CN102165914A - Method for cultivating cucumber seedlings for biological tests of blight - Google Patents
Method for cultivating cucumber seedlings for biological tests of blight Download PDFInfo
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- CN102165914A CN102165914A CN2011100077546A CN201110007754A CN102165914A CN 102165914 A CN102165914 A CN 102165914A CN 2011100077546 A CN2011100077546 A CN 2011100077546A CN 201110007754 A CN201110007754 A CN 201110007754A CN 102165914 A CN102165914 A CN 102165914A
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- fusarium wilt
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- 238000012360 testing method Methods 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title abstract description 16
- 244000299906 Cucumis sativus var. sativus Species 0.000 title 1
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Classifications
-
- Y02P60/216—
Landscapes
- Cultivation Of Plants (AREA)
Abstract
The invention provides a method for cultivating cucumber seedlings for biological tests of blight, which comprises the following steps of: immersing prepared cucumber seeds into a container in which sterile water is filled, putting the container on a constant-temperature shaker, and performing shake culture at the temperature of 25 DEG C and at the rotate speed of 90 revolutions/minute; stopping shaking until the cucumber seeds in the container germinate and sprouts grow to 0.5 centimeter; and sowing the germinated cucumber seeds in a seedling raising disc with a disinfected cultivation substrate, and putting the seedling raising disc into a 25 DEG C artificial climate chamber for culturing, wherein the photoperiod in the culturing process comprises 12 hours of illumination and 12 hours of darkness, the cucumber seeds are poured by nutrient solution, and the seedlings can be used for the biological tests of the blight after about 20 days. The method has the advantages that the seedling raising time can be shortened, the rate of germination is high, and due to the adoption of substrate cultivation, the cucumber seedlings are not influenced by factors such as soil microorganisms, soil structures and the like, so that the seedlings are uniform in growth.
Description
[technical field]
The present invention relates to a kind of biological breeding method, relate in particular to a kind of breeding method that is used for the cucumber seedling of fusarium wilt biologicall test.
[background technology]
The fusarium wilt of crops is a kind of destructive fungi soil-borne disease, and Fusarium oxysporum (Fusarium oxysporum Sch1) is its main pathogen.This evil of being critically ill is in extensive range, comprising: (1) economic crops: cotton, tung oil tree, castor-oil plant, flax etc.; (2) solanaceous crops: tomato, pepper, eggplant, tobacco, potato etc.; (3) melon crop: cucumber, joint melon, wax gourd, watermelon, pumpkin, custard squash, muskmelon, sponge gourd, white melon, snake melon, balsam pear, bottle gourd, Hami melon etc.; (4) legume: soybean, kidney bean, cowpea, crotalaria mucronata, pea etc.; (5) flower plant: carnation, tulip, zinnia, flameray gerbera, iris, narcissus, China aster, silk tree, lily, Indian fig, Herba Andrographitis, poinsettia, turfgrass etc.; (6) woods fruit plant: pine tree, acacia rachii, citrus, sea-buckthorn, metasequoia, Clematis plant, pear tree etc.; (7) other plant: lettuce, wheat, garlic, water chestnut, asparagus, ginger, banana etc.
In China south, take place the most serious with cucurbits fusarium wilt in the crop fusarium wilt; In northern China, take place the most serious with cotton wilt.Fusarium wilt all can take place in the time of infertility of plant, in case morbidity just is difficult to eradicate, along with the increase in continuous cropping time, the incidence of disease is serious all the more.Since the nineties in 20th century, China's facilities vegetable melon crop area constantly increases, because the special microclimate that forms in the facility of relative closure had both helped vegetable growth, also become the hotbed that various damage by disease and insect grow, continuous cropping all the year round in addition, make that the quantity of pathogen constantly increases in the soil, the fusarium wilt of facility cultivation vegetables takes place more and more heavier.Relevant result of study shows: the crop fusarium wilt generally causes underproduction 20-30%, and serious field piece can reach 50-80%, even total crop failure, has become limiting China's crop production development key factor.
Selecting disease-resistant variety for use is one of the most frequently used method of control crop fusarium wilt, and disease-resistant evaluation work is the important step of carrying out the anti-blight breeding.In the pathogenic mensuration of wilt, cucumber seedling is one of a kind of the most frequently used material, and not only aequum is big, and very strict to the requirement of seedling.Adopted field soil to grow seedlings mostly in the past, cultivation, the seedling that adopts this method to bring out, the susceptible situation of inoculation back plant is difficult to accurately judge or determine, because field soil composition complexity, soil structure and microbial population easily change, be difficult to guarantee the uniformity of each test, and the microorganism in the soil also may participate in the pathogenic effects of pathogenic bacteria, cause biologicall test the failure of an experiment, influence the process of breeding for disease resistance work, in addition, required time of traditional seedling-cultivating method is longer, generally be no less than 30 days, therefore, seek a kind of aseptic relatively and fast the means of growing seedlings be essential.
[summary of the invention]
Technical problem to be solved by this invention is to provide a kind of breeding method that is used for the cucumber seedling of fusarium wilt biologicall test, not only can shorten seedling raise period, emergence rate height, and adopt matrix to cultivate the influence that can not be subjected to factors such as edaphon and soil structure, make the growth of seedling uniformity.
The present invention solves the problems of the technologies described above by the following technical programs: a kind of breeding method that is used for the cucumber seedling of fusarium wilt biologicall test, it comprises the steps: earlier the cucumber seeds of getting ready to be dipped in the container that fills sterile water, and this container placed carry out shaken cultivation on the constant temperature shaking table, temperature is set at 25 ℃, rotating speed 90r/min, treat that cucumber seeds in the container germinates and bud grows to the failure of oscillations behind the 0.4-0.6cm, then the cucumber seeds that germinates is broadcast in the seedling-cultivating tray that fills sterile cultivation matrix, place 25 ℃ climatic cabinate to cultivate this seedling-cultivating tray then, photoperiod in the incubation is 12h illumination, the 12h dark, with nutrient solution pouring, cultivate after 20-22 days and seedling can be used for the fusarium wilt biologicall test.
Further, described cultivation matrix is made up of by 1: 3 volume ratio perlite and peat.
Further, described nutrient solution is the nutrient solution of commercially available common flowers, its component is: superphosphate 800mg/L, potassium nitrate 700mg/L, nitrate of lime 700mg/L, magnesium sulfate 280mg/L, iron sulfate 120mg/L, boric acid 0.6mg/L, manganese sulphate 0.6mg/L, zinc sulphate 0.6mg/L, copper sulphate 0.6mg/L, ammonium sulfate 0.6mg/L, prepare with water.
Beneficial effect of the present invention is: compare traditional field soil and grow seedlings, not only can shorten seedling raise period, emergence rate height, and adopt matrix to cultivate the influence that can not be subjected to factors such as edaphon and soil structure, make the growth of seedling uniformity, and then improve the reliability and the accuracy of follow-up fusarium wilt biologicall test.
[embodiment]
The breeding method that the present invention is used for the cucumber seedling of fusarium wilt biologicall test comprises the steps: earlier the cucumber seeds of getting ready to be dipped in the container that fills sterile water, and this container placed carry out shaken cultivation on the constant temperature shaking table, temperature is set at 25 ℃, rotating speed 90r/min, treat that cucumber seeds germination and bud in the container grow to 0.4-0.6cm (about shaken cultivation 24h) the back failure of oscillations, then the cucumber seeds that germinates is broadcast in the seedling-cultivating tray that fills sterile cultivation matrix, place 25 ℃ climatic cabinate to cultivate this seedling-cultivating tray then, photoperiod in the incubation is 12h illumination, the 12h dark, with nutrient solution pouring, cultivate after 20-22 days and seedling can be used for the fusarium wilt biologicall test.
Wherein, described cultivation matrix is made up of by 1: 3 volume ratio perlite and peat; Described nutrient solution is the nutrient solution of commercially available common flowers, its component is: superphosphate 800mg/L, potassium nitrate 700mg/L, nitrate of lime 700mg/L, magnesium sulfate 280mg/L, iron sulfate 120mg/L, boric acid 0.6mg/L, manganese sulphate 0.6mg/L, zinc sulphate 0.6mg/L, copper sulphate 0.6mg/L, ammonium sulfate 0.6mg/L, prepare with water.
Below be one embodiment of the present of invention and utilize this embodiment to cultivate the cucumber seedling that obtains and carry out the biologicall test of fusarium wilt.
1. be used for the cultivation of the cucumber seedling of fusarium wilt biotinylated biomolecule mensuration
(1) material: cucumber seeds, cultivate matrix, nutrient solution
(2) concrete steps of breeding method
Earlier the cucumber seeds of getting ready is dipped in the 150ml triangular flask that fills the 20ml sterile water, and triangular flask placed carry out shaken cultivation on the constant temperature shaking table, to promote seed sprouting and to improve seed germination rate, and temperature is set at 25 ℃, rotating speed 90r/min, treat that cucumber seeds germination and bud in the container grow to about 0.5cm (about shaken cultivation 24h) back failure of oscillations, then the cucumber seeds that germinates is broadcast in the seedling-cultivating tray that fills sterile cultivation matrix, place 25 ℃ climatic cabinate to cultivate this seedling-cultivating tray then, photoperiod in the incubation is 12h illumination, the 12h dark, and with the pouring of the nutrient solutions of commercially available common flowers, can cultivate the cucumber seedling that can be used for the fusarium wilt biologicall test after 22 days, compare traditional field soil seedling-cultivating method, the present invention has not only shortened about 1/3 on the cultivation time, and the cucumber seedling seedling shape unanimity that obtains is easy to select.
2. the biologicall test of fusarium wilt
(1) for the preparation that tries fusarium wilt pathogen spore suspension: earlier placing PS liquid nutrient medium (potato 200g for examination fusarium wilt pathogen (Fusarium oxysporum f.sp.cucumerinum), sucrose 20g) in, and this medium placed on the shaking table cultivated 7 days, cultivation temperature is set at 28 ℃, rotating speed 170r/min, then the bacterium liquid of cultivating the back gained is used double gauze elimination mycelia, at last filtrate is mixed with 1 * 10
7The spore suspension of CFU/ml is standby.
(2) following two kinds of inoculation method: a. pouring bacterination process is generally taked in the biologicall test of fusarium wilt: the cucumber seedling of selecting the growing way homogeneous, be transplanted into the polypots head after after extracting gently its main root and 2 coring roots being blocked, and every strain root pours into 10ml fusarium wilt pathogen spore suspension; B. soak the root bacterination process: select the cucumber seedling of growing way homogeneous, extract the back gently and clean root, then its main root and 2 coring roots are blocked in the back immersion spore suspension, be transplanted into behind the 2h in the polypots head that fills the aseptic substrate that waters clear water with sterile water.
Each inoculation method all is provided with control group and experimental group simultaneously, the cucumber seedling after gained survives in the experimental group employing 1, and the cucumber seedling that control group adopts traditional field seedling raising to obtain, and the seedling management measure of experimental group and control group is identical.Day by day observe the growth and the infection conditions of cucumber seedling in two groups, and the susceptible situation of cucumber seedling in each group after 30 days relatively.
The result shows, the inoculation pathogen is after 30 days, and the fusarium wilt incidence of disease of cucumber seedling is all below 60% in two control groups, and the morbidity performance is quite inconsistent, is difficult to provide unified the evaluation; And no matter experimental group is to adopt the pouring inoculation method or soak the root inoculation method, the fusarium wilt incidence of disease of its cucumber seedling all reaches 100%, not only highly susceptible, and susceptible performance is consistent, and then shows that the cucumber seedling that obtains according to the inventive method can satisfy the requirement of biologicall test and breeding for disease resistance fully.
Claims (3)
1. breeding method that is used for the cucumber seedling of fusarium wilt biologicall test, it is characterized in that: comprise the steps: earlier the cucumber seeds of getting ready to be dipped in the container that fills sterile water, and this container placed carry out shaken cultivation on the constant temperature shaking table, temperature is set at 25 ℃, rotating speed 90r/min, treat that cucumber seeds in the container germinates and bud grows to the failure of oscillations behind the 0.4-0.6cm, then the cucumber seeds that germinates is broadcast in the seedling-cultivating tray that fills sterile cultivation matrix, place 25 ℃ climatic cabinate to cultivate this seedling-cultivating tray then, photoperiod in the incubation is 12h illumination, the 12h dark, with nutrient solution pouring, cultivate after 20-22 days and seedling can be used for the fusarium wilt biologicall test.
2. the breeding method that is used for the cucumber seedling of fusarium wilt biologicall test as claimed in claim 1 is characterized in that: described cultivation matrix is made up of by 1: 3 volume ratio perlite and peat.
3. the breeding method that is used for the cucumber seedling of fusarium wilt biologicall test as claimed in claim 1, it is characterized in that: described nutrient solution is the nutrient solution of commercially available common flowers, its component is: superphosphate 800mg/L, potassium nitrate 700mg/L, nitrate of lime 700mg/L, magnesium sulfate 280mg/L, iron sulfate 120mg/L, boric acid 0.6mg/L, manganese sulphate 0.6mg/L, zinc sulphate 0.6mg/L, copper sulphate 0.6mg/L, ammonium sulfate 0.6mg/L, prepare with water.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100077546A CN102165914B (en) | 2011-01-14 | 2011-01-14 | Method for cultivating cucumber seedlings for biological tests of blight |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100077546A CN102165914B (en) | 2011-01-14 | 2011-01-14 | Method for cultivating cucumber seedlings for biological tests of blight |
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| Publication Number | Publication Date |
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| CN102165914A true CN102165914A (en) | 2011-08-31 |
| CN102165914B CN102165914B (en) | 2012-05-02 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107047009A (en) * | 2017-04-25 | 2017-08-18 | 张家口市农业科学院 | A kind of plantation cultural method of millet Albino Seedling |
| CN109006436A (en) * | 2018-08-31 | 2018-12-18 | 佛山科学技术学院 | A kind of breeding method of cucumber seedling |
| CN110547191A (en) * | 2019-08-28 | 2019-12-10 | 铜仁万山九丰现代农业科技有限公司 | Seed cultivation method for improving salt tolerance of common head cabbage |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107047009A (en) * | 2017-04-25 | 2017-08-18 | 张家口市农业科学院 | A kind of plantation cultural method of millet Albino Seedling |
| CN107047009B (en) * | 2017-04-25 | 2020-01-31 | 张家口市农业科学院 | Planting and culturing method for millet albino seedlings |
| CN109006436A (en) * | 2018-08-31 | 2018-12-18 | 佛山科学技术学院 | A kind of breeding method of cucumber seedling |
| CN110547191A (en) * | 2019-08-28 | 2019-12-10 | 铜仁万山九丰现代农业科技有限公司 | Seed cultivation method for improving salt tolerance of common head cabbage |
| CN110547191B (en) * | 2019-08-28 | 2021-03-09 | 铜仁万山九丰现代农业科技有限公司 | Seed cultivation method for improving salt tolerance of common head cabbage |
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| CN102165914B (en) | 2012-05-02 |
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Effective date of registration: 20231225 Address after: 350000 pudang village, Fuzhou suburb, Fujian Province Patentee after: Institute of Resources, Environment, and Soil Fertilizers, Fujian Academy of Agricultural Sciences Address before: 350000 No. 54, 247, Fuzhou, Fujian Patentee before: AGRICULTURAL BIORESOURCES INSTITUTE OF FUJIAN ACADEMY OF AGRICULTURAL SCIENCES |