CN105409557B - The method that capsicum systemic disease resistance is induced using phosphite as exciton - Google Patents
The method that capsicum systemic disease resistance is induced using phosphite as exciton Download PDFInfo
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- CN105409557B CN105409557B CN201510902052.2A CN201510902052A CN105409557B CN 105409557 B CN105409557 B CN 105409557B CN 201510902052 A CN201510902052 A CN 201510902052A CN 105409557 B CN105409557 B CN 105409557B
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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
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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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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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
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Abstract
The present invention relates to method and the application that capsicum systemic disease resistance is induced using phosphite as exciton, this method is the preparation for carrying out phosphite first；Secondly, Hot Pepper Seedling is produced；It is finally the detection of phosphorous acid exciton induction pepper disease resistance defense response efficiency, the phosphite prepared is handled into above-mentioned cultured Hot Pepper Seedling with spray-on process, after induction processing 48 hours, then with phytophthora blight of pepper zoospore stop liquid inoculation etiolated seedling, seedling incidence is observed after 35 days, to determine its effect to induction disease resistance of plant.The above method can be adapted to the phosphite of Field information with time saving and energy saving preparation, the detection of defense responses activation is carried out to it simultaneously, fill up in domestic and international pepper disease resistance Journal of Sex Research and done the understanding that enhances disease resistance of exciton activation defense response using phosphite, be significant.
Description
Technical field
The present invention relates to technical field of plant protection, discloses and induces capsicum system disease-resistant using phosphite as exciton
The method of property.
Background technology
Capsicum epidemic disease is the crushing fungal disease generally occurred in a kind of world wide, and it is first in 1918 in the U.S.
New Mexico is found, causes the phytophthora blight of pepper of capsicum epidemic diseasePhytophthora capsiciLeonian is Leon
Leonian names in nineteen twenty-two, and it belongs to pathogenic oomycetes.Phytophthora capsici is invalid in soil with egg spore or chlamydospore
Survived the winter in body, the several months can be survived even more for a long time, host range is wide, mainly infects capsicum, tomato, eggplant, cucumber, pumpkin
Etc. a variety of important vegetables.Capsicum epidemic disease open country and protecting field can occur, and the sick prevalence causes capsicum loss serious, and one
As diseased plant rate be 15-30%, up to more than 80% when serious.Since nineteen eighty, capsicum epidemic disease is sent out in China vegetable cultivation region
The course of disease is subsisted exacerbation, has had a strong impact on the economic benefit of vegetable cultivation industry, and the vegetable safety production to China causes huge
Economic loss.Therefore, height extensive concern of the phytophthora blight of pepper by various countries phytopathologist and breeding scholar.In recent years
It was found that after the chemical substance such as salicylic acid is absorbed by plant, the disease resistance of host plant can be strengthened, reduce the generation of disease.
Phosphite has been developed to agricultural chemicals or fertilizer is used for the field control of multiple diseases, but its real anti-interpretation of the cause, onset and process of an illness
System research is not understood also.By the research of the understanding to activating pepper disease resistance defense response using phosphite, can go deep into
Understand diseases prevention mechanism of the phosphite to phytophthora blight of pepper and searched out as target related to the diseases prevention mechanism in capsicum
Disease-resistant gene, the studies above is understands fully that mechanism of action of the phosphite in pepper disease resistance defense response provides basis.Pass through solution
The molecular mechanism of the pepper disease resistance defense response of phosphite activation is analysed, can be that breeding expert finds new resistance locus, most
The outburst for effectively control epidemic disease proposes new strategy eventually.But current research shows, phosphite is defended in pepper disease resistance
The missing of molecular mechanism parsing in reaction limits our further investigations to its mechanisms of control wheat scab.
The content of the invention
It is an object of the present invention to provide the method that capsicum systemic disease resistance is induced using phosphite as exciton, i.e., with capsicum
Seedling is experiment material, is used as by phosphite and excites subprocessing to be followed by kind of phytophthora blight of pepper to detect induction disease resisting effect.
The present invention's is as follows using the method and applying step of phosphite activation pepper disease resistance defense response：
(1) the preparation of phosphite is with 1 by the phosphorous acid of purity ＞ 95% and potassium hydroxide：1 grade uses again, first with water-soluble
The g of phosphorous acid 1 is solved, then dissolves the g of potassium hydroxide 1 on its basis, by above-mentioned solution constant volume to 1000 mL, deployed phosphorous
The pH value of hydrochlorate is 6.2 or so, and the Tween-20 that addition accounts for total volume fraction 2 ‰ can be applied directly；
(2) pepper seed is 30% sodium hypochlorite through volume fraction（Shanghai traditional Chinese medicines Cas#:7681-52-9）After surface sterilization,
Rinsed well again with sterile purified water, kind is scalded in 55 DEG C~58 DEG C water-baths after 20 minutes, seed is seeded in underlay two layers
In the enamel tray of gauze (cm of 30 * 50), then cultivated 5-7 days under the conditions of dark lucifuge in 28 DEG C of incubators, until
Elongated capsicum etiolated seedling is grown, removes illumination box culture 2-3 days；
Phosphite exciton induction pepper disease resistance defense response efficiency detection method be, first, by what is prepared
Phosphite handles above-mentioned cultured Hot Pepper Seedling with spray-on process, and after induction is handled 48 hours, then phytophthora blight of pepper moves about
Spore stops liquid spray inoculation, and incidence is observed after 3-5 days, to determine its to the effect that induces disease resistance of plant.
Phytophthora blight of pepper zoospore stop liquid preparation method to take 0.3cm × 0.3cm-1cm × 1cm phytophthora blight of pepper
4 pieces are inoculated on V8 flat boards, and 25 DEG C of growths produce a large amount of sporangiums in 10 days, and above-mentioned 4 DEG C of placement 15min of flat board are subsequently placed in
Phytophthora blight of pepper zoospore is obtained after 30min, vortex oscillation 30 seconds is the liquid that stops at 25 DEG C.
V8 flat boards are 10% V8 nutrient solutions：100mL V8 vegetable juice adds 0.2g CaCO3Afterwards, under 3000 revs/min from
The min of the heart 5, supernatant constant volume is taken to 1000ml, the agar powder of addition cumulative volume 1.5%.
The present invention solves the problems, such as to activate pepper disease resistance defense response using ite, is that examination material is examined with above-mentioned Hot Pepper Seedling
The effect of phosphorous acid induction defense responses is surveyed, with experimental period is short, is not influenceed by external environmental condition, experimental repeatability
Well, inoculation Symptoms are obvious, are easy to the advantages that classification and Germicidal efficacy.The pepper disease resistance activated by parsing phosphite is prevented
The molecular mechanism of reaction is defended, can be that breeding expert finds new resistance locus, the final outburst for effectively control epidemic disease proposes
New strategy.But current research shows, the missing of extension set mechanism parsing of the phosphite in pepper disease resistance defense response
Limit our further investigations to its mechanisms of control wheat scab.
Embodiment
According to the g of phosphorous acid 1：Potassium hydroxide 1g weight ratio prepares the mL of phosphite solution 1000, pepper seed warp
Induction produces Hot Pepper Seedling after sterilization, finally detects phosphite exciton induction pepper disease resistance defense response efficiency.
Embodiment 1:
According to the g of phosphorous acid 1：Potassium hydroxide 1g weight first dissolves the g of phosphorous acid 1 than preparing phosphite solution with water,
Dissolve the g of potassium hydroxide 1 on its basis again, by above-mentioned solution constant volume to 1000 mL, the pH value of deployed phosphite is
6.2 or so, addition accounts for the Tween-20 that total volume fraction is 2 ‰ and can directly applied；Platform green pepper 106 and U.S.'s Christian era green pepper seed warp
The sodium hypochlorite of 30% volume（Shanghai traditional Chinese medicines Cas#:7681-52-9）After surface sterilization, then rinsed well with sterile purified water,
Kind is scalded in 55 DEG C of water-baths after 20 minutes, seed is seeded in the enamel tray of two layers of gauze of underlay (cm of 30 * 50), so
Cultivated 5 days under the conditions of dark lucifuge in 28 DEG C of incubators afterwards, until growing elongated capsicum etiolated seedling, remove illumination cultivation
Case culture 2 days；Phosphorous acid is that first, will prepare as the detection method of exciton induction pepper disease resistance defense response efficiency
Phosphite above-mentioned cultured Hot Pepper Seedling is handled with spray-on process, after induction processing 48 hours, then swum with phytophthora blight of pepper
Zoospore stops liquid（107Individual/mL）Spray inoculation Hot Pepper Seedling, seedling incidence is observed after 3 days, to determine it to inducing Genes For Plant Tolerance
The effect of characteristic of disease, while control is used as using clear water of spraying.
The stop preparation method of liquid of phytophthora blight of pepper zoospore is put down to take 4 pieces of 1cm × 1cm phytophthora blight of pepper to be inoculated in V8
On plate, 25 DEG C of growths produce a large amount of sporangiums in 10 days, and above-mentioned 4 DEG C of placement 15min of flat board are subsequently placed at 25 DEG C after 30min
Obtain phytophthora blight of pepper zoospore, vortex oscillation 30 seconds is the liquid that stops.
V8 flat boards are 10% V8 nutrient solutions：100mL V8 vegetable juice adds 0.2g CaCO3Afterwards, under 3000 revs/min from
The min of the heart 5, supernatant constant volume is taken to 1000ml, add 1.5wt.% agar powder.
It is 75.67% that experimental result statistical analysis, which finds that phosphite induces disease-resistant effect,.
Embodiment 2
According to the g of phosphorous acid 1：Potassium hydroxide 1g weight first dissolves the g of phosphorous acid 1 than preparing phosphorous acid solution with water, then
The g of potassium hydroxide 1 is dissolved on its basis, and by above-mentioned solution constant volume to 1000 mL, the pH value of deployed phosphite is
6.2, addition accounts for the Tween-20 that total volume fraction is 2 ‰ and can directly applied；Red sun and spring peppery No. 19 seeds are through 30% chlorine
After sour sodium surface sterilization, then rinsed well with sterile purified water, scalded in 58 DEG C of water-baths kind after 20 minutes, seed is seeded in
In the enamel tray of two layers of gauze of underlay (cm of 30 * 50), then 7 are cultivated under the conditions of dark lucifuge in 28 DEG C of incubators
My god, until growing elongated capsicum etiolated seedling, remove illumination box culture 3 days；Phosphorous acid exciton induction pepper disease resistance is prevented
Defending the detection method of reaction efficiency is, first, the phosphite prepared is handled into above-mentioned cultured Hot Pepper Seedling with spray-on process,
After induction processing 48 hours, then stopped liquid with phytophthora blight of pepper zoospore（107Individual/mL）Etiolated seedling is inoculated with, is observed after 4 days
Seedling incidence, to determine its effect to induction disease resistance of plant, while control is used as using clear water of spraying.
Phytophthora blight of pepper zoospore stop liquid preparation method to take 4 pieces of 0.3cm × 0.3cm phytophthora blight of pepper to be inoculated in
On V8 flat boards, 25 DEG C of growths produce a large amount of sporangiums in 10 days, and above-mentioned 4 DEG C of placement 15min of flat board are subsequently placed at 25 DEG C
Phytophthora blight of pepper zoospore is obtained after 30min, and vortex oscillation 30 seconds is the liquid that stops.
V8 flat boards are 10% V8 nutrient solutions：100mL V8 vegetable juice adds 0.2g CaCO3Afterwards, under 3000 revs/min from
The min of the heart 5, supernatant constant volume is taken to 1000ml, addition cumulative volume 1.5wt.% agar powder.
It is 78.53% that experimental result statistical analysis, which finds that phosphite induces disease-resistant effect,.
The above results show, activate the defense responses of capsicum as exciton by using phosphite, can be significantly
The resistance against diseases of Hot Pepper Seedling is improved, this is significant to the anti-disease mechanism research and resistance breeding research for promoting capsicum.
The foregoing is only presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, it should all belong to the covering scope of the present invention.
Claims (1)
1. the method for capsicum systemic disease resistance is induced using phosphite as exciton, it is characterised in that:Following several respects:
(1) the preparation of ite is with 1 by the phosphorous acid of purity ＞ 95% and potassium hydroxide：1 grade uses again, first dissolves phosphorous acid with water
1 g, then the g of potassium hydroxide 1 is dissolved on its basis, by above-mentioned solution constant volume to 1000 mL, the pH of deployed phosphite
It is worth for 6.2, adds and account for the Tween-20 of cumulative volume 2 ‰ and mix, it is stand-by；
(2) pepper seed is rinsed well after volume fraction is 30% sodium hypochlorite surface sterilization, then with sterile purified water, at 55 DEG C
Kind is scalded in~58 DEG C of water-baths after 20 minutes, seed is seeded in the enamel pallet of two layers of gauze of soil underlay, Ran Hou
Cultivated 5-7 days under the conditions of dark lucifuge in 28 DEG C of incubators, until growing elongated capsicum etiolated seedling, remove illumination box
Culture 2-3 days；
(3) by step（1）The phosphite prepared handles above-mentioned cultured Hot Pepper Seedling, induction processing 48 hours with spray-on process
Afterwards, then stopped liquid spray inoculation with phytophthora blight of pepper zoospore, incidence is observed after 3-5 days, to determine it to induction
The effect of disease resistance of plant；
Phytophthora blight of pepper zoospore stop liquid preparation method to take 0.3cm × 0.3cm-1cm × 4 pieces of 1cm phytophthora blight of pepper
It is inoculated on V8 flat boards, 25 DEG C of growths produce a large amount of sporangiums in 10 days, by above-mentioned 4 DEG C of placement 15min of flat board, are subsequently placed in 25
Phytophthora blight of pepper zoospore is obtained after 30min, vortex oscillation 30 seconds is the liquid that stops at DEG C；
V8 flat boards are 10% V8 nutrient solutions：100mL V8 vegetable juice adds 0.2g CaCO3Afterwards, 5 are centrifuged under 3000 revs/min
Min, supernatant constant volume is taken to 1000ml, the agar powder of addition cumulative volume 1.5%.
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| CN106613555A (en) * | 2016-12-29 | 2017-05-10 | 福建省农业科学院植物保护研究所 | Method for inducing pepper against pestilence by using benzothiadiazole as exciton |
| CN110089326A (en) * | 2019-03-08 | 2019-08-06 | 贵州民族大学 | A kind of adjuvant and the method using its raising masson pine disease-resistant performance |
| CN110564667B (en) * | 2019-09-25 | 2023-02-28 | 云南省烟草农业科学研究院 | Method for efficiently suspending and concentrating zoospores of plant pathogenic oomycetes and application of zoospores |
| CN110896842B (en) * | 2019-12-27 | 2021-07-30 | 华中农业大学 | Efficient cultivation method of citrus virus-free etiolation stock and application thereof |
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