Screening method of solo-LTR toxic candidate strain with Inago2 in AvrPiz-t

An avrpiz-t, 1.avrpiz-t technology, applied in the field of molecular biology, can solve the problems of screening and identification primers and methods that do not retrieve virulent strains

Pending Publication Date: 2021-08-20
INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI
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Problems solved by technology

[0005] At present, there is no literature, primers or methods for the screening and identification of primers and methods

Method used

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  • Screening method of solo-LTR toxic candidate strain with Inago2 in AvrPiz-t
  • Screening method of solo-LTR toxic candidate strain with Inago2 in AvrPiz-t
  • Screening method of solo-LTR toxic candidate strain with Inago2 in AvrPiz-t

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Embodiment Construction

[0030] 1. Steps and verification of specific primer design

[0031] Primer design: compare the sequence of the upstream region of the insertion site of the AvrPiz-t gene promoter between the rice blast strains containing the insertion and those without the insertion, and select a short oligonucleotide sequence (with a GC content of about 50%) in the conserved region as the front primer; Then a conserved oligonucleotide sequence (with a GC content of about 50%) is selected downstream of the gene coding region (CDS), and its reverse complementary sequence is used as a back primer. Finally, compare whether the primer sequences before and after are easy to form primer dimers, and those that are easy to form primer dimers are redesigned. The positions of the designed front and rear primers include the upstream promoter region and CDS region of the insertion site. Verification: Submit the designed primer sequence to the sequencing company for synthesis, and then use the synthesized...

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Abstract

The invention belongs to the field of molecular biology, and particularly relates to a primer and method for rapidly identifying a toxic strain of a soo-LTR fragment of an Inago2 retrotransposon inserted in a magnaporthe oryzae avirulence gene AvrPiz-t by using a specific primer. The invention develops a pair of specific primers for the first time, and the primers can rapidly detect whether the soo-LTR fragment of the Inago2 retrotransposon is inserted into a promoter region of the magnaporthe oryzae gene AvrPiz-t or not, so that a guarantee is provided for rapid screening of toxic candidate strains, a technical support is provided for dynamic change detection of field strains, guidance information is provided for rice blast resistance breeding and comprehensive prevention and control of rice blast, and materials are provided for development of rice blast pathogenesis research. The primers are strong in specificity and high in sensitivity, can quickly screen out toxic candidate strains of magnaporthe oryzae causing pathopoiesis to IRBLzt-T, and have important scientific research and application values.

Description

technical field [0001] The invention belongs to the field of molecular biology, and specifically relates to a method for rapidly identifying toxic strains by using specific primers to determine whether the avirulent gene AvrPiz-t of rice blast fungus is inserted into the solo-LTR fragment of the Inago2 retrotransposon. Background technique [0002] Rice blast is one of the most devastating fungal diseases in rice production. The use of resistance gene (Resistance gene, R) is the most economical and eco-friendly measure to control rice blast. However, due to the high and rapid mutation of Magnaportheoryzae, rice resistant varieties "lose" resistance after 3-5 years of planting. The interaction between rice resistance gene (R) and rice blast fungus avirulence gene (Avr) conforms to Flor's gene-to-gene theory, that is, when two genes exist at the same time, they can interact to trigger the resistance response of rice. Conversely, the lack of either gene could not trigger an i...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/686C12Q2600/142C12Q2600/156C12Q2565/125
Inventor 李进斌王群陆琳何成兴
Owner INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI
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