CN106879452A - A kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility - Google Patents

A kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility Download PDF

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Publication number
CN106879452A
CN106879452A CN201710126914.6A CN201710126914A CN106879452A CN 106879452 A CN106879452 A CN 106879452A CN 201710126914 A CN201710126914 A CN 201710126914A CN 106879452 A CN106879452 A CN 106879452A
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buckwheat
pollination
rataria
sweet
seedling
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房世平
胡丹
秦国鹏
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Qingyang Dunbo Technology Development Co Ltd
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Qingyang Dunbo Technology Development Co Ltd
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • 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
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods 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
    • 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
    • A01H4/001—Culture apparatus for tissue culture
    • 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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  • Life Sciences & Earth Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Environmental Sciences (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Soil Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Cultivation Of Plants (AREA)

Abstract

The invention provides a kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility, belong to biotechnology breeding field.Comprise the following steps：1) Juvenile stage；2) material plantation；3) hybridization pollination；4) rataria is disinfected；5) rataria inoculated and cultured；6) training tissue culture seedling；7) seedling replanting.Beneficial effects of the present invention：Using the compatibility for repeating pollination raising pollen and column cap, using young embryo rescue techniques affiliation buckwheat interspecific hybridization rataria agensis farther out is overcome after pollination and cannot obtain the problem of Hybrids F1 plant, the merit of bitter buckwheat is transferred in sweet buckwheat simultaneously, the germplasm of sweet buckwheat is effectively improved, important positive role is played to enriching the hereditary basis of sweet buckwheat kind, optimization breeding technique.

Description

A kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility
Technical field
The invention belongs to biotechnology breeding field, specifically a kind of side for overcoming sweet buckwheat and bitter buckwheat interspecies sterility Method.
Background technology
Buckwheat belongs to buckwheat crop, and China is that buckwheat cultivates most long country, early in B.C. 3-5 centuries, 《Legendary god of farming's book》In " eight paddy grow a piece " just have the record that buckwheat grows.Fagopyrum cultigen is main by 2 kinds, i.e., sweet buckwheat and Bitter buckwheat, sweet buckwheat and bitter buckwheat have nutritive value and medical value very high, and it is most promising that it is referred to as 21 century by nutritionist Pollution-free food, whole body is precious, and nutritive value and medical value are high, containing the bioflavonoid that many other crops are not contained Beneficial to the material of health, there is flavonoids rutin etc. inoxidizability, anti-lipid peroxidation to react, maintenance vasodilation, Fragility of blood vessels is reduced, the functions such as angiocardiopathy generation and cancer-resisting is prevented, while also antiallergy, diuresis, antibechic, drop The effects such as blood fat, hypotensive, enhancing coronary blood flow.
Sweet buckwheat belongs to typical cross pollinated plant, and selfing is not affine, and bitter buckwheat belongs to typical self pollination crop, will be sweet Buckwheat and bitter buckwheat are hybridized, and Cross infertility is caused because rataria stops development, and cannot obtain the first generation of hybrid.Hybrid rescue culture Culture technique as tissue cultures a key areas, be the effective ways for overcoming after fertilization maldevelopment at present, in plant This important effect is played in breeding.At present, the interspecific hybridization of sweet buckwheat and bitter buckwheat does not succeed, the interspecific hybridization of sweet buckwheat and bitter buckwheat Technology page has no report, in order to overcome this phenomenon, widens the genetic resources of sweetness and bitterness buckwheat, improves its breediness, overcomes its miscellaneous Hand over infertility very urgent.
The content of the invention
It is an object of the present invention to provide a kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility.
In order to achieve the above object, the present invention uses following technical scheme.
1) Juvenile stage：Female parent selects more late-maturing sweet buckwheat kind, and male parent is from the excellent bitter buckwheat kind of Comprehensive Traits, essence The seed of full no disease and pests harm is selected in case plantation；
2) material plantation：Growth characteristics and flowering habit interval sowing according to Parent, to ensure the florescence of two parents Meet；
3) hybridization pollination：When the equal Post flowering of Parent, the maternal flower that selection is just bloomed, has been opened seed that removal has been tied Flower and the flower opened, bagging isolation, listing mark, in the next morning 9：00-10:Bloomed on 00 selection paternal plant Just luxuriant flower, removes little Hua together with bennet, opens maternal sulphur paper bag, and hand-held little Hua takes smearing female parent is small, enters Row pollination, sulphur paper bag is put after pollination to female parent, seals set sack, continuous pollination 4-5 days, marks Parent title and hybridization Date；
4) rataria is disinfected：3-6 days after sweet buckwheat and bitter buckwheat pollination, take the seed after hybridization and carry out disinfection, use 70% wine Essence sterilization 30-60s, 0.1% mercuric chloride sterilization 10-15min, then with sterile water wash 4-5 times, sterilizing blotting paper is placed on by seed Upper suck dry moisture；
5) rataria inoculated and cultured：On superclean bench, rataria is inoculated in root media, blake bottle is placed in temperature Spend for 25-26 DEG C, relative humidity be 65-70%, light application time for 12h constant incubator in cultivate；
6) training tissue culture seedling：Treat that rataria is taken root to the small after seedling of 8-10cm, open culture medium bottleneck, hardening 48-72h Afterwards, the culture medium that seedling running water rinses residual is taken out from blake bottle, is put into test tube, add running water, originally water logging Do not have seedling root, hardening treatment is carried out again, test tube is placed on temperature for 25-26 DEG C, relative humidity are 65-70%, illumination Time for 12h illumination cultivation room in cultivate 3-5d；
7) seedling replanting：In seedling replanting after hardening to the nutritive cube for filling nutrient matrix, film, every 3 day will be covered Sprinkling 1/10MS a great number of elements solution 100ml, until after plant survives, throw off film, in greenhouse routinely sweet buckwheat cultivate into Row management.
Above-mentioned steps 5) described in root media be improvement MS culture medium+0.2mg/L NAA+0.5mg/LIAA+ 0.4mg/L IBA+0.2mg/L activated carbon+0.1mg/L uniconazole Ps, a great number of elements and trace element of the MS culture mediums of improvement subtract Half, other elements content is normal.
Above-mentioned steps 7) described in the composition of nutrient matrix be by weight：Garden mould 40%+ plant ash 20%+ peat 20% After the mixing of+vermiculite 10%+ bone meal 5%+ rotten woods bits 5%, sterilizing.
Advantageous Effects of the invention：Using the compatibility for repeating pollination raising pollen and column cap, using children after pollination Embryo rescue techniques overcome affiliation buckwheat interspecific hybridization rataria agensis farther out and cannot obtain Hybrids F1 plant Problem, while the merit of bitter buckwheat is transferred in sweet buckwheat, the germplasm of sweet buckwheat is effectively improved, to enriching sweet buckwheat kind Hereditary basis, optimization breeding technique play important positive role.
Specific embodiment
Further make detailed elaboration to technical scheme with reference to embodiment, but it is not all of the invention in pairs Scope is limited.
Embodiment 1
1) Juvenile stage：Female parent selects more late-maturing sweet buckwheat kind tenth of the twelve Earthly Branches buckwheat 2, and male parent is from the excellent bitter buckwheat of Comprehensive Traits Kind Jiujiang bitter buckwheat, the seed of selected full no disease and pests harm is in case plantation；
2) material plantation：Growth characteristics and flowering habit interval sowing according to Parent, male parent are broadcast than the maternal 10d that shifts to an earlier date Kind, to ensure the flower synchronization of two parents；
3) hybridization pollination：When the equal Post flowering of Parent, the maternal flower that selection is just bloomed, has been opened seed that removal has been tied Flower and the flower opened, bagging isolation, listing mark, in the next morning 9：00-10:Bloomed on 00 selection paternal plant Just luxuriant flower, removes little Hua together with bennet, opens maternal sulphur paper bag, and hand-held little Hua takes smearing female parent is small, enters Row pollination, sulphur paper bag is put after pollination to female parent, seals set sack, continuous pollination 4 days, marks Parent title and hybridization day Phase；
4) rataria is disinfected：3 days after sweet buckwheat and bitter buckwheat pollination, take the seed after hybridization and carry out disinfection, use 70% alcohol Sterilization 30s, 0.1% mercuric chloride sterilization 10min, then with sterile water wash 4 times, seed are placed on sterilizing blotting paper and blot water Point；
5) rataria inoculated and cultured：On superclean bench, rataria is inoculated in root media, blake bottle is placed in temperature Spend for 25 DEG C, relative humidity are that 65%, light application time is culture in the constant incubator of 12h；
6) training tissue culture seedling：Treat that rataria is taken root to the small after seedling of 8cm, culture medium bottleneck is opened, after hardening 48h, from training Taking-up seedling running water rinses the culture medium for remaining in supporting bottle, is put into test tube, adds running water, originally water submerged seedling root Portion, carries out hardening treatment again, by test tube be placed on temperature be 25 DEG C, relative humidity be illumination that 65%, light application time is 12h 3d is cultivated in culturing room；
7) seedling replanting：In seedling replanting after hardening to the nutritive cube for filling nutrient matrix, film, every 3 day will be covered Sprinkling 1/10MS a great number of elements solution 100ml, until after plant survives, throw off film, in greenhouse routinely sweet buckwheat cultivate into Row management.
Embodiment 2
1) Juvenile stage：Female parent selects more late-maturing sweet buckwheat kind tenth of the twelve Earthly Branches buckwheat 2, and male parent is from the excellent bitter buckwheat of Comprehensive Traits Kind Jiujiang bitter buckwheat, the seed of selected full no disease and pests harm is in case plantation；
2) material plantation：Growth characteristics and flowering habit interval sowing according to Parent, male parent are broadcast than the maternal 15d that shifts to an earlier date Kind, to ensure the flower synchronization of two parents；
3) hybridization pollination：When the equal Post flowering of Parent, the maternal flower that selection is just bloomed, has been opened seed that removal has been tied Flower and the flower opened, bagging isolation, listing mark, in the next morning 9：00-10:Bloomed on 00 selection paternal plant Just luxuriant flower, removes little Hua together with bennet, opens maternal sulphur paper bag, and hand-held little Hua takes smearing female parent is small, enters Row pollination, sulphur paper bag is put after pollination to female parent, seals set sack, continuous pollination 5 days, marks Parent title and hybridization day Phase；
4) rataria is disinfected：6 days after sweet buckwheat and bitter buckwheat pollination, take the seed after hybridization and carry out disinfection, use 70% alcohol Sterilization 60s, 0.1% mercuric chloride sterilization 15min, then with sterile water wash 5 times, seed are placed on sterilizing blotting paper and blot water Point；
5) rataria inoculated and cultured：On superclean bench, rataria is inoculated in root media, blake bottle is placed in temperature Spend for 25 DEG C, relative humidity are that 70%, light application time is culture in the constant incubator of 12h；
6) training tissue culture seedling：Treat that rataria is taken root to the small after seedling of 10cm, culture medium bottleneck is opened, after hardening 72h, from training Taking-up seedling running water rinses the culture medium for remaining in supporting bottle, is put into test tube, adds running water, originally water submerged seedling root Portion, carries out hardening treatment again, by test tube be placed on temperature be 25 DEG C, relative humidity be illumination that 70%, light application time is 12h 5d is cultivated in culturing room；
7) seedling replanting：In seedling replanting after hardening to the nutritive cube for filling nutrient matrix, film, every 3 day will be covered Sprinkling 1/10MS a great number of elements solution 100ml, until after plant survives, throw off film, in greenhouse routinely sweet buckwheat cultivate into Row management.
Embodiment 3
1) Juvenile stage：Female parent selects more late-maturing sweet buckwheat kind tenth of the twelve Earthly Branches buckwheat 2, and male parent is from the excellent bitter buckwheat of Comprehensive Traits Kind Jiujiang bitter buckwheat, the seed of selected full no disease and pests harm is in case plantation；
2) material plantation：Growth characteristics and flowering habit interval sowing according to Parent, male parent are broadcast than the maternal 12d that shifts to an earlier date Kind, to ensure the flower synchronization of two parents；
3) hybridization pollination：When the equal Post flowering of Parent, the maternal flower that selection is just bloomed, has been opened seed that removal has been tied Flower and the flower opened, bagging isolation, listing mark, in the next morning 9：00-10:Bloomed on 00 selection paternal plant Just luxuriant flower, removes little Hua together with bennet, opens maternal sulphur paper bag, and hand-held little Hua takes smearing female parent is small, enters Row pollination, sulphur paper bag is put after pollination to female parent, seals set sack, continuous pollination 5 days, marks Parent title and hybridization day Phase；
4) rataria is disinfected：4 days after sweet buckwheat and bitter buckwheat pollination, take the seed after hybridization and carry out disinfection, use 70% alcohol Sterilization 45s, 0.1% mercuric chloride sterilization 12min, then with sterile water wash 4 times, seed are placed on sterilizing blotting paper and blot water Point；
5) rataria inoculated and cultured：On superclean bench, rataria is inoculated in root media, blake bottle is placed in temperature Spend for 25 DEG C, relative humidity are that 70%, light application time is culture in the constant incubator of 12h；
6) training tissue culture seedling：Treat that rataria is taken root to the small after seedling of 9cm, culture medium bottleneck is opened, after hardening 60h, from training Taking-up seedling running water rinses the culture medium for remaining in supporting bottle, is put into test tube, adds running water, originally water submerged seedling root Portion, carries out hardening treatment again, by test tube be placed on temperature be 25 DEG C, relative humidity be illumination that 70%, light application time is 12h 3d is cultivated in culturing room；
7) seedling replanting：In seedling replanting after hardening to the nutritive cube for filling nutrient matrix, film, every 3 day will be covered Sprinkling 1/10MS a great number of elements solution 100ml, until after plant survives, throw off film, in greenhouse routinely sweet buckwheat cultivate into Row management.
Rescue culture is carried out to the rataria after parent's tenth of the twelve Earthly Branches buckwheat 2 and the pollination of Jiujiang bitter buckwheat, rescue culture is carried out after finding pollination 4d Efficiency highest.
Number of seedling after the redemption of the rataria of table 1
Cross combination Number of days after pollination Rescue rataria number Obtain seedling number
No. 2 × Jiujiang of tenth of the twelve Earthly Branches buckwheat bitter buckwheat 3 50 12
No. 2 × Jiujiang of tenth of the twelve Earthly Branches buckwheat bitter buckwheat 4 50 20
No. 2 × Jiujiang of tenth of the twelve Earthly Branches buckwheat bitter buckwheat 5 50 16
No. 2 × Jiujiang of tenth of the twelve Earthly Branches buckwheat bitter buckwheat 6 50 13

Claims (3)

1. a kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility, it is characterised in that use following steps：
1) Juvenile stage：Female parent selects more late-maturing sweet buckwheat kind, and male parent is selected full from the excellent bitter buckwheat kind of Comprehensive Traits The seed of full no disease and pests harm is in case plantation；
2) material plantation：Growth characteristics and flowering habit interval sowing according to Parent, to ensure the flower synchronization of two parents；
3) hybridization pollination：When the equal Post flowering of Parent, the maternal flower that selection is just bloomed, has been bloomed piece at seed that removal has been tied And the flower opened, bagging isolation, listing mark, in the next morning 9：00-10:Bloomed on 00 selection paternal plant positive luxuriant The flower of Sheng, removes little Hua together with bennet, opens maternal sulphur paper bag, and hand-held little Hua takes smearing female parent is small, is awarded Powder, sulphur paper bag is put after pollination to female parent, seals set sack, continuous pollination 4-5 days, marks Parent title and hybridization day Phase；
4) rataria is disinfected：3-6 days after sweet buckwheat and bitter buckwheat pollination, take the seed after hybridization and carry out disinfection, disappeared with 70% alcohol Malicious 30-60s, 0.1% mercuric chloride sterilization 10-15min, then with sterile water wash 4-5 time, suction on sterilizing blotting paper are placed on by seed Solid carbon dioxide point；
5) rataria inoculated and cultured：On superclean bench, rataria is inoculated in root media, blake bottle is placed in into temperature is 25-26 DEG C, relative humidity be 65-70%, light application time for 12h constant incubator in cultivate；
6) training tissue culture seedling：Treat that rataria is taken root to the small after seedling of 8-10cm, open culture medium bottleneck, after hardening 48-72h, from The culture medium that seedling running water rinses residual is taken out in blake bottle, is put into test tube, add running water, originally water submerged seedling Root, carries out hardening treatment again, and test tube is placed on into temperature for 25-26 DEG C, relative humidity are that 65-70%, light application time are 3-5d is cultivated in the illumination cultivation room of 12h；
7) seedling replanting：Film in seedling replanting after hardening to the nutritive cube for filling nutrient matrix, will be covered, is sprayed within every 3 days 1/10MS a great number of elements solution 100ml, until after plant survives, throwing off film, routinely sweet buckwheat cultivation is managed in greenhouse Reason.
2. a kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility according to claim 1, it is characterised in that step 5) root media described in is the MS culture medium+0.2mg/L NAA+0.5mg/L IAA+0.4mg/L IBA+0.2mg/ of improvement L activated carbon+0.1mg/L uniconazole Ps, a great number of elements and trace element of the MS culture mediums of improvement halve, and other elements content is just Often.
3. a kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility according to claim 1, it is characterised in that step 7) composition of the nutrient matrix described in is by weight：Garden mould 40%+ plant ash 20%+ peat 20%+ vermiculite 10%+ bone meal After the mixing of 5%+ rotten woods bits 5%, sterilizing.
CN201710126914.6A 2017-03-06 2017-03-06 A kind of method for overcoming sweet buckwheat and bitter buckwheat interspecies sterility Pending CN106879452A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109042305A (en) * 2018-09-11 2018-12-21 宝鸡市农业科学研究院 A kind of selection of bitter buckwheat kind
CN110419401A (en) * 2019-09-04 2019-11-08 山西省农业科学院农作物品种资源研究所 A kind of method for creating of easy shelling bitter buckwheat germplasm

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109042305A (en) * 2018-09-11 2018-12-21 宝鸡市农业科学研究院 A kind of selection of bitter buckwheat kind
CN109042305B (en) * 2018-09-11 2021-09-10 宝鸡市农业科学研究院 Breeding method of tartary buckwheat variety
CN110419401A (en) * 2019-09-04 2019-11-08 山西省农业科学院农作物品种资源研究所 A kind of method for creating of easy shelling bitter buckwheat germplasm
CN110419401B (en) * 2019-09-04 2023-09-22 山西省农业科学院农作物品种资源研究所 Preparation method of tartary buckwheat germplasm easy to unshelling

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Application publication date: 20170623