iaam gene expression vector and method for cultivating drought-resisting corn by root specific expression iaam

A gene expression and carrier technology, applied in genetic engineering, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of plant dwarfing, achieve the effects of increasing water absorption, improving drought resistance, and increasing yield

Inactive Publication Date: 2017-03-08
BEIJING AGRO BIOTECH RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are certain deficiencies in transcription factor genes, which often bring relatively large "side effects" while enhancing drought resistance, such as causing plants to become shorter

Method used

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  • iaam gene expression vector and method for cultivating drought-resisting corn by root specific expression iaam
  • iaam gene expression vector and method for cultivating drought-resisting corn by root specific expression iaam
  • iaam gene expression vector and method for cultivating drought-resisting corn by root specific expression iaam

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Experimental program
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Effect test

Embodiment 1

[0037] An iaaM gene expression vector, the vector comprising the promoter EXP18-D shown in SEQ ID NO. 1 and the iaaM gene shown in SEQ ID NO. 3.

[0038] The PGM-35Sbar universal vector has been constructed in the laboratory (Wang Xiaohan et al. Anhui Agricultural Sciences 2012, 40: 12367-12370), the rice root-specific promoter EXP18D replaces the UBQI promoter at the HindIII / SpeI site, and the screening is as shown in SEQ ID NO.2. The marker gene CP4EPSP shown replaces the marker Bar gene between the vector SmaI / SacI, and the target gene iaaM is located between the SpeI / NotI restriction sites. The vector was verified to be correct by sequencing.

[0039] The construction method of the vector includes the following steps:

[0040] 1) PCR amplifies the rice promoter EXP18D, and clones the promoter EXP18D into the vector pMD18-T to obtain the vector pMD18-T: EXP18D;

[0041] 2) Clone the key auxin synthase gene iaaM from Agrobacterium, and clone iaaM into the vector pMD18-T to obtain th...

Embodiment 2

[0046] A method of cultivating drought-resistant corn by expressing the iaaM gene specifically in the root system:

[0047] Source of iaaM gene: According to the cDNA sequence of Agrobacterium gene BAA76346.1 (NCBI sequence number), it was synthesized by Shanghai Shenggong Biological Co., Ltd.

[0048] 1. Vector construction:

[0049] The PGM-35Sbar universal vector has been constructed in the laboratory (Wang Xiaohan et al. Anhui Agricultural Sciences 2012, 40: 12367-12370), the rice root-specific promoter EXP18D replaces the UBQI promoter at the HindIII / SpeI site, and the screening is as shown in SEQ ID NO.2. The marker gene CP4EPSP shown replaces the marker Bar gene between the vector SmaI / SacI, and the target gene iaaM is located between the SpeI / NotI restriction sites. The vector was verified to be correct by sequencing.

[0050] Build steps:

[0051] 1) PCR amplifies the rice promoter EXP18D, and clones the promoter EXP18D into the vector pMD18-T to obtain the vector pMD18-T: EXP...

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Abstract

The invention discloses an iaam gene expression vector and a method for cultivating drought-resisting corn by root specific expression iaaM; the vector includes a promoter EXP18-D and iaaM gene; the method includes steps of structuring iaaM gene expression vector, verifying target gene iaaM by enzyme digestion reaction, preparing linear DNA by a method of PCR (polymerase chain reaction) plasmid amplification, and transforming the linear DNA to the gene group of corn selfing line C92 gene group through a pollen tube path way; through the root specific promoter EXP18-D, auxin is driven to compound specific expression of key enzyme gene iaaM in the corn root; thus the root growth and development are promoted, and the drought resistance is improved.

Description

Technical field [0001] The invention belongs to the field of plant genetic engineering, and specifically relates to an iaaM gene expression vector and a method for cultivating drought-resistant corn by specifically expressing the iaaM gene in the root system. Background technique [0002] The root system plays a vital role in the growth and development of plants, which is mainly reflected in absorbing water and mineral elements from the soil, fixing the plant and forming a symbiotic relationship with the surrounding microorganisms to improve the nutrient utilization efficiency of the plant And resilience. In the process of evolution, the root system has gradually evolved from a very simple structure such as rhizomes into specialized organs with complex structures, which also reflects the importance of the root system to plants. In some cases, genes that regulate root structure can also increase crop utilization efficiency of nitrogen, phosphorus, and water, thereby increasing yi...

Claims

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Application Information

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IPC IPC(8): C12N15/52C12N15/82A01H5/00
CPCC12N9/00C12N15/8227C12N15/8273
Inventor 张中保吴忠义李向龙张春
Owner BEIJING AGRO BIOTECH RES CENT
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