A kind of endophytic bacteria of Atractylodes macrocephala and its application
An endophytic bacteria, Atractylodes macrocephala technology, applied in bacteria, applications, biocides, etc., can solve the problems of Atractylodes root rot that have not been reported.
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Embodiment 1
[0027] The endophytic bacteria are isolated and obtained according to the following steps: take the rhizome of Zhejiang Yuqian healthy fresh Atractylodes Rhizoma Rhizoma Rhizoma Rhizome and wash it under clean water to remove surface impurities. Cut rhizomes of appropriate size, wrap them in gauze and carry out three-step surface disinfection treatment, 75% ethanol solution for 1 min, 1% sodium hypochlorite solution for 3 min, and 75% ethanol solution for 30 s. Disinfect and take it out, add quartz sand and 20mL PBS buffer, fully grind to a slurry, take 100μL of supernatant and spread it on the surface of NA medium, and place it in a mold incubator at 28°C for 2-3 days in dark. Purify the endophytic bacteria of Atractylodes macrocephala by streaking method, observe regularly until a single colony is isolated, and finally obtain the endophytic bacterial strain AM201 of the present invention, such as figure 1 As shown, the overall color of the colony is yellowish-white, and the ...
Embodiment 2
[0030] Take the cryopreservation tube of the AM201 strain, and under aseptic conditions, pick an appropriate amount of bacteria into NB medium, and culture it for 48 hours at 26°C, in the dark, and at 180rpm to reach the OD of the bacteria solution 600 =0.6~1.0; the three kinds of pathogenic fungi of Atractylodes macrocephala root rot were made into bacterial blocks with a sterile puncher with a diameter of 5mm, respectively connected to the center of the freshly prepared PDA medium, and three equidistant ones were placed at the center of the circle 2cm Use a sterile filter paper piece (6 mm in diameter), and use a pipette gun to draw 5 μL of the bacterial suspension to soak the filter paper piece. 5 μL NB culture solution was used as a negative control, and each treatment was repeated 3 times. Put it into a mold incubator, and measure the relative bacteriostatic rate after cultivating in a dark environment at 26±2°C for 5-7 days. The results showed that AM201 endophytic bact...
Embodiment 3
[0032] Take the cryopreservation tube of the AM201 strain, under aseptic conditions, pick a small amount of bacteria with an inoculation needle, insert it into the sterilized NB liquid medium, and cultivate it in the dark at 26°C, 65% humidity, and 180rpm for 48 hours; further Under sterile conditions, draw the bacterial suspension and adjust the bacterial suspension to OD 600 = 0.6~1.0, the diseased plants of Atractylodes macrocephala were applied by root irrigation, the application amount was 5mL / time / 3d, and continued for 21 days, and an equal volume of distilled water was used as the control group, n=30. Investigate and count the number of Atractylodes macrocephala diseased plants, incidence rate, damage area, and death rate of diseased plants, detect the number of pathogenic bacteria with blood counting method, and calculate the disease index (DI) = [∑ (number of disease-level plants × representative value) / sum of the number of plants × the highest level of disease inciden...
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