Pseudomonas lini and application thereof in biological control of purple spot of green Chinese onion
A technology for Pseudomonas and purpura, which is applied to Pseudomonas flax and its application field in the biocontrol of purpura disease on green onions, can solve the problems of drug resistance and environmental pollution of pathogenic bacteria of purpura disease on green onions, and meets the requirements of prevention and treatment. The demand of green onion purpura, easy colonization, and the effect of strong antagonism
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Embodiment 1
[0019] Example 1 Isolation and identification of Pseudomonas lini X85
[0020] The antagonistic bacteria strain of scallion purple spot disease pathogen, numbered X85, was isolated, screened and cultivated from the surface of scallion purple spot disease occurrence field in Xinye County, Henan Province; it was inoculated on NA medium plate, 30°C, and cultivated for 24 hours; the colony was milky white , translucent, smooth surface, regular edges ( figure 1 ); microscopically, the bacterium is rod-shaped, with blunt round ends, and the size is 1.5-1.7 μm in length and 0.4-0.6 μm in width ( figure 2 ).
[0021] 16S rDNA identification:
[0022] Pick a small amount of single colony, put it in a PCR tube, add 20 μL of deionized water, treat at 95°C for 10 minutes, and use it as a PCR template after cooling for PCR amplification. The primers used for amplification were general primers 27F and 1492R for bacterial 16S rDNA sequence.
[0023] The PCR amplification system is 20 μL...
Embodiment 2
[0027] Embodiment 2 identification X85 is to the inhibitory action of scallion purple spot pathogenic bacteria (confrontation culture)
[0028] Carry out confrontation culture between X85 and the pathogenic bacteria of green onion purple spot disease (isolated from the diseased green onion leaves):
[0029] The X85 was inoculated onto the PDA plate by the spot method, and the scallion purple spot pathogenic bacteria cake (diameter 5mm) was inoculated 2cm away from the inoculation point; 3 replicates and a blank control were set. Cultivate at 25°C for 7 days, record the width of the inhibition zone, and judge the inhibitory strength against the pathogenic bacteria of scallion purple spot according to the width and growth of the inhibition zone. Such as Figure 4 As shown, the antibacterial bandwidth of X85 indicates that it has a high inhibitory efficiency to the growth of the pathogenic bacteria of scallion purple spot.
Embodiment 3
[0030] Example 3 Identification of the inhibitory effect of X85 on the scallion purple spot pathogen
[0031] Inoculate the purified X85 single colony on Pseudomonas basal medium (bovine muscle tissue digest 16.0g, enzymatically hydrolyzed casein 10.0g, K 2 SO 4 10.0 g, magnesium chloride hexahydrate MgCl 2 ·6H 2 O 1.4g, agar 11.0g, pH=7.1±0.2), after culturing at 30°C for 48h, insert 100mL Pseudomonas basal liquid medium (bovine muscle tissue digest 16.0g, enzymatically hydrolyzed casein 10.0g 、K 2 SO 4 10.0g, MgCl 2 ·6H2O 1.4g, pH=7.0±0.2) in a 250mL Erlenmeyer flask, cultured with shaking at 30°C and 200r / min for 24h. The bacterial suspension was centrifuged at 12000 rpm, and then filtered through a bacterial filter with a pore size of 0.22 μm.
[0032] Test group: Take 1mL of the filtrate and add it to 19mL of PDA medium at about 45°C, mix well, and after the medium is cooled, add the pathogenic bacteria cake of scallion purple spot disease (diameter 5mm);
[0033]...
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