Method for measuring pathogenicity of pear fire blight bacteria by means of in-vitro water culture approach for fragrant pear branches
A pear fire blight and pathogenicity technology, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microbial determination/inspection, etc. Reliable, easy to operate, stable in pathogenicity
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Embodiment 1
[0025] Example 1 The impact of different inoculation methods on isolated branches on the pathogenicity evaluation of pathogenic bacteria
[0026] 1. Preparation of E. amylovora inoculum: the E. amylovora strain E.a 0001, E.a 0019 and E.a 0087 was inoculated on NA+5% sucrose medium, activated at 28.5°C for 48 hours, picked a single colony and inoculated in NA+5% sucrose medium, cultured with shaking at 28.5°C and 160 rpm / min for 12 hours, and measured OD 600 The bacterial solution with a value of about 1.2 was diluted with sterile water to a concentration of 10 8CFU / mL was used as the inoculum.
[0027] 2. Preparation of isolated branches: Select the newborn young branches of the Korla fragrant pear, which is extremely susceptible to pear fire blight, as the test material, cut the test branches up to the stem section of about 30 cm, disinfect it with 75% alcohol, and then 1 / 3 into a conical flask filled with 0.5‰NaCl sterilized saline.
[0028] 3. Inoculation: In this...
Embodiment 2
[0040] Example 2 Determination of pathogenicity of E. amylovora pathogenicity and identification of host selection
[0041] 1. Preparation of E. amylovora inoculum: the E. amylovora strain E.a 0001, E.a 0017、 E.a 0055 were respectively inoculated on NA + 5% sucrose medium, activated at 28.5°C for 48 hours, picked a single colony and inoculated in NA + 5% sucrose medium, cultured with shaking at 28.5°C and 160 rpm / min for 12 hours, and measured OD 600 The bacterial solution with a value of about 1.2 was diluted with sterile water to a concentration of 10 8 CFU / ml was used as the inoculum.
[0042] 2. Preparation of isolated branches: choose pear fire blight susceptible varieties Xiangli, Dangshan pear, black sour pear, Du pear, Xinli No.7, yellow sour pear, brown Juju pear, Hong'anjiu, Huochengdong yellow pear, The newborn young shoots of Kuqa Amut were used as the test materials. The stems of the test branches up to about 30cm were sterilized with 75% alcohol, and 1 / 3...
Embodiment 3
[0049] Embodiment 3: the pathogenicity determination of different bacterial strains of amygdalus blight
[0050] 1. Preparation of E. amylovora inoculum: Inoculate 10 strains of E. amylovora from different sources in Table 1 on NA+5% sucrose medium, culture at 28.5°C for 48 hours to activate, pick a single colony and inoculate on NA +5% sucrose culture solution, shake culture at 28.5℃, 160rpm / min for 12h, measure OD 600 The bacterial solution with a value of about 1.2 was diluted with sterile water to a concentration of 10 8 CFU / ml was used as the inoculum.
[0051] 2. Preparation of isolated branches: select the new branches of the Korla fragrant pear, a high-susceptibility variety of pear fire blight, as the test branches, cut the test branches to a stem section of about 30 cm, disinfect with 75% alcohol, and insert 1 / 3 of them In a Erlenmeyer flask filled with 0.5‰NaCl sterilized saline.
[0052] 3. Inoculation: Use a sterilized scalpel to cut a 5mm hole in the leaf axil...
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