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Rice brown planthopper resistance gene bph9 and its molecular markers and applications

A technology of anti-BPH and molecular markers, applied in application, genetic engineering, plant gene improvement, etc., can solve problems such as difficult operation, increased production cost, and non-target biological poisoning

Active Publication Date: 2015-10-14
WUHAN UNIV
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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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  • Rice brown planthopper resistance gene bph9 and its molecular markers and applications
  • Rice brown planthopper resistance gene bph9 and its molecular markers and applications
  • Rice brown planthopper resistance gene bph9 and its molecular markers and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0162] Example 1 Positional cloning of Bph9 gene and development of linked molecular markers

[0163] 1. Preliminary positioning results of Bph9

[0164] Bph9-containing F 2 Populations, 93-11 and Pokkali (IRGC 108921) were all from the National Crop Germplasm Conservation Center of the Institute of Crop Science, Chinese Academy of Agricultural Sciences, and used the CTAB method (Murray MG&Thompson, 1980 Rapid isolation of high-molecular-weight plant DNA.Nucleic Acids Res 8 :4321-4325) to extract parent and F 2 Genomic DNA of each individual plant in the population. each F 2 A single plant obtains the corresponding F 2:3 family lineage. To identify F 2 To locate the N. lugens resistance phenotype of each individual plant in the population, the seedling group method was used to investigate the resistance performance of each individual plant of the F2:3 family (see figure 1 ) to F 2:3 Family resistance level represents F 2 BPH-resistant phenotype of individual plants. ...

Embodiment 2

[0221] Example 2 Functional Verification and Application of Bph9 Gene

[0222] 1. Construction of Genetic Transformation Vectors

[0223] (1) Construction of transformation vectors for the Bph9 gene's own promoter, its own complete ORF, and its own transcription terminator region. The vector used was pCAMBIA1301 (purchased from the Center for the Application of Molecular Biology to International Agriculture in Australia), and the pCAMBIA1301 vector was double-digested with SalI and EcoRI to connect the foreign fragment into the SalI and EcoRI sites of pCAMBIA1301.

[0224] According to the screening and sequencing results of the Fosmid genome library, the 3026 bp fragment obtained from the Fosmid clone where the Bph9 gene is located was double-digested with SalI and NcoI as the promoter region of the Bph9 gene (the NcoI site is the translation initiation codon of the Bph9 gene).

[0225] The full-length cDNA clone of the Bph9 gene was digested with NcoI and XhoI to obtain a ...

Embodiment 3

[0234] Example 3 Verification of Molecular Markers

[0235] 1. Materials and methods

[0236] 1.1 Materials: BPH-resistant parent Pokkali (IRGC 108921, containing brown planthopper-resistant gene Bph9), brown planthopper-susceptible rice variety Yangdao 6 (93-11) and F produced by crossing Pokkali and Yangdao 6 23 The family, 93-11 and Pokkali are all from the National Crop Germplasm Conservation Center of the Institute of Crop Science, Chinese Academy of Agricultural Sciences.

[0237] Molecular marker primers: InD2, RM28466, RM28438, InD28450, InD28453, InD14, InD28432, RM28481 and RM28486, the nucleotide sequences of which are respectively shown in SEQ ID No. 4-21.

[0238] 1.2 Method

[0239]Genomic DNA was extracted from rice samples by CTAB extraction. Sample DNA was amplified with primers InD2, RM28466, RM28438, InD28450, InD28453, InD 14, InD28432, RM28481 and RM28486, respectively. 10 μl system. The 10 μl reaction system includes: 10×PCR buffer, 1.0 μl; 10 mM dNT...

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Abstract

The present invention provides a rice brown planthopper resistance gene bph9 which has a nucleotide sequence shown as SEQ ID NO: 1 and a cDNA sequence shown as SEQ ID NO: 2, and the encoding protein and use thereof. The present invention also provides molecular markers of bph9 and use of the markers for screening brown planthopper resistance.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and specifically relates to a brown planthopper-resistant rice gene Bph9, and also relates to a molecular marker of the gene and the application of the gene and the molecular marker in breeding brown planthopper-resistant rice and rice seeds. Background technique [0002] Rice is an important food crop, which is the staple food for more than half of the world's people. At the same time, 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 g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/63C12N1/21C12N5/10C12Q1/68C12N15/82C12N15/84A01H5/00C12N15/11
CPCC07K14/415A01H1/04A01H5/10C12N15/8286Y02A40/146
Inventor 何光存陈荣智王洋荆胜利祝莉莉杜波
Owner WUHAN UNIV
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