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Clone and functional analysis of corn anti-fusarium graminearum stem rot gene ZmAuxRP1

A disease resistance, DNA molecule technology, applied in genetic engineering, plant genetic improvement, angiosperm/flowering plants, etc., can solve problems such as increased resistance measurement error, difficulty in corn stalk rot, etc.

Active Publication Date: 2016-01-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the extremely complex traits of corn stalk rot, which are greatly affected by the environment, resulting in increased errors in resistance determination, it is very difficult to carry out research on the genetic basis of corn stalk rot

Method used

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  • Clone and functional analysis of corn anti-fusarium graminearum stem rot gene ZmAuxRP1
  • Clone and functional analysis of corn anti-fusarium graminearum stem rot gene ZmAuxRP1
  • Clone and functional analysis of corn anti-fusarium graminearum stem rot gene ZmAuxRP1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Embodiment 1, the acquisition of ZmAuxRP1 gene

[0083] 1. Discovery of the ZmAuxRP1 gene

[0084] 1. Positioning

[0085] F 1 , F 1 Backcross with Y331 to get BC 1 f 1 , using BC 1 f 1 Segregation groups were initially located. On the basis of initial mapping, select the recombinant individual plants covering the QTL-qRfg2 mapping interval, and further backcross to obtain BC 2 f 1 , until BC 3 f 1 , in BC 3 f 1 In the population of , use the molecular marker screening at both ends of the initial positioning interval to obtain a single plant containing the entire initial positioning donor piece. Using this single plant, this experiment developed a segregation population for fine positioning ( figure 1 ).

[0086] The entire fine-mapped population includes: 983 strains of BC were obtained in 2008 5 f 1 groups (from 15 BC 4 f 1 Exchange single plant), 1888 strains BC in 2009 6 f 1 groups (from 28 BC 5 f 1 Exchange single plant), in 2010 a total of 2...

Embodiment 2

[0129] Example 2, Functional Verification of ZmAuxRP1 Gene

[0130] The gene ZmAuxRP1 shown in sequence 1 is artificially synthesized, the cDNA of the gene is sequence 2, the protein encoded by the gene is named ZmAuxRP1, and the amino acid sequence of the protein is sequence 3.

[0131] 1. RNAi interference to verify the function of the ZmAuxRP1 gene

[0132] 1. Construction of RNAi interference vector

[0133] In order to construct the RNAi interference vector. Specific primers capable of amplifying the conserved exon sequence of the candidate gene ZmAuxRP1 were designed, and PstI-EcoRI and BamHI-XhoI restriction sites were added at both ends of the primers (Table 5).

[0134] Take 1145 fresh samples and use the RNA extraction kit of Tiangen Company to extract total RNA, and then use the reverse transcription kit of Invitrogen Company to obtain cDNA. Using the cDNA as a template, PCR amplification was performed with the primers shown in Table 5 to obtain a 210 bp DNA frag...

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Abstract

The invention discloses clone and functional analysis of corn anti-fusarium graminearum stem rot gene ZmAuxRP1. The invention provides a protein. The protein is following a) or b): a) a protein consisting of amino acid sequences shown in sequence 2 in a sequence table; b) a protein derived from (a) substituting and / or deleting and / or adding one or more amino acid residual groups to the amino acid sequence shown in the sequence 2 in the sequence table and having the same function as (a). The new gene ZmAuxRP1 is discovered, transgenic corn is obtained by interfering the ZmZuxRP1 gene expression in wild corn, and compared with the non-interfered wild ZmAuxRP1, the corn stem rot resistance can be improved; by virtue of over-expression experiment, the interfered ZmAuxRP1 gene expression can be used for the paddy rice to resist the corn stem rot disease.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the cloning and functional analysis of a corn Fusarium graminearum stem rot resistance gene ZmAuxRP1. Background technique [0002] Corn (Zeamays L.) is an important food crop for both grain and feed, as well as an important industrial raw material. Corn is the largest crop in the world. In 2007, the planting area of ​​corn in my country surpassed that of rice. In 2010, the total output of corn surpassed that of rice. It has also become my country's largest food crop worthy of the name. It occupies a pivotal position in my country's food security system. [0003] Corn stalk rot is a serious soil-borne disease in corn production, which occurs in many countries and regions, such as South Africa, India, Canada, the United States, the United Kingdom, Nepal, the Philippines and Australia in Asia. Corn stalk rot has a great impact on corn yield. It was reported in the UK that during 1973-...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29A01H5/00
CPCC07K14/415C12N15/8282
Inventor 徐明良张东峰钟涛叶建荣朱芒张倩倩
Owner CHINA AGRI UNIV
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