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Corn pathogen induced promoter

A technology of corn sheath blight and promoter, applied in the field of plant genetic engineering, can solve the problems of quality decline, yield reduction and the like

Inactive Publication Date: 2014-01-29
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Corn is one of the most important food crops. Fungal diseases and bacterial diseases of corn will reduce yield and quality

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Sequence analysis of maize GRMZM2G011006 gene promoter pGRMZM2G011006

[0030] First, according to the GRMZM2G011006 gene sequence information in the database, determine the sequence 2011 bp upstream of its initiation codon ATG as its promoter, and its nucleotide sequence is shown in SEQ ID NO.1;

[0031] Select the upstream 2011bp sequence of the maize B73GRMZM2G011006 gene, and use PLACE (Higo K, Ugawa Y, Iwamoto M, Korenaga T. Plant cis-acting regulatory DNA-elements (PLACE). Nucl Acids Res1999, 27:297-300.) software for maize GRMZM2G011006 The promoter nucleotide sequence (SEQ ID NO.1 in the sequence listing) was sequence analyzed.

[0032] The above software was used to search and predict the cis-acting elements in the GRMZM2G011006 gene promoter pGRMZM2G011006. As a result, the promoter contained many homologous sequences with known eukaryotic cis-acting elements. Among them, the first base of the GRMZM2G011006 cDNA sequence is defined as the transcript...

Embodiment 2

[0038] Example 2 Constructing the recombinant vector of GRMZM2G011006 promoter sequence and GUS gene and transforming rice Zhonghua 11

[0039] Taking corn B73DNA as a template, using a forward primer, its sequence is shown in SEQ ID NO.2 (5′-AAG CTGCAG GCCCGAGGAAGACTAATACTAG-3′, the underlined base is the restriction endonuclease PstI recognition site) and reverse primer, its sequence is shown in SEQ ID NO.3 (5′-AAG GTC GAC TGCTCCCCAAATCCAAAC-3ˊ, the underlined base is the restriction endonuclease SalI recognition site) to amplify the promoter fragment;

[0040]The PCR reaction program is as follows: pre-denaturation at 94°C for 5min, denaturation at 94°C for 40s, annealing at 62°C for 40s, extension at 72°C for 2min, 35 cycles of reaction, post-extension at 72°C for 7min,

[0041] A fragment of the sequence SEQ ID NO.1 is obtained, and recognition sites for restriction endonucleases PstI and SalI are respectively added at both ends of the sequence. After the reaction, th...

Embodiment 3

[0043] Example 3 Detecting the induced response of pGRMZM2G011006 transgenic plants to pathogenic bacteria

[0044] The rice Zhonghua 11 transgenic seedlings obtained in Example 2 of the T1 generation grown in the greenhouse for about 3 weeks were subjected to various treatments. The rice seedlings were inoculated with rice bacterial blight pathogen Xanthomonas PXO99, rice bacterial streak RS105 and maize sheath blight YWK-196, and no inoculation was set as the control. The intensity of GUS activity of different transgenic lines before and after inoculation was detected. Among them, bacterial blight and thin streak bacteria were cultured on PSA medium at 28°C for 2 days, resuspended in sterile water, inoculated by injection, and the leaves of the plants were taken 1 day after inoculation of pathogenic bacteria for analysis; corn sheath blight bacteria were cultured on PDA medium Cultivate at 25°C for 2 days, and inoculate the leaves with a puncher to inoculate the leaves, and...

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Abstract

The invention relates to the technical field of plant genetic engineering, and provides a corn pathogen induced promoter which can be activated by rhizoctonia solani kahn, rice bacterial blight pathogen and rice bacterial leaf streak pathogen. After the promoter is verified by truncation, the two sections including -1721bp to -1601bp and -1276bp to -1181bp are confirmed to be response elements induced by pathogenic bacteria. The corn pathogen induced promoter can be used for building a plant expression vector which is used for improving the resistance of plants to various diseases by a plant genetic engineering technology.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, and provides a maize pathogen-induced promoter pGRMZM2G011006, which can be used to improve plant disease resistance and other beneficial production traits through plant genetic engineering technology. Background technique [0002] Plant diseases caused by fungi, bacteria, viruses and nematodes have a huge impact on crop production. Plants have two different resistance mechanisms: one is the passive defense mechanism, which refers to some resistance-related morphological structures, such as epidermis and hard cell wall (Brady JD, Fry S. Formation of di-isodityrosine and loss of isodityrosine in the cell walls of tomato cell-suspension cultures treated with fungal elicitors or H 2 o 2 . Plant Physiol 1997, 115:87-92.). The other is active defense, which is caused by the recognition and interaction between resistance genes and pathogenic bacteria. The elicitor encoded by the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/84A01H5/00
Inventor 储朝晖丁新华李宁卫书佟周文彗
Owner SHANDONG AGRICULTURAL UNIVERSITY
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