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Oedaleus decorus asiaticus lactase-phlorizin hydrolase (LPH) as well as coding gene and application thereof

A technology of encoding gene and hydrolase, which is applied in the field of asiatic locust lactase-phlorizin hydrolase LPH and its encoding gene and application, which can solve the problems of lagging disease and insect pest control

Inactive Publication Date: 2018-10-02
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This all shows the disadvantages of the relative lag in the control of crop diseases and insect pests at the present stage.

Method used

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  • Oedaleus decorus asiaticus lactase-phlorizin hydrolase (LPH) as well as coding gene and application thereof
  • Oedaleus decorus asiaticus lactase-phlorizin hydrolase (LPH) as well as coding gene and application thereof
  • Oedaleus decorus asiaticus lactase-phlorizin hydrolase (LPH) as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1, Acquisition of Lactase-Phlorizin Hydrolase LPH and Its Encoding Gene

[0073] 1. Extraction of total RNA from P. asiaticus

[0074] use The RNA isolation reagent was used to extract the RNA from the tissue samples of P. trocarpus. Specific steps are as follows:

[0075] 1) Put a 2mL homogenizer in an oven at 160°C for 3 hours, then cool to room temperature for later use.

[0076] 2) Put the homogenizer on ice, add 1mL RNA isolation reagent and 100-200 mg of P. asiaticus tissue, ground.

[0077] 3) Transfer the homogenate to a 1.5mL centrifuge tube and let stand at room temperature for 5min. Centrifuge at 13000r for 5min at 4°C.

[0078] 4) Transfer the supernatant to a clean 1.5mL centrifuge tube, add 200μL chloroform, and vortex for 15s. Place at room temperature for 5 minutes. Centrifuge at 13000r for 10min at 4°C.

[0079] 5) Transfer 400 μL of supernatant to a new 1.5 mL centrifuge tube, add 200 μL of chloroform, and vortex for 30 seconds. Pla...

Embodiment 2

[0112] Example 2, the dsRNA of the asiatic locust lactase-phlorizin hydrolase gene LPH and its application in pest control

[0113] 1. Synthesis of dsRNA

[0114] Using T7RiboMAX TM The Express RNAi System Kit synthesizes dsRNA. Specific steps are as follows:

[0115] 1) Synthesis of dsRNA primers

[0116] Design primers based on the cloned gene fragments, amplify the target fragment to about 600bp, and introduce T7 promoter at the 5' end of the primers. The primer sequences are as follows: LPH-2F: 5'-GGATCCTAATACGACTCACTATAGGACGGCACCTCCTTGTTCTTG-3'; LPH-2R: 5'-GGATCCTAATACGACTCACTATAGGACTTCTTCGGGCTCAACCAC-3'.

[0117] 2) Preparation of DNA template

[0118] The bacterial liquid plasmid was extracted with a kit, and the plasmid containing the gene fragment (recombinant vector in Example 1) was used as a template, and LPH-2F and LPH-2R were used for PCR amplification to obtain the target fragment containing the T7 promoter sequence.

[0119] The PCR reaction system is as...

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Abstract

The invention discloses oedaleus decorus asiaticus lactase-phlorizin hydrolase (LPH) as well as a coding gene and application thereof. The oedaleus decorus asiaticus LPH is cloned from oedaleus decorus asiaticus at first, and a primer is designed for the oedaleus decorus asiaticus LPH, so as to synthesize a dsRNA for interfering with an oedaleus decorus asiaticus LPH gene; and the dsRNA is introduced into the oedaleus decorus asiaticus through an injection method to perform RNA interfering (RNAi) on the oedaleus decorus asiaticus LPH. A result shows that after the dsRNA is injected into the oedaleus decorus asiaticus, the survival rate, the growth rate, the weight and the overall expressiveness are obviously reduced, which indicates that the oedaleus decorus asiaticus LPH plays an important role in an insect growth process. A new method is provided for molecular regulation and control of pests, and a theoretical basis is also provided for deeply understanding an insect growth mechanismand preparing a new biological pesticide preparation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the lactase-phloridzin hydrolase LPH of the Asian carcus and its coding gene and application. Background technique [0002] my country is a large agricultural country, so the agricultural economy is an important part of the national economy. However, due to the impact of crop diseases and insect pests, the harvest of crops has been greatly reduced, and the agricultural ecological environment and normal growth have been seriously affected. Harmonious development of society and Sustainable development is extremely unfavorable. Traditional pest control is mostly based on emergency control and chemical control, which relies too much on chemical pesticides. All these show the disadvantages of relatively lagging behind the control of crop diseases and insect pests at the present stage. The control of pests and diseases is a long process, which needs to be carried out through c...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12N9/38C12N15/56C12N15/85A01K67/033
CPCA01K67/0339A01K2227/706A01K2267/02C12N9/2402C12N9/2471C12N15/85C12Y302/01023C12Y302/01108
Inventor 张泽华崔伯阳黄训兵郝昆农向群王广君涂雄兵王峰赵海龙芭芭拉侯赛因张
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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