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Reference genes in growth, development and infection stages of peronophythora litchi, and primers and application thereof

A technology of Phytophthora lychee downy mildew and internal reference gene, applied in application, microorganism, genetic engineering and other directions, can solve problems such as hindering the molecular biology research of lychee frost blight, and achieve the effect of important application value

Active Publication Date: 2020-12-15
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the development and screening of internal reference genes of P. litchi is blank, which seriously hinders the molecular biology research of P. litchi. If the growth and development and pathogenic mechanism of P. litchi are to be studied, the reliable internal reference gene identification is Critical

Method used

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  • Reference genes in growth, development and infection stages of peronophythora litchi, and primers and application thereof
  • Reference genes in growth, development and infection stages of peronophythora litchi, and primers and application thereof
  • Reference genes in growth, development and infection stages of peronophythora litchi, and primers and application thereof

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Embodiment 1

[0026] First, by analyzing the transcriptome of P. litchi, we obtained 942 genes with relatively stable expression in the transcriptome data of P. litchi. Further, we screened 4 genes with higher expression levels among these 942 genes. Mold genes (PlACTIN, PLGAPDH, PlrpS5, and Pl006400, whose nucleotide sequences are shown in sequence as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, and SEQ ID NO.4) were further verified by real-time fluorescence Quantitative PCR method was used to measure the expression level changes of these genes in the stages of germination and infection of Pythora peronospora litchi, sporangia, zoospores, restospores, and restospores, and to compare the expression stability of selected genes. details as follows:

[0027] 1. Cultivation of Phytophthora peronosa of litchi. Inoculate P. litchi on carrot solid medium and culture at 28°C for one week until the culture dish is covered.

[0028] 2. Collect the samples of mycelia, sporangia, zoospores, restospores, a...

Embodiment 2

[0056] According to the method of the embodiment of the present invention 1, take P1006400 as internal reference gene, carried out transcript level analysis to Lychee downy mildew PlrpL13, the result is as follows Figure 9 As shown, the expression level of PlrpL13 gene was relatively high in the germination stage of P. litchi peronospora mycelia, sporangia, zoospores, restospores and restospores, but not in the infection stage.

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Abstract

The invention discloses reference genes in growth, development and infection stages of peronophythora litchi, and primers and application thereof. Four peronophythora litchi genes PlACTIN, PLGAPDH, PlrpS5 and Pl006400 are selected, the expression quantity changes of the genes in the germination and infection stages of peronophythora litchi hyphae, sporangia, zoospores, resting spores and resting spores are measured by a real-time fluorescence quantitative PCR method, the stability of the expression quantity of the selected genes is compared, and the result shows that the Pl006400 and PlACTIN can be used as the reference genes with stable expression quantity. The specific primer primers of the reference genes Pl006400 and PlACTIN can be widely applied to the gene expression study in the development and infection stages of peronophythora litchi, the blank of lack of reliable reference genes of peronophythora litchi is filled, and important application values are realized.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an internal reference gene in the growth and development and infection stages of Phytophthora litchie and its primers and application. Background technique [0002] Litchi frost blight caused by Peronophythora litchii is the most serious disease on litchi. It occurs in major production areas such as Guangdong and Guangxi in my country, and the annual incidence rate is about 30%-90%. During the growing period of litchi, the fruit will rot and cause fruit drop, which will also affect the preservation of litchi after picking, and cause a large number of litchi to fall ill during transportation, seriously restricting the development of litchi industry in China. [0003] Real-time fluorescent quantitative PCR has the advantages of high sensitivity, high specificity and high accuracy, and has been used in many research and application fields such as molecular biology, medicine, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/31C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6851C07K14/37C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 孔广辉冯迪南李雯黄佳敏连帅利习平根李敏慧姜子德
Owner SOUTH CHINA AGRI UNIV
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