CN108719068A - A kind of implantation methods of selenium-rich peanuts - Google Patents
A kind of implantation methods of selenium-rich peanuts Download PDFInfo
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- CN108719068A CN108719068A CN201810584947.XA CN201810584947A CN108719068A CN 108719068 A CN108719068 A CN 108719068A CN 201810584947 A CN201810584947 A CN 201810584947A CN 108719068 A CN108719068 A CN 108719068A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/40—Fabaceae, e.g. beans or peas
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of implantation methods of selenium-rich peanuts, belong to peanut cultivation technical field, include the following steps：（1）Pre-treatment；（2）Inoculation；（3）Laser irradiation；（4）Culture；（4）Transplanting.The peanut of implantation methods output of the present invention is safe and nontoxic, harmless, bioavilability is high, and breeding potential is high, and peanut quality is good, is rich in selenium element, very useful to health, has good market popularization value.
Description
Technical field
The invention belongs to peanut cultivation technical fields, and in particular to a kind of implantation methods of selenium-rich peanuts.
Background technology
Selenium is micronutrient element necessary to a kind of humans and animals, has anti-oxidant, anticancer, improves immunity of organisms, protects
Shield heart is dirty, protection liver, prevents the multiple pharmacological effects such as myopia and cataract, removing toxic substances, anti-aging and enhancing fecundity, quilt
It is described as " Promethean fire ", " heart patron saint ", " king of anticancer ", " Uther Lightbringer " and " long-lived element ".The health of selenium and human body
There is close contact, is currently known human diseases related with selenium deficiency up to more than 40 and plants, including cardiovascular and cerebrovascular disease, glycosuria
Disease, autoimmune disease, Keshan disease, Kaschin-Beck disease etc..With deepening continuously for medicine and Nutritional studies, people are increasingly
It recognizes the physiological action of selenium in human body and maintains the importance of health.Selenium is included in people by Chinese Soclety of Nutrition within 1988
Micronutrient element necessary to body, it is 50 ~ 250ug/d to recommend adult's intake, and Food Selenium highest amount safe for ingestion is
400ug/d.However, China has 72% area to be in low selenium or selenium deficiency area, provinces and cities of 13, China were done by Chinese Soclety of Nutrition
Investigation, the results showed that, it is 26 ~ 32ug/d, the minimum intake 50ug/ recommended with Chinese Soclety of Nutrition at selenium intake for each person
D is far apart.Therefore it need to meet the needs to selemium nutrition element containing the higher food of selenium by edible, this is ensured as China
The common practice of resident's health.
In Se accumulation approach, plant transformation is a kind of very common approach, the method for existing production selenium-rich peanuts
There are mainly two types of, one is selenium-enriched fertilizer after peanut cultivation, is imposed, fertilizer humidity itself is larger, is easy to grow many bacteriums,
It is serious to grow germ, pathogen is brought to them again while causing peanut to apply fertilizer, not only without promoting peanut growth,
Their normal developments are hindered instead, and peanut is very low to the bioavilability of selenium in selenium-enriched fertilizer；Another is to pass through
Selenium-rich seed-soaking liquid carries out seed soaking to peanut, and such method selenium-rich rate is very low, equally also results in the waste of resource；In addition
Existing most of selenium-rich are mostly addition sodium selenites to obtain selenium-rich peanuts fruit, but sodium selenite toxicity is big, safe range
Narrow, bioavailability is low, and additive amount can excessively lead to peanut lesion, and addition is very few to be unable to get selenium-rich peanuts.
Invention content
The purpose of the present invention is for existing problem, provide a kind of implantation methods of selenium-rich peanuts.Pass through this method
The peanut of plantation is safe and nontoxic, harmless, bioavilability is high, and breeding potential is high, and the peanut quality of output is good, economic benefit
Height has good market popularization value.
The present invention is achieved by the following technical solutions：
A kind of implantation methods of selenium-rich peanuts, include the following steps：
（1）Pre-treatment：By peanut be placed in tap water down-flow water rinse 30-40min, then by peanut be placed in superclean bench into
Row disinfection, sterilizing, after take out shelled peanut, be cut into 30-40mm3Fritter it is spare；
（2）Inoculation：By step（1）In fritter shelled peanut be placed in selenium-rich culturing base, every bottle inoculation 2-4 block fritter shelled peanuts,
They are close to equidistantly, and then sealed membrane seals culture bottle；
（3）Laser irradiation：By step（2）In culture bottle carry out laser irradiation 3-7min；
（4）Culture：By step（3）Culture bottle after middle laser irradiation, which is placed in incubator, to be cultivated, incubation time 31-
48d obtains peanut tissue culture seedlings；
（5）Transplanting：Opening steps（4）The sealed membrane of middle culture bottle, culture bottle is placed in and is placed at room temperature for 3-5d, then takes out tissue culture
In transplantation of seedlings to plantation field.
Further, the step（1）The specific procedure of middle disinfection, sterilizing, includes the following steps：
A. the preparation of sterilized solution：Weigh 3-5 parts of dodecyl dihydroxy ethyl glycine betaines, 4-6 parts of Aloe Vera Gels, 75% alcohol of 6-8 parts
It is collectively disposed in sterile triangular flask, triangular flask is placed on magnetic stirring apparatus, the rotating speed of magnetic stirring apparatus is 150-180rpm, temperature
Degree is 39-43 DEG C, and the time of stirring is 35-45min；
B. sterilized solution made from operation a being placed in high-pressure steam sterilizing pan and is sterilized, the temperature of sterilizing is 120-122 DEG C,
The time of sterilizing is 16-20min；
C. take out operation b in sterilize after sterilized solution be placed in superclean bench be cooled to room temperature it is spare；
D. in superclean bench, peanut is placed in the beaker of 500ml, it is that peanut is total that parts by weight are then added into beaker
During which the sterilized solution that the operation c of 7-8 times of parts by weight is obtained, sterilization time 15-21min constantly shake beaker with wrist；
E. it takes out the peanut that operation d is obtained and is placed in sterile water and impregnate 12-14min, then take out peanut with sterile water wash 6-
It is spare after 8min.
Preferably, the step（2）The preparation of middle selenium-rich culturing base, includes the following steps：
A. it measures 500-600 parts of deionized water to be placed in large beaker, by boiling to boiling, then weighs 6-8 parts of agar, 10-30
Part white sugar, 2-3 parts of Se-enriched yeasts, 1.5-1.7 parts of ammonium nitrate, 1.8-2 parts of potassium nitrate, 0.16-0.18 parts of potassium dihydrogen phosphates,
0.36-0.38 parts of magnesium sulfate, 0.0007-0.0009 parts of potassium iodide, 0.005-0.007 parts of boric acid, 0.021-0.023 parts of sulfuric acid
Manganese, 0.0085-0.0087 part zinc sulfate, 0.0002-0.0003 parts of sodium molybdates, 0.00002-0.00003 parts of copper sulphate,
0.00002-0.00003 parts of cobalt chlorides, 0.035-0.036 parts of disodium ethylene diamine tetraacetates, 0.026-0.028 parts of ferrous sulfate,
1-2 parts of multi-vitamins, 0.001-0.002 parts of glycine are added sequentially in large beaker, after each component is completely dissolved, are weighed
0.43-0.45 parts of calcium chloride are added in large beaker, are continued after heating 3-5min, and deionized water is added into large beaker and is settled to
1L obtains selenium-rich culturing based sols；
B. the selenium-rich culturing based sols that a is obtained will be operated to be dispensed into culture bottle, every bottle of packing 20-30ml, then sealed membrane seals
It is good, all culture bottles are placed in high-pressure steam sterilizing pan and are sterilized, the temperature of sterilizing is 120-122 DEG C, sterilizing
Time is 18-20min；
C. the selenium-rich culturing base operated after sterilizing in b is placed in superclean bench, the temperature with culture bottle is cooled to 50-60
DEG C when, measure 0.002-0.003 part molar concentrations as the 6- benzyls aminoadenine and 0.001-0.003 parts of molar concentrations of 1mol/L
It is added in each culture bottle for 2, the 4- dichlorphenoxyacetic acids of 1.5mol/L, shakes up stand for standby use.
Preferably, the step（3）The laser intensity of middle laser irradiation is 400-500W/cm2。
Preferably, the step（4）The illumination condition of middle culture is daily illumination 16-20h, dark 4-8h, the temperature of culture
Degree is 25-29 DEG C.
The present invention has the following advantages compared with prior art：
The present invention is to carry out tissue cultures to peanut seed using the totipotency of cell, and selenium-rich substance is added to tissue cultures
Culture medium in, the method to produce selenium-rich peanuts plant, by way of bioconversion, selenium element is transferred to flower again
In fruit grown.The plant cell of the totipotency of a plant cell i.e. life, as long as having complete membranous system and nucleus, it
A whole set of hereditary basis for developing into an intact plant is had, it can be by dividing, breaking up again under the conditions of one is appropriate
An intact plant is generated, ensures that sterile environment is critically important in tissue culture procedures, existing technology, which is substantially, to be passed through
The chemical substances such as 75% alcohol, 95% alcohol, sodium hypochlorite, mercuric chloride carry out disinfection sterilizing to plant tissue, wherein 95% alcohol pair
The stimulation of tissue is very big, and processing time is difficult to hold, and careless slightly, cell will be killed, and sodium hypochlorite belongs to corrosives,
The worker of this product is contacted through commonly using hand, palm is largely perspired, and nail is thinning, and trichomadesis, sodium hypochlorite has sensitization, puts
The chlorine gone out is possible to cause to be poisoned, although mercuric chloride usage amount is very small, mercuric chloride to human body have carcinogenicity, mutagenicity,
Lead to the harm such as poisoning, and mercuric chloride is dealt with improperly, is polluted the environment, the constituent of sterilized solution of the present invention is dodecyl two
Ethoxy glycine betaine, Aloe Vera Gel and 75% alcohol, nontoxic, irritation is small, soluble easily in water, and to ph stability, bactericidal property is good, reed
Luxuriant growth glue can also active cell vigor, improve the inductivity of tissue cultures；Tissue culture propagating technology compares other modess of reproduction,
With occupying little space, not by area, season limit, cultivation cycle is short, there is no variations, can keep former all maternal heredity
Feature, small investment, high financial profit, modes of reproduction are more, try out advantage wide in variety；Selenium-rich is added in selenium-rich culturing base of the present invention
Yeast, Se-enriched yeast are exactly to be added selenium element during culture yeasts, and selenium has been absorbed and utilized in when yeast growth, makes selenium and ferment
Protein and polysaccharide organic in parent, which combine, is converted into Biological Selenium, to eliminate chemical selenium（Such as sodium selenite）To human body
Toxicity and stomach stimulation, enable that selenium is more efficient, be more safely absorbed by the body utilization；Although tissue culture technique at present
Comparative maturity, but there are still the low problem of inductivity, the culture bottle after inoculation is carried out the photograph of certain laser intensity by the present invention
It penetrates, Mitotic frequency can be accelerated, cell differentiation is promoted to be proliferated, increase the inductivity of tissue cultures, improve the yield of peanut, this
Outer laser stimulation accelerates cell to be enriched with the absorption of selenium element and other nutrients, makes the nutritional ingredient in culture medium
Bioavilability reaches maximum, adds 6- benzyls aminoadenine and 2 in selenium-rich culturing base in addition, and 4- dichlorphenoxyacetic acids promote thin
Born of the same parents' division, differentiation, accelerate vine growth and development.The peanut of implantation methods output of the present invention is safe and nontoxic, harmless, biological utilisation
Degree is high, and breeding potential is high, and peanut quality is good, is rich in selenium element, very useful to health, has good marketing
Value.
Specific implementation mode
Embodiment 1
A kind of implantation methods of selenium-rich peanuts, include the following steps：
（1）Pre-treatment：Peanut is placed in tap water down-flow water and rinses 30min, then peanut is placed in superclean bench and is carried out
Disinfection, sterilizing, after take out shelled peanut, be cut into 30mm3Fritter it is spare；
（2）Inoculation：By step（1）In fritter shelled peanut be placed in selenium-rich culturing base, every bottle inoculation 2 pieces of fritter shelled peanuts, it
Close to equidistant, then sealed membrane seals culture bottle；
（3）Laser irradiation：By step（2）In culture bottle carry out laser irradiation 3min；
（4）Culture：By step（3）Culture bottle after middle laser irradiation, which is placed in incubator, to be cultivated, incubation time 31d,
Obtain peanut tissue culture seedlings；
（5）Transplanting：Opening steps（4）The sealed membrane of middle culture bottle, culture bottle is placed in and is placed at room temperature for 3d, then takes out tissue-cultured seedling
It is transplanted in plantation field.
Further, the step（1）The specific procedure of middle disinfection, sterilizing, includes the following steps：
A. the preparation of sterilized solution：3 parts of dodecyl dihydroxy ethyl glycine betaines, 4 parts of Aloe Vera Gels, 6 part of 75% alcohol is weighed to set jointly
In sterile triangular flask, triangular flask is placed on magnetic stirring apparatus, the rotating speed of magnetic stirring apparatus is 150rpm, and temperature is 39 DEG C,
The time of stirring is 35min；
B. sterilized solution made from operation a is placed in high-pressure steam sterilizing pan and is sterilized, the temperature of sterilizing is 120 DEG C, sterilizing
Time be 16min；
C. take out operation b in sterilize after sterilized solution be placed in superclean bench be cooled to room temperature it is spare；
D. in superclean bench, peanut is placed in the beaker of 500ml, it is that peanut is total that parts by weight are then added into beaker
During which the sterilized solution that the operation c of 7 times of parts by weight is obtained, sterilization time 15min constantly shake beaker with wrist；
E. it takes out the peanut that operation d is obtained and is placed in sterile water and impregnate 12min, then take out peanut with sterile water wash 6min
It is spare afterwards.
Preferably, the step（2）The preparation of middle selenium-rich culturing base, includes the following steps：
A. it measures 500 parts of deionized water to be placed in large beaker, by boiling to boiling, then weighs 6 parts of agar, 10 parts of white sugar, 2
Part Se-enriched yeast, 1.5 parts of ammonium nitrate, 1.8 parts of potassium nitrate, 0.16 part of potassium dihydrogen phosphate, 0.36 part of magnesium sulfate, 0.0007 part of iodate
Potassium, 0.005 part of boric acid, 0.021 part of manganese sulfate, 0.0085 part of zinc sulfate, 0.0002 part of sodium molybdate, 0.00002 part of copper sulphate,
0.00002 part of cobalt chloride, 0.035 part of disodium ethylene diamine tetraacetate, 0.026 part of ferrous sulfate, 1 part of multi-vitamins, 0.001 part
Glycine is added sequentially in large beaker, after each component is completely dissolved, is weighed 0.43 part of calcium chloride and is added in large beaker, after
After continuous heating 3min, into large beaker, addition deionized water is settled to 1L, obtains selenium-rich culturing based sols；
B. the selenium-rich culturing based sols that a is obtained will be operated to be dispensed into culture bottle, every bottle of packing 20ml, then sealed membrane is sealed,
All culture bottles are placed in high-pressure steam sterilizing pan and are sterilized, the temperature of sterilizing is 120 DEG C, and the time of sterilizing is
18min；
C. the selenium-rich culturing base operated after sterilizing in b is placed in superclean bench, the temperature with culture bottle is cooled to 50 DEG C
When, 6- benzyls aminoadenine and 0.001 part of molar concentration that 0.002 part of molar concentration of measurement is 1mol/L are the 2 of 1.5mol/L,
4- dichlorphenoxyacetic acids are added in each culture bottle, shake up stand for standby use.
Preferably, the step（3）The laser intensity of middle laser irradiation is 400W/cm2。
Preferably, the step（4）The illumination condition of middle culture is daily illumination 16h, and the temperature of dark 8h, culture are 25
℃。
Embodiment 2
A kind of implantation methods of selenium-rich peanuts, include the following steps：
（1）Pre-treatment：Peanut is placed in tap water down-flow water and rinses 35min, then peanut is placed in superclean bench and is carried out
Disinfection, sterilizing, after take out shelled peanut, be cut into 35mm3Fritter it is spare；
（2）Inoculation：By step（1）In fritter shelled peanut be placed in selenium-rich culturing base, every bottle inoculation 3 pieces of fritter shelled peanuts, it
Close to equidistant, then sealed membrane seals culture bottle；
（3）Laser irradiation：By step（2）In culture bottle carry out laser irradiation 5min；
（4）Culture：By step（3）Culture bottle after middle laser irradiation, which is placed in incubator, to be cultivated, incubation time 39d,
Obtain peanut tissue culture seedlings；
（5）Transplanting：Opening steps（4）The sealed membrane of middle culture bottle, culture bottle is placed in and is placed at room temperature for 4d, then takes out tissue-cultured seedling
It is transplanted in plantation field.
Further, the step（1）The specific procedure of middle disinfection, sterilizing, includes the following steps：
A. the preparation of sterilized solution：4 parts of dodecyl dihydroxy ethyl glycine betaines, 5 parts of Aloe Vera Gels, 7 part of 75% alcohol is weighed to set jointly
In sterile triangular flask, triangular flask is placed on magnetic stirring apparatus, the rotating speed of magnetic stirring apparatus is 165rpm, and temperature is 41 DEG C,
The time of stirring is 40min；
B. sterilized solution made from operation a is placed in high-pressure steam sterilizing pan and is sterilized, the temperature of sterilizing is 121 DEG C, sterilizing
Time be 18min；
C. take out operation b in sterilize after sterilized solution be placed in superclean bench be cooled to room temperature it is spare；
D. in superclean bench, peanut is placed in the beaker of 500ml, it is that peanut is total that parts by weight are then added into beaker
During which the sterilized solution that the operation c of 7.5 times of parts by weight is obtained, sterilization time 18min constantly shake beaker with wrist；
E. it takes out the peanut that operation d is obtained and is placed in sterile water and impregnate 13min, then take out peanut with sterile water wash 7min
It is spare afterwards.
Preferably, the step（2）The preparation of middle selenium-rich culturing base, includes the following steps：
A. 550 parts of deionized water is measured to be placed in large beaker, by boiling to boiling, then weigh 7 parts of agar, 20 parts of white sugar,
2.5 parts of Se-enriched yeasts, 1.6 parts of ammonium nitrate, 1.9 parts of potassium nitrate, 0.17 part of potassium dihydrogen phosphate, 0.37 part of magnesium sulfate, 0.0008 part
Potassium iodide, 0.006 part of boric acid, 0.022 part of manganese sulfate, 0.0086 part of zinc sulfate, 0.00025 part of sodium molybdate, 0.000025 part of sulphur
Sour copper, 0.000025 part of cobalt chloride, 0.0355 part of disodium ethylene diamine tetraacetate, 0.027 part of ferrous sulfate, 1.5 parts of compound dimension lifes
Element, 0.0015 part of glycine are added sequentially in large beaker, after each component is completely dissolved, is weighed 0.44 part of calcium chloride and are added to
In large beaker, continue after heating 4min, deionized water is added into large beaker and is settled to 1L, obtains selenium-rich culturing based sols；
B. the selenium-rich culturing based sols that a is obtained will be operated to be dispensed into culture bottle, every bottle of packing 25ml, then sealed membrane is sealed,
All culture bottles are placed in high-pressure steam sterilizing pan and are sterilized, the temperature of sterilizing is 121 DEG C, and the time of sterilizing is
19min；
C. the selenium-rich culturing base operated after sterilizing in b is placed in superclean bench, the temperature with culture bottle is cooled to 55 DEG C
When, 6- benzyls aminoadenine and 0.002 part of molar concentration that 0.0025 part of molar concentration of measurement is 1mol/L are 1.5mol/L's
2,4- dichlorphenoxyacetic acids are added in each culture bottle, shake up stand for standby use.
Preferably, the step（3）The laser intensity of middle laser irradiation is 450W/cm2。
Preferably, the step（4）The illumination condition of middle culture is daily illumination 18h, and the temperature of dark 6h, culture are 27
℃。
Embodiment 3
A kind of implantation methods of selenium-rich peanuts, include the following steps：
（1）Pre-treatment：Peanut is placed in tap water down-flow water and rinses 40min, then peanut is placed in superclean bench and is carried out
Disinfection, sterilizing, after take out shelled peanut, be cut into 40mm3Fritter it is spare；
（2）Inoculation：By step（1）In fritter shelled peanut be placed in selenium-rich culturing base, every bottle inoculation 4 pieces of fritter shelled peanuts, it
Close to equidistant, then sealed membrane seals culture bottle；
（3）Laser irradiation：By step（2）In culture bottle carry out laser irradiation 7min；
（4）Culture：By step（3）Culture bottle after middle laser irradiation, which is placed in incubator, to be cultivated, incubation time 48d,
Obtain peanut tissue culture seedlings；
（5）Transplanting：Opening steps（4）The sealed membrane of middle culture bottle, culture bottle is placed in and is placed at room temperature for 5d, then takes out tissue-cultured seedling
It is transplanted in plantation field.
Further, the step（1）The specific procedure of middle disinfection, sterilizing, includes the following steps：
A. the preparation of sterilized solution：5 parts of dodecyl dihydroxy ethyl glycine betaines, 6 parts of Aloe Vera Gels, 8 part of 75% alcohol is weighed to set jointly
In sterile triangular flask, triangular flask is placed on magnetic stirring apparatus, the rotating speed of magnetic stirring apparatus is 180rpm, and temperature is 43 DEG C,
The time of stirring is 45min；
B. sterilized solution made from operation a is placed in high-pressure steam sterilizing pan and is sterilized, the temperature of sterilizing is 122 DEG C, sterilizing
Time be 20min；
C. take out operation b in sterilize after sterilized solution be placed in superclean bench be cooled to room temperature it is spare；
D. in superclean bench, peanut is placed in the beaker of 500ml, it is that peanut is total that parts by weight are then added into beaker
During which the sterilized solution that the operation c of 8 times of parts by weight is obtained, sterilization time 21min constantly shake beaker with wrist；
E. it takes out the peanut that operation d is obtained and is placed in sterile water and impregnate 14min, then take out peanut with sterile water wash 8min
It is spare afterwards.
Preferably, the step（2）The preparation of middle selenium-rich culturing base, includes the following steps：
A. it measures 600 parts of deionized water to be placed in large beaker, by boiling to boiling, then weighs 8 parts of agar, 30 parts of white sugar, 3
Part Se-enriched yeast, 1.7 parts of ammonium nitrate, 2 parts of potassium nitrate, 0.18 part of potassium dihydrogen phosphate, 0.38 part of magnesium sulfate, 0.0009 part of iodate
Potassium, 0.007 part of boric acid, 0.023 part of manganese sulfate, 0.0087 part of zinc sulfate, 0.0003 part of sodium molybdate, 0.00003 part of copper sulphate,
0.00003 part of cobalt chloride, 0.036 part of disodium ethylene diamine tetraacetate, 0.028 part of ferrous sulfate, part multi-vitamins, 0.002 part
Glycine is added sequentially in large beaker, after each component is completely dissolved, is weighed 0.45 part of calcium chloride and is added in large beaker, after
After continuous heating 5min, into large beaker, addition deionized water is settled to 1L, obtains selenium-rich culturing based sols；
B. the selenium-rich culturing based sols that a is obtained will be operated to be dispensed into culture bottle, every bottle of packing 30ml, then sealed membrane is sealed,
All culture bottles are placed in high-pressure steam sterilizing pan and are sterilized, the temperature of sterilizing is 122 DEG C, and the time of sterilizing is
20min；
C. the selenium-rich culturing base operated after sterilizing in b is placed in superclean bench, the temperature with culture bottle is cooled to 60 DEG C
When, 6- benzyls aminoadenine and 0.003 part of molar concentration that 0.003 part of molar concentration of measurement is 1mol/L are the 2 of 1.5mol/L,
4- dichlorphenoxyacetic acids are added in each culture bottle, shake up stand for standby use.
Preferably, the step（3）The laser intensity of middle laser irradiation is 500W/cm2。
Preferably, the step（4）The illumination condition of middle culture is daily illumination 20h, and the temperature of dark 4h, culture are 29
℃。
Comparative example 1
This comparative example 1 compared with Example 2, by step（1）Disinfection, sterilizing program in pre-treatment change conventional disappear into
Poison, sterilizing program, method and step all same in addition to this.
Comparative example 2
This comparative example 2 compared with Example 2, saves step（2）Laser irradiation, method and step all same in addition to this.
Control group 1
The method for tissue culture of existing peanut seed.
In order to compare the effect of the present invention, the flower that the quality that same plantation field is collected with a batch is almost consistent is selected
It is raw to be used as experimental subjects, then use respectively embodiment 2, comparative example 1, comparative example 2, control group 1 cultural method into
Row culture, experiment condition is consistent with environmental Kuznets Curves, the growth of during which observation culture plant and microbiological contamination situation, statistical experiment number
According to.
Specific experiment correction data is as shown in table 1 below：
Table 1
| Inductivity（%） | Contamination rate（%） | |
| Embodiment 2 | 94.7 | 1.3 |
| Comparative example 1 | 91.8 | 12.3 |
| Comparative example 2 | 78.5 | 4.7 |
| Control group | 81.6 | 12.1 |
Note：What the inductivity described in upper table 1 specifically counted is after tissue-cultured seedling induces successfully, and the inductivity before transplanting lures
What conductance specifically counted is the contamination rate before transplanting after tissue-cultured seedling induces successfully.
Upper table 1 is as can be seen that the present invention is to the method for tissue culture of peanut, and inductivity is high, and contamination rate is low, breeding potential
Height, cultivation cycle is short, high financial profit.
Control group 2
Application No. is the implantation methods of selenium-rich peanuts disclosed in 201711063608.9.
In order to compare the effect of the present invention, the flower that the quality that same plantation field is collected with a batch is almost consistent is selected
The 20kg peanuts of selection are uniformly divided into 4 groups, every group of peanut 5kg by raw 20kg as experimental subjects, then respectively with embodiment 2,
Comparative example 1, comparative example 2, control group 2 implantation methods cultivated, way to manage is completely the same, during which observation flower
Raw upgrowth situation, statistical experiment data.
Specific experiment correction data is as shown in table 2 below：
Table 2
| Peanut Se content（mg/kg） | Peanut yield kg | |
| Embodiment 2 | 0.156 | 875 |
| Comparative example 1 | 0.152 | 801 |
| Comparative example 2 | 0.143 | 726 |
| Control group | 0.132 | 698 |
Note：Peanut yield described in upper table 2 is the peanut yield collected after 5kg peanut cultivations in experiment.
Upper table 2 as can be seen that implantation methods output of the present invention peanut it is safe and nontoxic, harmless, bioavilability is high, and
And breeding potential is high, peanut quality is good, is rich in selenium element, very useful to health, has good market popularization value.
Claims (5)
1. a kind of implantation methods of selenium-rich peanuts, which is characterized in that include the following steps：
（1）Pre-treatment：By peanut be placed in tap water down-flow water rinse 30-40min, then by peanut be placed in superclean bench into
Row disinfection, sterilizing, after take out shelled peanut, be cut into 30-40mm3Fritter it is spare；
（2）Inoculation：By step（1）In fritter shelled peanut be placed in selenium-rich culturing base, every bottle inoculation 2-4 block fritter shelled peanuts,
They are close to equidistantly, and then sealed membrane seals culture bottle；
（3）Laser irradiation：By step（2）In culture bottle carry out laser irradiation 3-7min；
（4）Culture：By step（3）Culture bottle after middle laser irradiation, which is placed in incubator, to be cultivated, incubation time 31-
48d obtains peanut tissue culture seedlings；
（5）Transplanting：Opening steps（4）The sealed membrane of middle culture bottle, culture bottle is placed in and is placed at room temperature for 3-5d, then takes out tissue culture
In transplantation of seedlings to plantation field.
2. a kind of implantation methods of selenium-rich peanuts rice according to claim 1, which is characterized in that the step（1）
The specific procedure of middle disinfection, sterilizing, includes the following steps：
A. the preparation of sterilized solution：Weigh 3-5 parts of dodecyl dihydroxy ethyl glycine betaines, 4-6 parts of Aloe Vera Gels, 75% alcohol of 6-8 parts
It is collectively disposed in sterile triangular flask, triangular flask is placed on magnetic stirring apparatus, the rotating speed of magnetic stirring apparatus is 150-180rpm, temperature
Degree is 39-43 DEG C, and the time of stirring is 35-45min；
B. sterilized solution made from operation a being placed in high-pressure steam sterilizing pan and is sterilized, the temperature of sterilizing is 120-122 DEG C,
The time of sterilizing is 16-20min；
C. take out operation b in sterilize after sterilized solution be placed in superclean bench be cooled to room temperature it is spare；
D. in superclean bench, peanut is placed in the beaker of 500ml, it is that peanut is total that parts by weight are then added into beaker
During which the sterilized solution that the operation c of 7-8 times of parts by weight is obtained, sterilization time 15-21min constantly shake beaker with wrist；
E. it takes out the peanut that operation d is obtained and is placed in sterile water and impregnate 12-14min, then take out peanut with sterile water wash 6-
It is spare after 8min.
3. a kind of implantation methods of selenium-rich peanuts rice according to claim 1, which is characterized in that the step（2）Middle selenium-rich
The preparation of culture medium, includes the following steps：
A. it measures 500-600 parts of deionized water to be placed in large beaker, by boiling to boiling, then weighs 6-8 parts of agar, 10-30
Part white sugar, 2-3 parts of Se-enriched yeasts, 1.5-1.7 parts of ammonium nitrate, 1.8-2 parts of potassium nitrate, 0.16-0.18 parts of potassium dihydrogen phosphates,
0.36-0.38 parts of magnesium sulfate, 0.0007-0.0009 parts of potassium iodide, 0.005-0.007 parts of boric acid, 0.021-0.023 parts of sulfuric acid
Manganese, 0.0085-0.0087 part zinc sulfate, 0.0002-0.0003 parts of sodium molybdates, 0.00002-0.00003 parts of copper sulphate,
0.00002-0.00003 parts of cobalt chlorides, 0.035-0.036 parts of disodium ethylene diamine tetraacetates, 0.026-0.028 parts of ferrous sulfate,
1-2 parts of multi-vitamins, 0.001-0.002 parts of glycine are added sequentially in large beaker, after each component is completely dissolved, are weighed
0.43-0.45 parts of calcium chloride are added in large beaker, are continued after heating 3-5min, and deionized water is added into large beaker and is settled to
1L obtains selenium-rich culturing based sols；
B. the selenium-rich culturing based sols that a is obtained will be operated to be dispensed into culture bottle, every bottle of packing 20-30ml, then sealed membrane seals
It is good, all culture bottles are placed in high-pressure steam sterilizing pan and are sterilized, the temperature of sterilizing is 120-122 DEG C, sterilizing
Time is 18-20min；
C. the selenium-rich culturing base operated after sterilizing in b is placed in superclean bench, the temperature with culture bottle is cooled to 50-60
DEG C when, measure 0.002-0.003 part molar concentrations as the 6- benzyls aminoadenine and 0.001-0.003 parts of molar concentrations of 1mol/L
It is added in each culture bottle for 2, the 4- dichlorphenoxyacetic acids of 1.5mol/L, shakes up stand for standby use.
4. a kind of implantation methods of selenium-rich peanuts rice according to claim 1, which is characterized in that the step（3）Middle laser
The laser intensity of irradiation is 400-500W/cm2。
5. a kind of implantation methods of selenium-rich peanuts rice according to claim 1, which is characterized in that the step（4）Middle culture
Illumination condition be daily illumination 16-20h, dark 4-8h, the temperature of culture is 25-29 DEG C.
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