Rice transcription factor OsGTBP for regulation and control cis-type reaction element GT-1 and application thereof

A technology of rice transcription factor, GT-1, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems such as yield and quality decline of sheath blight

Inactive Publication Date: 2016-04-20
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Rice is an important food crop in the world, but s

Method used

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  • Rice transcription factor OsGTBP for regulation and control cis-type reaction element GT-1 and application thereof
  • Rice transcription factor OsGTBP for regulation and control cis-type reaction element GT-1 and application thereof
  • Rice transcription factor OsGTBP for regulation and control cis-type reaction element GT-1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Verification of the interaction between OsGTBP and the pathogenic induction element GT-1

[0036] Through the yeast one-hybrid system, we screened a protein with unknown function that interacts with pathogen-inducing elements. This protein contains a conserved WD40 domain and named it OsGTBP.

[0037] Firstly, the interaction was verified by the yeast screening system. Using rice IRBB13cDNA as a template, using a forward primer, its nucleotide sequence is shown in SEQ ID NO.5 (5′-AAG ATCGAT ATGGCCTCGCCGTCATC-3', the underlined base is the restriction endonuclease ClaI recognition site) and reverse primer, its nucleotide sequence is as shown in SEQIDNO.6 (5'-AAG GAGCTC CTCAGTGACATATGTCTTGATCAGG-3', the underlined base is the restriction endonuclease SacI recognition site) to amplify the OsGTBP fragment, the reaction procedure is as follows: pre-denaturation at 94°C for 5min, denaturation at 94°C for 40s, annealing at 55°C for 40s, extension at 72°C for 1min...

Embodiment 2

[0043] Example 2: OsGTBP activates GT-1 and regulates Os2H16 gene expression

[0044] Using the 35Smini sequence as a template, use a forward primer whose nucleotide sequence is shown in SEQ ID NO.11 (5'-GAAAAAGAAAAACGCAAGACCCTTCCTCTATAA-3') and a reverse primer whose nucleotide sequence is shown in SEQ ID NO.12 (5'-GGGACTGACCTACCCGGG- 3ˊ) Amplify the 35Smini fragment with GT-1 element, the reaction procedure is as follows: pre-denaturation at 94°C for 5min, denaturation at 94°C for 40s, annealing at 55°C for 40s, extension at 72°C for 30s, reaction for 35 cycles, and post-extension at 72°C for 7min;

[0045] After the reaction, the PCR product was detected by 1.0% agarose gel electrophoresis, the target fragment was recovered and purified, and the obtained PCR product was verified by sequencing. Tobacco leaves were co-injected with OsGTBP-Myc Agrobacterium GV3101, and fluorescence observation was performed 5 days later. Compared with the control, stronger fluorescence signals...

Embodiment 3

[0051] Example 3: Induced expression pattern of OsGTBP gene after inoculation of pathogenic bacteria

[0052] In order to detect the expression pattern of OsGTBP gene induced by pathogenic bacteria, we used quantitative RT-PCR technology to analyze the expression pattern of OsGTBP gene induced by bacterial blight PXO99 and sheath blight YWK196. Leaf-cutting method was used for the inoculation of bacterial blight to inoculate rice at the adult plant stage, and toothpick insertion method was used for inoculation of sheath blight. Forward primer used, its nucleotide sequence is shown in sequence table SEQIDNO.17, 5'-GCTCCGCCCAGGTCAAG-3', reverse primer, its nucleotide sequence is shown in sequence table SEQIDNO.18, 5'-TCCTTCAGACATGTTCCAATAATCC- 3'.

[0053] The analysis results showed that the OsGTBP gene could be induced by bacterial blight PXO99 and sheath blight YWK196 ( image 3 ), indicating that it is involved in the resistance response of rice to bacterial blight and she...

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Abstract

The invention relates to the technical field of plant gene engineering, in particular to new functional verification and application of a rice transcription factor OsGTBP for a regulation and control cis-type reaction element GT-1. Driving of a strong promoter and the transcription technology based on the RNA interference principle are utilized for transferring an excess expression vector and an RNAi carrier of an OsGTBP gene to a rice variety middle flower (11), and expression of the OsGTBP gene in rice is excessive or is inhibited. Resistance, to xanthomonas oryzae and rhizoctonia solani, of the genetically-transformed rice with the expression quantity of the OsGTBP gene remarkably increased can be enhanced obviously, resistance, to xanthomonas oryzae and rhizoctonia solani, of the genetically-transformed rice with the expression quantity of the OsGTBP gene remarkably decreased can be weakened obviously, and it proves that the OsGTBP gene plays an important role in xanthomonas oryzae and rhizoctonia solani resistance of the rice; meanwhile, tolerance, to drought, of the genetically-transformed rice with the expression quantity of the OsGTBP gene remarkably increased can be enhanced obviously, tolerance, to drought, of the genetically-transformed rice with the expression quantity of the OsGTBP gene remarkably decreased can be weakened obviously, and it proves that the OsGTBP gene also plays an important role in drought resistance.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to OsGTBP, a transcription factor regulating pathogen-inducing element GT-1, and its application in rice improvement, resistance to bacterial blight and sheath blight, and drought resistance. Background technique [0002] Plants have formed constitutive and inducible defense systems to resist the infection of pathogenic bacteria during the evolution process. The inducible system is controlled by two types of genes: disease resistance genes and disease resistance related genes. Most disease resistance genes are race-specific and their products recognize elicitors secreted by pathogenic bacteria. Resistance-associated genes are capable of initiating signal transduction leading to defense responses and most resistance-associated genes are race-nonspecific. [0003] So far, promoters of a large number of disease resistance-related genes have been identified in differ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8205C12N15/8273C12N15/8281C12N2800/60
Inventor 李宁储昭辉丁新华
Owner SHANDONG AGRICULTURAL UNIVERSITY
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