A pxcg2 strain of carnobacter maltia aromaticum of diamondback moth and its application
The technology of Carnobacterium aromaticum and diamondback moth is applied in the field of agricultural microorganisms to achieve the effects of good biological control potential and application prospects, and improving insecticidal activity.
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Embodiment 1
[0023] Example 1 Isolation and culture of strain PxCG2
[0024] (1) Preparation of selective separation medium:
[0025] LB medium (10g peptone, 5g yeast extract, 10g NaCl, 15g agar, dissolved in 1L sterile ddH2O water, adjust pH to 7.0);
[0026] Nutrient agar medium NA (3 g beef extract, 10 g peptone, 5 g NaCl, 15 g agar, dissolved in 1 L sterile ddH2O, adjusted to pH 7.0).
[0027] (2) Dissection and plating: Dissect the 3rd instar larvae of Plutella xylostella in an ultra-clean bench under sterile conditions and take the contents of the midgut, centrifuge to separate the supernatant and precipitate, and dilute to 5 concentration gradients, according to the dilution of 10-4 After -10-5 times, the NA plates were respectively coated, placed in a 30°C constant temperature incubator, and observed every 24 hours;
[0028] (3) Continuous purification and culture and photographing: After a single colony grows in the selective medium, first pick a single colony according to the c...
Embodiment 2
[0029] Example 2 Identification and phylogenetic analysis of strain PxCG2
[0030] 1. Traditional biological identification
[0031] (1) Morphological characteristics of colonies
[0032] From the morphological point of view, PxCG2 bacteria are round white colonies with smooth and glossy surface, single, paired or chain-like slender bacilli. Heterofermentation, mainly L(+)-lactic acid is produced from glucose. The optimum growth temperature is 30 °C, aerobic growth, and slow growth under anaerobic conditions.
[0033] (2) Determination of Physiological and Biochemical Characteristics of Carnotella maltata
[0034] Bacterial physiology and biochemical assays were carried out with reference to the method in the Manual for the Identification of Common Bacterial Systems, using bacterial microbiochemical reaction tubes. The test results of available carbon sources are shown in Table 1. Galactose, sorbitol, rhamnose, arabinose, xylose, melibiose, D-ribose, sucrose, and yellow pi...
Embodiment 3
[0043] Example 3 Synergistic effect of PxCG2 strain on Bt
[0044] 1. Removal of intestinal bacteria of diamondback moth
[0045] Prepare antibiotic solution: 1mg / mL ciprofloxacin; 1mg / mL levofloxacin; 2mg / mL metronidazole;
[0046] Methods for the elimination of intestinal bacteria of the diamondback moth, and the addition of antibiotics. Specific method: Collect and sterilize the egg card of diamondback moth: collect the egg card produced by the diamondback moth in its peak spawning period, first place it in 5‰ sodium hypochlorite disinfectant for 10 minutes, and then soak it in clean water twice, 5 times each time minutes, and finally dried with absorbent paper and placed in a worm box, and fed with feed after 24 hours;
[0047] Antibiotic feeding treatment: Weigh 4 g of diamondback moth artificial feed into a sterile petri dish, then add more than 333 μL of the prepared ciprofloxacin, levofloxacin and metronidazole antibiotic solutions, mix well and place them in a worm ...
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