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Application of cucumber photosynthetic system I reaction center N subunit protein CsPSI-N to resisting melon epidemic diseases

A technology of photosynthetic system and reaction center, applied in the field of plant molecular biology and plant genetic engineering, can solve the problem that the N subunit protein of cucumber photosynthetic system I reaction center has not yet been discovered, and achieve the effect of wide application prospects

Active Publication Date: 2021-12-14
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have shown that the photosynthetic system I reaction center subunit protein plays an important role in plant photosynthesis, but there is no report on the cucumber photosynthetic system I reaction center N subunit protein CsPSI-N in the resistance to melon blight

Method used

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  • Application of cucumber photosynthetic system I reaction center N subunit protein CsPSI-N to resisting melon epidemic diseases
  • Application of cucumber photosynthetic system I reaction center N subunit protein CsPSI-N to resisting melon epidemic diseases
  • Application of cucumber photosynthetic system I reaction center N subunit protein CsPSI-N to resisting melon epidemic diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] EXAMPLE 1 Expression vectors Cucumber CsPSI-N gene construct through the embodiment

[0093] Cucumber CsPSI-N genes belonging to the center of photosystem I N-subunit gene reactions. Cucumber CsPSI-N CDS full length gene (SEQ ID NO.2) to construct pGWB5 (pGWB5 The plasmid map figure 1 The illustrated), the specific position (the alternate switching by genetic recombination ccdB gene), to obtain overexpression vector (pGWB5-CsPSI-N) between the attR1 and attR2 35S promoter; the cucumber CsPSI-N non-conserved region of the gene CDS fragment (CDS108-307bp, SEQ IDNO.4) 200bp fragment and its complementary segment length built onto pK7GWIW (pK7GWIW the plasmid map figure 2 ), As shown in the specific location (exchange replaced by genetic recombination ccdB gene), to obtain a gene silencing vector (pK7GWIW-CsPSI-N) between the attR1 and attR2 35S promoter.

[0094] The amino acid sequence of cucumber CsPSI-N protein is (129aa):

[0095] MSSIGQSILMALAVTLNKFASSNVQSVQRNKATATATVSSPI...

Embodiment 2

[0102] Example 2 Construction of transient overexpression / gene silencing cucumber Model

[0103] (1) in Example 1 overexpression and silencing vector Agrobacterium vectors were GV3101, for 48h on respective inverted resistant medium.

[0104] (2) Pick a single colony were added to 4mL of LB medium containing the appropriate antibiotics and rifampicin, 28 ℃, 180rpm shaking bacteria 24h.

[0105] (3) 1: 100 ratio with fresh LB medium containing the appropriate antibiotics and rifampicin, 28 ℃, 180rpm shaking to OD600 of about 3.0 bacteria.

[0106] (4) 3000rpm centrifuged for 5 minutes to collect the cells, suspensions (10mM MES, 10mM MgCl 2 ) Cells were resuspended and adjusted to OD600 of about 0.4, 200mM acetosyringone was added.

[0107] (5) was allowed to stand at room temperature 3h.

[0108] (6) each pierced with a syringe needle at both sides of a main vein pinhole cucumber cotyledons.

[0109] (7) with a 1mL syringe with the same amount of bacterial suspension after stand...

Embodiment 3

[0121] Example 3 Resistance in Cucumber Experimental

[0122] Construction of successful CsPSI-N instantaneously Example 2 Overexpression of cucumber model, CsPSI-N gene silencing cucumber models and cucumber wild type, the cucumber seedlings after injection of darkness was 22 ℃ lighted culture 3 After 12h cucumber cotyledon days later, inoculated with cucumber and melons Phytophthora culture dish after 28 ℃ in darkness was 24h.

[0123] Depending on cucumber cotyledon treated lesion size to determine the resistance of the gene function of the disease, such attachment results Figure 5 Indicated.

[0124] The results are shown in Cucumber transient overexpression CsPSI-N gene (pGWB5-CsPSI-N) can be made resistant to cucumber disease than in control (Control) increased significantly in cucumber transient gene silencing CsPSI-N (pK7GWIW -CsPSI-N) on the cucumber cotyledon resistance to disease compared with control (control) significantly reduced. Statistics on the lesion area of ​​e...

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Abstract

The invention belongs to the field of plant molecular biology and plant genetic engineering, and discloses application of cucumber photosynthetic system I reaction center N subunit protein CsPSI-N to resisting melon epidemic diseases. It is the first time for the invention to find that the cucumber photosynthetic system I reaction center N subunit protein CsPSI-N has resistance to the melon epidemic diseases: plant cotyledons that overexpress the cucumber photosynthetic system I reaction center N subunit protein CsPSI-N are inoculated with phytophthora melonis, so that the lesion area is obviously reduced, and plant cotyledons that silence the cucumber photosynthetic system I reaction center N subunit protein CsPSI-N are inoculated with the phytophthora melonis, so that the lesion area is obviously increased; it can be seen that the cucumber photosynthetic system I reaction center N subunit protein CsPSI-N plays a very important role in resisting the melon epidemic diseases of plants, can be used for resisting the melon epidemic diseases, preparing products for resisting the melon epidemic diseases and cultivating transgenic plants for resisting the melon epidemic diseases, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the fields of plant molecular biology and plant genetic engineering, and in particular relates to the application of CsPSI-N, an N subunit protein of cucumber photosynthetic system I reaction center, in resisting melon blight. Background technique [0002] Melon blight is a worldwide oomycete disease. The pathogen is Phytophthora melonis, which causes the leaves, stems, and fruits of cucumbers, pumpkins, wax gourds, and bitter melons to turn brown and cause death or dead seedlings. The disease occurs in areas where melons are grown in my country. In the north, the occurrence of summer and autumn open field cultivation and spring protected sweet melons is heavier, and in the south, the occurrence is heavier in spring and summer and autumn with high temperature and humidity. The entire growth period and various parts of melons can be damaged. The damaged growth point of the seedlings and the base of the tender stems are water-so...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/34
CPCC07K14/415C12N15/8282
Inventor 王瑞吴廷全杜虎林毓娥徐晓美金庆敏杨晓珊
Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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