-09-05 | 三峡大学 | Culture in vitro for fragrant fruit tree |
| CN104255459A (en) * | 2014-09-12 | 2015-01-07 | 南京通泽农业科技有限公司 | Rapid propagation method of lindera communis |
| CN108770689A (en) * | 2018-05-02 | 2018-11-09 | 上海应用技术大学 | A kind of propagation method of Emmenopterys henryi seedling |
-
2020
- 2020-07-07 CN CN202010645483.6A patent/CN111758569B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101027974A (en) * | 2007-04-10 | 2007-09-05 | 三峡大学 | Culture in vitro for fragrant fruit tree |
| CN104255459A (en) * | 2014-09-12 | 2015-01-07 | 南京通泽农业科技有限公司 | Rapid propagation method of lindera communis |
| CN108770689A (en) * | 2018-05-02 | 2018-11-09 | 上海应用技术大学 | A kind of propagation method of Emmenopterys henryi seedling |
Non-Patent Citations (5)
| Title |
|---|
| MENGGE ZHANG等: "Micropropagation via multiple shoot induction in Emmenopterys henryi Oliv.", 《EUROPEAN JOURNAL OF HORTICULTURAL SCIENCE》 * |
| 张梦鸽等: "香果树愈伤组织诱导及抗褐化研究", 《应用技术学报》 * |
| 彭祥等: "珍稀濒危植物香果树高频再生体系的建立", 《分子植物育种》 * |
| 熊丹等: "珍稀植物香果树植株再生体系的建立", 《北京林业大学学报》 * |
| 韦小丽等: "香果树组织培养技术研究", 《种子》 * |
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