Application of pipecolic acid as plant disease-resistant activator in preventing and treating apple fruit diseases
阅读说明：本技术 哌啶酸作为植物抗病激活剂在防治苹果果实病害中的应用 (Application of pipecolic acid as plant disease-resistant activator in preventing and treating apple fruit diseases ) 是由 王彩霞 孙子豪 韩春雪 李晓鹏 练森 李保华 于 2021-01-12 设计创作，主要内容包括：本发明公开了哌啶酸作为植物抗病激活剂在防治苹果果实病害中的应用,属于植物诱导抗病性技术领域。所述哌啶酸用于增强植物体内防御酶的活性和提高抗病基因的表达,所述的防御酶为PAL、PPO、POD和SOD,所述的抗病基因为PR1和PR5。浓度为0.2mmol·L～(-1)～0.5mmol·L～(-1)的哌啶酸溶液可用于诱导苹果果实抗病性,防治苹果果实病害。本发明的哌啶酸溶液本身无离体的杀菌或抑菌活性,在活体条件下才能诱发植物抗病性,诱导苹果果实产生的抗病性具有持效期长和抗病谱广特点,且环保安全,不宜导致病原菌产生抗药性；使用方法简单,浓度低,用量少。(The invention discloses application of piperidine acid as a plant disease-resistant activator in preventing and treating apple fruit diseases, and belongs to the technical field of plant induced disease resistance. The pipecolic acid is used for enhancing the activity of defense enzymes in plants and improving the expression of disease-resistant genes, wherein the defense enzymes are PAL, PPO, POD and SOD, and the disease-resistant genes are PR1 and PR 5. The concentration is 0.2 mmol.L ‑1 ～0.5mmol·L ‑1 The piperidine acid solution can be used for inducing apple fruit disease resistance and preventing and treating apple fruit diseases. The piperidine acid solution disclosed by the invention has no in-vitro bactericidal or bacteriostatic activity, can induce the disease resistance of plants under the living condition, has the characteristics of long lasting period and wide disease resistance spectrum for inducing the disease resistance of apple fruits, is environment-friendly and safe, and is not suitable for causing pathogenic bacteria to generate drug resistance; simple application method, low concentration and low dosage.)
1. The application of the plant disease-resistant activator on the apple fruits is characterized in that the effective component of the plant disease-resistant activator is piperidine acid.
2. The use as claimed in claim 1, wherein the pipecolic acid is used to enhance the activity of defense enzymes in plants such as PAL, PPO, POD and SOD and to increase the expression of disease-resistant genes such as PR1 and PR 5.
3. Use according to claim 1 or 2, wherein the pipecolic acid is prepared in aqueous solution at the time of use.
4. Use according to claim 3, wherein the piperidine acid solution is applied to the apple fruit by spraying or drenching.
5. Use according to claim 3, wherein the concentration of the piperidine acid solution is 0.2 mmol-L-1～0.5mmol·L-1。
6. The use of any one of claims 1 to 5, wherein the use is for inducing disease resistance in apple fruits and controlling apple diseases.
7. The use according to claim 6, wherein the apple disease is ring rot, gray mold, anthracnose, spot disease or moldy heart disease of apples caused by pathogenic bacteria.
8. The use according to claim 7, wherein the ring rot pathogenic bacteria of apple is Staphylococcus aureus (Botryosphaeria dothidea), Botrytis cinerea (Botrytis cinerea), anthracnose pathogenic bacteria is Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), Sphaerotheca fuliginea pathogenic bacteria is Acremonium sclerotiorum (Acremonium sclerogen), Mycoplasma corenum pathogenic bacteria is Trichosporon (Trichothecium roseum), Alternaria alternata (Alternaria Alternaria altena) and Fusarium sp.
9. A method for inducing disease resistance of apple fruit is characterized in that a sprayer is used for spraying the surface of the apple fruit with the concentration of 0.5 mmol.L-1Piperidine acid solution.
10. A method for inducing disease resistance of apple fruit is characterized in that the apple fruit is added with a concentration of0.2mmol·L-1Soaking in piperidine acid solution for 5-10 min.
Technical Field
The invention belongs to the technical field of plant induced disease resistance, and particularly relates to application of piperidine acid as a plant disease resistance activator in prevention and treatment of apple fruit diseases.
Background
The apple is an important economic crop and export fruit in China, the cultivation area and the yield are in the top of the world, however, the export rate of the apple is less than 3 percent of the total yield, the market share in the world is extremely incompatible with the status of the big apple production countries in China, and the apple diseases, particularly the fruit diseases, are an important factor causing the phenomenon. Diseases caused by fungal infection in various links of apple production bring huge losses to people. Apple fruit diseases not only cause fruit rot in the growing period, but also cause fruit rot in the processes of storage, preservation and transportation after picking, thereby causing serious economic loss.
The apple fruit diseases mainly comprise: apple ring rot (Botryosphaeria dothidea), gray mold (Botrytis cinerea), anthracnose (Colletotrichum gloeosporioides), dot disease (Acremonium sclerotium), and moldy heart disease (Trichothecium roseum or Alternaria alternata or Fusarium spp). At present, bagging protection is mainly used for preventing and controlling apple fruit diseases in production, and due to the fact that the cost of bagging apples rises year by year, bag-free cultivation of apples becomes a necessary trend, but the fruit diseases are the key problems to be solved firstly in bag-free cultivation of apples. The control of apple postharvest diseases mainly depends on a method of soaking fruits by chemical agents, but the problems of chemical agent residues of chemical bactericides, easy generation of resistance of pathogenic bacteria to the chemical agents, serious environmental pollution and the like are serious day by day, and various bactericides are limited or forbidden to use. Therefore, the search for safe and effective control measures capable of replacing chemical bactericides has become an urgent problem to be solved in the control of diseases of fruits during apple production and postharvest storage.
Many researches show that the method controls diseases by exciting the defense system of plants and is an important way for developing efficient, safe and green pesticides. Plant disease-resistant activators (Plant activators) are compounds capable of inducing plants to generate disease resistance, and various commercialized disease-resistant activators are applied to crops such as tomatoes, grapes and tobaccos at present, and achieve remarkable disease prevention effects. The induced resistance of plants starts from the recognition of signal substances from pathogenic bacteria by the plants, and leads to a series of resistance reactions of the plants by activating various signal pathways in the plants, such as salicylic acid, jasmonic acid, ethylene and the like, wherein the resistance reactions comprise the increase of the expression level of defense genes, the increase of the activity of defense enzymes, the accumulation of defense compounds and the like. A large number of researches show that the induction resistance is utilized to control plant diseases, has the advantages of long resistance duration, wide disease resistance spectrum, no environmental pollution and the like, and is considered as a new strategy and a feasible way for preventing and controlling the plant diseases.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides application of the piperidine acid serving as a plant disease-resistant activator in preventing and treating apple fruit diseases. The invention provides a method for inducing disease resistance of apple fruits aiming at the problems of serious harm and difficult control of apple fruit diseases, pesticide residue caused by chemical agents, pathogenic bacteria resistance and the like in the prior agricultural production, and the method can be used for effectively controlling the apple fruit diseases. Therefore, the using frequency and the using amount of the chemical agent are reduced, the harm to the environment and the influence on the human health are weakened, the occurrence of pathogenic bacteria resistant and drug-resistant strains of apple fruit diseases can be delayed, and the development of the apple industry towards a safe, efficient and sustainable direction is realized.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the invention discloses an application of a plant disease-resistant activator in apple fruits, wherein the effective component of the plant disease-resistant activator is piperidine acid.
On the basis of the scheme, the pipecolic acid is used for enhancing the activity of defense enzymes in plants and improving the expression of disease-resistant genes, wherein the defense enzymes are PAL, PPO, POD and SOD, and the disease-resistant genes are PR1 and PR 5.
On the basis of the scheme, when in use, the piperidine acid is prepared into an aqueous solution.
On the basis of the scheme, the piperidine acid solution is applied to apple fruits in a spraying or soaking mode.
On the basis of the above schemeThe concentration of the piperidine acid solution is 0.2 mmol.L-1～0.5mmol·L-1。
On the basis of the scheme, the method is used for inducing disease resistance of apple fruits and preventing and treating apple diseases.
On the basis of the scheme, the apple diseases are apple ring spot, gray mold, anthracnose, spot disease or moldy core disease caused by pathogenic bacteria.
On the basis of the scheme, the ring rot pathogenic bacteria of the apple is staphylococcus (Botryosphaeria dothidea), the Botrytis cinerea (Botrytis cinerea), the anthracnose pathogenic bacteria are Colletotrichum gloeosporioides, the speck pathogenic bacteria are sclerotinia sclerotiorum (Acremonium sclerotium), the heart disease pathogenic bacteria are Trichothecium roseum (Trichothecium roseum), Alternaria alternata (Alternaria alternata) and Fusarium sp.
Based on the same conception, the invention also discloses a method for inducing the disease resistance of the apple fruits, and the sprayer is used for spraying the apple fruits with the concentration of 0.5 mmol.L-1Piperidine acid solution.
Based on the same conception, the invention also discloses another method for inducing the disease resistance of apple fruits, which is to ensure that the concentration of the apple fruits is 0.2 mmol.L-1Soaking in piperidine acid solution for 5-10 min.
The invention has the beneficial effects that:
the compound piperidine acid provided by the invention can induce plant disease resistance, belongs to a plant disease resistance activator, and has the following advantages compared with the traditional pesticide:
(1) the plant has no in-vitro sterilization or bacteriostasis function, and can induce the immune system of the plant to resist the attack of diseases under the condition of living bodies;
(2) the disease resistance generated by induction has long-lasting effect and broad-spectrum characteristics, the apple fruits treated by the piperidine acid have obvious prevention effect on pathogenic bacteria of apple ring spot, gray mold, anthracnose, spot and moldy core, and the prevention and control effects on the apple ring spot and the gray mold of the apple fruits treated for 30 days are respectively 75.5% and 82.8%, so that the prevention and control effects are obvious;
(3) the disease resistance generated by the induction of the piperidine acid belongs to the physiological metabolic reaction of plant organisms, is safe to the environment and is not suitable for generating drug resistance;
(4) the use method is simple, the concentration is low, and the dosage is less;
(5) the effect of preventing and treating the apple fruit diseases is remarkable, the highest prevention and treatment effect can reach 80.51 percent, and the economic benefit is very high;
(6) the use of the pipecolic acid can also improve the activity of defense enzymes PAL, PPO, POD and SOD in apple fruits and can induce the expression level of disease-resistant related genes PR1 and PR5 to be obviously increased.
Drawings
FIG. 1 shows the effect of pipecolic acid of different concentrations on the growth of hyphae and spore germination of ring rot of apple, wherein A is the spore germination rate of ring rot of apple, and B is the colony diameter of ring rot of apple;
FIG. 2 shows the time-dependent changes of the effect of pipecolic acid on the control of pathogenic bacteria of ring rot of apple
FIG. 3 shows the time-dependent changes of the control effect of pipecolic acid on apple gray mold pathogens;
FIG. 4 shows that the piperidine acid soaking treatment improves the resistance of apple fruits to diseases, A is a control for inoculating apple ring rot pathogenic bacteria, and B is the control effect of the piperidine acid soaking treatment on apple ring rot; c is a control group inoculated with apple gray mold pathogenic bacteria, and D is the control effect of the piperidine acid soaking treatment on apple gray mold;
FIG. 5 Effect of pipecolic acid on defensive enzyme activity in apple fruit;
FIG. 6 shows the effect of pipecolic acid on the expression of disease resistance related genes in apple fruits.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the contents in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
The invention relates to a method for inducing disease resistance of apple fruits, which comprises the steps of applying piperidine acid aqueous solution with effective concentration to the apple fruits, and exciting the disease resistance of the fruits after absorption, thereby improving the disease resistance of the apple fruits.
The piperidine acid of the present invention has the following structure:
example 1
1.1 Effect of pipecolic acid on the growth of apple pathogenic hyphae
PDA medium was prepared, and an aqueous piperidine acid solution was added thereto to give a final concentration of 0.5 mmol.L-1The activated apple ring rot pathogen plasmodiophora viticola (Botryosphaeria dothidea), Botrytis cinerea (Botrytis cinerea), Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), Colletotrichum sclerotiorum (Acremonium sclerotium), myceliophthora sclerotiorum (Trichothecium roseum), Sclerotium roseum (Trichothecium roseum), Alternaria alternata (Alternaria alternata) and Fusarium (Fusarium spp.) were inoculated into the PDA medium, respectively. The above bacteria are all conventional pathogenic bacteria in the field, are cultured in dark at a constant temperature of 25 ℃ for 3-5 days, and the diameter of a bacterial colony is measured.
The PDA culture medium comprises the following components in percentage by weight: peeling potatoes, weighing 200g, cutting into small pieces, boiling in water for 15-20 min, filtering with four layers of gauze, adding 20g of glucose and 15g of agar powder, fixing the volume to 1000mL, keeping the pH value natural, and sterilizing with high-pressure steam at 121 ℃ for 20 min.
The results showed that 0.5 mmol. multidot.L-1The piperidine acid has no obvious influence on the colony diameter of the pathogenic bacteria, which shows that the compound at the concentration has no inhibition effect on the hypha growth of the pathogenic bacteria.
1.2 Effect of Piperidinic acid on spore Germination of apple pathogens
Respectively preparing spore suspension of the pathogenic bacteria of apple fruit diseases with concentration of 105each.mL-1Separately, an aqueous piperidine acid solution was added to each of the spore suspensions to give a final concentration of 0.5 mmol. multidot.L-1And (5) observing the spore germination condition for 12 hours under the condition of 25 ℃.
The results show that: 0.5 mmol. L-1The pipecolic acid has no obvious influence on the germination of the pathogenic bacteria spores, which indicates that the compound with the concentration has no inhibiting effect on the germination of the pathogenic bacteria spores.
The effect of pipecolic acid on mycelial growth and spore germination of physalospora piricola is shown in fig. 1.
Example 2
The control effect of the pipecolic acid on apple fruit diseases is as follows:
the solution was disposed at 0.5 mmol. multidot.L-1Piperidine acid aqueous solution, apple ring rot pathogen plasmopara viticola (Botryosphaeria dothidea), gray mold pathogen Botrytis cinerea (Botrytis cinerea), Colletotrichum anthracnose (Colletotrichum gloeosporioides), mycosphaerella sclerotinia (Acremonium sclerotium), moldy heart disease pathogen trichotheca roseum (Trichothecium roseum), Alternaria alternata (Alternaria alternata) and Fusarium (Fusarium spp.) were activated and cultured on PDA medium for 3d, and a cake with a diameter of 5mm was punched at the edge of the colony for later use.
Selecting healthy Fuji apples of uniform size, washing with running water, sterilizing with 75% ethanol, air drying at room temperature, stabbing at equator of apple with diameter of 5.0mm and depth of 3.0mm, and cutting 3 wounds per fruit, and mixing the above 0.5 mmol/L-1Spraying the piperidine acid water solution onto the fruit until water drops, air drying at room temperature, and inoculating the pathogenic bacteria cake at the puncture wound. And 3d and 5d after inoculation, respectively observing and measuring the diameter of the lesion spots, and calculating the prevention and treatment effect.
The control effect is (control group lesion diameter-treatment group lesion diameter)/control group lesion diameter multiplied by 100%.
The control effect of the piperidine acid on the apple fruit diseases is shown in table 1, and the piperidine acid aqueous solution has remarkable control effect on 7 apple fruit diseases, the control effect is over 46 percent, and the highest control effect can reach 80.51 percent, which shows that the plant activator has good broad-spectrum disease resistance and remarkable control effect.
TABLE 1 Effect of pipecolic acid on controlling apple fruit diseases
Example 3
The effect of the piperidine acid on control varies with the time interval between uses:
after treating apple fruit according to the method of example 2 to make a wound, 0.5 mmol.L-1Uniformly spraying the piperidine acid aqueous solution on the fruits until water drops, airing at room temperature, respectively inoculating apple ring rot (Botryosphaeria dothidea) and Botrytis cinerea (Botrytis cinerea) fungus cakes at 12h, 24h, 48h, 72h and 96h after spraying, and measuring the change of the effect of preventing and controlling the piperidine acid along with the use interval time.
The results are shown in FIGS. 2 and 3: the pathogenic bacteria are inoculated at different time intervals, the control effect is obviously different, wherein the time intervals are 48-72 hours, the control effect is the best, and the control effect is over 75 percent.
Example 4
The piperidine acid soaking treatment improves the resistance of apple fruits to diseases:
the solution was disposed at 0.2 mmol. multidot.L-1And (3) picking apples by using a piperidine acid aqueous solution, soaking the apples in the aqueous solution for 5-10 min, naturally drying the apples at room temperature, and storing the apples at 4 ℃ in the dark under the condition of relative humidity of 95%. And after storing for 30 days, inoculating the ring rot of apple and gray mold to the wound, measuring the diameter of the lesion spot after inoculating for 3 days, and calculating the prevention and treatment effect.
The control effect is (control group lesion diameter-treatment group lesion diameter)/control group lesion diameter multiplied by 100%.
The results are shown in FIG. 4: 0.2 mmol. L-1Soaking apple fruit in piperidine acid water solution for 30d, and inoculating pathogenic bacteria of ring rot and Botrytis cinerea respectively to obtain the final productMoreover, the apple fruits still have obvious control effect on pathogenic bacteria of the ring spot and the gray mold of the apples, and the control effect on the pathogenic bacteria of the ring spot and the gray mold of the apples is 75.5 percent and 82.8 percent respectively.
Example 5
The disease resistance mechanism of the fruit of the apple induced by the pipecolic acid is as follows:
treating fruits according to the method of example 2, disinfecting and air drying the surfaces of the fruits, stabbing at the equator of the apple, uniformly spraying 0.5 mmol.L of the fruit with the diameter of about 5.0mm and the depth of about 3.0mm at 3-4 wounds on each fruit-1Piperidine acid aqueous solution, sterile water treated fruit as control. Sampling 0, 12, 24, 48, 72, 96, 120 and 144h after treatment by taking the puncture as the center, and determining the change trend of defense-related enzyme activity and disease-resistant related genes in fruit tissues.
The results are shown in FIGS. 5 and 6: 0.5 mmol. L-1After the apple fruits are treated by the piperidinic acid, the activities of Phenylalanine Ammonia Lyase (PAL), polyphenol oxidase (PPO), Peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues are obviously improved compared with a control, and the expression levels of disease course related Protein (PRs) genes PR1 and PR5 are obviously up-regulated, which indicates that the piperidinic acid can induce the resistance of the apple fruits to diseases by improving the activity of defensive enzymes and the expression of disease-resistant genes in the fruits.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.