Isaria javanica strain and application thereof
A coryneform spore and strain technology, applied in the field of microorganisms, can solve the problem of few research data on Corynebacterium javanica, and achieve the effects of large application potential, high insecticidal activity, and high parasitic ability.
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Embodiment 1
[0017] The acquisition of embodiment 1 bacterial strain:
[0018] Common thrips adults infected with I. javanica collected from six major cowpea producing areas in Hainan Province were surface-sterilized with 75% alcohol for 30 seconds, and then rinsed with sterile water three times after surface disinfection with 0.1% mercuric solution for 30 seconds. Dry them on sterilized filter paper, put the dried common thrips adults on the prepared SDAY medium, and put 5 adults in each dish; Observe, pick the growing mycelium to a new SDAY medium, and store it at 4°C after the mycelium is overgrown.
[0019] Said SDAY medium (Sabouraud's dextrose agar medium) formula: 40g of glucose, 10g of peptone, 10g of yeast extract, 20g of agar, 1ml of egg yolk, 10ml of low-fat milk, and supplemented water to 1000ml. When the culture medium is at 45°C, add lactic acid, shake well and pour it into a 9cm-diameter petri dish for use.
Embodiment 2
[0020] Colony traits culture and morphological identification of Example 2 I. javanica strain
[0021] (1) Cultivation of colony traits: transfer the strains stored in the refrigerator at 4°C to a PDA plate with a diameter of 9 cm, activate at 27°C for 5 days, use a puncher with a diameter of 6 mm to cut off the bacterial block at the edge of the colony, and transfer to a new SDAY Plates were cultured in the dark at 27°C and repeated 3 times. From the 2nd day, the colony shape, color and colony growth were recorded until the colonies covered the plate.
[0022] The colony grows slowly on the PDA medium. After 12 days of culture, the colony morphology is shown in figure 1 , the diameter of the colony reaches 3.3-3.4cm, the front of the colony is cotton-like, the aerial hyphae are developed, white, and after 10 days, it becomes white-gray, and the back is initially white, and after 10 days, it becomes light yellow.
[0023] (2) Morphological identification of bacterial strains...
Embodiment 3
[0024] Embodiment 3 studies on the molecular biology of I. javanica BS-1:
[0025] The PCR amplification of the 16SrRNA of I. javanica strain of the present invention can be carried out by conventional methods in the art. The DNA of I. javanica was extracted by the kit method, and the primers used for amplification were general primers, which were synthesized by Shanghai Sangon Bioengineering Service Co., Ltd.
[0026] Universal PCR amplification primers:
[0027] ITS1-F: 5'-CTTGGTCATTTAGAGGAAGT-3'
[0028] ITS4-R: 5'-CCTCCGCTTATTGATATGC-3'
[0029] PCR reaction system (25 μl): template DNA 1 μL, 2×Eco Taq PCR SuperMix (Tiangen) 12.5 μL, upstream and downstream primers 1 μL, add deionized water to 25 μL. Reaction conditions: pre-denaturation at 94°C for 4min; 35 cycles at 94°C for 1min, 30s at 55°C, and 90s at 72°C; extension at 72°C for 10min. The PCR product was detected by 1% agarose gel electrophoresis, the results are shown in figure 2 ; The PCR amplification produc...
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