CN116369205A - Selenium-enriched wheat breeding method - Google Patents
Selenium-enriched wheat breeding method Download PDFInfo
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Classifications
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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
-
- 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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
-
- 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
-
- 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
- A01H4/002—Culture media for tissue culture
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- Biotechnology (AREA)
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- Cell Biology (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Cereal-Derived Products (AREA)
Abstract
The invention provides a selenium-rich wheat breeding method, and relates to the technical field of crop breeding. The selenium-enriched wheat breeding method comprises the following steps: s1, preprocessing wheat seeds: selecting wheat seeds which grow in the wild for 3-4 years and have excellent properties, washing the wheat seeds with clear water, drying the wheat seeds, putting the wheat seeds into a refrigerator for refrigeration, soaking and cleaning the wheat seeds in a sterile environment, and reserving the obtained wheat seeds for later use; s2, induction culture: and (2) placing the wild wheat seeds obtained in the step (S1) into an induction culture medium, and culturing for 50-65 days under the conditions that the culture temperature is 22-25 ℃, the air relative humidity is 70-80%, the illumination time is 8 hours/day and the illumination intensity is 1500-3000 Lux, thereby obtaining young wheat seedlings. By the breeding method, the wild wheat seeds can absorb more selenium element, and the ingestion of other nutrient substances is greatly increased, so that the selenium content of seedlings can be kept stable and continuously increased, and then the high-quality selenium-enriched wheat is used for secondary hybridization, so that the yield and quality of the selenium-enriched wheat are improved.
Description
Technical Field
The invention relates to the technical field of crop breeding, in particular to a selenium-enriched wheat breeding method.
Background
Selenium circularly affects human health in biological spheroidization, and researches show that in areas with relatively lack of selenium element content and relatively low bioavailability in environments such as rock, soil, water source and the like, insufficient selenium intake of human body can excite toxicity of human micro virus B19 (HPVB 19) to cause the disease of large bone, so that selenium is one of the indispensable intake elements of human body, and the selenium has remarkable effects in preventing AIDS, enhancing anticancer capability, improving immune function of human body and the like, and is often called as' life fire.
Selenium-rich wheat, namely wheat containing rich selenium elements, can improve human immunity, promote lymphocyte proliferation and synthesis of antibodies and immunoglobulins, meanwhile, selenium is taken as a trace element necessary for human body, can participate in synthesis of various selenium-containing enzymes and selenium-containing proteins in the human body after selenium is taken, is beneficial to human health, and besides the selenium-rich wheat, selenium-rich rice, selenium-rich agaric, 100 micrograms of selenium-rich corn flour and the like belong to selenium-rich agricultural products, so that the way of taking selenium by people is effectively ensured.
Based on the analysis, wheat is one of main grains of human beings, and selenium-enriched wheat is one of ways of taking trace elements by people, the existing cultivation technique of selenium-enriched wheat is also mature and complete, but the selenium-enriched wheat is not balanced and stable due to the difference of different areas and regions due to the growth and planting conditions of wheat, and the yield and quality of wheat are greatly different, so in order to provide high-quality selenium-enriched wheat seedlings, a selenium-enriched wheat breeding method is proposed by a person skilled in the art.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a selenium-enriched wheat breeding method, which solves the problems that the existing selenium-enriched wheat breeding method cannot balance and stabilize selenium-containing elements in seedlings and the yield and quality of wheat are lower.
In order to achieve the above purpose, the invention is realized by the following technical scheme: a selenium-rich wheat breeding method comprises the following steps:
s1, preprocessing wheat seeds:
selecting wheat seeds which grow in the wild for 3-4 years and have excellent properties, washing the wheat seeds with clear water, drying the wheat seeds, putting the wheat seeds into a refrigerator for refrigeration, soaking and cleaning the wheat seeds in a sterile environment, and reserving the obtained wheat seeds for later use;
s2, induction culture:
placing the wild wheat seeds obtained in the step S1 into an induction culture medium, and culturing for 50-65 days under the conditions that the culture temperature is 22-25 ℃, the air relative humidity is 70-80%, the illumination time is 8 hours/day and the illumination intensity is 1500-3000 Lux, so as to obtain young wheat seedlings;
s3, proliferation culture:
selecting young wheat seedlings which are cultured in an induction culture medium for 55-60 days, transferring the wheat seedlings into a proliferation culture medium, and culturing for 60-70 days under the culture condition of 2000-4000 Lux of illumination intensity under the condition of the same temperature and humidity state as the induction culture and with the illumination time of 10 hours/day;
s4, rooting culture:
transferring the young wheat seedlings subjected to proliferation culture in the step S3 into a rooting culture medium, and culturing for 30-40 days under the same temperature and humidity state as induction culture by matching with the illumination condition of the proliferation culture medium to obtain selenium-containing wheat seedlings;
s5, hybridization breeding:
transplanting and sowing the selenium-containing wheat seedlings obtained in the step S4, taking the first-period wheat seeds harvested after sowing as female parent, selecting high-quality selenium-rich wheat seeds as male parent, carrying out hybridization breeding with the female parent, carrying out second-generation hybridization on the obtained hybrid wheat seeds and the female parent, and sowing and maturing to obtain the selenium-rich wheat seeds.
Preferably, in the step S1, wheat seeds are put into a refrigerator with the temperature of 2-7 ℃ for refrigeration for 8-14 h after being washed.
Preferably, in the step S1, the method for soaking and cleaning in a sterile environment of wheat seeds comprises: soaking in 75% concentration alcohol solution for 40-70 s, washing with sterile water for 1-2 times, soaking in Tween 20-containing sodium hypochlorite for 5-9 min, washing with sterile water for 2-6 times, and repeating the steps for 1-2 times to complete the sterilizing treatment of wheat seed.
Preferably, the sodium hypochlorite solution containing tween 20 comprises tween 20 with a volume concentration of 0.05-0.1% and sodium hypochlorite solution with a volume concentration of 0.2-0.25%.
Preferably, in the step S2, the induction medium is composed of the following components in mass volume concentration: 1/2MS basic culture medium, 0.4mg/L NAA, 0.2mg/L ferric salt, 0.1-0.5 mg/L selenium amino acid fertilizer liquid, 30g/L sucrose and 3g/L agar; the pH value of the induction culture medium is 5.8-6.0.
Preferably, in the step S3, the proliferation medium is composed of the following components in mass volume concentration: MS basic culture medium, 10-20 g/L sodium selenite, 8-15 g/L sodium selenate, 1.0g/L hydrolyzed casein, 0.4mg/L Kt; the pH value of the proliferation culture medium is 5.8-6.0.
Preferably, in the step S4, the rooting medium is composed of the following components in mass volume concentration: WR4 basic culture medium, 1/2MS culture medium, 0.4g/L CH, 0.3mg/L NAA, 0.7mg/L sodium naphthalene acetate and 0.3-0.6 mg/L selenium mineral powder; the pH value of the rooting culture medium is 5.8-6.0.
Preferably, in the step S5, the high-quality selenium-enriched wheat seeds are any one of agricultural large 3753, selenium-enriched Y8, triticale No. 76 and black gem No. 2.
The invention provides a selenium-enriched wheat breeding method. The beneficial effects are as follows:
1. the invention optimizes the culture of the wild wheat seeds from the induction stage to the proliferation stage, and then to the rooting stage and the hybridization breeding stage, and particularly the operation method of the proliferation culture stage and the rooting culture stage of the wild wheat seeds, so that the selenium element absorbed by the wild wheat seeds is more, and simultaneously, other absorbed nutrient substances are greatly increased, so that the selenium element in the seedlings keeps stable and continuous rising state, and then the high-quality selenium-enriched wheat is matched for secondary hybridization, thereby improving the yield and quality of the selenium-enriched wheat.
2. According to the invention, the wild wheat seeds are put into a refrigerator for refrigeration, so that dormancy of the seeds can be broken, germination of the seeds is promoted, and the wheat seeds can be promoted to quickly absorb moisture by soaking with sterile water, so that bioactive substances in the wheat seeds are activated, germination speed of the wheat seeds is accelerated, and cultivation speed is accelerated.
3. According to the invention, the amino acid selenium fertilizer liquid is added into the induction culture medium, so that not only can the amino acid selenium fertilizer liquid provide better growth nutrition for wheat seeds, but also a foundation is laid for the selenium content of the wheat seeds, and then the sodium selenite and the sodium selenate are added into the proliferation culture medium to provide basic fertilizers for the wheat seeds, so that the development of the wheat seeds is further accelerated, and finally, the sodium naphthylacetate in the rooting culture medium can promote the growth of wheat seedlings and leaves and stems, and in proper concentration and illumination, rooting and the growth of wheat seedlings and the growth of the leaves and stems can be promoted by being matched with nutrient substances in the rooting culture medium.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1:
the embodiment of the invention provides a selenium-enriched wheat breeding method, which comprises the following steps:
s1, preprocessing wheat seeds:
selecting wheat seeds which grow in the wild for 3-4 years and have excellent properties, washing the wheat seeds with clear water, drying the wheat seeds, putting the wheat seeds into a refrigerator for refrigeration, soaking and cleaning the wheat seeds in a sterile environment, and reserving the obtained wheat seeds for later use;
further, the wheat seeds are put into a refrigerator with the temperature of 2-7 ℃ for refrigeration for 8 hours after being washed.
Further, the method for soaking and cleaning wheat seeds in the sterile environment comprises the following steps: soaking in 75% concentration alcohol solution for 40-70 s, washing with sterile water for 1-2 times, soaking in Tween 20-containing sodium hypochlorite for 5-9 min, washing with sterile water for 2-6 times, and repeating the steps for 1-2 times to complete the sterilizing treatment of wheat seed.
Specifically, the sodium hypochlorite solution containing tween 20 comprises tween 20 with the volume concentration of 0.05-0.1% and sodium hypochlorite solution with the volume concentration of 0.2-0.25%.
S2, induction culture:
placing the wild wheat seeds obtained in the step S1 into an induction culture medium, and culturing for 50-65 days under the conditions that the culture temperature is 22-25 ℃, the air relative humidity is 70-80%, the illumination time is 8 hours/day and the illumination intensity is 1500-3000 Lux, so as to obtain young wheat seedlings;
further, the induction medium consists of the following components in mass volume concentration: 1/2MS basic culture medium, 0.4mg/L NAA, 0.2mg/L ferric salt, 0.1mg/L selenium amino acid fertilizer liquid, 30g/L sucrose and 3g/L agar; the pH of the induction medium was 5.8.
S3, proliferation culture:
selecting young wheat seedlings which are cultured in an induction culture medium for 55-60 days, transferring the wheat seedlings into a proliferation culture medium, and culturing for 60-70 days under the culture condition of 2000-4000 Lux of illumination intensity under the condition of the same temperature and humidity state as the induction culture and with the illumination time of 10 hours/day;
further, the proliferation medium consists of the following components in mass volume concentration: MS basic culture medium, 10g/L sodium selenite, 8g/L sodium selenate, 1.0g/L hydrolyzed casein, 0.4mg/L Kt; the pH of the proliferation medium was 5.8.
S4, rooting culture:
transferring the young wheat seedlings subjected to proliferation culture in the step S3 into a rooting culture medium, and culturing for 30 days under the same temperature and humidity state as induction culture by matching with the illumination condition of the proliferation culture medium to obtain selenium-containing wheat seedlings;
further, the rooting medium consists of the following components in mass volume concentration: WR4 minimal medium, 1/2MS medium, 0.4g/L CH, 0.3mg/L NAA, 0.7mg/L sodium naphthalene acetate, 0.3mg/L selenium mineral powder; the pH value of the rooting culture medium is 5.8.
S5, hybridization breeding:
transplanting and sowing the selenium-containing wheat seedlings obtained in the step S4, taking the first-period wheat seeds harvested after sowing as female parent, selecting high-quality selenium-rich wheat seeds as male parent, carrying out hybridization breeding with the female parent, carrying out second-generation hybridization on the obtained hybrid wheat seeds and the female parent, and sowing and maturing to obtain the selenium-rich wheat seeds.
Further, the high-quality selenium-enriched wheat seeds are any one of agricultural large 3753, selenium-enriched Y8, black wheat No. 76 and black gem No. 2.
Example 2:
the same features as those of embodiment 1 are not repeated, and the different features are as follows: in step S2 of this embodiment: the addition amount of the amino acid selenium fertilizer liquid in the induction culture medium is 0.5mg/L.
Example 3:
the same features as those of embodiment 1 are not repeated, and the different features are as follows: in step S3 of this embodiment: the addition amount of sodium selenite in the proliferation medium is 15g/L, and the addition amount of sodium selenate is 10g/L.
Example 4:
the same features as those of embodiment 1 are not repeated, and the different features are as follows: in step S3 of this embodiment: the addition amount of sodium selenite in the proliferation medium is 20g/L, and the addition amount of sodium selenate is 15g/L.
Example 5:
the same features as those of embodiment 1 are not repeated, and the different features are as follows: in step S4 of this embodiment: the addition amount of selenium mineral powder in the rooting culture medium is 0.6mg/L.
Comparative example 1:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: in step S3 of this comparative example: the addition amount of the amino acid selenium fertilizer liquid in the induction culture medium is 0mg/L.
Comparative example 2:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: in step S4 of this comparative example: the addition amount of sodium selenite in the proliferation medium is 0g/L.
Comparative example 3:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: in step S4 of this comparative example: the addition amount of sodium selenate in the proliferation medium is 0g/L.
Comparative example 4:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: in step S4 of this comparative example: the addition amount of sodium selenite in the proliferation medium is 0g/L, and the addition amount of sodium selenate is 0g/L.
Comparative example 5:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: in the step S4 in this comparative example: the addition amount of selenium mineral powder in the rooting culture medium is 0mg/L.
Comparative example 6:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: in the step S1 in this comparative example: the wheat seeds are put into a refrigerator with the temperature of 2-7 ℃ for refrigeration for 15h after being washed.
Comparative example 7:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: in step S1 of this comparative example: the wild wheat seeds after washing and drying in the pretreatment stage of wheat seeds are not put into a refrigerator for refrigeration.
Comparative example 8:
the same features as those of embodiment 1 will not be described in detail, and the different features are as follows: step S1 in this comparative example: during the pretreatment stage of wheat seeds, wild wheat seeds are not subjected to aseptic soaking treatment.
The results of the statistical and analytical treatments of examples 1 to 5 and comparative examples 1 to 8, as well as the effects of selenium content, were shown in Table 1, on the seedling growth time (calculated as rooting culture growth period, induction culture and proliferation culture as germination period, not included):
table 1:
from examples 1 to 3 and comparative examples 1 to 8, it can be obtained that: after the wild wheat seeds are subjected to pretreatment, induction culture, multiplication culture and rooting culture, the whole breeding and selenium content are continuously increased, MS basic culture medium, 20g/L sodium selenite, 15g/L sodium selenate, 1.0g/L hydrolyzed casein and 0.4mg/L Kt are used as the optimal conditions during the culture of the multiplication culture medium, the whole selenium content of the cultivated seedlings is 28 percent, meanwhile, the rapid breeding is carried out through different culture mediums of induction culture, multiplication culture and rooting culture and illumination culture, and then the rapid breeding is carried out with high-quality selenium-enriched wheat seeds for hybridization, and the yield and quality of the selenium-enriched wheat are improved through second-generation hybridization.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. A selenium-rich wheat breeding method is characterized in that: the method comprises the following steps:
s1, preprocessing wheat seeds:
selecting wheat seeds which grow in the wild for 3-4 years and have excellent properties, washing the wheat seeds with clear water, drying the wheat seeds, putting the wheat seeds into a refrigerator for refrigeration, soaking and cleaning the wheat seeds in a sterile environment, and reserving the obtained wheat seeds for later use;
s2, induction culture:
placing the wild wheat seeds obtained in the step S1 into an induction culture medium, and culturing for 50-65 days under the conditions that the culture temperature is 22-25 ℃, the air relative humidity is 70-80%, the illumination time is 8 hours/day and the illumination intensity is 1500-3000 Lux, so as to obtain young wheat seedlings;
s3, proliferation culture:
selecting young wheat seedlings which are cultured in an induction culture medium for 55-60 days, transferring the wheat seedlings into a proliferation culture medium, and culturing for 60-70 days under the culture condition of 2000-4000 Lux of illumination intensity under the condition of the same temperature and humidity state as the induction culture and with the illumination time of 10 hours/day;
s4, rooting culture:
transferring the young wheat seedlings subjected to proliferation culture in the step S3 into a rooting culture medium, and culturing for 30-40 days under the same temperature and humidity state as induction culture by matching with the illumination condition of the proliferation culture medium to obtain selenium-containing wheat seedlings;
s5, hybridization breeding:
transplanting and sowing the selenium-containing wheat seedlings obtained in the step S4, taking the first-period wheat seeds harvested after sowing as female parent, selecting high-quality selenium-rich wheat seeds as male parent, carrying out hybridization breeding with the female parent, carrying out second-generation hybridization on the obtained hybrid wheat seeds and the female parent, and sowing and maturing to obtain the selenium-rich wheat seeds.
2. The selenium-enriched wheat breeding method according to claim 1, wherein the method comprises the following steps: in the step S1, wheat seeds are put into a refrigerator with the temperature of 2-7 ℃ for refrigeration for 8-14 h after being washed.
3. The selenium-enriched wheat breeding method according to claim 1, wherein the method comprises the following steps: in the step S1, the method for soaking and cleaning wheat seeds in a sterile environment comprises the following steps: soaking in 75% concentration alcohol solution for 40-70 s, washing with sterile water for 1-2 times, soaking in Tween 20-containing sodium hypochlorite for 5-9 min, washing with sterile water for 2-6 times, and repeating the steps for 1-2 times to complete the sterilizing treatment of wheat seed.
4. A selenium-enriched wheat breeding method according to claim 3, wherein: the Tween 20-containing sodium hypochlorite solution comprises Tween 20 with the volume concentration of 0.05-0.1% and sodium hypochlorite solution with the volume concentration of 0.2-0.25%.
5. The selenium-enriched wheat breeding method according to claim 1, wherein the method comprises the following steps: in the step S2, the induction culture medium consists of the following components in mass volume concentration: 1/2MS basic culture medium, 0.4mg/LNAA, 0.2mg/L ferric salt, 0.1-0.5 mg/L amino acid selenium fertilizer liquid, 30g/L sucrose and 3g/L agar; the pH value of the induction culture medium is 5.8-6.0.
6. The selenium-enriched wheat breeding method according to claim 1, wherein the method comprises the following steps: in the step S3, the proliferation medium consists of the following components in mass volume concentration: MS basic culture medium, 10-20 g/L sodium selenite, 8-15 g/L sodium selenate, 1.0g/L hydrolyzed casein, 0.4mg/LKt; the pH value of the proliferation culture medium is 5.8-6.0.
7. The selenium-enriched wheat breeding method according to claim 1, wherein the method comprises the following steps: in the step S4, the rooting culture medium consists of the following components in mass volume concentration: WR4 basic culture medium, 1/2MS culture medium, 0.4g/LCH, 0.3mg/LNAA, 0.7mg/L sodium naphthalene acetate, 0.3-0.6 mg/L selenium mineral powder; the pH value of the rooting culture medium is 5.8-6.0.
8. The selenium-enriched wheat breeding method according to claim 1, wherein the method comprises the following steps: in the step S5, the high-quality selenium-enriched wheat seeds are any one of agricultural large 3753, selenium-enriched Y8, black wheat No. 76 and black gem No. 2.
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| CN108207631A (en) * | 2018-01-22 | 2018-06-29 | 浙江东来天然生物制品有限公司 | A kind of selenium-enriched officinal dendrobium stem seedling tissue culture method for improving Se content |
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| CN105011020A (en) * | 2015-08-05 | 2015-11-04 | 南京贝杉国际贸易有限公司 | Wheat seedling powder high in biological selenium content and preparation method thereof |
| CN108207631A (en) * | 2018-01-22 | 2018-06-29 | 浙江东来天然生物制品有限公司 | A kind of selenium-enriched officinal dendrobium stem seedling tissue culture method for improving Se content |
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