Spray vaccination method for Blumeria graminis f.sp.tritici

Abstract

The invention discloses a spray vaccination method for Blumeria graminis f.sp.tritici and belongs to the technical field of crop pathogenic bacterium vaccination. The spray vaccination method comprises the following steps: (1) expanding propagation of monospora strains; (2) collection of conidia; (3) conidial powder suspension by use of C5-18-perfluorinated alkyl (FluorinertTM FC-40), and preparation of an erysiphe spore suspension with 106 spores / ml; and (4) spray vaccination. Uniform spray vaccination of Blumeria graminis f.sp.tritici is guaranteed, accurate quantification is ensured, and such shortcomings as nonuniform vaccination, accurate quantification failure and the like of a shaking method are overcome to guarantee normal implementation of Blumeria graminis f.sp.tritici related study works. The spray vaccination method can be popularized in Blumeria graminis f.sp.tritici resistance identification and breeding units.

Description

technical field

[0001] The invention belongs to the application technical field of plant pathogenic obligate parasitic bacteria inoculation method, in particular to a wheat powdery mildew spray inoculation method. Background technique

[0002] Wheat powdery mildew is a disease of airborne transmission, mainly through the conidia of powdery mildew with the airflow, and the bacteria cannot grow into obligate parasites on the medium, and can only be transferred from generation to generation on fresh host plants . Therefore, when studying the law of disease occurrence, identifying the disease resistance of varieties and measuring the biological activity of pesticides, it is necessary to rely on manual inoculation to draw more reliable conclusions.

[0003] It has been reported that 13 powdery mildew species including cucumber, pumpkin, watermelon, gourd, pepper, tomato, strawberry, grape, apple, melon, rubber tree, tobacco and rose can be evaluated by spore suspension spray and...

Examples

Embodiment 1

[0019] Different spore mass concentration inoculation effect of embodiment 1

[0020] Cut the first leaf of the pre-prepared susceptible variety "Jinmai 47" to a length of 3 cm, select 10 leaves face up and put them on the agar fresh-keeping medium containing benzimidazole, and collect them on the ultra-clean bench for pre-propagation. Put fresh conidia that have propagated into a sterile 5mL centrifuge tube, draw 4mL of C5-18-perfluoroalkane (FC-40) solution into the centrifuge tube containing spore powder, and make an inoculum suspension. Use a Vortex shaker to oscillate for 30s to suspend the spores evenly in the solution, draw 10 μL of the suspension and adjust the spore concentration to 10 with a hemocytometer under a microscope 6 、10 5 、10 4 、10 3 and 10 2 Each / mL, as the inoculum for later use; at the same time, use the FC-40 solution without spores as a negative control to observe whether FC-40 damages the wheat leaves. After inoculation, place it in the growth ro...

Embodiment 2

[0022] Embodiment 2, different spray amount inoculation effect

[0023] The preparation of inoculated leaves and inoculum is the same as in Example 1.

[0024] The spray volumes were 3mL, 4mL and 5mL respectively under the Porter spray tower, and placed in the growth room after inoculation (17±1°C, 17-19h light / day) for 8-10 days. The observation results showed that there was no difference in the inoculation effect of different spray volumes, and all of them could fully morbidity.

Embodiment 3

[0025] Embodiment 3, different wheat varieties disease resistance identification effect

[0026] According to the inoculation effect of different spore mass concentrations and the requirements of variety disease resistance identification, the concentration of spores used for disease resistance identification is 1×10 6 individual / ml. Cut the first leaf of 30 wheat varieties prepared in advance and interrupt the length of 3cm, and the blade face is placed on the agar fresh-keeping medium containing benzimidazole, and the inoculum spore suspension is prepared according to the method in Example 1, and the spores are adjusted Concentration 1×10 6 individual / ml. Place the 30 wheat varieties to be tested under the Porter spray tower, take 4mL of the spore suspension for spray inoculation, and continue to place it under the spray tower for 1-2 minutes after inoculation to ensure sufficient settlement of the inoculated spores. After inoculation, place it in the growth room and culti...

technical field

[0001] The invention belongs to the application technical field of plant pathogenic obligate parasitic bacteria inoculation method, in particular to a wheat powdery mildew spray inoculation method. Background technique

[0002] Wheat powdery mildew is a disease of airborne transmission, mainly through the conidia of powdery mildew with the airflow, and the bacteria cannot grow into obligate parasites on the medium, and can only be transferred from generation to generation on fresh host plants . Therefore, when studying the law of disease occurrence, identifying the disease resistance of varieties and measuring the biological activity of pesticides, it is necessary to rely on manual inoculation to draw more reliable conclusions.

[0003] It has been reported that 13 powdery mildew species including cucumber, pumpkin, watermelon, gourd, pepper, tomato, strawberry, grape, apple, melon, rubber tree, tobacco and rose can be evaluated by spore suspension spray and...

Examples

Embodiment 1

[0019] Different spore mass concentration inoculation effect of embodiment 1

[0020] Cut the first leaf of the pre-prepared susceptible variety "Jinmai 47" to a length of 3 cm, select 10 leaves face up and put them on the agar fresh-keeping medium containing benzimidazole, and collect them on the ultra-clean bench for pre-propagation. Put fresh conidia that have propagated into a sterile 5mL centrifuge tube, draw 4mL of C5-18-perfluoroalkane (FC-40) solution into the centrifuge tube containing spore powder, and make an inoculum suspension. Use a Vortex shaker to oscillate for 30s to suspend the spores evenly in the solution, draw 10 μL of the suspension and adjust the spore concentration to 10 with a hemocytometer under a microscope 6 、10 5 、10 4 、10 3 and 10 2 Each / mL, as the inoculum for later use; at the same time, use the FC-40 solution without spores as a negative control to observe whether FC-40 damages the wheat leaves. After inoculation, place it in the growth ro...

Embodiment 2

[0022] Embodiment 2, different spray amount inoculation effect

[0023] The preparation of inoculated leaves and inoculum is the same as in Example 1.

[0024] The spray volumes were 3mL, 4mL and 5mL respectively under the Porter spray tower, and placed in the growth room after inoculation (17±1°C, 17-19h light / day) for 8-10 days. The observation results showed that there was no difference in the inoculation effect of different spray volumes, and all of them could fully morbidity.

Embodiment 3

[0025] Embodiment 3, different wheat varieties disease resistance identification effect

[0026] According to the inoculation effect of different spore mass concentrations and the requirements of variety disease resistance identification, the concentration of spores used for disease resistance identification is 1×10 6 individual / ml. Cut the first leaf of 30 wheat varieties prepared in advance and interrupt the length of 3cm, and the blade face is placed on the agar fresh-keeping medium containing benzimidazole, and the inoculum spore suspension is prepared according to the method in Example 1, and the spores are adjusted Concentration 1×10 6 individual / ml. Place the 30 wheat varieties to be tested under the Porter spray tower, take 4mL of the spore suspension for spray inoculation, and continue to place it under the spray tower for 1-2 minutes after inoculation to ensure sufficient settlement of the inoculated spores. After inoculation, place it in the growth room and culti...

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