Method for quantitatively and rapidly detecting bacterial wilt resistance of plants
By injecting the bacterium wilt solution into the plant leaves and measuring the difference between ETR and Fq/Fm, the problems of high cost, low efficiency and harm to plants in the prior art are solved, and rapid and accurate bacterium wilt resistance are achieved. Test.
Patent Information
- Application Number
- CN202411245416.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-06
- Publication Date
- 2024-12-20
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
The existing plant bacterium wilt resistance detection methods are costly and inefficient, and the traditional methods cause irreversible damage to plants and cannot meet the needs of rapid detection.
By injecting the bacterium wilt solution into the plant leaves, the difference between ETR and/or Fq/Fm before and after 2 hours of bacterium infusion was measured, and quantitative detection was performed using the plant live photosynthesis imaging system to determine the plant's bacterium wilt Resistance.
It realizes low-cost and rapid quantitative detection of plant bacterium wilt resistance, saves time and resources, does not affect the normal growth of plants in the later stage, and the detection results are accurate and sensitive.
Smart Images
Figure CN119164926A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of plant disease resistance detection, and in particular relates to a method for quantitatively and rapidly detecting plant bacterial wilt resistance. Background Art
[0002] Bacterial wilt is a common agricultural disease. The pathogen is Ralstonia solanacearum, a bacterium. In the early stage of bacterial wilt infection, there are no abnormal symptoms in the aboveground part of the plant. After a few days, the plant suddenly loses its vitality and the entire aboveground part withers. It recovers on cloudy days and in the morning and evening. However, it withers soon after and shows symptoms of wilt. This process progresses very rapidly. After the crop is infected by bacterial wilt, the fine roots of the plant first turn brown, and the young and tender tissues at the top of the upper part (such as young leaves, tender tips, and new leaves) wilt. Before the leaves turn green, yellow, and dry, the plant will quickly dehydrate, wither, and wilt. Cutting the diseased stems close to the ground, it can be found that the vascular tissue has browned, and white bacterial fluid is secreted from this part. In dry climates and high temperatures, the plant will wither and die within a few days, with only the leaves becoming lighter in color, and the entire stem and leaves remain green. Bacterial wilt is a major disease of Solanaceae plants such as tomatoes, eggplants, peppers, potatoes, and tobacco, causing serious losses.
[0003] Detecting the resistance of different plant varieties to bacterial wilt is very important for improving production efficiency and reducing losses caused by diseases. The existing method for detecting plant resistance to bacterial wilt is to pour bacterial wilt solution into the soil where the plants grow, thereby simulating the infection process of bacterial wilt and judging the resistance of different plant varieties. This detection method requires a large amount of bacterial solution and soil, which is costly. Due to the uneven distribution of bacterial solution, the root contact with the bacterial solution cannot be guaranteed to be uniform, and the detection efficiency is low. Since it takes more than several weeks for bacterial wilt to infect plants and show symptoms, it cannot meet the demand for rapid detection of resistance of different varieties. There is also a method of judging the resistance of varieties by picking tissues from specific parts of the plant for genetic testing several weeks after inoculation. This method also requires picking specific plant tissues such as roots for resistance gene expression detection, which will cause irreversible and huge damage to the plants, or even death. Summary of the invention
[0004] In view of this, the object of the present invention is to provide a method for detecting plant bacterial wilt resistance, so as to achieve low-cost, short-time and high-efficiency quantitative detection of plant bacterial wilt resistance.
[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:
[0006] A method for detecting plant bacterial wilt resistance comprises the following steps: injecting bacterial wilt bacteria solution into plant leaves, and measuring the difference in ETR and / or Fq / Fm of the leaves injected with bacterial wilt bacteria before inoculation and 2 hours after inoculation; the smaller the difference, the better the plant's resistance to bacterial wilt.
[0007] As an embodiment, the bacterial wilt pathogenic bacteria liquid is obtained by suspending the bacterial wilt pathogenic bacteria in a MgCl2 solution, and the concentration of the MgCl2 solution is 10 mM.
[0008] As an embodiment, the OD of the bacterial wilt solution is 600 The value is 0.3-0.5, and the number of pathogens is 10 6 -10 8 cfu / ml.
[0009] As an embodiment, the injection is a quantitative injection, and the amount of the quantitative injection is 0.03-0.1 ml.
[0010] As an embodiment, the difference in ETR and / or Fq / Fm of leaves injected with bacterial wilt pathogens is measured before inoculation and within 2-6 hours of inoculation.
[0011] As an embodiment, the plant is a plant of the Solanaceae family.
[0012] Furthermore, the plant is any one or more of pepper, eggplant, tomato and tobacco.
[0013] As an implementation mode, the ETR and Fq / Fm are measured using a plant living photosynthesis imaging system, a chlorophyll fluorescence meter or a photosynthesis meter.
[0014] As an embodiment, the bacterial wilt pathogen is cultured in NA liquid culture medium, and the bacterial wilt pathogen is obtained after centrifugation.
[0015] Compared with the prior art, the present invention has the following beneficial effects:
[0016] The present invention provides a method for detecting plant resistance to bacterial wilt. By measuring the ETR and / or Fq / Fm difference of leaves injected with bacterial wilt bacteria solution before and after inoculation, the resistance of the plant to bacterial wilt is judged. The smaller the ΔETR and ΔFq / Fm values are, the better the plant's resistance to bacterial wilt is. The method of the present invention is fast in detection. After 2 hours of inoculation, the specific photosynthetic indexes of the plant are measured in vivo to achieve rapid screening of the resistance of different plant varieties to bacterial wilt. Compared with traditional detection methods, the present invention can quickly and quantitatively screen the resistance of different varieties to bacterial wilt in vivo at an early stage, saving cost and time, and does not affect the normal growth of the detected resistant plants in the later stage. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 These are the phenotypes of leaves under visible light and in vivo photosynthesis images of different pepper resistant varieties 24 hours after inoculation. 24hpi represents the phenotype of leaves under visible light 24 hours after inoculation; Fq / Fm(6hpi) represents the Fq / Fm status of leaves 6 hours after inoculation; ETR(6hpi) represents the ETR status of leaves 6 hours after inoculation.
[0018] Figure 2 It is the difference between the ETR value of leaves before inoculation and the ETR value of leaves 6 hours after inoculation of different pepper resistance varieties.
[0019] Figure 3 It is the difference between the Fq / Fm value of leaves before inoculation and the Fq / Fm value of leaves 6 hours after inoculation of different pepper resistance varieties.
[0020] Figure 4 The phenotypes of different pepper resistant varieties under visible light 6 days after root inoculation.
[0021] Figure 5 The figures are the phenotypes of eggplant and tobacco roots under visible light 6 days after inoculation and the in vivo photosynthesis images 6 hours after inoculation according to the method of the present invention. Fq / Fm(6hpi) represents the Fq / Fm status of the leaves 6 hours after inoculation; ETR(6hpi) represents the ETR status of the leaves 6 hours after inoculation. DETAILED DESCRIPTION
[0022] The present invention provides a method for detecting plant bacterial wilt resistance. A bacterial wilt bacteria solution is injected into plant leaves, and the differences in plant photosynthetic specific indicators ΔETR and ΔFq / Fm before and after inoculation are measured to achieve the purpose of quickly screening different varieties for bacterial wilt resistance.
[0023] The present invention suspends the bacterial wilt bacteria in a MgCl2 solution to obtain a bacterial wilt bacteria liquid. As some embodiments, the concentration of the MgCl2 solution of the present invention is 10mM. The bacterial wilt bacteria of the present invention adopt the existing bacterial wilt bacteria in the art, which can be purchased commercially or obtained by culture. As some embodiments, the present invention obtains the bacterial wilt bacteria by culturing a culture medium, and the culture medium can increase the number of bacterial wilt bacteria and maintain the activity of the bacteria. As an embodiment, the culture medium adopts NA culture medium, and the specific formula is: 3g beef extract, 10g glucose, 5g peptone, 0.5g yeast extract powder, and distilled water to 1L (adjust pH to 7.0). The present invention shakes and cultures the bacterial wilt bacteria in NA culture medium, and the culture conditions are 30°C and 260r / h. After culturing for a period of time, such as 24 hours, the culture solution is centrifuged, the supernatant on the top is poured out, and the resulting precipitated bacteria are suspended in a MgCl2 solution. As an embodiment, the suspension to liquid OD 600 The value is 0.3-0.5, and the number of pathogens is 10 6 -10 8cfu / ml. Those skilled in the art can adjust the concentration of the suspension of bacterial wilt pathogens according to the infection ability of bacterial wilt pathogens and the resistance level of plants, such as OD 600 The value is 0.4 and the number of pathogens is 10 7 cfu / ml, easy to inoculate.
[0024] The present invention injects bacterial wilt bacteria suspended in MgCl2 solution into plant leaves for infecting plants. In order to improve the accuracy and precision of the measurement, the present invention uses MgCl2 solution as a control and injects it into the leaves of the same node of the same plant variety to eliminate the interference of MgCl2 solution in the measurement of plant resistance level. The inoculation method of the present invention adopts quantitative injection. As an embodiment, the quantitative injection adopts a microinjector to accurately control the injection amount of bacterial wilt bacteria. As some embodiments, the amount of liquid quantitatively injected by the present invention is 0.03-0.1ml, and further 0.04-0.08ml. The present invention adopts quantitative injection to ensure the consistent inoculation amount of each plant, so that the measurement result is more accurate. In addition, extremely low injection volume can realize quantitative detection of plant resistance to bacterial wilt bacteria, with high sensitivity, greatly saving the amount of bacterial wilt bacteria, saving culture time and cost.
[0025] The present invention can determine the resistance of plants to bacterial wilt pathogens by measuring the difference in ETR and / or Fq / Fm between leaves injected with MgCl2 solution and leaves injected with bacterial wilt pathogen solution 2 hours after inoculation; the smaller the difference, the better the resistance of plants to bacterial wilt. Fq / Fm represents the parameter of the maximum photochemical efficiency of plant leaves under light conditions, reflecting the efficiency of light energy conversion by photosystem II (PSⅡ) of plants; ETR represents the photosynthetic electron transfer rate, reflecting the electron transfer rate of photosystem II (PSII) in plant photosynthesis. The present invention evaluates the response of plants to bacterial wilt pathogens by calculating and measuring the changes of these two indicators in a short period of time before and after inoculation. The present invention finds that ΔETR and ΔFq / Fm values can directly reflect the resistance level of plants to bacterial wilt, and the results are consistent with the results of traditional root inoculation, and the method of the present invention can achieve quantitative detection, which is more accurate in detecting the resistance level of plants. As an embodiment, the ETR value and / or Fq / Fm value are measured using a plant living photosynthesis imaging system, a chlorophyll fluorescence meter or a photosynthesis meter. In the embodiment of the present invention, a plant living photosynthesis imaging system is used to measure the ETR and Fq / Fm values of leaves before and after inoculation. As an embodiment, the photosynthesis measurement is selected between 2 and 6 hours after inoculation, which can achieve a rapid living body detection effect and obtain accurate and significant resistance level results.
[0026] In the present invention, the plant is a plant that can be infected by bacterial wilt pathogens, especially a plant of the Solanaceae family, such as pepper, eggplant, tomato and tobacco. The plant bacterial wilt resistance detection method of the present invention is applicable to any growth stage of the plant. As an embodiment, the resistance detection of bacterial wilt is carried out in the seedling stage of the plant. Selecting plant varieties of the same growth stage for detection is conducive to improving the accuracy of the detection.
[0027] Compared with the traditional root inoculation method which takes 6-8 days, the method of the present invention is fast and can obtain the resistance test results in as short as 2 hours. By calculating the photosynthetic specific indexes ΔETR and ΔFq / Fm values of plants before and after inoculation, the present invention can quickly and quantitatively screen the resistance of different varieties to bacterial wilt in vivo at an early stage, saving costs and time, and does not affect the normal growth of the tested resistant plants in the later stage.
[0028] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.
[0029] In the embodiment of the present invention, the plant living photosynthetic imaging system uses the multifunctional plant photosynthetic phenotype imaging measurement system PlantExplorer produced by Huinored (Beijing) Technology Co., Ltd. PRO+ .
[0030] The bacterial wilt strain in the embodiment of the present invention is derived from strains preserved in the laboratory of the Institute of Plant Protection, Fujian Academy of Agricultural Sciences.
[0031] Example 1
[0032] Identification of resistance to bacterial wilt in pepper varieties
[0033] Add bacterial wilt bacteria (or single colony) to NA liquid medium, and culture at 30℃, 200r / min shaking for more than 24 hours until the medium becomes turbid. NA liquid medium: 3g beef extract, 10g glucose, 5g peptone, 0.5g yeast extract powder, add distilled water to 1L (add NaOH solution to adjust pH to 7.0), and autoclave at 121℃ for 20min.
[0034] The bacterial culture was centrifuged at 6000 r / min, and the supernatant was discarded to obtain the bacterial precipitate. The precipitated bacterial cells were resuspended in 10 mM MgCl2 solution to an OD of 600 The value is 0.3 and the number of pathogens is 10 6 cfu, and obtain a suspension of bacterial wilt pathogen.
[0035] Four pepper varieties were selected and named pepper 1, pepper 2, pepper 3 and pepper 4, and their resistance levels to bacterial wilt were determined. The specific method is as follows:
[0036] At the 4-6 leaf stage of pepper, select pepper seedlings of the same growth, and inject 0.05 ml of bacterial wilt suspension into the leaves of the same node of each pepper variety. Use the plant photosynthesis imaging system to measure the ETR and Fq / Fm values of the leaves of the injected plants before and 6 hours after inoculation, and calculate the difference ΔETR and ΔFq / Fm. The smaller the ΔETR and ΔFq / Fm values, the better the resistance of the variety. At the same time, inject the same amount of MgCl2 solution as a control to eliminate the interference of MgCl2 solution.
[0037] The results are shown in Table 1 and Figure 1-3 From the results, we can see that pepper 1 has the smallest ΔETR and ΔFq / Fm values, and has the best resistance to bacterial wilt; followed by pepper 2 and pepper 4, and pepper 3 has the largest ΔETR and ΔFq / Fm values, and has poor resistance to bacterial wilt. This is consistent with the conclusion of the traditional root inoculation method of comparative examples 1 and 2 ( Figure 4 ).
[0038] Table 1 Statistics of Fq / Fm and ETR values of pepper varieties
[0039] Chili 1 Chili 2 Chili 3 Chili 4 ETR-0h 47 40.3 46 45.8 ETR-6h 44.1 36.6 31.2 33.2 ΔETR 2.9 3.7 14.8 12.6 Fq / Fm-0h 0.549 0.466 0.531 0.533 Fq / Fm-6h 0.515 0.425 0.362 0.387 ΔFq / Fm 0.034 0.041 0.169 0.146
[0040] Example 2
[0041] Identification of resistance to bacterial wilt in eggplant varieties
[0042] The bacterial wilt pathogen (or a single colony) is added to NA liquid culture medium for cultivation in the same manner as in Example 1.
[0043] The bacterial culture was centrifuged at 6000 r / min, and the supernatant was discarded to obtain the bacterial precipitate. The precipitated bacterial cells were resuspended in 10 mM MgCl2 solution to an OD of 600 The value is 0.4 and the number of pathogens is 10 7 cfu, and obtain a suspension of bacterial wilt pathogen.
[0044] Two eggplant varieties were selected, named Eggplant 1 and Eggplant 2, and their resistance levels to bacterial wilt were determined. The specific method is as follows:
[0045] At the 4-6 leaf stage of eggplant seedlings, select eggplant seedlings of the same growth, and inject 0.05 ml of bacterial wilt suspension into the leaves at the same node of each eggplant variety. Use the plant living photosynthesis imaging system to measure the ETR and Fq / Fm values of the leaves of the injected plants before and 6 hours after inoculation, and calculate the difference ΔETR and ΔFq / Fm. The smaller the ΔETR and ΔFq / Fm values, the better the resistance of the variety. At the same time, inject the same amount of MgCl2 solution as a control to eliminate the interference of MgCl2 solution.
[0046] The results are shown in Table 2 and Figure 5From the results, we can see that the ΔETR and ΔFq / Fm values of eggplant 1 are smaller than those of eggplant 2, indicating that eggplant 1 has good resistance to bacterial wilt, which is consistent with the conclusion of the traditional root inoculation method of comparative examples 1 and 2.
[0047] Table 2 Statistics of Fq / Fm and ETR values of eggplant varieties
[0048] Eggplant 1 Eggplant 2 ETR-0h 40.2 45 ETR-6h 38.4 42.1 ΔETR 1.8 2.9 Fq / Fm-0h 0.048 0.053 Fq / Fm-6h 0.042 0.035 ΔFq / Fm 0.006 0.018
[0049] Example 3
[0050] Identification of tobacco varieties' resistance to bacterial wilt
[0051] The bacterial wilt pathogen (or a single colony) is added to NA liquid culture medium for cultivation in the same manner as in Example 1.
[0052] The bacterial culture was centrifuged at 6000 r / min, and the supernatant was discarded to obtain the bacterial precipitate. The precipitated bacterial cells were resuspended in 10 mM MgCl2 solution to an OD of 600 The value is 0.5, and the number of pathogens is 10 8 cfu, and obtain a suspension of bacterial wilt pathogen.
[0053] Two tobacco varieties were selected, named tobacco 1 and tobacco 2, and their resistance levels to bacterial wilt were determined. The specific method is as follows:
[0054] At the 5-7 leaf stage of tobacco, select seedlings of tobacco varieties with consistent growth, and inject 0.05 ml of bacterial wilt suspension into the leaves at the same node of each tobacco variety. Use the plant living photosynthesis imaging system to measure the ETR and Fq / Fm values of the leaves of the plants injected with the bacterial solution before and 6 hours after inoculation, and calculate the difference ΔETR and ΔFq / Fm. The smaller the ΔETR and ΔFq / Fm values, the better the resistance of the variety. At the same time, inject the same amount of MgCl2 solution as a control to eliminate the interference of MgCl2 solution.
[0055] The results are shown in Table 3 and Figure 5 From the results, we can see that the ΔETR and ΔFq / Fm values of tobacco 1 are smaller than those of tobacco 2, indicating that tobacco 1 has good resistance to bacterial wilt, which is consistent with the conclusion of the traditional root inoculation method of comparative examples 1 and 2.
[0056] Table 3 Statistics of Fq / Fm and ETR values of tobacco varieties
[0057] Tobacco 1 Tobacco 2 ETR-0h 36.7 37 ETR-6h 35.3 34.6 ΔETR 1.4 2.4 Fq / Fm-0h 0.044 0.0486 Fq / Fm-6h 0.043 0.0462 ΔFq / Fm 0.001 0.0024
[0058] Example 4
[0059] Identification of resistance to bacterial wilt in tomato varieties
[0060] The bacterial wilt pathogen (or a single colony) is added to NA liquid culture medium for cultivation in the same manner as in Example 1.
[0061] The bacterial culture was centrifuged at 6000 r / min, and the supernatant was discarded to obtain the bacterial precipitate. The precipitated bacterial cells were resuspended in 10 mM MgCl2 solution to an OD of 600 The value is 0.4 and the number of pathogens is 10 7 cfu, and obtain a suspension of bacterial wilt pathogen.
[0062] Two tomato varieties were selected, named Tomato 1 and Tomato 2, and their resistance levels to bacterial wilt were determined. The specific method is as follows:
[0063] At the 4-6 leaf stage of tomatoes, select seedlings of tomato varieties with consistent growth, and inject 0.1 ml of bacterial wilt suspension into the leaves at the same node of each tomato variety. Use the plant living photosynthesis imaging system to measure the ETR and Fq / Fm values of the leaves of the plants injected with the bacterial solution before and 2 hours after inoculation, and calculate the difference ΔETR and ΔFq / Fm. The smaller the ΔETR and ΔFq / Fm values, the better the resistance of the variety. At the same time, inject the same amount of MgCl2 solution as a control to eliminate the interference of MgCl2 solution.
[0064] The results are shown in Table 4. From the results, it can be seen that the ΔETR and ΔFq / Fm values of Tomato 1 are smaller than those of Tomato 2, indicating that Tobacco 1 has a good ability to resist bacterial wilt, which is consistent with the conclusion of the traditional root inoculation method of Comparative Examples 1 and 2.
[0065] Table 4 Statistics of Fq / Fm and ETR values of tomato varieties
[0066] Tomato 1 Tomato 2 ETR-0h 53.7 56.4 ETR-6h 44.6 49.1 ΔETR 9.1 5.3 Fq / Fm-0h 0.068 0.057 Fq / Fm-6h 0.035 0.045 ΔFq / Fm 0.033 0.012
[0067] Comparative Example 1
[0068] Traditional root immersion inoculation method
[0069] Add the bacterial wilt strain (or single colony) to the NA liquid medium, and culture at 30°C with shaking at 200r / min for 24 hours (or more) until the medium becomes turbid to obtain the bacterial wilt solution. The NA liquid medium includes: 3g beef extract, 10g glucose, 5g peptone, 0.5g yeast extract powder, add distilled water to 1L (add NaOH solution to adjust the pH to 7.0), and autoclave at 121°C for 20min.
[0070] Select seedlings of plant varieties with consistent growth. Pull them out of the seedling tray, rinse off the root matrix with tap water, immerse them in the bacterial wilt solution for 15 minutes, and then transplant them into sterilized soil (new matrix). After inoculation, spray water to clean the leaves, water them every day to keep the soil moist, and keep the soil temperature at 30℃.
[0071] The disease level survey was started on the third day after transplanting, and was conducted every three days for a total of 3-4 times. The resistance level of the plant to bacterial wilt was determined based on the disease level results.
[0072] Grading standards: Grade 0: healthy plants;
[0073] Level 1: 1 leaf withered plant;
[0074] Level 2: Plants with 2-3 leaves withering;
[0075] Level 3: Most of the leaves are withered;
[0076] Level 4: Plants are withered, dead or about to die.
[0077] Comparative Example 2
[0078] Traditional root injection inoculation method
[0079] The preparation method of the bacterial wilt pathogen solution is the same as that of Comparative Example 1.
[0080] Select seedlings of plant varieties with consistent growth. Before inoculation, cut a 3-5 cm deep groove 1 cm from one side of the seedling with a knife to cut off the roots. Use a 25 ml syringe to quantitatively take the bacterial solution and inject the bacterial solution on the cut side. 20 seedlings (row length 50 cm) are injected with 50 ml. After inoculation, spray water to clean the leaves, water every day to keep the soil moist, and the soil temperature is 30 ℃. To promote bacterial infection, 28 days after inoculation, the roots can be cut off and inoculated again 1 cm away from the base of the seedling roots.
[0081] The disease level survey was started on the 3rd day after inoculation, and was conducted every 3 days for 3-4 times in total. The grading standard was the same as that of comparative example 1. The resistance level of the plant to bacterial wilt was determined based on the disease level results.
[0082] It can be seen from Examples 1-4 and Comparative Examples 1 and 2 that the traditional root inoculation method (root immersion inoculation or root wounding infusion inoculation) requires a large amount of bacterial solution, the amount of bacteria in contact with the root is difficult to control, and can only achieve qualitative detection, the determination of plant resistance level is not accurate, and the detection time is long, which has a destructive effect on plant growth. The present invention can achieve quantitative detection of living plants, and the detection can be carried out 2 hours after inoculation. The process is fast, the amount of inoculation is small, the resistance level is accurately determined, the sensitivity is good, and it does not affect the normal growth of the detected resistant plants in the later stage.
[0083] Comparative Example 3
[0084] Effects of other photosynthesis parameters on identification of resistance to bacterial wilt in pepper varieties
[0085] The pepper varieties with different resistance to bacterial wilt in Example 1 were used, and the method for preparing the bacterial wilt suspension was the same as in Example 1. The specific method is as follows:
[0086] At the 4-6 leaf stage of pepper, select pepper seedlings of the same growth and inject 0.05 ml of bacterial wilt suspension into the leaves at the same node of each pepper variety. Use the plant photosynthesis imaging system to measure the Fv / Fm value of the leaves of the plants injected with the bacterial solution before and 6 hours after inoculation. At the same time, inject the same amount of MgCl2 solution as a control to eliminate the interference of MgCl2 solution.
[0087] Fv / Fm reflects the potential maximum photosynthetic capacity of plants. This comparative example measured the Fv / Fm values of plant leaves before and after inoculation with bacterial wilt pathogens, and found that there was no obvious regularity in the changes in the Fv / Fm values of plant leaves before and after inoculation, and there was no significant correlation with the conclusions of the traditional root inoculation method of comparative examples 1 and 2 for identifying plant resistance to bacterial wilt pathogens, and it cannot be used to detect the evaluation of plant resistance to bacterial wilt.
[0088] Table 5 Statistics of Fv / Fm values of pepper varieties
[0089] Chili 1 Chili 2 Chili 3 Chili 4 Fv / Fm-0h 0.550 0.524 0.544 0.537 Fv / Fm-6h 0.506 0.476 0.674 0.653 ΔFv / Fm 0.044 0.048 -0.13 -0.116
[0090] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A method for detecting plant resistance to bacterial wilt, characterized in that: The method comprises the following steps: injecting bacterial wilt solution into plant leaves, and measuring the difference in ETR and / or Fq / Fm of the leaves injected with bacterial wilt before inoculation and 2 hours after inoculation; the smaller the difference, the better the resistance of the plant to bacterial wilt.
2. The method for detecting plant bacterial wilt resistance according to claim 1, characterized in that: The bacterial wilt pathogen solution is obtained by suspending the bacterial wilt pathogen in a MgCl2 solution, and the concentration of the MgCl2 solution is 10 mM.
3. The method for detecting plant bacterial wilt resistance according to claim 1 or 2, characterized in that: The OD of the bacterial wilt solution 600 The value is 0.3-0.5, and the number of pathogens is 10 6 -10 8 cfu / ml.
4. The method for detecting plant bacterial wilt resistance according to claim 1, characterized in that: The injection is a quantitative injection.
5. The method for detecting plant bacterial wilt resistance according to claim 4, characterized in that: The amount of the quantitative injection is 0.03-0.1 ml.
6. The method for detecting plant bacterial wilt resistance according to claim 1, characterized in that: The difference in ETR and / or Fq / Fm of leaves injected with bacterial wilt pathogens was measured before inoculation and within 2-6 hours of inoculation.
7. The method for detecting plant bacterial wilt resistance according to claim 1, characterized in that: The plant is a plant of the Solanaceae family.
8. The method for detecting plant bacterial wilt resistance according to claim 1 or 7, characterized in that: The plant is any one or more of pepper, eggplant, tomato and tobacco.
9. The method for detecting plant bacterial wilt resistance according to claim 1 or 6, characterized in that: The ETR and Fq / Fm are measured using a plant living photosynthesis imaging system, a chlorophyll fluorescence meter or a photosynthesis meter.
10. The method for detecting plant resistance to bacterial wilt according to claim 2, characterized in that: The bacterial wilt pathogen is cultured in NA liquid culture medium, and bacterial wilt pathogen bodies are obtained after centrifugation.