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

[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...

[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...

[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...