CN103238634B - Application of amaranthus in plant disease prevention and treatment - Google Patents
Application of amaranthus in plant disease prevention and treatment Download PDFInfo
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Abstract
The invention provides an application of amaranthus in plant disease prevention and treatment, wherein amaranthus leaves are taken as the raw material and extracted by an organic solvent to obtain an extract, acidovorax avenae subsp.citrulli, xanthomonas axonopodis pv. Citri and rhizoctonia solani are selected as sites of action, and research on indoor bacteriostasis effect and in-vitro and pot culture control effects is performed to find that the amaranthus leaf extract has an inhibition effect on acidovorax avenae subsp.citrulli, xanthomonas axonopodis pv. Citri and rhizoctonia solani; and the in-vitro and pot culture tests also indicate that the amaranthus leaf extract has certain prevention and treatment effects on acidovorax avenae subsp.citrulli, xanthomonas axonopodis pv. Citri and rhizoctonia solani; and all the facts show that the amaranthus has prevention and treatment effects on the plant diseases in addition to edible and medical values. The research discovers and expands the functions of the vegetable, and simultaneously provides a new train of thought and a new way for studying and developing biopesticide for preventing and treating the plant diseases.
Description
Technical field
The invention belongs to vegetables application, be specifically related to the preventive and therapeutic effect of a kind of three-coloured amaranth to plant disease, more particularly, relate to the preventive and therapeutic effect of a kind of three-coloured amaranth to bacterial fruit blotch of watermelon, citrus bacterial canker disease and rice sheath blight disease.
Background technology
Three-coloured amaranth is the herbaceous plant of Amaranthaceae Amaranthus, is mainly edible, but its anti-oxidant, anticancer, liver-protective medical function constantly excavates.In recent years, along with three-coloured amaranth research deepen continuously, find three-coloured amaranth extract to food saprophytic bacteria as: detection of Salmonella (Salmonella spp.), Escherichia coli (E.coli), staphylococcus aureus (S.aureus), Listeria (L.monocytogenes) etc. are inhibited.Seed extract is inhibited to fungies such as lichee sour germ (Geotrichum candidum), Fusarinm solani (Fusrium solani), penicillium chrysogenum (Penicillium chryso-genum), aspergillus candidus (Aspergillus candidus).But, have no research report about the inhibitory action of three-coloured amaranth to phytopathogen and to the preventive and therapeutic effect of plant disease.Utilize plant exploitation biopesticide, not only safety, efficient, low toxicity, pollution-free, and can solve the problems such as the chemical pesticide construction cycle is long, and expense is high, pesticide resistance, and be the focus of current novel pesticide initiative research, there is vast potential for future development.The various plants such as existing cloves, peppermint, ginkgo, fragrant camphor tree, garlic, onion, ginger, Aztec marigold, the root of herbaceous peony have the report of bacteriostatic activity to phytopathogen, but there are no the report of three-coloured amaranth.
Acidovorax Avenae Subsp (Acidovorax avenae subsp.citrulli Willems) mainly infects watermelon seedlings and fruit, and field relies on seed-borne fungi to propagate, by the watermelon of this infection process almost without any value.Therefore, do the prevention and control of bacterial fruit blotch of watermelon well, can ensure the safety in production of watermelon.On producing, can take to strengthen the measures such as quarantine, aseptic seed, Rational Irrigation, plantation disease-resistant variety, crop rotation, chemical control.The advantage such as save time because chemical control has, laborsaving, quick and easy, is still the Main Means of control Acidovorax Avenae Subsp.At present, mainly rely on copper agent or antibiotic, as the Kocide SD of: copper agent, goodly grant copper and tetracycline, protect streptomycin, chain tetracycline etc. more.But the easy contaminated environment of chronic administration copper agent, destroys the ecological balance, and excessive administration of antibiotics is also easily induced germ generation drug resistance or pesticide resistance, reduction control efficiency.
Citrus bacterial canker disease is the bacterial disease of a kind of serious harm oranges and tangerines branch, leaf and the fruit that are caused by carpetweed Xanthomonas campestris pv.citri (Xanthomonas axonopodis pv.citri).This disease all has generation in some oranges and tangerines producing regions of China, is especially injured seriously with provinces (district) such as Guangdong, Guangxi, Hunan, Fujian, causes fallen leaves, spike top, weakens tree vigo(u)r, and serious caused shedding, affects orange yield and fruit quality.Because this disease is quarantine disease, developed country adopts large area to destroy elimination method by fire more at present, but this kind of method cost is very high, implements more difficult in China's oranges and tangerines producing region, the control of citrus bacterial canker disease remains the major issue of Citrus Industry, can not get effective radical cure.China epidemic-stricken area is taking chemical control as main, is aided with the comprehensive preventive health measures of strengthening clear garden of rich water quality management, control shoot, winter etc., and however, canker still occurs every year.At present, prevent and treat the conventional dose of canker mainly taking copper agent as main, include bordeaux mixture, Kocide SD, copper oxychloride, burnt ammonia copper, can kill etc., but the long-term use of copper agent causes the accumulation of copper in soil and environment, causes the pollution to environment.Therefore, explore the new method of preventing and treating very necessary.
Rice sheath blight disease (Rhizoctonia solani) is one of key factor affecting increasing production of rice, and the loss being caused by it can reach 10-30%.In recent years, along with the popularization of high yield, of short stem, many tillers breeding and the directly improvement of the culture technique such as rice transplanting, cause the generation of banded sclerotial blight day by day serious.On producing, the main chemical pesticide control that adopts of past, but because easily causing the problems such as residue of pesticide, contaminated environment, the destruction ecological balance, mainly use now jinggangmeisu, but long-term, a large amount of, single administration jinggangmeisu, can cause sheath blight fungus to develop immunity to drugs, and European Union also classify jinggangmeisu as dangerous medicament.At present, utilize natural plant active substance to prevent and treat rice sheath blight disease and become the focus of research.
In the variety comparative test of vegetables land for growing field crops, the inventor observes, and disease in various degree all can occur tens kinds of vegetables in process of growth, has three-coloured amaranth only and is free from germs and infects.According to this phenomenon, inventor infers in three-coloured amaranth may contain the material that some suppress germ.Therefore, the present invention has carried out the preventive and therapeutic effect research of three-coloured amaranth to bacterial fruit blotch of watermelon, citrus bacterial canker disease, rice sheath blight disease, is intended to find the new function of three-coloured amaranth controlling plant diseases; And provide new way and new approaches for biological prevention and control plant disease.
Summary of the invention
Technical problem to be solved by this invention is: for the technical blank of three-coloured amaranth controlling plant diseases, provide the application in a kind of three-coloured amaranth control of plant disease, be specially the application in control bacterial fruit blotch of watermelon, citrus bacterial canker disease and rice sheath blight disease.
In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is: the application of a kind of three-coloured amaranth in controlling plant diseases.Particularly, this plant disease is bacterial fruit blotch of watermelon, citrus bacterial canker disease or rice sheath blight disease.
The present invention is taking three-coloured amaranth as raw material, three-coloured amaranth leaf extracted with organic solvent, obtains three-coloured amaranth leaf extract.This organic solvent can be ethyl acetate, absolute ethyl alcohol or acetone.Certainly, no matter adopt which kind of technology, as long as can obtain three-coloured amaranth leaf extract at once, in the present invention, just as an example of ethyl acetate example, the preparation of three-coloured amaranth leaf extract is described, detailed process is as follows:
Get three-coloured amaranth leaf clean after, dry to water content be 0.5-3.5%, be crushed to 100-120 order fine powder with the rotating speed of 8000-12000r/min; Adopt again analysis pure ethyl acetate to carry out lixiviate to three-coloured amaranth leaf powder, three-coloured amaranth leaf powder disposable adding, ethyl acetate divides 3 times and adds, after the ratio that at every turn adds 10-20mL ethyl acetate in 1g three-coloured amaranth leaf powder adds, on shaking table at 20-25 DEG C with 90-120r/min jolting 24-36h, filter, collect filtrate, merge 3 times filtrate; Gained filtrate is inserted to rotation concentration and evaporation instrument, is that 40-43 DEG C, condensation temperature are that 8-10 DEG C and rotating speed are under 80-120r/min at 0.09-0.1MPa pressure, bath temperature, is concentrated into paste.
Three-coloured amaranth leaf extract of the present invention is inhibited to Acidovorax Avenae Subsp, c itrus canker germ and Rhizoctonia solani Kuhn; And in vitro and pot experiment also shows that three-coloured amaranth leaf extract has certain control efficiency to bacterial fruit blotch of watermelon, citrus bacterial canker disease and rice sheath blight disease.Show that three-coloured amaranth has the function of controlling plant diseases; Realization of the present invention simultaneously, provides scientific basis for developing three-coloured amaranth leaf natural active matter and research and development neoformation agricultural chemicals.
Brief description of the drawings
Fig. 1 is the inhibition figure of three-coloured amaranth leaf extract to Acidovorax Avenae Subsp.
Wherein: 1 represents ethyl acetate extract; 2 represent ethyl acetate.
Fig. 2 is the inhibition figure of three-coloured amaranth leaf extract to c itrus canker germ.
Wherein: 1 represents ethyl acetate extract; 2 represent ethyl acetate.
Fig. 3 is the inhibition figure of three-coloured amaranth leaf extract to Rhizoctonia solani Kuhn.
Wherein: 1 represents ethyl acetate extract; 2 represent ethyl acetate.
Embodiment
Below in conjunction with concrete test and embodiment, technical scheme of the present invention is described further.
Watermelon Acidovorax avenae subsp (Pseudomonas pseudoalcaligenes subsp.citrulli), c itrus canker germ (Xanthomonas axonopodis pv.citri), the Rhizoctonia solani Kuhn (Rhizoctonia solani) in following examples, used are provided by Agricultural University Of Hunan's plant pathology laboratory.The activation of Acidovorax Avenae Subsp and c itrus canker germ bacterial classification is: adopt oese, scraping 3 is encircled bacterial classification access beef extract-peptone slant medium (NA), cultivates 24h at 28 DEG C; The activation of Rhizoctonia solani Kuhn is: bacterium cake is directly seeded to (PDA) on potato dextrose agar, cultivates 72h at 25-28 DEG C.
The preparation (describing as extracting solvent taking ethyl acetate) of embodiment 1 three-coloured amaranth leaf extract
Get after three-coloured amaranth leaf is cleaned, dry to water content be 3%, under normal temperature, be crushed to 100-120 order fine powder through high speed disintegrator with the rotating speed of 8000-12000r/min, obtain three-coloured amaranth leaf powder; Get 10g three-coloured amaranth leaf powder, add and analyze pure ethyl acetate 180mL, at 20-25 DEG C, with 90-120r/min jolting 30h on shaking table, filter, collect filtered fluid; Filter residue rejoins ethyl acetate 180mL, at 20-25 DEG C, with 90-120r/min jolting 30h on shaking table, filters, and collects filtered fluid; Filter residue so repeats 1 time, merges 3 filtered fluids and is extract; Gained filtrate is inserted to rotation concentration and evaporation instrument, is that 40-43 DEG C, condensation temperature are that 8-10 DEG C and rotating speed are under 80-120r/min at 0.09-0.1MPa pressure, bath temperature, is concentrated into paste, obtains extract.
The inhibitory action of embodiment 2 three-coloured amaranth leaf extracts to Acidovorax Avenae Subsp and c itrus canker germ
Get 1g three-coloured amaranth leaf extract and add analysis pure ethyl acetate 5-10mL dissolving, obtain active matter to be measured.Adopt plate punch method to detect bacteriostasis.Respectively by Acidovorax Avenae Subsp suspension and c itrus canker germ suspension (1 × 10
8cfu/mL) 60 μ L evenly coat NA flat board, then, at the central side perforating of flat board (φ=5mm), inject active matter to be measured, punch and inject ethyl acetate for contrast in the central opposite side symmetry of flat board, every hole 50 μ L, cultivate 24-48h, observe antibacterial situation for 28 DEG C.Each processing repeats for 3 times.Through observation shows that, three-coloured amaranth leaf extract is all inhibited to Acidovorax Avenae Subsp and c itrus canker germ, sees Fig. 1 and Fig. 2, and contrast ethyl acetate is to two kinds of equal unrestraint effects of germ.Illustrate in three-coloured amaranth leaf and have Substance, can be used to prevent and treat phytobacterial disease.
The inhibitory action of embodiment 3 three-coloured amaranth leaf extracts to Rhizoctonia solani Kuhn
Get 1g three-coloured amaranth leaf extract and add analysis pure ethyl acetate 5-10mL dissolving, obtain active matter to be measured.Adopt the inhibition activity of punch method Detection and Extraction thing to Rhizoctonia solani Kuhn.Rhizoctonia solani Kuhn is put by central authorities at PDA flat board, and in the punching of 1cm place, both sides, a side is injected active matter to be measured, and opposite side injects ethyl acetate and compares, and every processing repeats for 3 times, is placed in 25-28 DEG C of incubator, cultivates 48h, observes and has or not antibacterial phenomenon.Through observation shows that, three-coloured amaranth extract is inhibited to Rhizoctonia solani Kuhn, sees Fig. 3, and contrast ethyl acetate is to sheath blight fungus unrestraint effect.Illustrate in three-coloured amaranth leaf and have Substance, also can be used to prevent and treat fungal diseases of plants.
Antibacterial activity and Detection of Stability that embodiment 4 different solvents dissolve three-coloured amaranth leaf extract
Get the mixture 5-10mL that 1g three-coloured amaranth extract adds respectively absolute ethyl alcohol, sterile water, sterile water and absolute ethyl alcohol, obtain three-coloured amaranth leaf extract dissolved matter.Adopt plate punch method to detect bacteriostatic activity and the stability of three-coloured amaranth extract dissolved matter.Respectively by Acidovorax Avenae Subsp suspension and c itrus canker germ suspension (1 × 10
8cfu/mL) 60 μ L evenly coat NA flat board, then in dull and stereotyped central authorities' punchings (φ=5mm), in 50 μ L extract hand-hole to be measured, and respectively in another is dull and stereotyped taking the mixture of absolute ethyl alcohol, sterile water, sterile water and absolute ethyl alcohol as contrast, cultivate 24-72h for 28 DEG C, each processing repeats for 3 times, observes and measure 24h, 48h, 96h inhibition zone size, to detect bacteriostatic activity and the stability of three-coloured amaranth leaf extract dissolved matter.
Results and analysis: three-coloured amaranth leaf extract dissolved matter is all inhibited to Acidovorax Avenae Subsp and c itrus canker germ, and different solvents contrasts Acidovorax Avenae Subsp and c itrus canker germ unrestraint effect.Known in conjunction with following table 1 and table 2, different solvents dissolved matter is from big to small successively to the inhibition activity of Acidovorax Avenae Subsp and c itrus canker germ: the mixture > sterile water > absolute ethyl alcohol of sterile water and absolute ethyl alcohol.Along with the prolongation of time, different solvents dissolved matter weakens gradually to the inhibition of Acidovorax Avenae Subsp and c itrus canker germ, wherein, 24h fungistatic effect is best, antibacterial circle diameter maximum, the fungistatic effect of 48h takes second place, a little less than the fungistatic effect of 96h, but sustainable 1 month of fungistatic effect, illustrates that it has stability.
The inhibition of table 1 different solvents dissolved matter to watermelon Acidovorax avenae subsp
The inhibition of table 2 different solvents dissolved matter to c itrus canker germ
Example 5 different solvents dissolve the Antifungi Activity and stabill of three-coloured amaranth leaf extract
Get the mixture 5-10mL that 1g three-coloured amaranth leaf extract adds respectively absolute ethyl alcohol, sterile water, sterile water and absolute ethyl alcohol, obtain three-coloured amaranth leaf extract dissolved matter.Adopt plate punch method to detect bacteriostatic activity and the stability of three-coloured amaranth leaf extract dissolved matter.Respectively activated Rhizoctonia solani Kuhn bacterium cake is placed in to the dull and stereotyped central authorities of PDA, then in the flat board symmetrical punching of central bilateral symmetry (φ=5mm), wherein dissolved matter to be measured is injected in a hole, another hole is injected corresponding solvent and is compared, 50 μ L are injected in every hole, and each processing repeats for 3 times, cultivate 24-72h for 28 DEG C, observe and measure 24h, 48h, the antibacterial bandwidth size of 96h, to detect bacteriostatic activity and the stability of three-coloured amaranth leaf extract dissolved matter.
Results and analysis: three-coloured amaranth leaf extract dissolved matter is inhibited to Rhizoctonia solani Kuhn, and different solvents is to Rhizoctonia solani Kuhn unrestraint effect.Known in conjunction with following table 3, different solvents dissolved matter is from big to small successively to the inhibition activity of Rhizoctonia solani Kuhn: the mixture > sterile water > absolute ethyl alcohol of sterile water and absolute ethyl alcohol.Along with the prolongation of time, different solvents dissolved matter weakens gradually to the inhibition of sheath blight fungus, and wherein, 24h fungistatic effect is best, antibacterial circle diameter maximum, and the fungistatic effect of 48h takes second place, a little less than the fungistatic effect of 96h.
The inhibition of table 3 different solvents dissolved matter to Rhizoctonia solani Kuhn
The potted plant preventive effect of example 6 three-coloured amaranth leaf extracts to bacterial fruit blotch of watermelon
Pot experiment is chosen in Agricultural University Of Hunan's teaching and scientific research building illumination box carries out, first the watermelon seedlings cultivating is transplanted in plastic basin, after transplanting 10d, adopt stem's stab inoculation Acidovorax Avenae Subsp, keep the temperature inside the box 29-32 DEG C on daytime, evening 21-24 DEG C, humidity is 90%, so that the watermelon seedling in different basin is evenly fallen ill.Potted plant experiment is divided into 5 concentration: 5,10,20,40,80mg/mL three-coloured amaranth leaf extract, make medicament control treatment with 0.6mg/mL streptomycin sulphate, and taking clear water as blank.Establish altogether three processing: 1. inoculate after germ 1h, spray three-coloured amaranth leaf extract; 2. inoculate after germ 7d, spray three-coloured amaranth leaf extract; 3. inoculate after germ 14d, spray three-coloured amaranth leaf extract.Each test sprays with micro pressure formula watering can, taking squirt leave dual sides just not lower as spending.Every processing repeats 3 times.5 watermelon seedlings of every processing, test needs altogether 5 concentration × 3 to process × 3 processing of × 5 watermelon seedling+2 of × 3 repetitions contrast and repeats × 5 watermelon seedling=315 watermelon seedlings × 3 times.
After spraying three-coloured amaranth leaf extract, a situation arises for 10-15d investigation records bacterial fruit blotch of watermelon.Being calculated as follows of severity Scaling standard, disease index and preventive effect:
Bacterial fruit blotch of watermelon blade grade scale:
0 grade: do not fall ill;
1 grade: on sick leaf, have fragmentary scab, scab accounts for below 1% of leaf area;
2 utmost points: have a small amount of scab on sick leaf, scab accounts for the 2%-5% of leaf area;
3 grades: on sick leaf, Lesion number is medium, scab accounts for the 6%-20% of leaf area;
4 grades: on sick leaf, Lesion number is more, scab accounts for the 21%-40% of leaf area;
5 grades: on sick leaf, have a large amount of scabs, scab accounts for the more than 41% of leaf area.
Computational methods:
The pot experiment effect of three-coloured amaranth leaf extract to bacterial fruit blotch of watermelon:
From following table 4, the protection effect of using after three-coloured amaranth leaf extract is followed successively by: after inoculation germ 1h, spray after three-coloured amaranth leaf extract > inoculation germ 7d, spray after three-coloured amaranth leaf extract > inoculation germ 14d, spray three-coloured amaranth leaf extract, its protection effect is respectively: 63.43-82.06%, 65.81-81.03%, 65.66-80.31%.Along with the rising of three-coloured amaranth leaf extract concentration, protection effect strengthens gradually, and when three-coloured amaranth leaf extract concentration is during at 40-80mg/mL, protection effect preferably reaches 80.04-82.06%, a little more than the preventive effect 74.4-79.3% of contrast medicament streptomycin.When three-coloured amaranth leaf extract concentration is during at 5-20mg/mL, three-coloured amaranth leaf extract protection effect 63.43-79.38%.Significant difference between variance analysis shows to manage everywhere.Result of the test show three-coloured amaranth leaf extract to the protection effect of watermelon fruit blotch all more than 60%, in the time that three-coloured amaranth extract concentrations is 80mg/mL, protection effect reaches as high as 82.06%, illustrates that three-coloured amaranth leaf extract has the potentiality that are developed to biopesticide.Meanwhile, three-coloured amaranth leaf extract control bacterial fruit blotch of watermelon should be used at the initial stage of a disease, and more early effect is better.
The potted plant control effect of table 4 variable concentrations three-coloured amaranth leaf extract to watermelon fruit blotch
Note: the numerical value in table is mean value 3 times; Different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
The in vitro preventive effect of example 7 three-coloured amaranth leaf extracts to citrus bacterial canker disease
In incubator, carry out, adopt injection that the c itrus canker germ having activated is seeded on oranges and tangerines excised leaf, put into enamel basin moisturizing with cover.Enamel basin is put incubator, temperature 29-32 DEG C on daytime, evening 21-24 DEG C so that the morbidity of Citrus leaf in different basin.Experiment is divided into 5 concentration: 5,10,20,40,80mg/mL three-coloured amaranth leaf extract, make medicament control treatment with 0.6mg/mL streptomycin sulphate, and taking clear water as blank.Establish altogether three processing: 1. inoculate after germ 1h, spray three-coloured amaranth leaf extract; 2. inoculate after germ 7d, spray three-coloured amaranth leaf extract; 3. inoculate after germ 14d, spray three-coloured amaranth leaf extract.Each test sprays with micro pressure formula watering can, taking squirt leave dual sides just not lower as spending.Every processing repeats 3 times.5 Citrus leafs of every processing, test needs altogether 5 concentration × 3 to process × 3 processing of × 5 Citrus leaf+2 of × 3 repetitions contrast and repeats × 5 Citrus leaf=315 Citrus leafs × 3 times.
After spraying three-coloured amaranth leaf extract, a situation arises for 10-15d investigation records citrus bacterial canker disease.Being calculated as follows of severity Scaling standard, disease index and preventive effect:
Citrus bacterial canker disease grade scale:
0 grade: anosis;
1 grade: every leaf (really) has 1-5 of scabs;
3 grades: every leaf (really) has 6-10 of scabs;
5 grades: every leaf (really) has 11-15 of scabs;
7 grades: every leaf (really) has 16-20 of scabs;
9 grades: every leaf (really) has scab more than 21;
Computational methods:
Disease index=[Σ (the sick number of sheets × number of levels at different levels) × 100]/(investigating the total number of sheets × highest level numerical value)
The in vitro preventive effect of three-coloured amaranth leaf extract to citrus bacterial canker disease:
From following table 5, the protection effect of using after three-coloured amaranth leaf extract is followed successively by: after inoculation germ 14d, spray after three-coloured amaranth leaf extract > inoculation germ 1h, spray after three-coloured amaranth leaf extract > inoculation germ 7d, spray three-coloured amaranth leaf extract.Its protection effect is respectively: 69.53-79.08%, 64.89-77.07%, 62.35-76.44%.Along with the rising of three-coloured amaranth leaf extract concentration, protection effect is better.When three-coloured amaranth leaf extract concentration is during at 40-80mg/mL, protection effect preferably reaches 72.56-79.08%, substantially suitable with the preventive effect 77.07-81.21% that contrasts medicament streptomycin.When three-coloured amaranth leaf extract concentration is during at 5-20mg/mL, three-coloured amaranth leaf extract protection effect 64.89-77.5%.Variance analysis shows significant difference between the processing of variable concentrations three-coloured amaranth leaf extract.When three-coloured amaranth leaf extract concentration is 80mg/mL, protection effect is up to 79.08%.
The in vitro control effect of table 5 variable concentrations three-coloured amaranth leaf extract to citrus bacterial canker disease
Note: the numerical value in table is mean value 3 times; Different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
The potted plant preventive effect of example 8 three-coloured amaranth leaf extracts to rice sheath blight disease
Pot experiment is chosen in illumination box carries out, first the rice seedling cultivating is transplanted in plastic basin, adopt oese by cultured Rhizoctonia solani Kuhn pure culture biscuits involvng inoculation to paddy rice base portion leaf sheath, keep canopy temperature 29-32 DEG C on daytime, evening 21-24 DEG C, humidity is 90%, so that the paddy rice in different basin evenly falls ill.Experiment is divided into 5 concentration: 5,10,20,40,80mg/mL three-coloured amaranth leaf extract, make medicament control treatment with 75mg/mL jinggangmeisu, and taking clear water as blank.Establish altogether three processing: 1. inoculate after germ 1h, spray three-coloured amaranth leaf extract; 2. inoculate after germ 7d, spray three-coloured amaranth leaf extract; 3. inoculate after germ 14d, spray three-coloured amaranth leaf extract.Each test sprays with micro pressure formula watering can, taking squirt leave dual sides just not lower as spending.Every processing repeats 3 times.Every processing 5 strain rice plants, test needs altogether 5 concentration × 3 to process × 3 repetition × 5 strain rice plant+2 × 3 processing of a contrast and repeats × 5 strain rice plant=315 strain rice plants × 3 times.
After spraying three-coloured amaranth leaf extract, a situation arises for 10-15d investigation records rice sheath blight disease.Being calculated as follows of severity Scaling standard, disease index and preventive effect:
Rice sheath blight disease grade scale is as follows:
0 grade of plant leaf sheath and blade have no symptom;
1 grade of rice strain base portion has the fragmentary scab of minority;
3 grades of scabs extend to down 3 leaves (sword-like leave is for falling 0 leaf);
5 grades of scabs extend to down 2 leaves;
7 grades of scabs extend to down 1 leaf;
9 grades of scabs extend to sword-like leave or complete stool is withered.
Computational methods:
The pot experiment effect of three-coloured amaranth leaf extract to rice sheath blight disease:
From following table 6, the protection effect of using after three-coloured amaranth leaf extract is followed successively by: after inoculation germ 14d, spray after three-coloured amaranth leaf extract > inoculation germ 7d, spray after three-coloured amaranth leaf extract > inoculation germ 1h, spray three-coloured amaranth leaf extract.Along with the raising of three-coloured amaranth leaf extract concentration, its protection effect is also better, and wherein, in the time that concentration is 80mg/mL, preventive effect the best is 74.36-79.72%, substantially suitable with the control efficiency of jinggangmeisu.In the time that three-coloured amaranth leaf extract concentration is 5-40mg/mL, the protection effect of three-coloured amaranth leaf extract is lower than streptomycin sulphate, but still exceedes 60%, and three-coloured amaranth leaf extract of this explanation low concentration also has certain control efficiency to rice sheath blight disease.Variance analysis shows significant difference between the processing of variable concentrations three-coloured amaranth leaf extract.
The potted plant control effect of table 6 variable concentrations three-coloured amaranth leaf extract to rice sheath blight disease
Note: the numerical value in table is mean value 3 times; Different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
Effect assessment:
Three-coloured amaranth leaf extract is all inhibited to Acidovorax Avenae Subsp, c itrus canker germ and Rhizoctonia solani Kuhn, and in vitro and pot experiment also has certain control efficiency, three-coloured amaranth leaf extract is respectively 80.04-82.06%, 72.56-79.08%, 74.36-79.72% to the best control efficiency of bacterial fruit blotch of watermelon, citrus bacterial canker disease, rice sheath blight disease, all a little more than contrast medicament or suitable.Illustrate that three-coloured amaranth leaf extract not only has preventive and therapeutic effect to phytobacterial disease, and fungal diseases of plants is had to preventive and therapeutic effect, effective object wide spectrum.Having exploitation becomes the potentiality of broad-spectrum biological agricultural chemicals.In addition, observe according to system, duration of test has no poisoning and occurs, and each test reagent has no adverse effects to plant growth, and three-coloured amaranth leaf extract is the good biological active matter of control bacterial fruit blotch of watermelon, citrus bacterial canker disease and rice sheath blight disease.
Claims (4)
1. the application of three-coloured amaranth in controlling plant diseases, wherein, the application of the ethyl acetate extract that refers to three-coloured amaranth in controlling plant diseases; This plant disease is bacterial fruit blotch of watermelon, citrus bacterial canker disease or rice sheath blight disease; The ethyl acetate extract of this three-coloured amaranth is, after getting three-coloured amaranth leaf and cleaning, to shine dry grinding, then adopts and analyze pure ethyl acetate three-coloured amaranth leaf powder is carried out to lixiviate, and gained filtrate is inserted rotation concentration and evaporation instrument, is concentrated into paste and obtains.
2. the application of three-coloured amaranth as claimed in claim 1 in controlling plant diseases, wherein, described employing is analyzed pure ethyl acetate and three-coloured amaranth leaf powder is carried out to lixiviate is referred to three-coloured amaranth leaf powder disposable adding, ethyl acetate divides 3 times and adds, after the ratio that at every turn adds 10-20mL ethyl acetate in 1g three-coloured amaranth leaf powder adds, on shaking table at 20-25 DEG C with 90-120r/min jolting 24-36h, filter, collect filtrate, merge 3 times filtrate.
3. three-coloured amaranth, in the application suppressing in phytopathogen, wherein, refers to that the ethyl acetate extract of three-coloured amaranth is in the application suppressing in phytopathogen; This phytopathogen is Acidovorax Avenae Subsp, c itrus canker germ or Rhizoctonia solani Kuhn; The ethyl acetate extract of this three-coloured amaranth is, after getting three-coloured amaranth leaf and cleaning, to shine dry grinding, then adopts and analyze pure ethyl acetate three-coloured amaranth leaf powder is carried out to lixiviate, and gained filtrate is inserted rotation concentration and evaporation instrument, is concentrated into paste and obtains.
4. three-coloured amaranth as claimed in claim 3 is in the application suppressing in phytopathogen, wherein, described employing is analyzed pure ethyl acetate and three-coloured amaranth leaf powder is carried out to lixiviate is referred to three-coloured amaranth leaf powder disposable adding, ethyl acetate divides 3 times and adds, after the ratio that at every turn adds 10-20mL ethyl acetate in 1g three-coloured amaranth leaf powder adds, on shaking table at 20-25 DEG C with 90-120r/min jolting 24-36h, filter, collect filtrate, merge 3 times filtrate.
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