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Protein for stimulating hypersensitive response of plants and encoding genes of protein

An allergic reaction and protein technology, which is applied in the fields of plant genetic improvement, plant products, genetic engineering, etc., can solve the problem of undiscovered disease-resistant immune response XopP protein, etc., and achieve the effect of important practical value

Inactive Publication Date: 2017-06-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] After searching the literature of the prior art, it is found that there is no report on the XopP protein and the corresponding coding gene xopP that can stimulate the plant to produce a disease-resistant immune response in the phytopathogenic Xanthomonas

Method used

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  • Protein for stimulating hypersensitive response of plants and encoding genes of protein
  • Protein for stimulating hypersensitive response of plants and encoding genes of protein
  • Protein for stimulating hypersensitive response of plants and encoding genes of protein

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1, phytopathogenic Xanthomonas xopP gene prokaryotic expression vector construction

[0033] The present embodiment is obtained from the Xoc RS105 bacterial strain of the rice stripe bacterium, Xoo PXO99 of the rice bacterial blight bacterium A The xopP gene in bacterial strain and Xac 29-1 strain DNA of X. citri was used as a template, and primers xopP1-F / R (SEQ ID NO.3 and NO.4) and xopP2-F / R (SEQ ID NO.5 and NO. 6) Perform PCR amplification, and the PCR amplification conditions are: 94°C / 4min+(94°C / 30sec+55°C / 30sec+72°C / 2min)×32 cycles+72°C / 10min.

[0034] Using the primer Xopp1-F whose sequence is shown in SEQ ID NO.5 and the primer Xopp1-R whose sequence is shown in SEQ ID NO.6, through PCR amplification, it can be obtained from rice stripe Xoc RS105 strain, rice bacterial blight Germ Xoo PXO99 AThe xopP1 gene is amplified from the genomic DNA of the bacterial strain and Xac 29-1 strain of Xacillus citri, the nucleotide sequence of which is shown in SE...

Embodiment 2

[0040] Example 2, XopP Observation of Anaphylaxis on Tobacco Mediated by Agrobacterium

[0041] Activate the Agrobacterium strains EH105 (XopP1) and EH105 (XopP2) to be determined on a fresh plate, EH105 (EV) as a negative control, and EH105 (Hpa1) as a positive control, respectively transferred to the liquid medium of LB For mutants or zygotes, the corresponding antibiotics should be added, shake culture at 28°C for about 12-16 hours, and the bacterial concentration should reach 10 8 CFU / mL; collect the bacteria by low-speed centrifugation (600 = 0.6. Select moderately growing tobacco from the greenhouse and leave it at room temperature for 24 hours. Use a sterile needle to gently poke the inoculation port on the back of the leaf (be careful not to break the leaf), draw the bacterial solution into a needle-free sterile syringe, and mark it on the leaf Injection range, observe the phenotype after about 48 hours, and take pictures. In this example, Sansheng tobacco is Nicotia...

Embodiment 3

[0042] Embodiment 3, XopP stimulates the determination of the minimum concentration of tobacco allergic reaction

[0043] In this example, XopP adjusts different concentration values ​​and inoculates Sansheng tobacco respectively to determine the minimum HR excitation concentration. At the same time, inject tobacco with the same concentration of Hpa1 as a positive control, and Agrobacterium with an empty plasmid as a negative control. After 48 hours, observe the tobacco Allergic reaction to leaves. It was found that the minimum concentration of XopP to stimulate HR was 0.1 mg / ml (see figure 2 ).

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Abstract

The invention relates to a protein for stimulating hypersensitive response of plants and encoding genes of the protein, and belongs to the field of biotechnologies. XopP1 amino acid sequences of the protein for stimulating the hypersensitive response of the plants are shown as SEQ ID NO.2, and xopP1 nucleotide sequences of the encoding genes of the protein are shown as SEQ ID NO.1; XopP2 amino acid sequences of the protein are shown as SEQ ID NO.4, and xopP2 nucleotide sequences of the encoding genes of the protein are shown as SEQ ID NO.3. The content of cysteine in amino acid sequences of the two encoding genes is lower than 1%, the isoelectric point of the protein is 8.8, the protein is weakly alkaline and contains abundant aliphatic amino acid such as alanine and leucine and abundant alkaline amino acid such as serine and arginine, the molecular weight of the protein is 7.9 kD approximately, and the total content of the alanine and the leucine can reach 25% at least. Hypersensitive response can be carried out by tobaccos. Compared with the prior art, the protein and the encoding genes have the advantages that xopP genes can be used as gene resources for plant disease resistance breeding, and XopP proteins can be used as protein medicines for stimulating the disease resistance of the plants.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a protein and a gene thereof that stimulate plants to produce allergic reactions. Background technique [0002] The resistance of plants to pathogen infection is manifested in two aspects: non-host and host resistance. Gram-negative plant pathogenic bacteria produce hypersensitive response (Hypersensitive response, HR) on non-host plants is determined by certain elicitors in the pathogenic bacteria. This kind of HR is an anti-disease immune response, and the corresponding proteins that stimulate HR are called harpin-like proteins. The harpin-like proteins that have been identified from Gram-negative plant pathogenic bacteria include HrpZ, HrpN, HrpW, PopA1, Hpa1, HopAK1 and HopP1, and it is clear that HrpW and Hpa1 in plant pathogenic Xanthomonas stimulate non-host Plants produce HR harpin-like proteins. These harpin proteins have common characteristics: hydrophilic, g...

Claims

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Application Information

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IPC IPC(8): C07K14/195C12N15/31C12N15/10C12N1/21C12N15/82A01H5/00
CPCC07K14/195C12N15/8281
Inventor 陈功友蔡璐璐杨阳阳陈晓斌邹丽芳黄坤炫
Owner SHANGHAI JIAO TONG UNIV
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