CN103460971B - Method for improving transplanting survival rate of trichosanthes kirilowii tissue culture seedlings - Google Patents
Method for improving transplanting survival rate of trichosanthes kirilowii tissue culture seedlings Download PDFInfo
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- 230000004083 survival Effects 0.000 title claims abstract description 20
- 210000001519 tissues Anatomy 0.000 title claims abstract description 17
- 240000006023 Trichosanthes kirilowii Species 0.000 title abstract description 6
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- 241000196324 Embryophyta Species 0.000 claims abstract description 20
- 230000000844 anti-bacterial Effects 0.000 claims abstract description 8
- 239000000758 substrate Substances 0.000 claims abstract description 8
- 240000004013 Trichosanthes cucumerina Species 0.000 claims description 53
- 235000008326 Trichosanthes anguina Nutrition 0.000 claims description 52
- 238000000338 in vitro Methods 0.000 claims description 25
- 239000011159 matrix material Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 230000036536 Cave Effects 0.000 claims description 8
- 239000003415 peat Substances 0.000 claims description 8
- 239000002985 plastic film Substances 0.000 claims description 8
- 239000004576 sand Substances 0.000 claims description 8
- 229910052902 vermiculite Inorganic materials 0.000 claims description 8
- 235000019354 vermiculite Nutrition 0.000 claims description 8
- 239000010455 vermiculite Substances 0.000 claims description 8
- 229920003023 plastic Polymers 0.000 claims description 7
- 238000004659 sterilization and disinfection Methods 0.000 claims description 7
- JNPZQRQPIHJYNM-UHFFFAOYSA-N Carbendazim Chemical group C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 claims description 6
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- XSQUKJJJFZCRTK-UHFFFAOYSA-N urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 6
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- 229910052570 clay Inorganic materials 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 244000037666 field crops Species 0.000 claims description 3
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- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 claims description 2
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- 241000345998 Calamus manan Species 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
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- 240000005308 Juniperus chinensis Species 0.000 description 1
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Classifications
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- Y02P60/216—
Abstract
The invention discloses a method for improving the transplanting survival rate of trichosanthes kirilowii tissue culture seedlings. According to the method, the antibacterial ability of the trichosanthes kirilowii tissue culture seedlings is enhanced by proportioning different substrate varieties with different ratios, seedling-hardening and domesticating processing, transplanting and managing in a restoring growing period after the transplanting. By the method disclosed by the invention, the trichosanthes kirilowii tissue culture seedlings are basically suitable for the external large-land growth environment, the plant root system is thick and strong, the new sprouts are good in growing vigor, the rattans are robust, and the transplanting survival rate can achieve 80-95% which is greatly improved compared with the survival rate of 10-20% of the traditional method. The method is convenient to operate, low in price and suitable for processing in batches and industrialized production, and has excellent economical efficiency and social benefits.
Description
Technical field
The invention belongs to plant tissue culture nursery field, specifically relate to hardening, substrate preparation, transplanting and the management after transplanting to improve the method for snakegourd plantlet in vitro transplanting survival rate.
Background technology
Snakegourd [Trichosanthes kirilowii Maxim] is Curcurbitaceae snake gourd herbaceous perennial vine plant, and another name Snakegourd Fruit is a kind of dioecian plant, is China's tradition parts of generic medicinal plants.Snakegourd whole body is all precious, and its fruit, root all can be used as medicine, and seed claims semen trichosanthis, and root, root of Chinese trichosanthes, has clearing heat and eliminating phlegm, relieving chest stiffness to dissipate mass, moisturizes the effect such as laxation, detumescence and apocenosis.Edible Chinese juniper beach wormwood seed is nutritious, and having significant protective effect to hypertension, high fat of blood, high cholesterol, acute myocardial ischemia, is middle-aged and old first-selected green health care foods.Meanwhile, Seeds of Trichosanthes kirilowii can improve body immune function, and has weight reducing, beauty functions.Melon seeds outward appearance brown after frying is gorgeous, seed benevolence is full, mouthfeel profit is continuous, crisp and fragrant is special, and enjoying endless aftertastes of food, is described as " king of melon seeds ".
In recent years, the research of snakegourd tissue culture technology was many, mainly contained the research of snakegourd quick propagating technology; Snakegourd tissue cultures and the research of non-test-tube plantlet fast breeding technique; The research of plant regeneration in snakegourd tissue cultures; Snakegourd shoot tip meristem virus-free culture and Plantlet Differentiation etc.Current snakegourd tissue culture technology has tended to ripe, but the snakegourd manually large-area China that is planted in also is in the starting stage, development speed is relatively slow, main cause is that snakegourd plantlet in vitro transplanting survival rate is low, especially individual month of the 1-2 of the firm bottle outlet of plantlet in vitro, mainly because plantlet in vitro is large in the growing environment difference of the environment of culturing room and transplanting, have a strong impact on the transplanting survival rate of snakegourd plantlet in vitro, failed to make group culturation rapid propagating technology be applied to production practices.
At present, R&D institution and enterprise is not almost had to carry out correlative study for the hardening of snakegourd plantlet in vitro, domestication, transplanting medium.By carrying out the hardening of snakegourd plantlet in vitro, domestication, transplanting medium research, this technology can substantially increase the transplanting survival rate of snakegourd plantlet in vitro.
Summary of the invention
The object of the invention is to there are provided a kind of method improving snakegourd plantlet in vitro transplanting success rate, the antibacterial ability of snakegourd plantlet in vitro can be strengthened, make it better adapt to external environment, the survival rate of snakegourd plantlet in vitro can be improved.
For achieving the above object, the present invention takes following technical measures:
The technical solution adopted in the present invention is by substrate preparation and process, hardening, transplanting and restoration ecosystem period management, makes snakegourd plant little by little adapt to extraneous natural environment,
Improve a method for snakegourd plantlet in vitro transplanting survival rate, its step is as follows
(1) tissue cultures:
Conventionally carry out the tissue cultures of snakegourd seedling in indoor, in the present invention, snakegourd prescription of rooting medium is MS+0.2 (mg/L) NAA.
(2) in-bottle seeding:
Annual late March to late June chooses snakegourd height of seedling 3-5cm, leaf 2-3 sheet, the aseptic bottle seedling of root 3-5 bar is put in the green house under natural environment, band bottle hardening 5-7d, then open and cultivate bottle cap hardening, in blake bottle, add appropriate distilled water submergence snakegourd root, keep every day air humidity to be greater than 70%, after uncork, place 3-5d.
(3) the outer hardening of bottle:
Substrate preparation and process: with the mixture of river sand, vermiculite, peat for hardening matrix, and described hardening matrix is by bactericide disinfection, then sabot.
Described river sand is 4-10 order, apparent density 2000-3000kg/m
3, bulk density 1000-2000kg/m
3, modulus of fineness is less than 3, and clay content is less than 2%, and its granularity is between 2.5mm-5mm.
Described vermiculite is 4-10 order, and specification is 2-4mm, and density is 2.4-2.7g/cm
3.
Described peat proportion is 1.20-1.60, and water content is 70-85%, the content of organic matter more than 30%, content of humic acid more than 30%, and pH value is 5-6.5.
In described matrix, the volume of mixture of river sand, vermiculite and peat is than being 1-2:1-2:1-2.
Described matrix bactericide is carbendazim, and the carbendazim that every 1 cubic metre of matrix sprays 10L 800 times of liquid carries out disinfection.
The vinyl disc of described dress seedling is 72 hole seedling culture hole plates, circle 72 holes (6 × 12) cave dish: upper bore: 40mm, lower bore: 21mm, highly: 45mm, and volume: 35ml, cave dish size: 532mm × 278mm.
Transplant: after uncork is placed, with tweezers the plant in tissue culture bottle is pressed from both sides out gently, with running water, the medium on plant is rinsed well, and then hang 1h (hang environment: 15-25 DEG C, air humidity remains more than 75%), move to and be equipped with in the plastics cave dish of sterilization matrix, each cavities is planted a strain, and root system stretches as far as possible, compresses matrix when planting as far as possible, water permeable by the mode that watering pot sprays after plantlet in vitro is transplanted, keep air humidity at 70-80%; Grow after young leaves and Xin Gen until snakegourd seedling, front 5-7d humid control, at 70-80%, reduces humidity later gradually, and the scope that the humidity of average every day declines is 1%-5%, until identical with canopy outer land for growing field crops ambient humidity.
The method of described controlled humidity, the conventional method that available cultivation time control is wet or matrix first time water permeable after, directly put into wide 1.2m, long 10m, high 0.5m transparent plastic film cover shed, shed surrounding soil covers and compacting film, every morning, 8:00-9:00 rod beat shed gently, allow the globule formed in shed plastic film drop onto in seedling-cultivating tray, utilize water cycle to keep canopy humidity and can using water wisely.
(3) restoration ecosystem period management: grow after young leaves and Xin Gen until snakegourd seedling, every 5-7d adds and uses the urea that mass volume ratio is 0.05%-0.1%, and plant grows to about 10cm, root system and matrix can agglomerating deviate from time, Field planting can be carried out by placing.
Shading suitable during whole hardening, make intensity of illumination be the 35%-40% of natural daylight, intensity of illumination controls at 0.6-1.0 ten thousand lux.
Bottle outer hardening and restoration ecosystem period management during, soil humidity remains on 40-60%.
Compared with prior art, the invention has the advantages that:
Make public for the first time a kind of method improving snakegourd plantlet in vitro transplanting survival rate, the method carries out by different substrates kind the antibacterial ability that different proportion proportioning, hardening acclimation, transplanting and the restoration ecosystem period management after transplanting strengthen snakegourd group training aseptic seedling, make it better adapt to external environment, the survival rate of snakegourd plantlet in vitro can be improved.The method is easy to operate, and expense is low, is suitable for batch process and factorial praluction, has good business efficiency and social benefit.
Hardening off method of the present invention has done very large improvement, the antibacterial ability of seedling and adaptive faculty are had very large improvement, the plantlet in vitro of the substrate culture of being tamed by hardening adapts to extraneous grown in field environment substantially, plant root is sturdy, young sprout growing way is good, rattan is healthy and strong, and its transplanting survival rate can reach 80%-95%.Be greatly improved compared with the 10%-20% survival rate of conventional method.
Accompanying drawing explanation
Fig. 1 is the snakegourd effect photo of a kind of snakegourd plantlet in vitro transplanting method of the present invention after 3 months;
Matrix in figure is matrix of the present invention.
Fig. 2 is the snakegourd effect photo of a kind of conventional snakegourd plantlet in vitro transplanting method after 3 months;
Embodiment
Below in conjunction with embodiment, the invention will be further described.
Embodiment 1:
A kind of method improving snakegourd plantlet in vitro transplanting survival rate:
(1) tissue cultures:
Conventionally carry out in indoor snakegourd seedling tissue cultures (bibliography: Yang Lina, Liu Jie, Tao Jianmin. the tissue cultures of snakegourd and Fast-propagation [J]. Jiangsu's agriculture science, 2008,15 (4): 89-90.),
In the present embodiment, snakegourd prescription of rooting medium is MS+0.2 (mg/L) NAA.
(2) in-bottle seeding: on March 20th, 2013, by 50 young plant height 3-5cm, leaf 2-3 sheet, the snakegourd aseptic seedling of root 3-5 bar is put into hardening 7d in green house, then open and cultivate bottle cap hardening, in blake bottle, add appropriate distilled water submergence snakegourd root, keep air humidity to be greater than 70%, place 3d.
(3) the outer hardening of bottle
Substrate preparation and process: carry out proportioning for 1:1:2 with river sand, vermiculite, peat volume ratio, load in plastics cave dish, carry out matrix disinfect with carbendazim, and the carbendazim that every 1 cubic metre of matrix sprays 10L 800 times of liquid carries out disinfection.
Described river sand is 4-10 order, apparent density 2000-3000kg/m
3, bulk density 1000-2000kg/m
3, modulus of fineness is less than 3, and clay content is less than 2%, and its granularity is between 2.5mm-5mm.
Described vermiculite is 4-10 order, and specification is 2-4mm, and density is 2.4-2.7g/cm
3.
Described peat proportion is 1.20-1.60, and water content is 70-85%, the content of organic matter more than 30%, content of humic acid more than 30%, and pH value is 5-6.5.
Transplant: transplant on March 30th, 2013, with tweezers the plant in tissue culture bottle is pressed from both sides out gently during transplanting, with running water, the medium on plant is rinsed well, and then hang 1h and carry out transplanting (hang environment: temperature 15-25 DEG C, air humidity remains more than 75%), move in the plastics cave dish that is equipped with through sterilizing matrix, each cavities is planted a strain, root system stretches as far as possible, compresses matrix as far as possible, waters permeable after transplanting by the mode that watering pot sprays.Then put into wide 1.2m, long 10m, high 0.5m transparent plastic film cover shed, shed surrounding soil covers and compacting film, every morning, 8:00-9:00 rod beat shed gently, the interior globule formed of shed plastic film is allowed to drop onto in seedling-cultivating tray, utilize water cycle to keep canopy humidity and can using water wisely, keep air humidity at 70-80%.After 15d, plant grows young leaves and Xin Gen, then the plastic film at shed two is first opened, air humidity in shed is made to control at 70-80%, again plastic film is opened completely after 5d, every day reduces humidity gradually, the scope that the humidity of average every day can decline is 1%-5%, until identical with canopy outer land for growing field crops ambient humidity.
(3) restoration ecosystem period management: after 15 days, snakegourd seedling grows young leaves and Xin Gen, then every 6d adds snakegourd seedling and uses the urea that mass volume ratio is 0.05%-0.1%, and after 30d, plant grows to about 10cm, root system and matrix can agglomerating deviate from time, namely placing carries out Field planting.
Shading suitable during whole hardening, make intensity of illumination be the 35%-40% of natural daylight, intensity of illumination controls at 0.6-1.0 ten thousand lux.
The shading black sunshade net of shading rate 50%.
Bottle outer hardening and restoration ecosystem period management during, soil humidity remains on 40-60%.
In the plant of having refined seedling, choose 50 strains carry out field planting, totally 42 strains survive, and survival rate is 84%.
Embodiment 2:
Conventional snakegourd plantlet in vitro transplanting method:
With embodiment 1, its difference is: plantlet in vitro is placed on natural conditions lower refining seedling 5-7d, then opens and cultivates bottle cap again after hardening 3-5d, be directly transplanted to large Tanaka and go.Choose 50 strains plant by above-mentioned refining in the plant of seedling, totally 8 strains survive, survival rate only 16%.
Above-described embodiment does not limit the present invention in any form, and the present invention is also not limited to above-mentioned citing.The technical scheme that the mode that the use of the art is equal to replacement or equivalent transformation obtains, all drops in protection scope of the present invention.
Claims (2)
1. improve a method for snakegourd plantlet in vitro transplanting survival rate, its step is as follows:
(1) tissue cultures:
Conventionally carry out the tissue cultures of snakegourd seedling in indoor, snakegourd prescription of rooting medium is MS+0.2mg/LNAA;
(2) in-bottle seeding:
Annual late March to late June chooses height of seedling 3-5cm, leaf 2-3 sheet, the blake bottle snakegourd seedling of root 3-5 bar is put in the green house under natural environment, band bottle hardening 5-7d, then open and cultivate bottle cap hardening, in blake bottle, add appropriate distilled water submergence snakegourd root, keep every day air humidity to be greater than 70%, after uncork, place 3-5d;
(3) the outer hardening of bottle:
Substrate preparation and process: with the mixture of river sand, vermiculite, peat for hardening matrix, and described hardening matrix is by bactericide disinfection, then sabot;
Described river sand is 4-10 order, apparent density 2000-3000kg/m
3, bulk density 1000-2000kg/m
3, modulus of fineness is less than 3, and clay content is less than 2%, and its granularity is between 2.5mm-5mm;
Described vermiculite is 4-10 order, and specification is 2-4mm, and density is 2.4-2.7g/cm
3;
Described peat proportion is 1.20-1.60, and water content is 70-85%, the content of organic matter more than 30%, content of humic acid more than 30%, and pH value is 5-6.5;
In described hardening matrix, the volume ratio of river sand, vermiculite and peat is 1-2:1-2:1-2;
Described bactericide is carbendazim, and the carbendazim that every 1 cubic metre of matrix sprays 10L 800 times of liquid carries out disinfection;
The vinyl disc of dress seedling is 72 hole seedling culture hole plates, circle 72 holes (6 × 12) cave dish: upper bore: 40mm, lower bore: 21mm, highly: 45mm, and volume: 35ml, cave dish size: 532mm × 278mm;
Transplant: after uncork is placed, pressed from both sides out by the plant in blake bottle with tweezers, rinsed well by the medium on plant with running water, and then hang 1h, hang environment: 15-25 DEG C, air humidity remains more than 75%; Move to and be equipped with in the plastics cave dish of the matrix of sterilization, each cavities is planted a strain, and root system stretches, and compresses matrix when planting, and waters permeable after plantlet in vitro is transplanted by the mode that watering pot sprays, and keeps air humidity at 70-80%; Grow after young leaves and Xin Gen until snakegourd seedling, front 5-7d air humidity controls, at 70%-80%, to reduce humidity gradually later, and the scope that the humidity of average every day declines is 1%-5%, until identical with canopy outer land for growing field crops ambient humidity;
(4) restoration ecosystem period management: grow after young leaves and Xin Gen until snakegourd seedling, every 5-7d adds and uses the urea that mass volume ratio is 0.05%-0.1%, and plant grows to 10cm, root system and matrix can agglomerating deviate from time, Field planting can be carried out by placing;
Shading suitable during whole hardening, make intensity of illumination be the 35%-40% of natural daylight, intensity of illumination controls at 0.6-1.0 ten thousand lux;
Bottle outer hardening and restoration ecosystem period management during, soil humidity remains on 40-60%.
2. the method for raising snakegourd plantlet in vitro transplanting survival rate according to claim 1, after transplanting during controlled humidity, matrix first time water permeable after, directly put into wide 1.2m, long 10m, high 0.5m transparent plastic film cover shed, shed surrounding soil covers and compacting film, every morning, 8:00-9:00 rod beat shed gently, allowed the globule formed in shed plastic film drop onto in seedling-cultivating tray, utilized water cycle to keep canopy humidity and can using water wisely.
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| CN104145700B (en) * | 2014-08-28 | 2016-07-20 | 广州白云山中一药业有限公司 | The cultural method of Radix Trichosanthis medical material |
| CN104521756B (en) * | 2014-12-24 | 2016-08-17 | 广西大学 | A kind of method producing Fructus Trichosanthis tissue cultured seedling |
| CN106417033B (en) * | 2016-11-10 | 2018-09-04 | 聊城大学 | A kind of Gaotang snakegourd rapid propagation method |
| CN108077022A (en) * | 2017-12-18 | 2018-05-29 | 长沙智博生物科技有限公司 | For the matrix of blueberry hardening and blueberry hardening off method |
| CN111226770B (en) * | 2020-03-07 | 2021-08-17 | 广州甘蔗糖业研究所湛江甘蔗研究中心 | Liquid seedling hardening method for improving transplanting survival rate of tissue culture seedlings of cinnamomum kanehirae |
| CN111345230A (en) * | 2020-04-29 | 2020-06-30 | 蒙树生态建设集团有限公司 | Seedling hardening method for iris tissue culture seedlings |
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| CN101496497A (en) * | 2008-08-09 | 2009-08-05 | 周小红 | Seedling exercising technique for test-tube plantlet with full sunshine and spray |
| CN101699989A (en) * | 2009-11-20 | 2010-05-05 | 杨保成 | Method for tissue culture and rapid propagation of Trichosanthes kirilowii Maxim |
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