CN106035059A - Method for artificially synthesizing novel anti-clubroot brassica napus material - Google Patents
Method for artificially synthesizing novel anti-clubroot brassica napus material Download PDFInfo
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- CN106035059A CN106035059A CN201610363713.3A CN201610363713A CN106035059A CN 106035059 A CN106035059 A CN 106035059A CN 201610363713 A CN201610363713 A CN 201610363713A CN 106035059 A CN106035059 A CN 106035059A
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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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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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
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
- A01H1/08—Methods for producing changes in chromosome number
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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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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
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Abstract
The invention discloses a method for artificially synthesizing a novel anti-clubroot brassica napus material. The method includes: taking anti-clubroot brassica oleracea as a female parent to hybridize with anti-clubroot pakchoi; after 15 days of pollination of a generation F1, selecting seeds for embryo rescue, using a B5 fluid medium containing 2% of sucrose to culture immature seeds, and after immature embryos emerge and send up cotyledons, transferring the embryos to the B5 fluid medium containing 2% of sucrose, using 0.1% colchicines in 3-4-leaf stage to treat for 2h to realize doubling germination, cleaning, transferring into soil, and enabling selfing of synthesized plants by means of artificial pollination or saltwater spraying in a flowering stage; in a germination stage of selfing seeds, adopting a clubroot P4 physiological race for inoculation while taking an infected material as a contrast, inoculating at the same time, taking out seedling roots after 35 days, cleaning, observing whether lumps exist or not, and determining that the synthesized plants are resistant to clubroot if no lump exists. An effective approach is provided for breeding of anti-clubroot brassica napus strains, and clubroot inoculation efficiency is improved.
Description
Technical field
The present invention relates to cabbage type rape synthetic technical field, specifically a kind of synthetic anti-clubroot Wild cabbage type
The method of Brassica campestris L new material.
Background technology
Clubroot is a kind of fungal disease caused by Brassica genus plasmodiophora brassicae (Plasmodiophora brassicae), existing
Being distributed in the whole world, Temperate Region in China incidence is particularly acute.Nineteen ninety-five, China's reported first Brassica campestris L clubroot, subsequently should
Sick spreading in whole nation large area, major part provinces, municipalities and autonomous regions all have a distribution at present, but be mainly distributed on Yunnan, Sichuan, Guizhou,
The ground such as Hubei, Anhui, Southern Shaanxi.This infection process and harm almost all of crucifer, particularly Brassica genus are planted
Thing, such as Chinese cabbage, Caulis et Folium Brassicae capitatae etc. (Piao etc., 2009).Brassica campestris L clubroot started to occur from seedling stage, mainly infects root, causes plant
Growth retardation, blade turns yellow, and plant is downgraded, and shows hydropenia, and time serious, seedling is withered.The susceptible tumor of Adult plant mostly occurs in side
Root and the bottom of main root, advanced stage tumours chaps and coarse, finally rots, had a strong impact on the normal absorption of moisture and nutrient,
Thus cause Brassica campestris L Severe Reduction or total crop failure.Brassica campestris L clubroot sickness rate in seedling stage reaches 17% according to research reports, Adult plant field
Average attack rate is 15%, typically results in underproduction 10%-20%, the serious field underproduction more than 50% (Tao Xuejun, 2015).At me
State, clubroot long-term hazard area about 320-400 ten thousand hectares, account for more than the 1/3 of crop in cruciferae cultivated area, be very popular year
Part occurs with hazard area up to 9,000,000 hectares (Wang Jing etc., 2008).
There is multiple biological strain in Cruciferae clubroot, in China, cabbage type rape clubroot cause of disease at least exists 2
Number, 3 biological strain types such as No. 4 and No. 13, wherein No. 4 microspecies exist widest in area general.Abroad, such as Canada, tool
There are at least 5 biological strains such as 2,3,5,6,8, the most predominantly No. 3 biological strains.Corresponding to different biological strains
Disease-causing gene is not quite similar, after the kind of the anti-clubroot in a certain area plants other region, usually due to biological strain
Changing and cause original anti-clubroot kind to become feeling the kind of clubroot, this brings to the preventing and treating of clubroot greatly
Difficulty.Additionally, due to more existing Cruciferae material resistances are more single, after plantation, its resistance is gradually degenerated for many years
(Leboldus etc., 2012), therefore, are badly in need of cultivating the kind being polymerized multiple resistant genes.
Due to clubroot infectiousness is strong, tool low concentration causes a disease threshold value (Hatakeyama etc., 2013), spread speed is fast, pass
Broadcast approach many, and we are not enough etc. to the understanding of the life cycle of this pathogenic bacteria and pathogenic disastrous mechanisms so that clubroot becomes generation
Crushing soilborne fungal pathogens in the range of boundary.Traditional chemical method is utilized can not fundamentally to solve to prevent and treat clubroot
Brassica campestris L clubroot is propagated, and also brings the pollution of environment simultaneously, is unfavorable for the sustainable development of agricultural.The most many experts are formed
Common recognition, the effective ways solving Brassica campestris L clubroot are the new rape varieties cultivating anti-clubroot, are phased out the product not having resistance
Kind.But in cabbage type rape, the rapeseed germplasm of anti-clubroot extremely lacks at present, it is difficult to effectively be applied in rapeseed breeding,
And the nearly edge species Chinese cabbage of cabbage type rape, Caulis et Folium Brassicae capitatae etc. also exist the material of many anti-clubroots.Therefore, we are badly in need of research
The heredity of clubroot and the mechanism of action, formulate the Brassica campestris L material of anti-clubroot, cultivates the new rape variety of anti-clubroot, and in addition
Promote, alleviate the harm of China's Brassica campestris L clubroot.
At present, the inoculation method of the clubroot of most places is the most extensive, and even a lot of units are only reflected with Field inoculation
Fixed result is as judging the most anti-clubroot of plant, even if some mix soil inoculation in the bacterium solution that the most mostly uses of laboratory inoculation
Method, often occur individuality not inoculate successful phenomenon, thus give the anti-clubroot of germ plasm resource identify bring the biggest
Error, usually the prior year be accredited as anti-clubroot material and next year show as feel clubroot.Clubroot exists in recent years
China's Brassica campestris L main producing region is spread rapidly, has had a strong impact on the production of China's Brassica campestris L, is therefore badly in need of cultivating the oil with anti-clubroot
Dish new varieties.But owing to the resource of clubroot anti-in China's cabbage type rape is the most rare, and its nearly edge species Chinese cabbage, Caulis et Folium Brassicae capitatae,
There is the material of anti-clubroot in black mustard, therefore the present invention intends the method using distant hybridization, utilizes Chinese cabbage and resisting in Caulis et Folium Brassicae capitatae
The germ plasm resource of clubroot, the brassica napus of the anti-clubroot of synthetic, grope one and synthesize anti-clubroot Wild cabbage type
The method of napus lines, makes up the shortage of China's Brassica Napus clubroot resource.Carry out clubroot inoculation technique simultaneously
Research, improves the inoculation efficiency of clubroot, and the Resistance Identification for clubroot provides good technology to ensure.Therefore, it is badly in need of former
Some clubroot inoculation methods are improved, and improve clubroot and inoculate successfully efficiency.
Summary of the invention
The method that it is an object of the invention to provide a kind of synthetic anti-clubroot cabbage type rape new material, to solve
The problem proposed in above-mentioned background technology.
For achieving the above object, the present invention provides following technical scheme:
The method of a kind of synthetic anti-clubroot cabbage type rape new material, with the Caulis et Folium Brassicae capitatae of anti-clubroot for female parent with anti-
The Plantula Brassicae chinensis hybridization of clubroot, after F1 generation is pollinated 15 days, selects seed and carries out rescue culture, utilize the B5 liquid of the sucrose containing 2%
The immature seed of culture medium culturing, when growing cotyledon after rataria occurs, moves to the B5 solid medium containing 2% sucrose by embryo
On, 3-4 utilizes the Colchicine process of 0.1% to double for 2 hours during the leaf phase, moves in soil, utilize stripping after rinsed clean
The method of flower bud pollination or Spraying in the flowering stage saline makes the plant selfing of synthesis, and the seed of selfing, at germination period, utilizes clubroot P4
Biological strain inoculates, and simultaneously using susceptible material as comparison, inoculates simultaneously；After 35 days, taking out the root of seedling, clean observation is
No have lump, does not has the plant of lump explanation synthesis to have clubroot resistance.
As the further scheme of the present invention: described Caulis et Folium Brassicae capitatae is the green offspring in cabbage variety century, described little Bai
Dish is the offspring of varieties of Brassica campestris four seasons green vegetable.
As the present invention further scheme: choose tender leaf at peduncle-growing period for rapeseed and utilize Ploidy Analyzer that plant is entered
Row Ploidy Identification, observes the fertility of plant during the florescence, final judgement doubles the most successful.
As the present invention further scheme: it is the most affine that selfing removed by described stripping flower bud selfing or Spraying in the flowering stage saline.
As the present invention further scheme: concrete inoculation method is as follows: by ripe seed accelerating germination on filter paper
After, root is soaked in the bacterium solution of the P4 biological strain of 107 spores/milliliter 2 hours, then transplant to containing weakly acidic summary
In wet substrate, the most every plumelet is inoculated in the 107 spores/milliliter bacterium solution to substrate of 5 milliliters, and transplanting plumelet is moved to greenhouse
In.
As the present invention further scheme: described Greenhouse day temperature is 22 DEG C, night temperatures is 18 DEG C.
Compared with prior art, the invention has the beneficial effects as follows: provide a selection-breeding anti-clubroot cabbage type rape
Strain effective way, improves clubroot inoculation efficiency.
Accompanying drawing explanation
Fig. 1 is the anti-clubroot cabbage type rape strain in the method for synthetic anti-clubroot cabbage type rape new material
Synthetic line figure.
Fig. 2 be synthetic anti-clubroot cabbage type rape new material method in clubroot Disease Resistance Identification schematic diagram.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Describe, it is clear that described embodiment is only a part of embodiment of the present invention rather than whole embodiments wholely.Based on
Embodiment in the present invention, it is every other that those of ordinary skill in the art are obtained under not making creative work premise
Embodiment, broadly falls into the scope of protection of the invention.
Refer to Fig. 1～2, in the embodiment of the present invention, the side of a kind of synthetic anti-clubroot cabbage type rape new material
Method, is the little of maternal and anti-clubroot with the Caulis et Folium Brassicae capitatae (coming from the offspring of cabbage variety-" century green ") of anti-clubroot
Chinese cabbage (coming from the offspring of varieties of Brassica campestris " four seasons green vegetable ") hybridizes, F1After generation pollination 15 days, select seed and carry out rescue culture,
Immature seed cultivated by the B5 fluid medium utilizing the sucrose containing 2%, when growing cotyledon after rataria occurs, is moved to by embryo
On B5 solid medium containing 2% sucrose, 3-4 utilizes the Colchicine process of 0.1% to double for 2 hours during the leaf phase, rinsing
Move to after Gan Jing in soil, choose tender leaf at peduncle-growing period for rapeseed and utilize Ploidy Analyzer plant to be carried out Ploidy Identification, during the florescence
Observing the fertility of plant, final judgement doubles the most successful.The method utilizing cabage-type rape or Spraying in the flowering stage saline makes synthesis
Plant selfing (remove selfing the most affine), the seed of selfing, at germination period, utilizes clubroot P4 biological strain inoculate, while with
Susceptible material, as comparison, is inoculated simultaneously, after 35 days, takes out the root of seedling, cleans and seen whether lump, do not have lump to say
The plant of bright synthesis has clubroot resistance.
Concrete inoculation method is as follows: by ripe seed on filter paper after accelerating germination, by root 107Spore/milliliter
Immersion 2 hours in the bacterium solution of P4 biological strain, then transplant to containing in weakly acidic substrate (the wettest), and the most every plumelet is inoculated
The 10 of 5 milliliters7In spore/milliliter bacterium solution to substrate, transplanting plumelet is moved to (22/18 DEG C, day/night) in greenhouse, first 3 days
Try not to water, if hydropenia occurring after 3 days, just spraying a small amount of moisture in right amount with watering can, only keeping the moistening of substrate, not
Open fire occurs.
The present invention utilizes Chinese cabbage and the anti-clubroot resource in Caulis et Folium Brassicae capitatae, the cabbage type rape of the anti-clubroot of synthetic, adopts
The method identified with indoor inoculation, adjusts original indoor inoculation parameter, inoculates seedling root at Germination period, uses simultaneously
Strict control measures, improve and inoculate successfully efficiency.
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of the spirit or essential attributes of the present invention, it is possible to realize the present invention in other specific forms.Therefore, no matter
From the point of view of which point, all should regard embodiment as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit requires rather than described above limits, it is intended that all by fall in the implication of equivalency and scope of claim
Change is included in the present invention.Should not be considered as limiting involved claim by any reference in claim.
Although moreover, it will be appreciated that this specification is been described by according to embodiment, but the most each embodiment only wraps
Containing an independent technical scheme, this narrating mode of description is only that for clarity sake those skilled in the art should
Description can also be formed those skilled in the art through appropriately combined as an entirety, the technical scheme in each embodiment
May be appreciated other embodiments.
Claims (6)
1. the method for a synthetic anti-clubroot cabbage type rape new material, it is characterised in that with the Caulis et Folium Brassicae capitatae of anti-clubroot
For the maternal Plantula Brassicae chinensis hybridization with anti-clubroot, F1After generation pollination 15 days, select seed and carry out rescue culture, utilize the sugarcane containing 2%
Immature seed cultivated by the B5 fluid medium of sugar, when growing cotyledon after rataria occurs, embryo moves to the B5 containing 2% sucrose
On solid medium, 3-4 utilizes the Colchicine process of 0.1% to double for 2 hours during the leaf phase, moves to soil after rinsed clean
In, utilize the method for cabage-type rape or Spraying in the flowering stage saline to make the plant selfing of synthesis, the seed of selfing, at germination period, utilizes
Clubroot P4 biological strain inoculates, and simultaneously using susceptible material as comparison, inoculates simultaneously；After 35 days, take out the root of seedling, wash
Only see whether lump, do not have the plant of lump explanation synthesis to have clubroot resistance.
The method of synthetic the most according to claim 1 anti-clubroot cabbage type rape new material, it is characterised in that institute
Stating the green offspring that Caulis et Folium Brassicae capitatae is cabbage variety century, described Plantula Brassicae chinensis is the offspring of varieties of Brassica campestris four seasons green vegetable.
The method of synthetic the most according to claim 1 anti-clubroot cabbage type rape new material, it is characterised in that
Peduncle-growing period for rapeseed is chosen tender leaf and is utilized Ploidy Analyzer that plant is carried out Ploidy Identification, observes the fertility of plant, finally during the florescence
Judgement doubles the most successful.
The method of synthetic the most according to claim 1 anti-clubroot cabbage type rape new material, it is characterised in that institute
State stripping flower bud selfing or the removal selfing of Spraying in the flowering stage saline is the most affine.
The method of synthetic the most according to claim 1 anti-clubroot cabbage type rape new material, it is characterised in that institute
The concrete method stating inoculation is as follows: by ripe seed on filter paper after accelerating germination, by root 107The P4 of spore/milliliter is raw
Soaking in the bacterium solution of reason microspecies 2 hours, then transplanting is to containing in the wettest weakly acidic substrate, and the most every plumelet inoculation 5 is in the least
10 risen7In spore/milliliter bacterium solution to substrate, transplanting plumelet is moved in greenhouse.
The method of synthetic the most according to claim 5 anti-clubroot cabbage type rape new material, it is characterised in that institute
The Greenhouse day temperature stated is 22 DEG C, and night temperatures is 18 DEG C.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108617494A (en) * | 2017-03-17 | 2018-10-09 | 四川省农业科学院作物研究所 | A kind of selection of Brassica Napus clubroot maintainer |
| CN109042291A (en) * | 2018-07-28 | 2018-12-21 | 湖南科技大学 | A method of novel vegetable is created using Chinese cabbage and wild cabbage distant hybridization |
| CN110999779A (en) * | 2019-12-13 | 2020-04-14 | 西北农林科技大学 | Method for synthesizing clubroot-resistant brassica napus by using polymerization breeding and biotechnology |
| CN114303516A (en) * | 2022-01-04 | 2022-04-12 | 沈阳农业大学 | Method for inoculating plasmodiophora brassicae and application of plasmodiophora brassicae in screening of plasmodiophora brassicae resistant breeding materials |
| CN115918517A (en) * | 2022-10-31 | 2023-04-07 | 云南省农业科学院园艺作物研究所 | Rapid breeding method of anti-clubroot bolting vegetable breeding material |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108617494A (en) * | 2017-03-17 | 2018-10-09 | 四川省农业科学院作物研究所 | A kind of selection of Brassica Napus clubroot maintainer |
| CN109042291A (en) * | 2018-07-28 | 2018-12-21 | 湖南科技大学 | A method of novel vegetable is created using Chinese cabbage and wild cabbage distant hybridization |
| CN110999779A (en) * | 2019-12-13 | 2020-04-14 | 西北农林科技大学 | Method for synthesizing clubroot-resistant brassica napus by using polymerization breeding and biotechnology |
| CN110999779B (en) * | 2019-12-13 | 2023-01-20 | 西北农林科技大学 | Method for synthesizing clubroot-resistant brassica napus by using polymerization breeding and biotechnology |
| CN114303516A (en) * | 2022-01-04 | 2022-04-12 | 沈阳农业大学 | Method for inoculating plasmodiophora brassicae and application of plasmodiophora brassicae in screening of plasmodiophora brassicae resistant breeding materials |
| CN114303516B (en) * | 2022-01-04 | 2022-12-13 | 沈阳农业大学 | Method for inoculating plasmodiophora brassicae and application of plasmodiophora brassicae in screening of plasmodiophora brassicae resistant breeding materials |
| CN115918517A (en) * | 2022-10-31 | 2023-04-07 | 云南省农业科学院园艺作物研究所 | Rapid breeding method of anti-clubroot bolting vegetable breeding material |
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