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Brown planthopper resistant gene Bph41 as well as encoding protein and application thereof

A technology for resistance to brown planthopper and protein, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of non-target biological poisoning, increased drug resistance of brown planthopper, increase production cost, etc., to reduce damage and improve brown planthopper resistance. Effect

Pending Publication Date: 2021-07-30
WUHAN UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the brown planthopper outbreaks mostly occur in the mature rice filling stage, when the rice plants are growing vigorously, it is very difficult to apply insecticides to the base of the rice plants.
In fact, due to the large-scale application of chemical insecticides year after year, the resistance of brown planthoppers has doubled, making the effect of pesticide control limited.
At the same time, the use of chemical pesticides to control brown planthoppers, on the one hand, increases the production cost of farmers, and on the other hand, chemical pesticides also cause environmental and ecological problems such as poisoning of non-target organisms, environmental and food pollution, etc.

Method used

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  • Brown planthopper resistant gene Bph41 as well as encoding protein and application thereof
  • Brown planthopper resistant gene Bph41 as well as encoding protein and application thereof
  • Brown planthopper resistant gene Bph41 as well as encoding protein and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Cloning and molecular marker development of embodiment 1Bph41 gene

[0040] 1. Genome-wide association analysis

[0041] 1,520 rice varieties were obtained from the Institute of Crop Science, Chinese Academy of Agricultural Sciences, located in 74 countries around the world, mainly including indica and japonica rice. The evolutionary trees of these rice varieties are as follows:figure 1 As shown, the regions with different gray levels represent different subpopulations, and different branches represent different types.

[0042] The present invention tests the brown planthopper resistance of 1520 rice varieties, the method is to measure the body weight of the brown planthopper female adults before feeding and after feeding for 48 hours, and then obtain the weight gain rate (WG) of the brown planthopper. For each rice variety, the average body weight gain rate of 12 N. lugens was used to indicate the resistance level. get as figure 2 The results shown are a histogram o...

Embodiment 2

[0054] Functional verification and application of embodiment 2Bph41 gene

[0055] 1. Construction of genetic transformation vector

[0056] Construction of Bph41 gene overexpression vector. The vector used is pCXUN (provided by Professor Wang Guoliang of Ohio State University in the United States). The pCXUN vector is cut with XcmI, and the foreign fragment can be directly connected after adding A.

[0057] The Bph41 gene was directly amplified by PCR, and then connected into the vector after adding A. After the sequence verification is correct, the obtained vector is the Bph41 gene overexpression vector, which is transformed into Agrobacterium EHA105. Pick a single clone for expansion and culture, and after PCR verification, add an equal volume of 50% glycerol to mix, and store at -70°C for later use.

[0058] 2. Genetic Transformation

[0059] The genetic transformation method mediated by Agrobacterium EHA105 (Hiei et al., 1994, Efficient transformation of rice (Oryza sa...

Embodiment 3

[0062] Example 3 Verification of Molecular Markers

[0063] 1. Materials and methods

[0064] 1.1 Materials: BPH-resistant parent G68 (containing the BPH-resistant gene Bph41), BPH-susceptible rice variety Nipponbare and 549 varieties of 3K rice.

[0065] Molecular marker primers: 68-75, the nucleotide sequences of which are shown in SEQ ID No.3-4 respectively.

[0066] 1.2 Method

[0067] Genomic DNA was extracted from rice samples by CTAB extraction. Sample DNA was amplified with primers 68-75. 50 μl system. 50μl reaction system includes: 2×Phanta Max Buffer, 25.0μl; 10mM dNTP, 1.0μl; ddH 2 O, 9.0 μl; 10 μM primer, 2.0 μl; Phanta Max Super-F DNA Polymerase, 1.0 μl, and 50 ng DNA template. The amplification reaction was carried out on a Bioer PCR instrument: 95°C for 3min; 95°C for 15s, 72°C for 60s, 30 cycles; 72°C for 5min. The amplified products of 68-75 were detected by Sanger sequencing in the base forms of G68 and Nipponbare at rs4_11033430, rs4_11033910 and rs4_...

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Abstract

The invention relates to an anti-brown planthopper gene Bph41 as well as a coding protein and application of the anti-brown planthopper gene Bph41. The brown planthopper resistant gene Bph41 comprises any one of the following nucleotide sequences: i) a nucleotide sequence as shown in SEQ ID NO: 1; or ii) a nucleotide sequence which is obtained by substituting, deleting and / or adding one or more nucleotides of the nucleotide sequence as shown in SEQ ID No.1 and is used for coding the same functional protein. The nilaparvata lugens resistant gene Bph41 is found through research; wherein the resistance of rice to nilaparvata lugens can be remarkably improved by transferring the gene into a common rice variety, so that the harm caused by nilaparvata lugens is reduced, and the purposes of yield increase and stable yield are achieved.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to an anti-brown planthopper gene Bph41 and its encoded protein and application. Background technique [0002] Rice is an important food crop, which is the staple food for more than half of the world's people. Since the fine genetic map and physical map of the rice genome have been completed, its transgenic technology is relatively easy, and it has collinearity with other grass crop genomes, so it is regarded as a model plant. With the completion of genome sequencing of various organisms including rice, human beings have entered the post-genome era. It has become the frontier field of life science to carry out functional genome research in an all-round way. Therefore, the study of rice functional genes is of great significance to socioeconomic development and biological research. [0003] Food security is a challenge faced by people all over the world. The two scientific...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12Q1/6895C12N15/11
CPCC07K14/415C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 何光存周聪杜波杨远柱陈荣智祝莉莉秦鹏王凯邓钊
Owner WUHAN UNIV
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