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Detection method and kit for anti-PPO inhibitor herbicide Amaranth retroflexi

A technology of retroflexus and herbicide, applied in the field of molecular biology, to achieve the effects of fast speed, low cost, high amplification efficiency and specificity

Active Publication Date: 2021-02-02
北京绿城堡农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The mechanism of resistance to PPO inhibitor herbicides of Amaranthus retroflexus has not been reported yet.

Method used

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  • Detection method and kit for anti-PPO inhibitor herbicide Amaranth retroflexi
  • Detection method and kit for anti-PPO inhibitor herbicide Amaranth retroflexi
  • Detection method and kit for anti-PPO inhibitor herbicide Amaranth retroflexi

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Effect test

Embodiment 1

[0025] The establishment of the detection method of embodiment 1 anti-PPO inhibitor class herbicide retroflexa

[0026] 1. The inventor has collected a large number of samples of Amaranthus retroflexus that are resistant to PPO inhibitor herbicides from different regions in my country, a total of 450 samples, and a total of 100 samples of Amaranthus retroflexus that are sensitive to PPO inhibitors herbicides. Informatics analysis found that Amaranthus retroflexus sensitive to PPO inhibitor herbicides (referred to as sensitive Amaranthus retroflexus) and Amaranthus retroflexus resistant to PPO inhibitors herbicides (referred to as resistant Amaranthus retroflexus) in SEQ ID The amino acid sequence described in NO.2 differs at two positions, and the 128th amino acid in the amino acid sequence described in SEQ ID NO.2 of 426 copies of resistant Amaranthus reflexae is mutated from arginine to glycine. And in the sensitive Amaranthus reflexae, the 128th amino acid in the amino acid s...

Embodiment 2

[0047] Embodiment 2 Application of detection method of the present invention

[0048] Collect samples suspected to have resistance to Amaranthus retroflexus, take about 200 mg of Amaranthus reflexae leaves and quickly add liquid nitrogen to grind, and use the CTAB method to extract genomic DNA, and the DNA concentration must be greater than 5 ng / μL. See Example 1 for the test method. Detecting primers shows that this primer has good specificity to the specificity of amplification Amaranthus reflexae PPX2 gene, and primer PPX2-1f / 1r amplifies PPX2 gene fragment and is about 1000bp ( figure 1 ), using the primers to amplify the resistant and sensitive Amaranthus retroflexus PPX2 genes, the PCR products were sequenced respectively, and the mutation sites were found by comparison ( image 3 ).

[0049] 12 suspected resistant Amaranthus retroflexi samples collected from different soybean fields in Heilongjiang and Inner Mongolia were detected according to the detection method of ...

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Abstract

The invention relates to the technical field of molecular biology, in particular to a detection method and a kit for anti-PPO inhibitor herbicide reflexa retroflexi. The invention provides a specific DNA fragment related to resistance to PPO inhibitor herbicides and a specific primer pair for detecting resistance to PPO inhibitor herbicides, the specific DNA fragment includes nucleosides The arginine at position 128 of the DNA fragment whose acid sequence is shown in SEQ ID NO.1 is mutated into glycine. The detection primer pair provided by the invention has higher amplification efficiency and specificity, and can efficiently, specifically and sensitively amplify the PPX2 gene fragment of A. The method has the advantages of simple operation, fast speed, low cost and high accuracy, and is suitable for popularization and application.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a detection method and a kit for anti-PPO inhibitor herbicide reflexa retroflexi. Background technique [0002] Protoporphyrinogen oxidase (PPO for short) is a key enzyme in plant photosynthesis. PPO inhibitor herbicides are herbicides that target the PPO enzyme. After this type of herbicide is sprayed to weeds, it hinders the synthesis of chlorophyll in the plastid and mitochondria of protoporphyrinogen IX, thereby causing a large amount of accumulation of protoporphyrinogen IX, and a large amount of protoporphyrinogen IX accumulated from the plastid And the mitochondria leak into the cytoplasm, transform into protoporphyrin IX, destroy the synthesis of chlorophyll, affect photosynthesis, make weeds wither and die. In my country, the PPO inhibitor herbicides that are widely used in the control of weeds in soybean fields mainly include: lactofen, acifluorfen, oxyfluor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 黄兆峰魏守辉黄红娟张朝贤周欣欣姜翠兰
Owner 北京绿城堡农业科技有限公司
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