Application of vdscp27 protein in enhancing plant resistance and inducing plant defense response
A plant resistance, vdscp27 technology, applied in the field of microbial protein application, can solve the problem of low induction efficiency, achieve obvious induction effect, broad application prospects, and improve plant resistance
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Embodiment 1
[0050] Example 1 VdSCP27 protein stimulates tobacco immune response
[0051] (1) VdSCP27 protein induces ROS response in tobacco leaves
[0052] Using a 1 mL syringe without a needle, inject about 20 μL of 100 nM VdSCP27 protein into 3-4 week-old tobacco leaves from the back of the leaves. Tris-HCl (20 nM, pH 8.0) at the same concentration was used as a negative control. After 6 hours of injection, the treated tobacco leaves were rinsed with deionized water, placed in DAB dyeing solution, incubated at 25°C in the dark for 8 hours, the dyeing solution was removed and the dyed tobacco leaves were rinsed with deionized water, and washed with 95 Decolorized with % ethanol, observed and photographed with a stereomicroscope.
[0053] Results: Tobacco leaves injected with VdSCP27 protein had a large amount of H in the injected area 2 o 2 Accumulation of tobacco leaves injected with Tris-HCl, injection area H 2 o 2 The accumulation is extremely weak, such as figure 1 shown.
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Embodiment 2
[0063] Example 2 VdSCP27 protein induces tobacco resistance to pathogenic bacteria
[0064] Using a 1 mL syringe without a needle, inject about 20 μL of 100 nM VdSCP27 protein into 4-week-old tobacco leaves from the back of the leaves. Tris-HCl (20 nM, pH 8.0) at the same concentration was used as a negative control. 12h-24h after injection, use a needle to pierce the injection area, and prepare 5 μL in advance, 5×10 6 The spores / mL spore suspension of different pathogenic bacteria to be tested was dropped directly above the eye of the needle, cultivated at 25°C and 80% humidity, and observed and measured the area of each lesion 3-5 days after inoculation. Experimental results such as Figure 4 shown. The inhibition of VdSCP27 protein on tobacco leaf lesions caused by various pathogenic bacteria is shown in Table 1.
[0065] Table 1 Inhibition of VdSCP27 protein on tobacco leaf lesions caused by various pathogenic bacteria
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[0067] The above results indicate...
Embodiment 3
[0068] Example 3 VdSCP27 protein induces resistance responses of different plants to Botrytis cinerea
[0069] The VdSCP27 protein solution with a concentration of about 100nM was used to spray or root-irrigate plants in different experimental groups. Each group of experiments was sprayed with 1-2mL of the above-mentioned protein solution on the leaves or 10-15mL of the above-mentioned protein solution, and the number of plants used in the experiment was 20. Tris-HCl (20 nM, pH 8.0) at the same concentration was used as a negative control. After about 24 hours of treatment, 20 mL of pre-prepared 5×10 6 The spores / mL Botrytis cinerea spore suspension was cultured at 25° C. and 80% humidity, and was inoculated for 30 days to observe and count the incidence of plants in each experimental group.
[0070] According to the incidence, the disease grades are divided into grade 0, grade 1, grade 2, grade 3, and grade 4. The incidence of different grades of disease is as follows:
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