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Phakopsora pachyrhizi molecular probe and application thereof

A molecular probe, soybean rust technology, applied in DNA preparation, DNA/RNA fragment, microbial determination/inspection, etc., can solve the problem of infected leaves infected with disease spots, etc., achieve high accuracy, simple operation, Observe the intuitive effect

Inactive Publication Date: 2012-06-06
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In 2002, American scientists developed soybean rust molecular probe primers based on the sequence of the fungal transcript spacer ITS region, which can effectively distinguish soybean rust from bacterial leaf spot. Needle inoculation volume is 10 5 Spores / mL are not detectable until 6 days after inoculation, and usually at this inoculation intensity, the infected leaves have obvious infection lesions

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  • Phakopsora pachyrhizi molecular probe and application thereof
  • Phakopsora pachyrhizi molecular probe and application thereof
  • Phakopsora pachyrhizi molecular probe and application thereof

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preparation example Construction

[0072] The preparation method of above-mentioned soybean rust molecular probe, it comprises the steps:

[0073] 1). According to the published soybean rust genome sequence information, a genome fragment consisting of 447 bases was selected from the soybean rust genome JGIAFNA-61H15: Phakopsora pachyrhizi clone, and no identical sequence was found by comparing the sequences on Genebank. The fragment of the source, thus inferring that it is a specific fragment of soybean rust, its sequence information is as follows:

[0074] caatatgcca tcagcaagca gtacaaaggg cgctccacac caatctcttt gtaaactgtt 60

[0075]ttaagagatt tgagggaaac agaagcaaca ctagattttg catttttacc ttttcctatt 120

[0076] gaacttggtg ttgaagtggt gtcttgactt tttctatggg cttgaaccat atttaatgct 180

[0077] atgagtacgt gaatgattct gtcaagaaat tcatcaagat ttttgaaagt atccatggca 240

[0078] gagtttttct gggtattttc atttccttca acgccctcta tagtagtat tgatttttga 300

[0079] ttcttgttga aagaaaagca agatgccaaa aatcctgcta aagcacattc aatagataaa ...

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Abstract

The invention relates to a phakopsora pachyrhizi molecular probe, a preparation method thereof and application of the phakopsora pachyrhizi molecular probe. The phakopsora pachyrhizi molecular probe is characterized by being formed by carrying out PCR (Polymerase Chain Reaction) amplification on an upstream primer and a downstream primer, and the sequence of the probe is formed by encoding a section of sequences in a phakopsora pachyrhizi genome; and the length of the segment is 447bp. The preparation method comprises the following steps of: (1) according to the information of an announced phakopsora pachyrhizi genome information, cloning and selecting 447 genome segments formed by basic groups from a phakopsora pachyrhizi genome JGIAFNA-61H15: Phakopsora pachyrhizi; and (2) designing a PCR amplification primer according to the sequence information, wherein the sequence of the upstream primer is 5'GTTATCAGCAAGCAGTACAA-3', and the sequence of the downstream primer is 5'GGGCCAATCTCTTGTTTGAA-3'. The invention has the characteristics of high accuracy, high sensitivity, no strict limit to sampling, no worry about pathogen cross infection and early diagnosis.

Description

technical field [0001] The invention relates to a soybean rust molecular probe and a preparation method thereof, as well as the application of the soybean rust molecular probe, belonging to the field of fungal molecular biology. Background technique [0002] Soybean rust caused by Phakopsora pachyrhizi (Basidomycetes, Urediales, Malampsoraceae, Phakopsora, Phakopsora pachyrhizi Sydow) was once a tropical and subtropical disease. The main disease of soybean production in the region. Since the beginning of the 21st century, soybean rust has been prevalent in Africa and the Americas, causing hundreds of thousands of tons of output loss every year. the third major disease. The control of soybean rust is mainly through chemical control, and the premise of chemical control is to be able to accurately predict the development of the disease. At present, the prediction of soybean rust is mainly based on conventional pathological research methods such as observing the symptoms of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12Q1/68C12N15/11C12N15/10
Inventor 单志慧沙爱华陈海峰陈李淼郝青南孙佃臣田星星周新安
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI