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.

Active Publication Date: 2018-11-23
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the current research on I. clavulinae at home and abroad mainly includes classification and identification, the determination of pathogenicity to insects, and the application of biological control. There are few research materials on I. japonicus.

Method used

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  • Isaria javanica strain and application thereof
  • Isaria javanica strain and application thereof
  • Isaria javanica strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to an Isaria javanica strain, which is classified as Isaria javanica, and is preserved in the China Center for Type Culture Collection with a deposit number of CCTCC M 2018241 onApril 28, 2018. The Isaria javanica strain is separated from ordinary large adult thrips infected with Isaria javanica, and can be used for controlling common thrips. A large number of biological experiments prove that the Isaria javanica strain can not only allow the second instar nymphs and adults of common thrips to parasitize, but also has an effect of controlling more than 80% of common large thrips.

Description

technical field [0001] The invention relates to a strain of I. javanica, which belongs to the technical field of microorganisms. Background technique [0002] The common giant thrips [Megalurothrips usitatus (Bagnall)] is an insect of the thrips subfamily Megalurothrips Bagnall in the order Thysanoptera. In China, the insect is distributed in Hainan, Taiwan, Shaanxi, Guangxi, Guizhou, Hubei and other provinces (regions), while abroad it is distributed in Japan, the Philippines, India, Sri Lanka, Australia, Malaysia, Fiji and other countries. Common thrips mainly damage legume plants. In recent years, this insect has become the main pest on cowpea in Hainan Province. Its nymphs and adults damage young shoots, flowers and young pods, causing growth stagnation, leaf shrinkage and deformity, and flower drop. Pod drop, etc., seriously cause cowpea production reduction, ranging from 20% to 30%, seriously up to 50% to 80%, or even extinction. When the thrips are severely damaged,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N63/04A01P7/04C12R1/645
CPCA01N63/30C12N1/145C12R2001/645
Inventor 范咏梅吴迪孔伟浩孙靖雨周倩应芳
Owner HAINAN UNIVERSITY
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