CN102239805A - Tissue-culture quick breeding method for iris tectorum - Google Patents
Tissue-culture quick breeding method for iris tectorum Download PDFInfo
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- 244000071493 Iris tectorum Species 0.000 title claims abstract description 11
- 238000009395 breeding Methods 0.000 title abstract 3
- 239000001963 growth medium Substances 0.000 claims abstract description 50
- 230000006698 induction Effects 0.000 claims abstract description 25
- 238000012258 culturing Methods 0.000 claims abstract description 24
- 238000000034 method Methods 0.000 claims abstract description 14
- 230000004083 survival effect Effects 0.000 claims abstract description 6
- 239000002609 medium Substances 0.000 claims description 19
- 238000002791 soaking Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 230000001954 sterilising effect Effects 0.000 claims description 14
- 239000011159 matrix material Substances 0.000 claims description 13
- 241000196324 Embryophyta Species 0.000 claims description 11
- 239000012883 rooting culture medium Substances 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 238000005520 cutting process Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- 229920001817 Agar Polymers 0.000 claims description 8
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 8
- 229930006000 Sucrose Natural products 0.000 claims description 8
- 239000008272 agar Substances 0.000 claims description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims description 8
- 239000003599 detergent Substances 0.000 claims description 6
- 229960002523 mercuric chloride Drugs 0.000 claims description 6
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000008223 sterile water Substances 0.000 claims description 6
- 239000002689 soil Substances 0.000 claims description 4
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 claims description 3
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 claims description 3
- 239000006013 carbendazim Substances 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 3
- 239000003415 peat Substances 0.000 claims description 3
- 239000002985 plastic film Substances 0.000 claims description 3
- 229920006255 plastic film Polymers 0.000 claims description 3
- 230000004069 differentiation Effects 0.000 abstract description 3
- 238000011081 inoculation Methods 0.000 abstract description 3
- 239000005556 hormone Substances 0.000 abstract description 2
- 229940088597 hormone Drugs 0.000 abstract description 2
- 238000004161 plant tissue culture Methods 0.000 abstract description 2
- 239000003242 anti bacterial agent Substances 0.000 abstract 1
- 229940088710 antibiotic agent Drugs 0.000 abstract 1
- 241000421576 Iris sp. ser. Hexagonae Species 0.000 description 8
- 238000012136 culture method Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 238000005286 illumination Methods 0.000 description 4
- 238000001784 detoxification Methods 0.000 description 3
- 241000167854 Bourreria succulenta Species 0.000 description 2
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- 239000003899 bactericide agent Substances 0.000 description 2
- 235000019693 cherries Nutrition 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 2
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical compound C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 description 2
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- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 description 1
- 229930191978 Gibberellin Natural products 0.000 description 1
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- 238000011109 contamination Methods 0.000 description 1
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- 238000011031 large-scale manufacturing process Methods 0.000 description 1
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- 239000000463 material Substances 0.000 description 1
- IQUFIVQRGAZYMZ-UHFFFAOYSA-N n-benzyl-7h-purin-6-amine Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CC=C1.N=1C=NC=2N=CNC=2C=1NCC1=CC=CC=C1 IQUFIVQRGAZYMZ-UHFFFAOYSA-N 0.000 description 1
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- 238000011069 regeneration method Methods 0.000 description 1
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- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
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Abstract
The invention discloses a tissue-culture quick breeding method for iris tectorum, belonging to the technical field of plant tissue culture. The method comprises the following technical steps: (1) preparing a culture medium; (2) culturing iris tectorum detoxified tissue-culture seedlings; and (3) hardening and transplanting the tissue-culture seedlings. With the method, through adjustment on the type and the content of culture medium hormones, addition of antibiotics and other technical measures, the pollution rate of a iris tectorum explant is obviously lowered, and an inoculation survival rate is above 90%; the induction differentiation rate of the explant can be obviously improved to 99% from 70-80% in the prior art; and the tissue-culture strong seedling rate of the iris tectorum is improved. The tissue-culture quick breeding method can be popularized and applied to landscaping and tissue culture enterprises.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a tissue culture method of Louisiana iris.
Background
Louisiana Iris is an aquatic floral plant of Iris of Iridaceae, native to Louisiana, USA. The plant has been widely applied in North America and Europe in some countries, the flowering phase is 4-5 months, the flower color is rich, the plant not only has the excellent ornamental properties of large flower, bright color, leaf flower combination and the like, but also has the characteristics of strong adaptability, strong cold resistance, strong drought resistance, extensive management and the like, and simultaneously has the advantages of evergreen and the like in the south area of the Yangtze river in China. At present, the market situation looks good, but the supply of high-quality nursery stocks is not in demand. The series of flowers are mainly propagated by plants and seeds in the past, so that the propagation speed is low, labor and time are wasted, and the market demand cannot be met; meanwhile, the germination rate of the plant seeds is low, the separation of the posterity of the seed propagation is serious, a plurality of varieties can not reproduce the characteristics of the original excellent varieties, and the plant division propagation is limited by seasons. Although there have been reports on tissue culture methods of louisiana iris in the prior art, for example, juxudong, tissue culture of aquatic evergreen hybrid iris, chinese floriculture, 2007, 10: 23-25; louisiana iris tissue culture and seedling growth rule preliminary study, Fujian forestry science and technology, 2009, 36 (3): 175 — 179); callus induction and bud differentiation in the process of tissue culture of Wu Yueyi Yan, Louisiana Iris, Zhejiang agricultural science, 2009, 1: 86-89; establishment of a giamling, louisiana iris rapid propagation system, scientific and technological reporting, 201026 (4): 518-522; however, the above techniques mainly have the following problems: the contamination rate is higher due to improper sterilization technology, so that the survival rate of explant inoculation is lower-basically 60-70%; the bud induction rate is lower-70-80% due to improper hormone concentration; therefore, a tissue culture and rapid propagation method of iris which can solve the technical defects, is high in speed and propagation coefficient is urgently needed in production, the problem that the seedling supply is not in short supply in the production practice of the Louisiana iris is solved, and the popularization and application work of the Louisiana iris is accelerated.
Disclosure of Invention
The invention aims to provide an iris tissue culture rapid propagation method which is simple in operation, low in pollution rate, high in plant regeneration power and excellent in seedling quality, aiming at the defects of high explant pollution rate and low bud induction rate in the existing iris tissue culture technology.
A tissue culture and rapid propagation method of iris comprises the following steps:
(1) the preparation of the culture medium comprises the components of a basic culture medium and the culture medium of each phase of tissue culture, and the weight and the volume of each liter are as follows:
1) basic culture medium: adopting LS basic culture medium, wherein white sugar 15-30g/L, agar 5-8g/L, and pH5.6-5.8;
2) bud induction medium: LS + Shannon No. 5-10ml/L +6-BA 0.3-1.0mg/L + NAA0.1-0.3 mg/L;
3) subculture multiplication medium: LS +6-BA 0.5-1.5mg/L + NAA 0.2-0.5 mg/L;
4) strong seedling culture medium: LS +6-BA 0.3-0.5mg/L and NAA 0.5-0.8 mg/L;
5) rooting culture medium: LS + NAA0.5-1.5mg/L + GA0.3-0.5 mg/L;
(2) and (3) culturing the virus-free tissue culture seedlings of the iris:
1) selection and sterilization of explants: taking tender underground buds of the robust iris plants, cutting off leaves and root systems above the basal parts, soaking for 8-10min by using 10% detergent, and washing for 1-2h by using running water; washing with sterile water on a clean bench, sterilizing with 70% ethanol for 30-60s, soaking in 0.1% mercuric chloride water solution for 8-15min, sterilizing, and washing with sterile water for 5-6 times; removing the stem tip tissue block, and cutting into 0.3-0.5cm3The small blocks are used as explants for standby;
2) and (3) bud induction culture: inoculating the stem tip explant to a bud induction culture medium, and carrying out induction culture for 15-20 days at the temperature of 25 +/-2 ℃, under the conditions of light intensity of 1500-;
3) bud multiplication culture: transferring the cluster buds to a subculture multiplication medium, and culturing for 15-25 days at 25 +/-2 ℃ under the conditions of light intensity of 1500-; according to the requirement of the number of the cluster buds, carrying out re-proliferation once every 10-20 days according to the same method;
4) strong seedling culture: transferring the subculture cluster buds to a strong seedling culture medium, and culturing for 20-30 days at 25 +/-2 ℃ under the conditions of light intensity of 1500-;
5) rooting culture: transferring the strong seedlings into a rooting culture medium, culturing for 15-30 days at 25 +/-2 ℃, with light intensity of 1500-;
(3) hardening and transplanting tissue culture seedlings:
1) hardening seedlings: transferring the tissue culture seedling with the length of 10-15cm and the length of more than or equal to 5 roots into a common greenhouse, placing for 1-3 days, opening the bottle cap, and exercising for 3-5 days under the conditions of 25-28 ℃, humidity of 70-80% and light intensity of 7000-;
2) transplanting and culturing: cleaning a root culture medium of the domesticated tissue culture seedlings, soaking roots of the domesticated tissue culture seedlings in 0.1% carbendazim solution for 30-60 seconds, transplanting the domesticated tissue culture seedlings into a mixed matrix of sandy soil, peat and yellow soil according to the volume ratio of 6: 3: 1, covering the mixed matrix with a plastic film, covering the mixed matrix with a shading net, culturing the mixed matrix under the conditions that the temperature is 28-30 ℃, the humidity is 80-95%, the light intensity is weak firstly and then strong, culturing the mixed matrix for 1 week under the condition that the light intensity is less than 10000lx, and gradually opening the film; after 2 weeks, performing conventional management, wherein the transplanting survival rate of the tissue culture seedlings reaches more than 95%; culturing for 1-2 months to obtain rhizoma Iridis Tectori seedling.
The formula of the culture medium comprises the components of a basic culture medium and the culture medium at each stage of tissue culture, and the weight and the volume of each liter of the components are as follows:
1) basic culture medium: LS culture medium, wherein white sugar 20g/L, agar 5.5g/L, pH5.8;
2) bud induction medium: LS + Shannon No. 7ml/L +6-BA0.5mg/L + NAA0.2 mg/L;
3) subculture multiplication medium: LS +6-BA1.0mg/L + NAA0.3 mg/L;
4) strong seedling culture medium: LS +6-BA0.3mg/L + NAA0.5 mg/L;
5) rooting culture medium: LS + NAA1.0mg/L + GA0.3 mg/L.
The invention has the beneficial effects that:
1) according to the invention, the underground tender shoots which just germinate are taken at the time of early spring in the late winter to obtain relatively clean explants, proper sterilization technology is combined, and a proper dosage (5-10ml/L) of broad-spectrum biological bactericide Shannon I is added into the initial generation culture medium, so that the inoculation survival rate of the explants is improved to more than 90% from the conventional 60-70%, and a large amount of sterile materials are obtained in a short period;
2) the invention accelerates the induced differentiation of the explant by proper 6-BA concentration (0.3-1.0mg/L), so that the bud induction rate reaches 99 percent, and is obviously improved compared with 70-80 percent generally reported in literature data; the propagation coefficient and the strong seedling rate of multiple buds are improved through proper combination of 6-BA (0.3-1.5mg/L) and NAA (0.2-0.5mg/L), and the application of comprehensive technologies such as screening the optimal rooting agent and the like is adopted, so that the propagation quantity of the tissue culture seedlings of the iris tectorum reaches more than 20 ten thousand plants per year, and the large-scale production of the iris tectorum seedlings is realized.
Detailed Description
The present invention is further illustrated in detail by the following examples, but the present invention is not limited thereto. Wherein,
6-BA: 6-Benzylaminopurine (6-Benzylaminopurine), Sigma import domestic split charging, analytically pure, purity 99.9%.
NAA: alpha-Naphthylacetic acid (1-naphylacetic acid), Sigma import domestic split charging, analytically pure, purity 99.5%.
GA3: gibberellin (Gibberellin A3) is imported and distributed at home, and is analytically pure with the purity of 90%.
Shannon No. one: broad-spectrum biological bactericide, provided by prant biotechnology limited.
Liquid detergent: libai detergent, general detergent, Guangzhou Libai Enterprise group Co.
Example 1: (tissue culture method of iris 1)
The method comprises the following steps:
(1) the preparation of the culture medium comprises the components of a basic culture medium and the culture medium of each stage of tissue culture, and the weight of each liter is as follows:
1) basic culture medium: LS culture medium, wherein white sugar 20g/L, agar 5.5g/L, pH5.8;
2) bud induction medium: LS + Shannon No. 7ml/L +6-BA0.5mg/L + NAA0.2mg/L;
3) subculture multiplication medium: LS +6-BA1.0mg/L + NAA0.3mg/L;
4) strong seedling culture medium: LS +6-BA0.3mg/L and NAA0.5mg/L;
5) rooting culture medium: LS + NAA1.0mg/L + GA0.3mg/L;
wherein the formula of the LS basic culture medium is as follows;
(2) and (3) culturing the virus-free tissue culture seedlings of the iris:
1) selection and sterilization of explants: taking young underground buds of the robust iris plants, cutting off leaves and root systems above the basal parts, soaking for 8-10min by using 10% Libai detergent, and washing for 1-2h by using running water; washing with sterile water on a clean bench, sterilizing with 70% ethanol for 45s, soaking in 0.1% mercuric chloride water solution for 10min, sterilizing, and washing with sterile water for 5-6 times; removing the stem tip tissue block, and cutting into 0.3cm3The small blocks are used as explants for standby;
2) and (3) bud induction culture: inoculating the stem tip explant to a bud induction culture medium, and carrying out induction culture for 15-20 days under the conditions of 25 +/-2 ℃, light intensity of 1500Lx and illumination of 12h/d until cluster buds are formed;
3) bud multiplication culture: transferring the cluster buds to a subculture multiplication medium, and culturing for 15-25 days at 25 +/-2 ℃ under the conditions of light intensity of 1500Lx and illumination of 12h/d until the subculture cluster buds are formed; according to the requirement of the number of the cluster buds, carrying out re-proliferation once every 10-20 days according to the same method;
4) strong seedling culture: transferring the subculture cluster buds to a strong seedling culture medium, and culturing for 20-30 days at 25 +/-2 ℃ under the conditions of light intensity of 1500Lx and illumination of 12h/d until the height of the seedlings is 6-8 cm;
5) rooting culture: transferring the strong seedlings into a rooting culture medium, culturing for 15-30 days at 25 +/-2 ℃, under the conditions of light intensity of 1500Lx and illumination of 12h/d until 1-5 root systems grow on the base part, thus obtaining the virus-free tissue culture seedlings of the iris tectorum;
(3) hardening and transplanting tissue culture seedlings:
1) hardening seedlings: transferring the tissue culture seedling with the length of 10-15cm and the length of more than or equal to 5 roots into a greenhouse, placing for 1-3 days, opening the bottle cap, and exercising the seedling for 3-5 days under the conditions of 25-28 ℃, humidity of 70-80% and light intensity of 7000-;
2) transplanting and culturing: cleaning a root culture medium of the domesticated tissue culture seedlings, soaking roots of the domesticated tissue culture seedlings in 0.1% carbendazim solution for 30-60 seconds, transplanting the domesticated tissue culture seedlings into a mixed matrix of sandy loam, peat and yellow loam according to the volume ratio of 6: 3: 1, covering the mixed matrix with a plastic film, covering the mixed matrix with a shading net, culturing the domesticated tissue culture seedlings for 1 week at the temperature of 28-30 ℃, the humidity of 80-95% and the light intensity of less than 10000lx in a weak and strong mode, and gradually opening the film; after 2 weeks, conventional management is carried out, and the transplanting survival rate of the tissue culture seedlings can reach more than 95%; culturing for 1-2 months to obtain rhizoma Iridis Tectori seedlings, and outplanting.
Example 2: (tissue culture method of iris tectorum 2)
In this example, the preparation of the medium in step (1): 15g/L of white sugar, 6g/L of agar and 5.7 of pHs in the basic culture medium; the bud induction culture medium is: LS + Shannon No. 5ml/L +6-BA0.3mg/L + NAA0.1 mg/L; the subculture multiplication medium comprises: LS +6-BA0.5mg/L + NAA0.2mg/L; the strong seedling culture medium comprises: LS +6-BA0.3mg/L + NAA 0.6 mg/L; the rooting culture medium comprises: LS + NAA0.8+ GA30.4 mg/L; step (2), cultivation of the Iris detoxification tissue culture seedlings: 1) selection and sterilization of explants: soaking tender shoots in 70% alcohol for 30s,soaking in 0.1% mercuric chloride water solution for 8min, removing stem tip tissue block, and cutting into 0.4cm pieces3Small blocks; the light intensity of each stage of bud induction, subculture multiplication, strong seedling and rooting culture is 2500 Lx; the rest of the steps are the same as the example 1.
Example 3: (Louisiana iris 'cherry' tissue culture method 3)
In this example, the preparation of the medium in step (1): 25g/L of white sugar, 7g/L of agar and 5.6 of pHs in a basic culture medium; the bud induction culture medium is: LS + Shannong No. 8ml/L +6-BA0.8mg/L + NAA0.2 mg/L; the subculture multiplication medium comprises: LS +6-BA0.8mg/L + NAA0.4mg/L; the strong seedling culture medium comprises: LS +6-BA0.4mg/L + NAA 0.7 mg/L; the rooting culture medium comprises: LS + NAA0.5+ GA30.5 mg/L; step (2), cultivation of the Iris detoxification tissue culture seedlings: 1) selection and sterilization of explants: soaking young bud in 70% alcohol for 50s, soaking in 0.1% mercuric chloride water solution for 12min, removing stem tip tissue block, and cutting into 0.45cm pieces3Small blocks; the light intensity of each stage of bud induction, subculture multiplication, strong seedling and rooting culture is 1800 Lx; the rest of the steps are the same as the example 1.
Example 4: (Louisiana iris 'cherry' tissue culture method 4)
In this example, the preparation of the medium in step (1): 30g/L of white sugar, 8g/L of agar and 5.7 of pHs in the basic culture medium; the bud induction culture medium is: LS + Shannon No. I10 ml/L +6-BA1.0mg/L + NAA0.3mg/L; the subculture multiplication medium comprises: LS +6-BA1.5mg/L + NAA0.5mg/L; the strong seedling culture medium comprises: LS +6-BA0.5mg/L + NAA0.8 mg/L; the rooting culture medium comprises: LS + NAA1.5+ GA30.3 mg/L; step (2), cultivation of the Iris detoxification tissue culture seedlings: 1) selection and sterilization of explants: soaking young bud in 70% alcohol for 60s, soaking in 0.1% mercuric chloride water solution for 15min, removing stem tip tissue block, and cutting into 0.5cm pieces3Small blocks; the light intensity of each stage of bud induction, subculture multiplication, strong seedling and rooting culture is 2000 Lx; the rest of the steps are the same as the example 1.
Claims (2)
1. A tissue culture and rapid propagation method of iris tectorum is characterized by comprising the following steps:
(1) the preparation of the culture medium comprises the components of a basic culture medium and the culture medium of each phase of tissue culture, and the weight and the volume of each liter are as follows:
1) basic culture medium: adopting LS basic culture medium, wherein white sugar 15-30g/L, agar 5-8g/L, and pH5.6-5.8;
2) bud induction medium: LS + Shannon No. 5-10ml/L +6-BA 0.3-1.0mg/L + NAA0.1-0.3 mg/L;
3) subculture multiplication medium: LS +6-BA 0.5-1.5mg/L + NAA 0.2-0.5 mg/L;
4) strong seedling culture medium: LS +6-BA 0.3-0.5mg/L and NAA 0.5-0.8 mg/L;
5) rooting culture medium: LS + NAA0.5-1.5mg/L + GA0.3-0.5 mg/L;
(2) and (3) culturing the virus-free tissue culture seedlings of the iris:
1) selection and sterilization of explants: taking tender underground buds of the robust iris plants, cutting off leaves and root systems above the basal parts, soaking for 8-10min by using 10% detergent, and washing for 1-2h by using running water; washing with sterile water on a clean bench, sterilizing with 70% ethanol for 30-60s, soaking in 0.1% mercuric chloride water solution for 8-15min, sterilizing, and washing with sterile water for 5-6 times; removing the stem tip tissue block, and cutting into 0.3-0.5cm3The small blocks are used as explants for standby;
2) and (3) bud induction culture: inoculating the stem tip explant to a bud induction culture medium, and carrying out induction culture for 15-20 days at the temperature of 25 +/-2 ℃, under the conditions of light intensity of 1500-;
3) bud multiplication culture: transferring the cluster buds to a subculture multiplication medium, and culturing for 15-25 days at 25 +/-2 ℃ under the conditions of light intensity of 1500-; according to the requirement of the number of the cluster buds, carrying out re-proliferation once every 10-20 days according to the same method;
4) strong seedling culture: transferring the subculture cluster buds to a strong seedling culture medium, and culturing for 20-30 days at 25 +/-2 ℃ under the conditions of light intensity of 1500-;
5) rooting culture: transferring the strong seedlings into a rooting culture medium, culturing for 15-30 days at 25 +/-2 ℃, with light intensity of 1500-;
(3) hardening and transplanting tissue culture seedlings:
1) hardening seedlings: transferring the tissue culture seedling with the length of 10-15cm and the length of more than or equal to 5 roots into a common greenhouse, placing for 1-3 days, opening the bottle cap, and exercising for 3-5 days under the conditions of 25-28 ℃, humidity of 70-80% and light intensity of 7000-;
2) transplanting and culturing: cleaning a root culture medium of the domesticated tissue culture seedlings, soaking roots of the domesticated tissue culture seedlings in 0.1% carbendazim solution for 30-60 seconds, transplanting the domesticated tissue culture seedlings into a mixed matrix of sandy soil, peat and yellow soil according to the volume ratio of 6: 3: 1, covering the mixed matrix with a plastic film, covering the mixed matrix with a shading net, culturing the mixed matrix under the conditions that the temperature is 28-30 ℃, the humidity is 80-95%, the light intensity is weak firstly and then strong, culturing the mixed matrix for 1 week under the condition that the light intensity is less than 10000lx, and gradually opening the film; after 2 weeks, performing conventional management, wherein the transplanting survival rate of the tissue culture seedlings reaches more than 95%; culturing for 1-2 months to obtain rhizoma Iridis Tectori seedling.
2. The method of claim 1, wherein the culture medium comprises the following components in terms of weight and volume per liter:
1) basic culture medium: LS culture medium, wherein white sugar 20g/L, agar 5.5g/L, pH5.8;
2) bud induction medium: LS + Shannon No. 7ml/L +6-BA0.5mg/L + NAA0.2 mg/L;
3) subculture multiplication medium: LS +6-BA1.0mg/L + NAA0.3 mg/L;
4) strong seedling culture medium: LS +6-BA0.3mg/L + NAA0.5 mg/L;
5) rooting culture medium: LS + NAA1.0mg/L + GA0.3 mg/L.
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102524082A (en) * | 2012-03-17 | 2012-07-04 | 常熟市尚湖农业生态园有限公司 | Method of nanometer tissue culture of iris ensata |
| CN102893861A (en) * | 2012-02-21 | 2013-01-30 | 张子学 | Technology for regenerating and recycling plant tissue medium polluted by undesired bacteria |
| CN102893872A (en) * | 2012-11-03 | 2013-01-30 | 云南省农业科学院花卉研究所 | Tissue culture method for domesticated seedlings of iris pallida |
| CN103202230A (en) * | 2013-04-22 | 2013-07-17 | 江苏省中国科学院植物研究所 | Rapid propagation method of red-seed iris |
| CN103348914A (en) * | 2013-06-28 | 2013-10-16 | 西藏自治区农牧科学院蔬菜研究所 | Plant cultivation method |
| CN103734020A (en) * | 2014-01-26 | 2014-04-23 | 上海上房园艺有限公司 | Tissue culture method for German iris tectorum |
| CN104303809A (en) * | 2014-11-12 | 2015-01-28 | 浙江采悠园林有限公司 | Container cultivation method for iris hexagonus hybrid |
| CN104604481A (en) * | 2015-01-08 | 2015-05-13 | 杭州嘉泰园艺有限公司 | Rapid propagation method of Louisiana irises |
| CN104839019A (en) * | 2015-04-29 | 2015-08-19 | 浙江农林大学 | Method for in-vitro rapid propagation of iris laevigata by using immature fruits |
| CN105104187A (en) * | 2015-07-24 | 2015-12-02 | 浙江省嘉兴市农业科学研究院（所） | Crocus sativus L. tissue cultured corm strengthening and rooting medium and tissue culture method |
| CN109006487A (en) * | 2018-09-30 | 2018-12-18 | 西南林业大学 | A kind of method for tissue culture of pale flag |
| CN109362563A (en) * | 2018-10-31 | 2019-02-22 | 杭州晨航环境工程有限公司 | The rapid propagation method of the red Rett of Louisiana's iris |
| CN110537491A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园艺有限公司 | method for rapid breeding of African dual-color wild iris through tissue culture |
| CN116649218A (en) * | 2023-06-28 | 2023-08-29 | 江苏省中国科学院植物研究所 | Efficient breeding method of Lissanamia iris |
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| CN102893861A (en) * | 2012-02-21 | 2013-01-30 | 张子学 | Technology for regenerating and recycling plant tissue medium polluted by undesired bacteria |
| CN102524082A (en) * | 2012-03-17 | 2012-07-04 | 常熟市尚湖农业生态园有限公司 | Method of nanometer tissue culture of iris ensata |
| CN102524082B (en) * | 2012-03-17 | 2014-03-05 | 常熟市润丰农业有限公司 | Method of nanometer tissue culture of iris ensata |
| CN102893872A (en) * | 2012-11-03 | 2013-01-30 | 云南省农业科学院花卉研究所 | Tissue culture method for domesticated seedlings of iris pallida |
| CN103202230A (en) * | 2013-04-22 | 2013-07-17 | 江苏省中国科学院植物研究所 | Rapid propagation method of red-seed iris |
| CN103348914A (en) * | 2013-06-28 | 2013-10-16 | 西藏自治区农牧科学院蔬菜研究所 | Plant cultivation method |
| CN103734020A (en) * | 2014-01-26 | 2014-04-23 | 上海上房园艺有限公司 | Tissue culture method for German iris tectorum |
| CN104303809A (en) * | 2014-11-12 | 2015-01-28 | 浙江采悠园林有限公司 | Container cultivation method for iris hexagonus hybrid |
| CN104604481A (en) * | 2015-01-08 | 2015-05-13 | 杭州嘉泰园艺有限公司 | Rapid propagation method of Louisiana irises |
| CN104839019A (en) * | 2015-04-29 | 2015-08-19 | 浙江农林大学 | Method for in-vitro rapid propagation of iris laevigata by using immature fruits |
| CN105104187A (en) * | 2015-07-24 | 2015-12-02 | 浙江省嘉兴市农业科学研究院（所） | Crocus sativus L. tissue cultured corm strengthening and rooting medium and tissue culture method |
| CN109006487A (en) * | 2018-09-30 | 2018-12-18 | 西南林业大学 | A kind of method for tissue culture of pale flag |
| CN109362563A (en) * | 2018-10-31 | 2019-02-22 | 杭州晨航环境工程有限公司 | The rapid propagation method of the red Rett of Louisiana's iris |
| CN110537491A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园艺有限公司 | method for rapid breeding of African dual-color wild iris through tissue culture |
| CN116649218A (en) * | 2023-06-28 | 2023-08-29 | 江苏省中国科学院植物研究所 | Efficient breeding method of Lissanamia iris |
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