CN101347096A - Method for breeding cotton with greensickness-resistance - Google Patents

Method for breeding cotton with greensickness-resistance Download PDF

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Publication number
CN101347096A
CN101347096A CNA2008102224923A CN200810222492A CN101347096A CN 101347096 A CN101347096 A CN 101347096A CN A2008102224923 A CNA2008102224923 A CN A2008102224923A CN 200810222492 A CN200810222492 A CN 200810222492A CN 101347096 A CN101347096 A CN 101347096A
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cotton
verticillium wilt
verticillium
disease
seed
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CN101347096B (en
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张永军
李修立
李方顺
吴孔明
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Institute of Plant Protection of Chinese Academy of Agricultural Sciences
Institute of Food Science and Technology of CAAS
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Institute of Plant Protection of Chinese Academy of Agricultural Sciences
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Abstract

The invention relates to a verticillium wilt resistant cotton breeding method, which comprises the following steps: (1) a disease nursery is established and the pathogenic seeds are annually updated; (2) the seeds are soaked, infected with pathogen and sowed; (3) the verticillium wilt resistant cotton material is bred. The method can also identify and screen out a cotton variety with compatible resistance by introducing a Bt gene. The variety which is accurately bred by the method and the breeding process provided by the invention has the advantages of strong and stable verticillium wilt resistance, high yield, wide applicability and remarkable economic benefits.

Description

The breeding method of cotton with greensickness-resistance
Technical field
The invention belongs to the crop breeding field, specifically, is a kind of breeding method of verticillium wilt resistance of cotton by same.
Background technology
Verticillium wilt is one of most important disease of cotton, and the title of cotton " cancer " is arranged, and nineteen thirty-five is imported China into.After the end of the eighties in last century, fusarium wilt was controlled, verticillium wilt rose to the first disease of cotton.Since the nineties in 20th century, this disease has spreaded all over each main product cotton region of China, and the diseased plant rate Chang Gaoda in grave illness cotton field is more than 80%, and production loss is serious.1993, verticillium wilt continuous large-area in 1995 and 1996 takes place, wherein national cotton verticillium wilt disaster area in 1993 reaches 4,000 ten thousand mu (266.67 ten thousand hm2), loss gined cotton 2,000,000 loads (100,000,000 kg), especially northern cotton region has many cotton fields the phenomenon that diseased plant in blocks is fallen leaves into the light stalk to occur, cause the underproduction significantly, so that total crop failure, the threat of verticillium wilt have become an outstanding problem of restriction Cotton Production.Verticillium wilt belongs to soil biography, vascular bundle diseases, and chemical control is difficult to prove effective, and cultivating disease-resistant variety is this sick main method of control, but the present domestic high verticillium wilt-resistant cotton kind of still not having.
Cotton is most important in the world economic crops, also is the important source of China's foreign exchange earning.China just begins cotton breeding as far back as the forties in 20th century, carries out crossbreeding after the fifties, and the cotton field disease was light at that time, basically according to output performance seed selection kind.The generation of cotton wilt increases the weight of gradually after the seventies, begun the anti-blight breeding, but breeding technique adopts the susceptible experimental field of nature to carry out disease resistance basically and identifies, because pathogen deficiency, the field morbidity is inhomogeneous, the time differences is big, and breeding parent and filial generation material qualification result are inaccurate, and material is elected or superseded blindness is bigger.Verticillium wilt takes place gradually seriously after the nineties, press for and cultivate the resisting verticillium kind, but disease resistance is identified with seed selection and is still adopted and the similar method of anti-blight, breeding efficiency is low, progeny material is selected inaccurate, and the improved variety disease resistance is poor, and disease-resistant variety output is lower.In the later stage nineties in last century, the soil of transgenic Bt cotton flower is in China large tracts of land commercialization plantation, though remarkable to target pest control actions such as cotton bollworms, transgenic cotton against pests in the market is resisting verticilliums not basically.Therefore set up important content and an urgent demand that the verticillium wilt resistance of cotton by same breeding technique becomes the cotton breeding technological innovation.
Summary of the invention
The verticillium wilt resistance of cotton by same breeding method that the purpose of this invention is to provide a kind of precise and high efficiency, strong operability.
Verticillium wilt resistance of cotton by same breeding method provided by the invention, it comprises the steps:
1, set up the disease garden: at first set up the sick garden of conventional verticillium wilt, the annual short flannel soil residue powder that has virulent verticillium wilt pathogen that adds in the sick garden of conventional verticillium wilt is with the soil mixing;
2, seed soaking microbiological contamination, sowing: verticillium wilt is taken place by serious cotton diseased plant pulverize, be soaked in water, filter and obtain bacteria suspension, fully soak cotton seed with bacteria suspension again, dry in the air to the seed hair and sow to the sick garden of verticillium wilt that step 1 is set up when white;
3, seed selection cotton with greensickness-resistance: the susceptible contrast of a highly susceptible and good stability is set, standard of perfection in conjunction with the greensickness-resistance evaluation, filter out disease-resistant plant, the verticillium wilt pathogen height is selected pressure effect parent and offspring thereof in sick garden again, identifies and filter out the cotton variety of colory resisting verticillium.
Wherein, in the step 1, the sick garden of conventional verticillium wilt be based upon strong pathogenicity fungus strain widely distributed, be subjected to that weather influence is little, summer 7, August mean temperature of air surpass time of 28 ℃ and be less than 20 days, help that verticillium wilt takes place and morbidity area uniformly; In the normal time, the susceptible check variety diseased plant rate of this area is more than 80%, and disease index is between 40.0-60.0.
Add the short flannel soil residue that has virulent verticillium wilt pathogen every year in the sick garden of conventional verticillium wilt and pulverize mixing as the bacterial classification that verticillium wilt keeps and upgrades, the short flannel soil residue that is used for the disease garden is no less than 1200g/m 2Soil, the residue that carries disease germs applies the field equably, turns over 2-3 time again, and it is even that germ and soil are mixed, and guarantees that disease garden soil germ quantity more than 200/g soil, slows down the decline in disease garden.
Short flannel soil residue powder is that the gross cottonseed that picks up from the serious different cottons producing region of verticillium wilt generation produces after the cotton gin lint, its by indoor germ cultivate, molecular probe detects, connect bacterium disease resistance standard in conjunction with cotton plant in the greenhouse, identify and confirm that it is rich in highly pathogenic germ.
In the step 2, the process of seed soaking microbiological contamination is pulverized for verticillium wilt is taken place serious cotton diseased plant, is soaked in water 48 hours, filters and obtains bacteria suspension, and again according to cotton seed: the w/v of bacteria suspension=1: 5-10 soaked cotton seed 24 hours.
In the step 3, susceptiblely be chosen in normal time disease index at the cotton variety more than 50.0 to correlating, as, can select " cotton No. 11 of Ji " or " Hubei Province chaste tree No. 1 " for use, or other height susceptible variety.The standard of perfection of greensickness-resistance evaluation is the regulation according to " cotton disease resistance worm property assessment technique standard-Di 5 parts: verticillium wilt ", with the cotton material of relative disease index<20 as anti-(anti-) verticillium wilt breeding material.High pressure select to be meant the disease-resistant cotton plants that in sick garden, will filter out again through interspecific cross, backcross or add hybridization again in sick garden, until obtaining the stable improved seeds of resisting verticillium for the seed selection offspring through Hainan.
Verticillium wilt resistance of cotton by same breeding method provided by the present invention, it also comprises in the cotton variety of the resisting verticillium that step 3 obtains, and imports the Bt gene, and then identifies and filter out the hold concurrently improved seeds of bollworm resisting of resisting verticillium.
Verticillium wilt resistance of cotton by same breeding method provided by the present invention has following advantage:
1, the bacterial classification that keeps and upgrade as verticillium wilt with the virulent short flannel soil residue powder that adds every year, after used year after year in the sick garden of soil disease, the soil content of molds increased, and distributed more to become evenly, and qualification result is also more reliable.
2, plant the new cotton variety that anti-(anti-) verticillium wilt such as cotton No. 2, No. 6, No. 8 and No. 1, planting, No. 5, planting, assorted No. 2 of planting are held concurrently bollworm resisting and high yield, high-quality in successively having bred by the inventive method, its seed selection process is accurate, anti-(anti-) verticillium wilt of the kind of breeding is stablized by force, the output height, wide adaptability, remarkable in economical benefits.
Embodiment
Following examples further specify content of the present invention, but should not be construed as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, modification or replacement to the inventive method, step or condition are done all belong to scope of the present invention.
Plant 372 seed selection in the embodiment 1 verticillium wilt-resistant cotton kind
The sick garden of conventional cotton verticillium wilt is set up in scientific research pilot scale base, Xinxiang, the Chinese Academy of Agricultural Sciences Institute of Plant Protection in 1993, mean temperature of air surpassed time of 28 ℃ and was less than 20 days this base summer 7, August, build the garden and add in the sick garden of conventional resisting verticillium then and pick up from serious different cottons producing region takes place verticillium wilt the virulent short flannel soil residue of gross cottonseed after processing and pulverize to mix as verticillium wilt and keep and novel bacterial more, the short flannel soil residue powder that is used for the disease garden is no less than 1200g/m 2Soil, the residue that carries disease germs applies the field equably, guarantees that disease garden soil germ quantity more than 200/gram soil, carries out above-mentioned bacterial classification every year and upgrade.
Low temperature drying is deposited after selecting the serious diseased plant of local verticillium wilt generation last year to pulverize mixing simultaneously, before sowing then, soaked 48 hours with well water, filter the back and obtain bacteria suspension, according to cotton seed: (the W: V of bacteria suspension=1: 5, w/v) ratio is soaked anti-(anti-) verticillium wilt and yielding ability and was showed preferably cotton seed 24 hours, dries in the air to cause the cottonseed hair and sow to the sick garden of verticillium wilt when white.
According to qualification result in the past, select anti-(anti-) property of more than 300 verticillium wilt preferably cotton material in sick garden, carry out the resisting verticillium seed selection, each cotton material repeats for 3 times, each 2 row that repeat, the long 6-10 rice of row, sub-district strain number is no less than 50 strains, plants by normal sowing time of cotton and field management mode, keep the suitable humidity in field, be beneficial to the generation of verticillium wilt.With the disease-resistant cotton plants that filters out in the sick garden through interspecific cross, backcross, identified in continuous 2 years to filter out the alternative materials that 12 disease resistances reach anti-(resist) grade, wherein best to plant 372 disease resistances in the plant.
The seed selection that embodiment 2 resisting verticilliums are held concurrently and planted cotton No. 2 in the bollworm resisting cotton kind
Nineteen ninety-five is with Mr. Guo Sandui of biotechnology institute of Chinese Academy of Agricultural Sciences clone and transformed Bt gene, by planting 372 among the pollen tube passage method importing embodiment 1, add numerous through Hainan, select 65 of individual plants that contain the bollworm resisting gene, the sick garden plantation in scientific research pilot scale base, the Chinese Academy of Agricultural Sciences Xinxiang in 1997, carrying out individual plant separates and the resisting verticillium seed selection, select 106 of pest-resistant resisting verticillium individual plants, miscegenation in 1998, select 100 individual plants, planted in sick garden in 1999, select and show preferably 12 of individual plants, advanced the trial test of people base in 2000, select 02,04,08 3 plants, calendar year 2001 is participated in the base varietal yield test, and it is stable to select bollworm resisting, resisting verticillium, high resistance to wilt good, yielding ability is good, colory 04 be name in plant cotton No. 2 (GK44), continue to participate in the base varietal yield test.Carried out transgenosis safe release and production application safety evaluation in 2002-2004.Participated in national Huanghe valley spring cotton variety regional test and production test in 2004-2005.Identify through Cotton Inst., Chinese Agricultural Academy, plant cotton No. 2 withered indexes 4.0 in 2004, the Huang index 11.1 that withers belongs to high anti-withered resisting verticillium kind, and withered index 3.8 in 2005, Huang index 14.8 that withers belongs to high anti-withered resisting verticillium type.Mill trial results performance volume increase in 2005 is extremely remarkable.2006 is careful trans Bt gene high resistance helicoverpa armigera of domestic first state and high resistance to wilt good, resisting verticillium kind by state careful (careful number of state: state examines cotton 2006001), has filled up the not blank of resisting verticillium of China's transgenic cotton against pests.This kind amount of increase in production is big, and economic results in society are remarkable.
The seed selection of the embodiment 3 high new cotton varieties of anti-verticillium wilt the " assorted No. 2 of planting "
Do and maternally carry out the quadrature reciprocal cross with numerous product based materials and match combination in a large number to plant cotton No. 2 (GK44) choosing system among the embodiment 2, simultaneously plant Hybrid and carry out variety comparative test with current master, 2003, from 50 combinations, determine 10 combinations preferably, 2004 on the basis of multiple spot variety comparative test, determine at last with high yield, disease-resistant, it is maternal planting cotton No. 2 (GK44) choosing in the pest-resistant strain, stability adaptability is strong, economical character is good, the good new 59-4 of new lines of fiber quality is that male parent is hybridized the anti-blight that the assembly seed selection becomes, anti-verticillium wilt, cotton newly makes up high-yield hybrid, name and be planting assorted No. 2, authorized by Henan Province in 2007.
Participated in national Huanghe valley regional test and production test in 2006-2007 continuously.Planting in 2006 No. 2 withered indexes 4.0 of mixing, Huang index 23.2 that withers belongs to high anti-withered anti-verticillium wilt kind, and withered index 5.5 in 2005, Huang index 25.2 that withers belongs to high anti-withered anti-verticillium wilt type.In May, 2008 is by the authorization of national crop varieties validation board.Planting is domestic first high resistance to wilt good, anti-verticillium wilt, bollworm resisting hybrid new breed by authorization assorted No. 2, has filled up the blank of high anti-withered, anti-(anti-) verticillium wilt new cotton variety of China's transgenosis hybridization Insect Resistant Cotton.

Claims (7)

1, a kind of method of cultivating verticillium wilt-resistant cotton, it comprises the steps:
(1) set up disease garden and every year novel bacterial more: at first set up the sick garden of conventional verticillium wilt, the annual short flannel soil residue powder that has virulent verticillium wilt pathogen that adds in the sick garden of conventional verticillium wilt is with the soil mixing;
(2) seed soaking microbiological contamination, sowing: verticillium wilt is taken place by serious cotton diseased plant pulverize, be soaked in water, filter and obtain bacteria suspension, fully soak cotton seed with bacteria suspension again, dry in the air to the seed hair and sow to the sick garden of verticillium wilt that step (1) is set up when white;
(3) seed selection cotton with greensickness-resistance: the susceptible contrast of a highly susceptible and good stability is set, standard of perfection in conjunction with the greensickness-resistance evaluation, filter out disease-resistant plant, the verticillium wilt pathogen height is selected pressure effect parent and offspring thereof in sick garden again, identifies and filter out the cotton variety of colory resisting verticillium.
2, the method for claim 1, it is characterized in that, the sick garden of conventional verticillium wilt described in the step (1) be based upon strong pathogenicity fungus strain widely distributed, be subjected to that weather influence is little, summer 7, August mean temperature of air surpass time of 28 ℃ and be less than 20 days, help that verticillium wilt takes place and morbidity area uniformly; In the normal time, the susceptible check variety diseased plant rate of this area is more than 80%, and disease index is between 40.0-60.0.
3, the method for claim 1, it is characterized in that, the soil of short flannel described in the step (1) residue powder is that the gross cottonseed that picks up from the serious different cottons producing region of verticillium wilt generation produces after the cotton gin lint, its by indoor germ cultivate, molecular probe detects, connect bacterium disease resistance standard in conjunction with cotton plant in the greenhouse, identify and confirm that it is rich in highly pathogenic germ.
As the arbitrary described method of claim 1-3, it is characterized in that 4, sick garden soil verticillium wilt pathogen viable count is more than 200/g soil.
5, the method for claim 1, it is characterized in that, seed soaking microbiological contamination process described in the step (2) is pulverized for verticillium wilt is taken place serious cotton diseased plant, be soaked in water 48 hours, filtration obtains bacteria suspension, and again according to cotton seed: the w/v of bacteria suspension=1: 5-10 soaked cotton seed 24 hours.
6, the method for claim 1, it is characterized in that, high pressure described in the step (3) select to be meant the disease-resistant cotton plants that in sick garden, will filter out again through interspecific cross, backcross or add hybridization again in sick garden, until obtaining the stable improved seeds of resisting verticillium for the seed selection offspring through Hainan.
7, the method for claim 1 is characterized in that, it also comprises in the cotton variety of the resisting verticillium that step (3) obtains, and imports the Bt gene, and then identifies and filter out the hold concurrently improved seeds of bollworm resisting of resisting verticillium.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103602743A (en) * 2013-11-20 2014-02-26 中国农业科学院农产品加工研究所 Method for detecting resistance of cotton plants to greensickness by molecular marker-assisted selection
CN104480189A (en) * 2014-11-21 2015-04-01 商丘师范学院 In vitro identification method for tomato resistance to verticillium wilt
CN108207613A (en) * 2017-12-29 2018-06-29 河南科技学院 A kind of Cotton Breeding Methods of resisting verticillium
CN109169127A (en) * 2018-09-27 2019-01-11 湖北省农业科学院经济作物研究所 A kind of Cotton Breeding Methods of resisting verticillium
CN110089296A (en) * 2019-04-12 2019-08-06 湖北省烟草科学研究院 The method for establishing tobacco black shank resistance identification sick nursery
CN110089379A (en) * 2019-04-12 2019-08-06 湖北省烟草科学研究院 For identifying the method for building up of burley tobaccos kind root black rot resistance sick nursery
CN110089295A (en) * 2019-04-12 2019-08-06 湖北省烟草科学研究院 The method for building up of tobacco bacterial wilt Resistance Identification sick nursery
CN112400646A (en) * 2020-11-23 2021-02-26 湖北省农业科学院经济作物研究所 Anti-verticillium wilt cotton breeding method

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103602743A (en) * 2013-11-20 2014-02-26 中国农业科学院农产品加工研究所 Method for detecting resistance of cotton plants to greensickness by molecular marker-assisted selection
CN103602743B (en) * 2013-11-20 2014-10-15 中国农业科学院农产品加工研究所 Method for detecting resistance of cotton plants to greensickness by molecular marker-assisted selection
CN104480189A (en) * 2014-11-21 2015-04-01 商丘师范学院 In vitro identification method for tomato resistance to verticillium wilt
CN108207613A (en) * 2017-12-29 2018-06-29 河南科技学院 A kind of Cotton Breeding Methods of resisting verticillium
CN109169127A (en) * 2018-09-27 2019-01-11 湖北省农业科学院经济作物研究所 A kind of Cotton Breeding Methods of resisting verticillium
CN110089296A (en) * 2019-04-12 2019-08-06 湖北省烟草科学研究院 The method for establishing tobacco black shank resistance identification sick nursery
CN110089379A (en) * 2019-04-12 2019-08-06 湖北省烟草科学研究院 For identifying the method for building up of burley tobaccos kind root black rot resistance sick nursery
CN110089295A (en) * 2019-04-12 2019-08-06 湖北省烟草科学研究院 The method for building up of tobacco bacterial wilt Resistance Identification sick nursery
CN112400646A (en) * 2020-11-23 2021-02-26 湖北省农业科学院经济作物研究所 Anti-verticillium wilt cotton breeding method

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