CN116491508A

CN116491508A - Application of chalcone compound in preventing and treating cotton wilt and preventing and treating method of cotton wilt

Info

Publication number: CN116491508A
Application number: CN202310412494.3A
Authority: CN
Inventors: 陈全家; 祖倩丽; 曲延英; 邓晓娟; 郑凯; 陈琴
Original assignee: Xinjiang Agricultural University
Current assignee: Xinjiang Agricultural University
Priority date: 2023-04-18
Filing date: 2023-04-18
Publication date: 2023-07-28

Abstract

The invention belongs to the technical field of biological control of cotton wilt, and particularly relates to application of chalcone compounds in control of cotton wilt and a control method of cotton wilt. The invention uses chalcone compound as the only active component, and the flat plate bacteriostasis test and the potting test prove that the chalcone can inhibit the growth of cotton fusarium wilt, reduce the disease index of the cotton fusarium wilt and prevent and treat the cotton fusarium wilt.

Description

Application of chalcone compound in preventing and treating cotton wilt and preventing and treating method of cotton wilt

Technical Field

The invention belongs to the technical field of biological control of cotton wilt, and particularly relates to application of chalcone compounds in control of cotton wilt and a control method of cotton wilt.

Background

Cotton wilt is one of two global cotton diseases and affects cotton production in cotton producing countries in the world to different degrees. Fusarium oxysporum wilt specialized fungi (Fusarium oxysporum. Sp. Vasinefectum) are pathogenic bacteria of cotton wilt, which survive easily under natural conditions, are capable of producing conidia, and are transmitted mainly through soil, seeds and fertilizers. Fusarium oxysporum only infects few plants such as cotton, okra, kenaf and the like, and can infect various plants such as wheat, solanaceae, leguminous plants, melons, tobacco and the like under the condition of inoculation in a greenhouse or a disease nursery. Cotton wilt is a typical vascular bundle disease and cotton can be affected throughout the entire growth phase. There are many types of disease symptoms, including bacterial wilt type, yellowing type, wrinkling type, yellow reticulate pattern, red leaf type, etc. in seedling stage; the bud stage includes wrinkled, spot-like, half-yellow, top-dead, stalk, etc. The disease often causes a large number of dead seedlings in the early stage of cotton, and leaves and bolls in the later stage fall off in a large number, even die.

Although cotton wilt is effectively controlled along with the planting of disease-resistant varieties, cotton wilt also has a rising trend along with the planting of insect-resistant cotton. Analyzing main reasons, and planting in large scale and centralization to cause rotation difficulty; the disease-resistant breeding period is longer, the workload is large, and the resistance is easy to lose; the chemical pesticide has obvious treatment effect on the disease plants and local soil in sporadic disease areas, but hardly achieves the effect of the vascular bundle parts of cotton plants, has poor effect and pollutes the environment; the chemical agent has high toxicity, most of the agents are volatile, have the characteristics of cancerogenesis, teratogenesis and mutation, and the pesticide enters into the human body for administration in a large amount through oral and respiratory tracts or direct contact, so that acute poisoning is easy to be caused, and the health of people and animals is seriously endangered.

Chalcone is an alpha, beta-unsaturated ketone, also called styryl phenyl ketone, is an important natural product, widely exists in medicinal plants such as liquorice, angelica keiskei, safflower and the like, and a conjugated system formed by double bonds and carbonyl groups is regarded as a pharmacophore of the chalcone, because the biological activity is reduced or even eliminated by removing the structure. The skeleton of chalcone is formed by connecting two aromatic rings with alpha, beta-unsaturated ketone, has great flexibility, exists in a plurality of conjugated forms in nature, can be combined with a plurality of receptors, has wide physiological activities such as anti-tumor, anti-inflammatory and anti-infection, and has no report on the use of chalcone for preventing and treating cotton wilt.

Disclosure of Invention

The invention aims to provide application of chalcone compounds in inhibiting cotton fusarium wilt, inhibiting the growth of hyphae of cotton fusarium wilt and preventing and treating cotton fusarium wilt.

In order to achieve the above object, the present invention provides an application of a chalcone compound in inhibiting fusarium wilt of cotton.

Preferably, the inhibition of fusarium wilt of cotton comprises inhibition of hypha growth of fusarium wilt of cotton.

Preferably, the fusarium wilt of cotton comprises fusarium oxysporum.

The invention also provides application of the chalcone compound in preventing and treating cotton wilt.

Preferably, 10mL of chalcone solution with the concentration of 5-10 mg/mL is applied to each cotton seedling root irrigation.

Preferably, the solvent of the chalcone solution is a 50% DMSO solution.

The invention also provides a method for preventing and treating cotton wilt, which comprises the step of applying the chalcone compound to cotton seedlings.

Preferably, the solvent of the chalcone solution is a 50% DMSO solution.

Preferably, the cotton seedling comprises two-leaf and one-heart cotton seedling.

The beneficial effects are that:

the invention uses chalcone compound as the only active component, and the flat plate bacteriostasis test and the potting test prove that the chalcone can inhibit the growth of cotton fusarium wilt, reduce the disease index of the cotton fusarium wilt and prevent and treat the cotton fusarium wilt.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below.

FIG. 1 is the results of a chalcone compound plate bacteriostasis experiment of example 1; wherein a is the result of the control group, b is the result of the test 1 group, c is the result of the test 2 group;

FIG. 2 is the results of a comparative example 1 flavanone compound plate bacteriostasis experiment; wherein a is the result of the control group, b is the result of the test 1 group, c is the result of the test 2 group;

FIG. 3 is a graph showing the results of a chalcone compound plate bacteriostasis test (filter paper method) of example 2; wherein a is a result of dripping 20 mu L of sterile distilled water, B is a result of dripping 20 mu L of a chalcone solution of 10mg/ml, c is a result of dripping 20 mu L of 50% DMSO, and A is a partial enlarged graph of B;

FIG. 4 is a graph showing the results of a comparative example 2 flavanone compound plate bacteriostasis test (filter paper method); wherein a is the result of dripping 20 mu L of sterile distilled water, b is the result of dripping 20 mu L of 30mg/ml flavanone solution, and c is the result of dripping 20 mu L of 50% DMSO;

FIG. 5 is the results of potting experiments with chalcone compounds of example 3; a is 06-146, B is DJ-07-136, C is Xinhai No. 14, D is NMGB-4.

Detailed Description

The invention provides application of chalcone compounds in inhibiting cotton fusarium wilt.

In the present invention, the inhibition of cotton fusarium wilt preferably includes inhibition of cotton fusarium wilt hypha growth; the fusarium oxysporum preferably comprises fusarium oxysporum.

The purity of the chalcone compound of the present invention is preferably 97%; the structural formula of the chalcone compound is as follows:

the invention has no strict requirement on the source of the chalcone compound, and the chalcone compound is purchased conventionally.

According to the invention, a plate bacteriostasis test is carried out, and the result of the plate bacteriostasis test shows that the low-concentration chalcone compound (5 mg/mL) can inhibit the growth of hyphae of the fusarium wilt of cotton, and the high-concentration chalcone compound (10 mg/mL) can completely inhibit the growth of hyphae of the fusarium wilt of cotton.

In the invention, preferably, 10mL of chalcone solution with the concentration of 5-10 mg/mL is applied to each cotton seedling root irrigation; the solvent of the chalcone solution is preferably a 50% DMSO solution. The cotton seedling of the present invention preferably comprises a two-leaf one-heart cotton seedling.

In the invention, preferably, 10mL of chalcone solution with the concentration of 5-10 mg/mL is applied to each cotton seedling root irrigation; the solvent of the chalcone solution is preferably a 50% DMSO solution. The cotton seedling preferably comprises two-leaf and one-core cotton seedling.

According to the invention, a potting test is carried out, and the result of the potting test shows that pathogenic bacteria are inoculated when two leaves of cotton seedlings are in one heart, chalcone is applied after inoculation for 3d, and the condition of the cotton wilt disease is reduced after treatment for 14d compared with that of a control group, so that the chalcone can reduce the occurrence of the cotton wilt disease and has a great application prospect.

For further explanation of the present invention, the application of the chalcone compounds provided in the present invention to control cotton wilt and the control method of cotton wilt will be described in detail with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.

Materials and reagents involved in the following examples

The seed material is a sea island cotton (Gossypium barbadense l.) resource material: sea island cotton 06-146 (anti-fusarium wilt), new sea No. 14 (fusarium wilt), sea island cotton DJ-07-136 (anti-fusarium wilt) and sea island cotton NMGB-4 (fusarium wilt);

the fusarium oxysporum strain used is fusarium oxysporum cotton, physiological race No. 7 (Feng Jie, chen he. Basic research outline of cotton fusarium wilt [ J ]. Cotton theory, 1994 (02): 65-69);

chalcone compounds were purchased from Shanghai Yuan Ye Biotechnology Co., ltd (CAS number: 94-41-7, cat number: S29931)

The sea island cotton 06-146 is an excellent strain cultivated by the economic crop research institute of Xinjiang agricultural academy of sciences by using a compound hybridization technology, and has the variety characteristics of: zero branching, high coat content, high resistance to type II wilt, see (Chai Yanjun. Island cotton F_ (2:5) population yield, wilt resistance genetic analysis, and QTL localization [ D ]. Xinjiang university of agriculture, 2013.).

The new sea No. 14 is a preferred line of the new sea No. 10 multiplied by the army sea No. 1 utilized by the agricultural science of the farmer, the variety characteristic is zero branch, the plant type is compact, and the resistance reaction of the fusarium wilt is S.

Island cotton DJ-07-136 was provided by Xinjiang crop genetic improvement and germplasm innovation key laboratory at Xinjiang university (Han Moli. Island cotton anti-wilt high-density linkage map construction and disease resistance gene excavation [ D ]. Xinjiang university, 2021.);

NMGB-4 is provided by Xinjiang crop genetic improvement and germplasm innovation key laboratory at Xinjiang university (Zhao Fuxiang. Island cotton drought resistance evaluation and drought resistance trait QTL location [ D ]. Xinjiang university, 2019.).

Example 1

Chalcone plate bacteriostasis experiment

1. The chalcone compound was weighed and dissolved in 50% DMSO solution to prepare chalcone solutions at concentrations of 5mg/ml and 10 mg/ml.

2. 5mg/ml chalcone solution and Fusarium oxysporum cotton (spore concentration 1X 10) ⁷ Mixing the materials according to the volume ratio of 1:1, standing for 24 hours, sucking 20 mu L of the mixed solution, dripping the mixed solution in the center of a PDA solid culture medium (20 mL), inverting the mixed solution to be cultured in a culture box at 28 ℃, and recording the culture box as a test group 1;

mixing chalcone solution 10mg/ml and Fusarium oxysporum (spore concentration 1×10) ⁷ Each ml) was mixed in a volume ratio of 1:1, left for 24 hours, and 20. Mu.L of the mixture was pipetted and dropped onto PDA solidCulture medium (20 mL) was placed in the center and inverted at 28℃incubator, and the culture was designated as test group 2;

sterile water and Fusarium oxysporum (spore concentration 1×10) ⁷ Each mL) was mixed at a volume ratio of 1:1, left for 24 hours, and 20. Mu.L of the mixture was pipetted into the center of PDA solid medium (20 mL), and incubated in an inverted 28℃incubator, designated as Control (CK).

3. After 3 days of culture in the test 1 group, the test 2 group and the control group, growth conditions of Fusarium oxysporum gossypii were observed, and the hypha growth inhibition rate was calculated according to the following formula, and the results are shown in FIG. 1 and Table 1.

Hypha growth inhibition (%) = colony diameter (CK) -colony diameter (treatment)/colony diameter (CK)

TABLE 1 growth of Fusarium oxysporum in different treatment groups of cotton

Treatment mode	Colony diameter (cm)	Inhibition ratio (%)
			Test 1 group	7.3	13.10％
Test 2 groups	0	100％
			Control group (CK)	8.4	/

As can be seen from FIG. 1 and Table 1, the test 1 group had a partial bacterial growth with a hypha growth inhibition of 13.10% compared with the control group, indicating that 5mg/ml chalcone solution was able to inhibit growth of Fusarium oxysporum cotton hypha; the test group 2 had no hypha growth and the hypha growth inhibition rate was 100%, indicating that the chalcone solution of 10mg/mL was able to completely inhibit the hypha growth of Fusarium oxysporum gossypii.

Comparative example 1

With example 1, the difference is that:

weighing flavanone compound (CAS number: 487-26-3, product number: B29177, purity of 98% or more) and dissolving in 50% DMSO solution to obtain flavanone solution with concentration of 15mg/ml and 30 mg/ml;

test 1 group: 15mg/ml flavanone solution and Fusarium oxysporum cotton (spore concentration 1×10) ⁷ Mixing the materials according to the volume ratio of 1:1, standing for 24 hours, sucking 20 mu L of the mixed solution, dripping the mixed solution in the center of a PDA solid culture medium (20 mL), and culturing in an inverted 28 ℃ incubator;

test 2 group: mixing 30mg/ml flavanone solution and Fusarium oxysporum (spore concentration 1×10) ⁷ Mixing the components according to the volume ratio of 1:1, standing for 24 hours, and sucking 20 mu L of the mixed solution to drop on a PDA solid medium (20 mL);

control group (CK): sterile water and Fusarium oxysporum (spore concentration 1×10) ⁷ Each mL) was mixed at a volume ratio of 1:1, left for 24 hours, and 20. Mu.L of the mixture was pipetted into the center of PDA solid medium (20 mL) and incubated in an inverted 28℃incubator.

After 3 days of incubation in test 1, test 2 and control groups, the growth of Fusarium oxysporum gossypii is shown in FIG. 2 and Table 2.

TABLE 2 growth of Fusarium oxysporum in different treatment groups

As can be seen from table 2 and fig. 2, compared with the control group, the hypha growth condition of the test 1 group is consistent with that of the control group, the hypha growth inhibition rate is 0.133%, and the flavanone solution with 15mg/ml can not inhibit the growth of fusarium oxysporum cotton hypha; compared with the control group, the hypha growth condition of the test group 2 is consistent with that of the control group, the hypha growth inhibition rate is 0.400%, and the flavanone solution with the concentration of 30mg/mL can not inhibit the growth of fusarium oxysporum of cotton.

Example 2

Chalcone plate bacteriostasis experiment (filter paper method)

1. Preparation of pathogen plates: 100. Mu.l of Fusarium oxysporum (spore concentration 1X 10) ⁷ The suspension was dropped onto PDA solid medium (20 ml), and the spore suspension was spread evenly on PDA solid medium (20 ml) using a spreading bar until PDA solid medium (20 ml) completely absorbed no-flow liquid of spore suspension, i.e. pathogen plate preparation was completed.

2. After the plates were dried, 3 pieces of sterile filter paper with a diameter of 10mm were randomly placed, 20. Mu.L of sterile distilled water was added dropwise to one piece of filter paper, 20. Mu.L of 50% DMSO was added dropwise to the other piece of filter paper, and 20. Mu.L of the chalcone solution of example 1 at a concentration of 10mg/ml was added dropwise to the other piece of filter paper, and the resultant was cultured in an incubator at 28℃for 3 days and then observed, and the results were shown in FIG. 3.

As can be seen from FIG. 3, the sterile filter paper sheet with the sterile distilled water and 50% DMSO dropped thereon was full of germs, while the sterile filter paper sheet with the chalcone solution dropped thereon at 10mg/ml was not full of germs, indicating that chalcone could inhibit the hyphal growth of cotton blight germs.

Comparative example 2

With example 1, the difference is that:

after the plates were dried, 3 pieces of sterile filter paper sheets having a diameter of 10mm were randomly placed, 20. Mu.L of sterile distilled water was dropped on one piece of filter paper, 20. Mu.L of 50% DMSO was dropped on the other piece of filter paper, 30. Mu.L of the flavanone solution of comparative example 2 was dropped on the other piece of filter paper, and the resultant was cultured in an incubator at 28℃for 3 days, and the results were shown in FIG. 4.

As can be seen from FIG. 4, the sterile filter paper sheet with the sterile distilled water and 50% DMSO was full of germs, while the sterile filter paper sheet with the flavanone solution of 30mg/ml was also full of germs, indicating that flavanone could not inhibit the growth of cotton blight germs.

Example 3

Chalcone potting anti-efficacy test

Sterilizing cotton seeds of sea island cotton 06-146 (anti-fusarium wilt), sea island cotton DJ-07-136 (anti-fusarium wilt), xinhai No. 14 (fusarium wilt) and NMGB-4 (fusarium wilt) with a black plastic package bag, packaging the cotton seeds by ventilation, placing the cotton seeds in a constant temperature incubator at 26 ℃, picking out the white cotton seeds the next day, planting the cotton seeds in plastic small bowls (with the diameter of 10 cm) with the soil proportion of 2 parts by mass of sterile soil and 1 part by mass of vermiculite, planting 5 cotton seeds in each small bowl, and culturing the cotton seeds in a cotton culture chamber with the photoperiod of 16h (light)/8 h (dark) and the temperature of 25 ℃ and the relative humidity of 60% -70%;

when cotton seedlings grow to two leaves and one center, fusarium oxysporum suspension (spore concentration 10) of cotton is used by root cutting and bacterium dipping method ⁷ Seed per mL) of cotton seedlings, 50mL per pot, 3d later, root irrigation with 10mg/mL chalcone solution of example 1, 50mL per pot root irrigation with 10mg/mL chalcone solution, sterile water treatment as control. Each treatment in the potting test was set up with 3 replicates of 30 plants each and the investigation was carried out 14 days after treatment. According to GBT 22101.4-2009 cotton insect resistance evaluation technical specification part 4: wilt was investigated. The survey grading criteria are as follows:

the main symptoms of cotton wilt in the seedling stage of the greenhouse are a wilt type and a yellow reticulate pattern, the leaves and the true leaves are wilted, the leaves are softened and drooping, the leaf margin starts to wither, the leaf veins are yellow, and the leaves wilt and the cotton plants die. The individual disease stage grading criteria are as follows:

level 0: cotton plants are healthy, have no diseased leaves and grow normally.

Stage 1: cotton plant 1-2 pieces She Bianhuang wilting.

2 stages: the cotton plant has 2 cotyledons and 1 true She Bianhuang wilting, and the veins are yellow reticulate.

3 stages: 2 cotyledons of cotton plant and 2 or more than 2 leaves of cotton plant are wilted with true She Bianhuang, and the veins are yellow reticulate pattern or wilted pattern; dwarfing or wilting cotton plants.

4 stages: all leaves of cotton plants are ill, and the cotton plants die.

Statistics of investigation results: and calculating the disease index of each variety according to the investigation result.

The index of disease is calculated in "DI" as follows:

disease Index (DI) = [ Σ (d) _c ×n _c )/(n _t ×4)]X 100, where d _c For the corresponding disease level, n _c For the number of plants of each disease grade, n _t The total plant number.

The calculation result is accurate to the two bits after the decimal point, and the result is shown in table 3 and fig. 5.

TABLE 3 Effect of chalcone solution root irrigation on cotton wilt disease index

As can be seen from Table 3 and FIG. 5, cotton seedlings irrigated with chalcone solution showed slower and lighter occurrence of symptoms such as leaf yellowing, wilting and leaf falling than cotton seedlings irrigated with sterile water as a control, indicating that chalcone had a certain control effect on cotton wilt.

From the above, it can be seen that the chalcone compound can inhibit the growth of cotton fusarium wilt, reduce the disease index of cotton fusarium wilt, and prevent and treat cotton fusarium wilt.

Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.

Claims

1. The application of chalcone compounds in inhibiting cotton fusarium wilt.

2. The use according to claim 1, wherein said inhibition of fusarium wilt of cotton comprises inhibition of mycelium growth of fusarium wilt of cotton.

3. The use according to claim 1 or 2, characterized in that the fusarium oxysporum comprises fusarium oxysporum (f.oxysporum).

4. The application of chalcone compounds in preventing and treating cotton wilt.

5. The method according to claim 4, wherein 10mL of chalcone solution of 5-10 mg/mL is applied to the root of each cotton seedling.

6. The use according to claim 5, wherein the solvent of the chalcone solution is a 50% DMSO solution.

7. A method for controlling cotton wilt comprising the step of applying a chalcone compound to cotton seedlings.

8. The method of claim 7, wherein 10mL of 5-10 mg/mL chalcone solution is applied to each cotton seedling root.

9. The use according to claim 8, wherein the solvent of the chalcone solution is a 50% DMSO solution.

10. The method of claim 7, wherein the cotton seedling comprises a two-leaf, one-heart cotton seedling.