CN112493132A - Method for improving gas microenvironment of plant tissue culture bottle seedlings and special culture medium - Google Patents
Method for improving gas microenvironment of plant tissue culture bottle seedlings and special culture medium Download PDFInfo
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- CN112493132A CN112493132A CN202011500483.3A CN202011500483A CN112493132A CN 112493132 A CN112493132 A CN 112493132A CN 202011500483 A CN202011500483 A CN 202011500483A CN 112493132 A CN112493132 A CN 112493132A
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- 239000001963 growth medium Substances 0.000 title claims abstract description 41
- 238000004161 plant tissue culture Methods 0.000 title claims abstract description 15
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- 239000008187 granular material Substances 0.000 claims abstract description 25
- 238000010521 absorption reaction Methods 0.000 claims abstract description 24
- 239000000463 material Substances 0.000 claims description 25
- 239000003795 chemical substances by application Substances 0.000 claims description 22
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 12
- 238000013268 sustained release Methods 0.000 claims description 11
- 239000012730 sustained-release form Substances 0.000 claims description 11
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 9
- 239000011347 resin Substances 0.000 claims description 9
- 229920005989 resin Polymers 0.000 claims description 9
- 229910052708 sodium Inorganic materials 0.000 claims description 9
- 239000011734 sodium Substances 0.000 claims description 9
- 230000002745 absorbent Effects 0.000 claims description 8
- 239000002250 absorbent Substances 0.000 claims description 8
- 229920001577 copolymer Polymers 0.000 claims description 8
- 229920001495 poly(sodium acrylate) polymer Polymers 0.000 claims description 8
- NNMHYFLPFNGQFZ-UHFFFAOYSA-M sodium polyacrylate Chemical compound [Na+].[O-]C(=O)C=C NNMHYFLPFNGQFZ-UHFFFAOYSA-M 0.000 claims description 8
- 239000004745 nonwoven fabric Substances 0.000 claims description 6
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 6
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 6
- HSEYYGFJBLWFGD-UHFFFAOYSA-N 4-methylsulfanyl-2-[(2-methylsulfanylpyridine-3-carbonyl)amino]butanoic acid Chemical compound CSCCC(C(O)=O)NC(=O)C1=CC=CN=C1SC HSEYYGFJBLWFGD-UHFFFAOYSA-N 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- RNIHAPSVIGPAFF-UHFFFAOYSA-N Acrylamide-acrylic acid resin Chemical compound NC(=O)C=C.OC(=O)C=C RNIHAPSVIGPAFF-UHFFFAOYSA-N 0.000 claims description 4
- 229920002126 Acrylic acid copolymer Polymers 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- LCQXXBOSCBRNNT-UHFFFAOYSA-K ammonium aluminium sulfate Chemical group [NH4+].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O LCQXXBOSCBRNNT-UHFFFAOYSA-K 0.000 claims description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 claims description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 abstract description 30
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 abstract description 10
- 230000004083 survival effect Effects 0.000 abstract description 10
- 230000005068 transpiration Effects 0.000 abstract description 10
- 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 abstract description 9
- 229930006000 Sucrose Natural products 0.000 abstract description 9
- 239000005720 sucrose Substances 0.000 abstract description 8
- 238000010672 photosynthesis Methods 0.000 abstract description 6
- 239000001569 carbon dioxide Substances 0.000 abstract description 5
- 230000029553 photosynthesis Effects 0.000 abstract description 5
- 230000009286 beneficial effect Effects 0.000 abstract description 4
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- 230000000694 effects Effects 0.000 abstract description 2
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 abstract description 2
- 239000011148 porous material Substances 0.000 abstract description 2
- 238000002360 preparation method Methods 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 15
- 239000002245 particle Substances 0.000 description 15
- 230000012010 growth Effects 0.000 description 13
- 239000007789 gas Substances 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- 229960004793 sucrose Drugs 0.000 description 8
- 229920001817 Agar Polymers 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 230000000243 photosynthetic effect Effects 0.000 description 4
- 230000029058 respiratory gaseous exchange Effects 0.000 description 4
- 238000002054 transplantation Methods 0.000 description 4
- 241001313857 Bletilla striata Species 0.000 description 3
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 3
- 239000005977 Ethylene Substances 0.000 description 3
- 241000220223 Fragaria Species 0.000 description 3
- 230000008020 evaporation Effects 0.000 description 3
- 238000001704 evaporation Methods 0.000 description 3
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 235000016623 Fragaria vesca Nutrition 0.000 description 2
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 238000005469 granulation Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- ZCTXEAQXZGPWFG-UHFFFAOYSA-N imidurea Chemical compound O=C1NC(=O)N(CO)C1NC(=O)NCNC(=O)NC1C(=O)NC(=O)N1CO ZCTXEAQXZGPWFG-UHFFFAOYSA-N 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 230000002786 root growth Effects 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 239000003104 tissue culture media Substances 0.000 description 2
- 229930192334 Auxin Natural products 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 description 1
- 239000004062 cytokinin Substances 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 230000009569 heterotrophic growth Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000003617 indole-3-acetic acid Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
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- 239000007787 solid Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
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Classifications
-
- 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
- A01H4/008—Methods for regeneration to complete plants
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Cultivation Of Plants (AREA)
Abstract
A method for improving the gas microenvironment of a plant tissue culture bottle seedling and a special culture medium, in particular to a method for regulating carbon dioxide and water in a plant tissue culture bottle. Inoculating the bud seedlings with CO after the culture medium is inoculated2The slow release granules are put into a bottle and spread on the surface of the culture medium, a moisture absorption bag is hung near the bottle mouth, and the bottle is covered for culture. The invention improves the CO in the bottle2The supply amount is beneficial to photosynthesis of the bottle seedlings, the humidity in the culture bottle is reduced, the cuticle of the leaf of the bottle seedlings is developed, the function of opening and closing pores is enhanced, the transpiration is favorably reduced, and the transplanting survival rate of the bottle seedlings is improved. The invention can reduce the usage amount of the sucrose in the culture medium, thereby reducing the pollution rate of the culture medium. CO of the invention2The slow release granule, the moisture absorption bag and the application method thereof have the characteristics of low cost, simple preparation, convenient application and obvious effect.
Description
Technical Field
The invention belongs to the technical field of plant biology, and relates to a plant tissue culture method, in particular to a method for regulating carbon dioxide and water in a plant tissue culture bottle and a special culture medium.
Background
The plant tissue culture technology has wide market prospect in seedling production, virus removal, germplasm improvement and the like. In traditional plant tissue culture, tissue culture seedling is cultured in sealed test tube or glass bottle due to overhigh humidity and CO in container2Deficiency causes that the cultured plantlet can not normally carry out photosynthesis, but has to depend on sugar in a culture medium for heterotrophic growth, which causes that the culture time is too long, the growth of the plantlet is slow, the growth vigor of the plantlet is weak, and the transplanting survival rate is low; the sugar in the culture medium can also promote the propagation of microorganisms and increase the pollution rate of plants [ Mamingjian, Song overwintering, tissue culture seedling sugar-free culture system based on environmental control, agricultural engineering report, 2009,25 (6): 192-]The details are as follows.
1. At present, the content of carbon source sucrose in a plant tissue culture medium is generally 3 percent, illumination and darkness alternate culture is carried out every day in the culture, and high-concentration CO in a bottle is caused by respiration in a dark period2At the beginningCO within 1-2 h of illumination2The concentration is from several thousand. mu.l.L-1Quickly drops to 70-90 mul.L-1This low concentration state is maintained throughout subsequent photoperiods "[ Guo Hai, et al2Application of gas in plant tissue culture and its application prospect, nuclear agriculture report 2004,18(5): 368-371]It approximates the average CO of plant photosynthesis2Offset point 50. mu.l.L-1Relative to atmospheric CO2Concentration 350. mu.l.L-1Much lower, the photosynthesis efficiency is very low.
2' the relative humidity in the incubator is close to 100%, so that the cuticle of the epidermis of the plantlet is not developed enough, the water potential during the acclimation is low, and the survival rate of the plantlet in the acclimation stage is reduced. In addition, the water potential of the air in the incubator is much higher than that outside the incubator, so that the transpiration amount of the plantlet is small and is unfavorable for the photosynthesis of the plantlet "[ Schering. the growth environment and regulation of tissue culture plantlet, 1990(3): 65-67 ]. The air vapor in the culture bottle comes from leaf transpiration and also from evaporation of the moisture of the culture medium agar to the space in the bottle.
3, the increase of the contents of auxin, cytokinin, indoleacetic acid, BA and KIN in the culture medium can promote the generation of ethylene and inhibit the differentiation and breeding of organs. And CO2Can inhibit the generation of ethylene, delay the senility of plants and promote the growth and development of plants. "[ Guo Hai, etc.. is as above]. Harmful gas is not easy to be emitted in the closed bottle, and the harmful gas can cause toxicity to the growth of the bottle seedlings.
The tissue culture seedling culture gas microenvironment automatic regulation system has the advantages of simple structure, convenient operation, easy implementation, and the like, and can not be widely applied to tissue culture seedling culture gas microenvironment automatic regulation systems in the aspects of open type and semi-open type, special equipment, multiple links and the like at present [ Xuxiong, et al, research and test of tissue culture seedling culture gas microenvironment automatic regulation systems, agricultural engineering bulletin, 2003, 19(6): 14-17.
(III) improving CO in the tissue culture bottle2The concentration and the reduction of the air moisture content have low requirement on control precision, which creates conditions for improving the gas microenvironment of the plant tissue culture seedling with low cost by simple operation without special equipment investment.
Disclosure of Invention
The invention mainly aims to provide a method for regulating carbon dioxide and humidity in air in a plant tissue culture bottle and a special culture medium.
The technical scheme provided by the invention is as follows:
a method for improving the gas microenvironment of tissue culture seedlings of plants is characterized in that CO is inoculated to a tissue culture medium after the seedlings are inoculated2The slow release granules are put into a bottle and spread on the surface of the culture medium, a moisture absorption bag is hung near the bottle mouth, and the bottle is covered for culture.
Said CO2Sustained release granules containing CO2Releasing agent and water-absorbing slow-release material capable of fully absorbing water, cold sterilizing prepared raw materials in advance, and CO2The weight ratio of the releasing agent aluminum ammonium sulfate to the sodium bicarbonate is 1: 10-20. The water-absorbing slow-release material is a material capable of releasing water after absorbing water, and comprises super absorbent resin, wherein the super absorbent resin comprises sodium acrylate-vinyl alcohol copolymer or acrylamide-acrylic acid copolymer. During granulation, the water-absorbing slow-release material is crushed and fully absorbs CO2Mixing the release agent and water-absorbing sustained-release material at a weight ratio of 25-50:1, granulating under sterile condition, and adding CO2The sustained-release granule is spherical, has a diameter of 0.3-0.8cm, and can be stored under aseptic condition. When in use, 2-10 granules are put in each 100ml of the volume of the culture flask.
A moisture absorption bag is made by filling sodium polyacrylate with filter paper or non-woven fabric, the appearance is nearly flat, the edge of the bag is attached with fine lines, and the bag is sterilized by conventional dry hot air (160-170 ℃ for 2 hours).
The invention also provides a special culture medium for improving the gas microenvironment of the plant tissue culture bottle seedlings, which is characterized in that CO is used as a nutrient medium2The slow release particles are spread on the surface of the culture medium, and 2-10 particles are put in each 100ml of the volume of the culture bottle. 1-4 moisture absorption bags are hung near the bottle mouth.
Said CO2Sustained release granules containing CO2Releasing agent and water-absorbing slow-release material with sufficient water absorption, CO2The weight ratio of the aluminum ammonium sulfate to the sodium bicarbonate of the releasing agent is 1: 10-20. The water absorption slow release material comprises super absorbent resin, and the super absorbent resin comprises sodium acrylate-vinyl alcohol copolymer or acrylamide-acrylic acid copolymer. CO 22Water absorption of releasing agent and water absorptionMixing the slow release material with CO at a weight ratio of 25-50:1 for granulation2The slow release granule is spherical and has a diameter of 0.3-0.8 cm.
The hygroscopic bag is made by filling sodium polyacrylate with filter paper or non-woven fabric, the appearance is nearly flat, and the edge of the bag is attached with fine lines.
The principle and the technical effect of the invention are as follows:
1.CO2the sustained release granule contains CO2A releasing agent and a water-absorbing slow-release material.
The water absorption slow release material has the function of balancing water, can absorb water under high humidity and can release water under low humidity.
CO2Mixing and granulating the release agent and the water-absorbing slow-release material with pre-water absorption to prepare CO2Sustained release granule, wherein the water-absorbing sustained release material fully absorbing water in the granule slowly releases CO2The releasing agent is the reaction medium of ammonium aluminum sulfate and sodium bicarbonate to generate CO2The particles permeate and are discharged into the air in the bottle, and when CO is accumulated in the particles2When the pressure reaches a certain value, the particles are cracked due to expansion, and CO in the particles is also generated2Is discharged out.
When the water absorption slow release material in the granules is insufficient in water, the water absorption slow release material can also absorb water vapor from the space in the bottle for supplying CO2The release agent reacts while reducing the humidity of the air in the bottle.
2. The high water-absorbing resin sodium polyacrylate packaged by the filter paper or the non-woven fabric is used for manufacturing the moisture absorption bag, the sodium polyacrylate has strong moisture absorption, can absorb a large amount of moisture in high-humidity air, and can automatically release the moisture when the air humidity is reduced to a certain degree, so that the air humidity is adjusted in a certain range.
3. Compared with the conventional tissue culture, the invention has lower humidity in the bottle, larger water vapor pressure difference between the water content of the agar of the culture medium and the space in the bottle, accelerates the evaporation of the water content of the agar, possibly causes insufficient water supply of root systems to influence the growth of bottle seedlings, so that CO (carbon monoxide) is generated2The slow release particles are spread on the surface of the culture medium to prevent the water evaporation on the surface of the culture medium to a certain degree.
4.CO2The slow release particles are spherical, and the spherical shape can ensure that the slow release particles have smaller contact area with the surface of the culture medium to prevent the slow water absorption thereinThe release material absorbs water directly from the agar medium. In addition CO2The slow release particles are tightly paved on the surface of the culture medium, and the spherical body can ensure that larger gaps are formed among the particles and between the particles and the bottle seedlings, so that the gas exchange between the solid culture medium and the space in the bottle is facilitated, and the internal oxygen supply of the culture medium is ensured to be sufficient to facilitate the root respiration.
The invention has the beneficial effects that (1) the invention improves the CO in the bottle2The invention reduces the humidity in the culture bottle, makes the cuticle of the leaf of the bottle seedling developed, enhances the function of opening and closing pores, and is beneficial to reducing the transpiration, so the invention can improve the survival rate of the bottle seedling transplantation. (2) The invention improves the CO in the bottle2The supply amount is bottle seedling photosynthesis, so that the supply of carbon source of the culture medium can be reduced, the use amount of cane sugar is reduced, and the pollution rate of the culture medium is reduced. (3) CO 22Can inhibit the generation of plant ethylene and reduce the bottle seedling poisoning and even death. (4) CO 22The slow release granule also increases CO in the bottle during the dark period (at night)2Concentration, inhibits the respiration of the bottle seedlings, reduces the respiration loss of organic matters and is beneficial to the growth and accumulation of the bottle seedlings. CO of the invention2The slow release granule and the moisture absorption bag have low cost, simple preparation and convenient application.
Detailed description of the invention
Example one
Production of CO2Sustained release granules containing CO2The release agent and the water-absorbing slow-release material are prepared by pre-crushing the raw materials and performing ultraviolet cold sterilization. CO 22The releasing agent is aluminum ammonium sulfate and sodium bicarbonate with the weight ratio of 1:12, and the water-absorbing slow-release material is sodium acrylate-vinyl alcohol copolymer powder which fully absorbs water. Introducing CO2The releasing agent and the water-absorbing sodium acrylate-vinyl alcohol copolymer are mixed according to the weight ratio of 30:1, and then 0.3 percent of imidazolidinyl urea preservative is added, and the mixture is mixed and granulated under the aseptic condition, and is in a spherical shape with the diameter of 0.4 cm. Freezing and storing under aseptic condition.
A square filter paper bag 2.5X 2.5cm filled with 1.3g of sodium polyacrylate was sealed to form a moisture-absorbing bag with a nearly flat appearance and a thin line attached to the edge of the bag. The mixture is sterilized by dry hot air (160-170 ℃) and maintained for 2 hours.
Conventional tissue culture (control group), volume of 200ml culture bottle (cylindrical, diameter about 6 cm), culture medium MS + NAA 0.2 mg/L + sucrose 3%, treatment group (invention) culture bottle and control, culture medium MS + NAA 0.2 mg/L + sucrose 1.5%.
Strawberry (A)Fragaria× ananassa) The bud seedling explant has no obvious root growth, the average fresh weight is 0.073 g/plant, the average height is 1.13 cm, the average leaf number is 2.1 pieces/plant, the conventional inoculation is carried out, 12-14 plants are inoculated in each bottle, and the conventional culture is carried out by covering a control group. The treatment group will have 12 CO particles2The slow release particles are put into a bottle, spread on the surface of a culture medium and arranged at intervals, 2 moisture absorption bags are hung at the position close to the bottle mouth, and the bottle is covered for culture.
Culturing for 30 days, hardening off the seedlings of the control group and the treatment group in a greenhouse for 5 days, transplanting the seedlings in the greenhouse into the soil, and performing conventional management in the greenhouse.
(II) results
During the culture period, the humidity in the control group bottle is 95-100%, and the humidity in the treatment group bottle is 65-80%. After 30 days of culture, the pollution rate of the culture bottles in the control group is 4.02 +/-0.38%, the pollution rate of the treatment group is 0%, the pollution rate is obviously reduced compared with that of the control group, and the bottle seedling growth indexes are shown in the table below.
Table. growth and transplanting condition of strawberry bottle seedling
The net photosynthetic rate of the bottle seedlings is higher in the treatment group, and the difference with the control is obvious. The average fresh weight of the stem leaves of the treated group is greatly improved compared with the control, and the average leaf number of the treated group is more than that of the control, so that the difference is obvious. The survival rate of the two groups of bottle seedlings is the same.
The average fresh weight of the root system of the treated group is heavier, the difference with the control is obvious, and the average root number of the treated group is obviously increased compared with the control. The transpiration rate treatment group was low and very significantly different from the control. After 1 month after the transplantation, the survival rate of the seedlings in the treatment group is remarkably improved compared with that in the control group.
(III) conclusion
The sucrose content of the culture medium of the treatment group is reduced by half, so that the propagation nutrition of microorganisms can be reduced, and the pollution of the culture medium can be reduced.
The invention increases the carbon dioxide supply in the bottle, improves the net photosynthetic rate of the stem leaves, and improves the growth amount of the stem leaves and the roots. The bottle seedlings grow in a lower humidity environment, the transpiration capability is reduced, transpiration water loss after the plants are transplanted can be reduced, and the bottle seedlings have developed root systems, so that the bottle seedling transplanting survival rate is improved compared with a control method.
Example two
Production of CO2Sustained release granules containing CO2The release agent and the water-absorbing slow-release material are prepared by pre-crushing the raw materials and performing ultraviolet cold sterilization. CO 22The weight ratio of the releasing agent aluminum ammonium sulfate to the sodium bicarbonate is 1:15, and the water-absorbing slow-release material is sodium acrylate-vinyl alcohol copolymer powder. Introducing CO2The releasing agent and the fully water-absorbing sodium acrylate-vinyl alcohol copolymer are mixed according to the weight ratio of 40:1, 0.3 percent of imidazolidinyl urea preservative is added, and the mixture is mixed and granulated under the aseptic condition and is in a spherical shape with the diameter of 0.7 cm. Freezing and storing under aseptic condition.
A square nonwoven fabric bag 4X 4cm was filled with 2.4g of sodium polyacrylate, and sealed to form a moisture-absorbing bag having a nearly flat appearance with a thin line attached to the edge of the bag, and was sterilized with dry hot air (160 ℃ C. to 170 ℃ C.) for 2 hours.
Conventional tissue culture (control group), blue vase with volume of 650ml (bottom diameter about 8cm, bottleneck diameter about 4 cm), culture medium 1/2MS + NAA 0.2 mg/L + sucrose 3%, culture medium 1/2MS + NAA 0.2 mg/L + sucrose 1.5%.
Bletilla striata (A)Bletilla striata) The bud seedling explant has no obvious root growth, the average fresh weight is 0.097 g/plant, the average height is 1.98 cm, the average leaf number is 2.7 pieces/plant, conventional inoculation is carried out, 16-18 plants are inoculated in each bottle, and a control group is covered and cultured conventionally. The treatment group will have 20 CO particles2Slowly-releasing granule is put into the bottle, and the tiling is on the culture medium surface, and the interval is arranged, hangs 3 moisture absorption bags near bottleneck department, covers the cultivation.
Culturing for 90 days, hardening off the seedlings of the control group and the treatment group in a greenhouse for 7 days, transplanting the seedlings in the greenhouse into the soil, and performing conventional management in the greenhouse.
(II) results
During the culture period, the humidity in the control group bottle is 100%, and the humidity in the treatment group bottle is 70-85%. After the culture in the bottle is carried out for 90 days, the pollution rates of the culture mediums of the control group and the treatment group are respectively 3.45 +/-0.39 percent and 1.78 +/-0.08 percent, the pollution rate of the treatment group is obviously reduced, and the growth indexes of the bottle seedlings are shown in the following table.
Table, growth and transplantation of bottle seedling of bletilla striata
The bottle seedling net photosynthetic rate treatment group is obviously increased compared with the control pole. The average fresh weight of the stems and leaves of the treated group is heavier, the difference with the control is extremely obvious, and the average leaf number of the treated group is more than that of the control, and the difference is obvious. The survival rate of the bottle seedlings is remarkably improved.
The average fresh weight of the root system of the treated group is heavier, the difference with the control is obvious, and the difference of the average root number and the two groups is not obvious. The transpiration rate treated group was significantly lower than the control. After 1 month after the transplantation, the survival rate of the seedlings in the treatment group is remarkably improved compared with that in the control group.
(III) conclusion
The sucrose content of the culture medium is reduced by half, the propagation nutrition of microorganisms is reduced, and the pollution of the culture medium is reduced.
The invention increases the carbon dioxide supply in the bottle, improves the net photosynthetic rate of the stem leaves, and improves the growth amount of the stem leaves and the roots. The bottle seedlings grow in a lower humidity environment, the transpiration capability is reduced, transpiration water loss after the plants are transplanted can be reduced, and the bottle seedlings have developed root systems, so that the bottle seedling transplanting survival rate is improved compared with a control method.
Claims (6)
1. A method for improving the gas microenvironment of plant tissue culture seedlings is characterized in that: inoculating the bud seedling explant with CO after the culture medium is inoculated2Spreading the slow-release granules on the surface of the culture medium in a bottle, hanging a moisture absorption bag at the position close to the bottle mouth, and covering the bottle for culture;
said CO2The sustained release granule contains CO2Releasing agent and water-absorbing slow-release material, CO2Mixing the release agent and the water-absorbing slow-release material which absorbs water sufficiently according to the weight ratio of 25-50:1, granulating, and putting 2-10 granules with the diameter of 0.3-0.8cm according to the volume of a culture bottle per 100 ml;
the hygroscopic bag is filled with sodium polyacrylate through filter paper or non-woven fabric, the appearance is nearly flat, fine lines are attached to the edge of the bag, and 1-4 bags are hung in each bottle.
2. The method of claim 1, wherein said CO is selected from the group consisting of CO, and mixtures thereof2The releasing agent is the mixture of aluminum ammonium sulfate and sodium bicarbonate according to the weight ratio of 1: 10-20;
the water-absorbing slow-release material comprises super absorbent resin, and the super absorbent resin comprises sodium acrylate-vinyl alcohol copolymer or acrylamide-acrylic acid copolymer.
3. The method of claim 1, wherein said CO is produced by improving the gas microenvironment of said tissue culture plantlets2The slow release granules are pre-sterilized by cold, and the moisture absorption bag is sterilized by dry and hot air.
4. The special culture medium for improving the gas microenvironment of the plant tissue culture bottle seedlings is characterized in that: CO is flatly laid on the surface of the culture medium of the conventional culture bottle2Slowly releasing granules, namely spreading 2-10 granules per 100ml of the volume of a culture bottle, and hanging 1-4 moisture absorption bags at the position close to the bottle mouth of the culture bottle;
said CO2The sustained release granule contains CO2Releasing agent and water-absorbing slow-release material, CO2The releasing agent and the water-absorbing slow-release material which fully absorbs water are in a spherical shape with the diameter of 0.3-0.8cm according to the weight ratio of 25-50: 1; the moisture absorption bag is filled with sodium polyacrylate through filter paper or non-woven fabric, and the appearance is nearly flat.
5. The special culture medium for improving the gas microenvironment of the tissue culture bottle seedlings of the plants as claimed in claim 4, wherein: said CO2The releasing agent is ammonium aluminum sulfate and bicarbonateThe sodium is mixed according to the weight ratio of 1:10-20, the water absorption slow release material comprises super absorbent resin, and the super absorbent resin comprises sodium acrylate-vinyl alcohol copolymer or acrylamide-acrylic acid copolymer.
6. The special culture medium for improving the gas microenvironment of the tissue culture bottle seedlings of the plants as claimed in claim 4, wherein: said production of CO2The slow release granule is prepared by cold sterilizing raw materials in advance, attaching fine lines to the edge of the moisture absorption bag, and performing dry heat sterilization.
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| US4683258A (en) * | 1984-05-28 | 1987-07-28 | Mitsui Toatsu Chemicals, Incorporated | Agent for absorbing and releasing water vapor |
| JPH01240183A (en) * | 1988-03-22 | 1989-09-25 | Toagosei Chem Ind Co Ltd | Culture atmosphere regulator |
| US6046243A (en) * | 1993-02-12 | 2000-04-04 | Bernard Technologies, Inc. | Compositions for sustained release of a gas |
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