CN102986464A - Method for preventing and treating watermelon wilt and promoting growth of watermelon seedlings - Google Patents
Method for preventing and treating watermelon wilt and promoting growth of watermelon seedlings Download PDFInfo
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- CN102986464A CN102986464A CN2012105389411A CN201210538941A CN102986464A CN 102986464 A CN102986464 A CN 102986464A CN 2012105389411 A CN2012105389411 A CN 2012105389411A CN 201210538941 A CN201210538941 A CN 201210538941A CN 102986464 A CN102986464 A CN 102986464A
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Abstract
The invention discloses a method for preventing and treating the watermelon wilt and promoting the growth of watermelon seedlings, comprising the following step: irrigating a rhamnolipid solution to the periphery of the roots of the watermelon seedlings, wherein the concentration of the rhamnolipid in the rhamnolipid solution is 0.9-1.1mg/mL. The method can be used for preventing the watermelon wilt effectively and promoting the growth of the watermelon seedlings.
Description
Technical field
The invention belongs to watermelon blight disease prevention and control field, be specifically related to a kind of novel watermelon blight control preparation and using method thereof, thereby effectively prevent and treat the generation of watermelon blight.
Background technology
Watermelon (Citrullus lanatus) belongs to the annual dicotyledon of Curcurbitaceae, and fruit is false berry.Watermelon originates in African equator and Mediterranean Region, is important horticultural crop in the world.China is the maximum in the world watermelon place of production, according to the World Food Programme (FAO) statistics, and China growth of watermelon area 2,210,000 hm2 in 2009, gross yield 6 820 ten thousand t account for respectively 58% and 68% of the world.Therefore, China's watermelon production occupies critical role in the world, and economic benefit is outstanding, provides the important channel for peasant household increases income.
Watermelon blight is one of main soil-borne vascular bundle disease due to being infected by Fusarium oxysporum f. sp. niveum (Fusariun oxysporum f.sp.niveum), and generation is all arranged all over the world.Fusarium is not only large and variable, and of a great variety, and can extensively be present in soil and the animals and plants organism.This disease is a kind of destructive disease, the especially more serious even total crop failure of continuous cropping plot morbidity.Watermelon blight has become the limiting factor that the restriction watermelon is produced in certain areas, production causes serious economic loss to watermelon.Yet watermelon blight there is no at present specific drug and can effect a radical cure, but this is because chlamydospore Survival for 10 Years in soil of watermelon blight germ.Germ is generally invaded from root system, causes that the vascular bundle browning festers, and causes melon seedling withered.In recent years, market is to the demand increase year after year of watermelon, and the growth of watermelon area enlarges thereupon, and watermelon blight is on the rise under the condition that crop rotation can't be implemented in full.2009, according to suburb of Beijing investigation, general diseased plant rate was 10% ~ 20%, and severe one reaches 80% ~ 90%, second crop soil even cause total crop failure.In the last few years, facility cultivating watermelon area expanding day, and the interior environment of facility is very beneficial for the Growth and reproduction of watermelon blight germ, causes the fusarium wilt incidence of disease to raise year by year, and exploring effectively preventing watermelon blight method is major issue anxious to be resolved in the current production.The upper people of at present production often adopt soil disinfectant to prevent soil-borne disease and obtain certain effect, but drug variety is various on the market, and some also has toxicity, and human body and environment are all had certain impact.Rhamnolipid is the very important biosurfactant of a class that is produced by pseudomonas aeruginosa (Pseudomonas aeruginosa).This surfactant has can significantly reduce the surface tension at interface, improve hydrophobicity, and toxicity little, be easy to the special physiological functions such as biological degradation, be widely used in the industrial circle such as biological prosthetic of oil, medicine, food, cosmetics, industrial pollutants, be counted as natural environment-friendly products.
Summary of the invention
The technical problem to be solved in the present invention provides a kind ofly can prevent and treat the method that watermelon blight can promote again the watermelon plant strain growth.
In order to solve the problems of the technologies described above, the invention provides and a kind ofly can prevent and treat the method that watermelon blight can promote again the watermelon plant strain growth, rhamnolipid solution is watered around the watermelon root, the concentration of rhamnolipid is 0.9 ~ 1.1mg/mL in the rhamnolipid solution.
Can prevent and treat the improvement that watermelon blight can promote again the method for watermelon plant strain growth as of the present invention: the concentration of rhamnolipid is 1.0mg/mL in the rhamnolipid solution.
The inventor finds that in the invention process concentration that rhamnolipid is filled with root is most important to the control of watermelon blight.The inventor finds that by research the rhamnolipid solution of variable concentrations has larger difference to the inhibition of Sclerotium rolfsii.
Filling root of the present invention waters rhamnolipid solution around the watermelon root exactly.Watermelon seedlings is carried out root irrigation, and generally filling with the root amount is 200 ~ 300ml/ strain.
In the invention process, the inventor uses first the PDA medium culture watermelon blight germ that contains the variable concentrations rhamnolipid under the condition that exsomatizes, the rhamnolipid concentration that contains in the PDA medium of variable concentrations rhamnolipid is respectively 0.1 g/L, 0.5 g/L, 1.0 g/L, 5.0 g/L, with the PDA medium that does not add rhamnolipid in contrast, purpose is that the result is compared to determine whether rhamnolipid has inhibitory action to withered germ of water-melon.The result shows that rhamnolipid can be grown by the establishment Sclerotium rolfsii, and concentration is larger, and it is better to suppress the pathogen growth result.When concentration was 1.0 mg/mL, inhibition reached 63.53%, and when concentration was 5.0 mg/mL, inhibition reached 96.96%(Fig. 1).Optimum concentration in vitro is 0.5g/L, and this moment, rhamnolipid presented utmost point significant difference to inhibitory action and the contrast of wilt.
The inventor finds that also rhamnolipid can effectively promote the activity of the disease-resistant relevant enzyme chitin of watermelon and dextranase.The inventor adopts variable concentrations rhamnolipid (concentration is: 0.1 g/L, 0.5 g/L, 1.0 g/L, 5.0 g/L) to fill with the method proof rhamnolipid of root to promoting the effect of watermelon chitinase and dextranase by pot experiment, the result shows, rhamnolipid can effectively promote the activity of the disease-resistant relevant enzyme chitin of watermelon and dextranase, thereby improves the disease resistance (Fig. 2) of plant itself.
For above-mentioned experimental result, in the invention process, the inventor proves further that by the method that pot experiment adopts variable concentrations rhamnolipid (concentration is set the same) to fill with root and inoculation rhamnolipid is to the control efficiency of watermelon blight.At first will carry out to the consistent watermelon seedlings of growth potential the rhamnolipid root irrigation of variable concentrations, fill with root and inoculate after three days, hinder the root processing under standardized in matrix with pocket knife before inoculation, then inoculate the wilt spore suspension, concentration is 10
6Individual/ml, inoculum concentration is the 20ml/ strain.About 13 days left and right sides plant begin morbidity after the inoculation, when beginning to fall ill record morbidity strain number, recording occurring continuously 6 days, and observe plant growth condition.When the choice experiment material, should select the consistent watermelon seedlings of growth potential as far as possible, remove sick watermelon seedlings plant.The result shows that in the pot experiment, rhamnolipid can effectively be prevented and treated the generation of watermelon blight, and concentration is larger, and control effect is better.The Pathogen of Fusarium Wilt inoculation is after 18 days, and when concentration was 1.0 mg/mL, the fusarium wilt preventive effect reached 75%, and when concentration was 5.0 mg/mL, the fusarium wilt preventive effect reached 95%(table 1).
Simultaneously, the present invention has also studied the impact on watermelon growing of using of variable concentrations rhamnolipid.The inventor has studied the impact of rhamnolipid on watermelon growing by the method that pot experiment adopts the variable concentrations rhamnolipid to fill with root, the result shows that low concentration rhamnolipid (0.5g/L, 1.0 g/L) promotes the growth of watermelon, (2.0 g/L, the 5.0g/L) growth (Fig. 3) of inhibition watermelon of height concentration rhamnolipid.
During the actual use of the present invention, rhamnolipid is configured to required concentration, watermelon seedlings is carried out root irrigation, generally filling with the root amount is 200 ~ 300ml/ strain.
The present invention has following features:
1), rhamnolipid is to come by carbon source through fermentation, and is relatively stable in the environment of different temperature, acid-base value and salinity;
2), adopt rhamnolipid solution to carry out root irrigation, easy and simple to handle;
3), during rhamnolipid ex vivo treatment Sclerotium rolfsii, concentration is larger, it is better to suppress the pathogen growth result.In actual the use, optimum concentration is 1.0g/L, and can also promote the growth of watermelon this moment when effectively preventing and treating watermelon blight.That is, the obvious contrast of the growth tendency of watermelon seedlings is all good with other processing under this concentration, and preventive and therapeutic effect also presents significant difference compared with the control.This result causes the problem of the underproduction that preferably solution is provided to fusarium wilt in the actual production, has preferably directive significance.
Description of drawings
Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.
Fig. 1 is that the rhamnolipid solution of variable concentrations is to the inhibition comparison diagram of watermelon blight pathogen growth;
Fig. 2 is that rhamnolipid concentration is on the comparison diagram that affects of watermelon chitinase and dextranase activity;
Fig. 3 is that rhamnolipid is on the comparison diagram that affects of watermelon growing.
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 the replacement method of the present invention, step or condition made all belong to scope of the present invention.
Embodiment 1: rhamnolipid is to the inhibition of watermelon blight pathogen
Experimental technique
1 experiment material
This experimental selection watermelon specialized form Sclerotium rolfsii is experimental subjects.This watermelon specialized form Sclerotium rolfsii (Fusarium oxysporum f. sp. niveum) can be available from Xinjiang Agricultural Sciences institute.
2 experimental techniques
Carry out successively following steps:
1), actication of culture:
With constant temperature activation under 28 ℃ of conditions of watermelon specialized form Sclerotium rolfsii access PDA medium three days.
The PDA medium: peeled potatoes 200g, glucose 20g, agar 20g, distilled water is settled to 1000ml.
2), wilt switching
Be that the card punch of 0.7cm was transferred on the rhamnolipid PDA medium that contains variable concentrations under 28 ℃ of conditions continuous culture three days with the wilt diameter of activation, measure the size of bacterial plaque every day.The concentration of rhamnolipid PDA medium is 0.1g/L, 0.5g/L, 1.0g/L, 5.0g/L.
Concrete steps are: cultivated three days under 28 ℃ of conditions of the germ after will transferring, carry out difference analysis with the size of right-angled intersection method measurement bacterial plaque to the result every day.
3 interpretations of result
Can be drawn by Fig. 1, the in vitro rhamnolipid has preferably inhibitory action to the watermelon blight germ, and along with the increase inhibitory action of rhamnolipid concentration obviously strengthens.When rhamnolipid concentration is 0.1 g/L, inhibitory action and the control medium of watermelon blight germ presented notable difference, almost be the growth that has fully suppressed the watermelon blight germ when concentration reaches 5.0 g/L.When concentration was 0.1g/L and 0.5g/L, difference was not clearly when cultivating one day and two days, but all presented compared with the control significant difference; When the concentration of rhamnolipid is 1.0g/L, can obviously find out inhibition.When rhamnolipid concentration was 5.0g/L, the size of bacterial plaque was all unchanged after three days, and the bacterial plaque color becomes transparence, showed that germ is killed.
Embodiment 2: rhamnolipid is to the facilitation effect of watermelon chitinase and dextranase activity
Experimental technique
1 experiment material
The variety of watermelon that this experiment is selected is " 8424 " variety of watermelon, selects simultaneously the watermelon specialized form Sclerotium rolfsii among the embodiment 1.
2 experimental techniques
Carry out successively following steps:
1), adopts the 10-15min that hots water treatment of seeds, constant temperature seed soaking about three hours, 31 ℃ of constant temperature vernalization sowing in 2-3 days.Select the consistent watermelon seedlings of growing way when transplanting seedlings, slow seedling 4-7 days of the rear high temperature of transplanting seedlings.Foregoing is usual manner.
2), the watermelon seedlings behind the slow seedling in the step 1) is carried out root irrigation with the rhamnolipid solution of variable concentrations, the concentration of rhamnolipid is respectively 0.1g/L, 0.5g/L, 1.0 g/L, 5.0 g/L, and the clear water filling root of its moderate in contrast.Filling with the root amount is the 250ml/ strain, and each is processed three times and repeats, and each repeats 8 strain seedling.Measure the activity of its dextranase and chitinase after 6 hours
3), process after the watermelon plant get second leaf, take by weighing weight after, add the phosphate buffer (PBS) of 3ml 0.1M pH5.0, add again 2% PVP and grind, centrifugal 10 minutes of 10000g gets supernatant, can survey the activity of enzyme.Chitinase and dextranase activity adopt Shanghai to grind GS-E0098 and the GS-E0150 kit measurement of territory commerce and trade Co., Ltd.
3 interpretations of result
Can be drawn by Fig. 2, the rhamnolipid of variable concentrations has obvious facilitation effect to watermelon chitinase and dextranase.Chitinase and dextranase are the important enzymes of Plant defense responses in the plant, are the signs of plant defense, and rhamnolipid can promote the activity of chitinase and dextranase, and the disease resistance response that rhamnolipid can activated plant is described.
Embodiment 3: root is filled with to the control efficiency of watermelon blight in the rhamnolipid field
Experimental technique
1 experiment material
Watermelon material and Pathogen of Fusarium Wilt that this experiment is adopted are the same.
2 experimental techniques
Carry out successively following steps:
1), adopts the 10-15min that hots water treatment of seeds, constant temperature seed soaking about three hours, 31 ℃ of constant temperature vernalization sowing in 2-3 days.Select the consistent watermelon seedlings of growing way when transplanting seedlings, slow seedling 4-7 days of the rear high temperature of transplanting seedlings.
2), the watermelon seedlings behind the slow seedling in the step 1) is carried out root irrigation with the rhamnolipid solution of variable concentrations, the concentration of rhamnolipid is respectively 0.5g/L, 1.0 g/L, 2.0 g/L, 5.0 g/L, and the clear water filling root of its moderate in contrast.Filling with the root amount is the 250ml/ strain, and each is processed three times and repeats, and each repeats 8 strain seedling.
3), the wilt of activation is transferred in the liquid B ilay medium, 100r shakes three days (72 hours) under 28 ℃ of constant temperatures in shaking table, with the number of blood cell counting plate at microscopically metering spore, and concentration is transferred to 10
6Individual/ml is for subsequent use.Bilay culture medium prescription: 1gKH
2PO
4, 0.2g starch, 1gKNO
3, 0.5gMgSO
47H
2O, 0.2g glucose, 0.5gKCl, 0.2g sucrose, water are settled to 1000ml.
4), afterwards inoculation of root irrigation three days, before inoculation, hinder root with pocket knife.Inoculum concentration is the 20ml/ strain.
5), keep hot and humid onset condition (30 ℃ of temperature, humidity about 60%) after the inoculation, note preventing the field diseases such as aphid, watered a seedling fertilizer every 3-4 days and guarantee that seedling grows up healthy and sound.
6), began morbidity in 13-15 days after the inoculation, from morbidity beginning statistics morbidity in continuous 6 days strain number, carry out at last interpretation of result.
3 interpretations of result
Rhamnolipid is very good to the field control effect of watermelon blight as can be seen from Table 1, and in the pot experiment, rhamnolipid can effectively be prevented and treated the generation of watermelon blight, and concentration is larger, and control effect is better.The Pathogen of Fusarium Wilt inoculation is after 18 days, and when concentration was 1.0 mg/mL, the fusarium wilt preventive effect reached 75%, and when concentration was 5.0 mg/mL, the fusarium wilt preventive effect reached 95%.
Table 1 rhamnolipid is to the watermelon blight incidence of disease and preventive effect result
Annotate: in the upper table: a-e represents significance analysis difference, and numerical value mark a is maximum with mark e difference
Embodiment 4: rhamnolipid is filled with root to the impact of watermelon growing
With the plant that above-mentioned inoculation was still survived after 10 days, observe.
The result is as follows:
Fig. 3 shows, does not adopt the watermelon that rhamnolipid is processed and Pathogen of Fusarium Wilt is inoculated dead, and the rhamnolipid of variable concentrations is processed and all effectively prevented and treated the watermelon blight generation; Low concentration rhamnolipid (0.5g/L, 1.0 g/L) promotes the growth of watermelon plant, and high concentration rhamnolipid (2.0 mg/mL, 5.0 mg/mL) suppresses the growth of watermelon plant.1.0 the watermelon growing situation that g/L concentration rhamnolipid is processed is best.
Embodiment 5,
Certain growth of watermelon ground, sickle-like bacteria content is in the soil of this planting site after measured: contain 743 sickle-like bacteria (Fusarium oxysporum f. sp. niveum) in every gram dry ground.Transplanted watermelon seedlings in above-mentioned planting site March 16; Select the consistent watermelon seedlings of growing way when transplanting seedlings, slow seedling 4-7 days of the rear high temperature of transplanting seedlings.Then be that the rhamnolipid solution of 1 g/L carries out root irrigation with concentration, fill with the root amount and be about the 250ml/ strain, as the rhamnolipid processed group; To fill with the in contrast group of equivalent clear water.
The morbidity of rhamnolipid processed group is evening relatively, begins to occur onset state May 18, begins occurred frequently in the first planting melon maturing stage; 3 batches of melons of watermelon results are respectively May 20, June 21, July 17.
The incidence of disease and the output of rhamnolipid processed group and control group are as shown in table 2 below.
Table 2
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (2)
1. can prevent and treat watermelon blight and can promote again the method for watermelon plant strain growth, it is characterized in that: rhamnolipid solution is watered around the watermelon root, and the concentration of rhamnolipid is 0.9 ~ 1.1mg/mL in the rhamnolipid solution.
2. according to claim 1ly can prevent and treat the method that watermelon blight can promote again the watermelon plant strain growth, it is characterized in that: the concentration of rhamnolipid is 1.0mg/mL in the rhamnolipid solution.
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| CN104789519A (en) * | 2015-04-29 | 2015-07-22 | 浙江传化生物技术有限公司 | Culture medium for in vitro fast growth of dendrobium officinale |
| CN107787973A (en) * | 2017-10-25 | 2018-03-13 | 盐城师范学院 | Application of the rhamnolipid in terms of plant salt stress is alleviated |
| CN107996227A (en) * | 2017-12-21 | 2018-05-08 | 潘卫剑 | A kind of watermelon blight control method |
| CN111134125A (en) * | 2020-01-20 | 2020-05-12 | 浙江大学 | Biological pesticide and plant growth regulating complexing agent and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104789519A (en) * | 2015-04-29 | 2015-07-22 | 浙江传化生物技术有限公司 | Culture medium for in vitro fast growth of dendrobium officinale |
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| CN107787973A (en) * | 2017-10-25 | 2018-03-13 | 盐城师范学院 | Application of the rhamnolipid in terms of plant salt stress is alleviated |
| CN107787973B (en) * | 2017-10-25 | 2020-11-17 | 盐城师范学院 | Application of rhamnolipid in relieving plant salt stress |
| CN107996227A (en) * | 2017-12-21 | 2018-05-08 | 潘卫剑 | A kind of watermelon blight control method |
| CN111134125A (en) * | 2020-01-20 | 2020-05-12 | 浙江大学 | Biological pesticide and plant growth regulating complexing agent and preparation method thereof |
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Application publication date: 20130327 |