Application of yellow mosaic virus P2 protein and gene thereof in cultivation of wheat resistant to yellow mosaic disease
A protein and wheat technology, which can be used in applications, angiosperms/flowering plants, introduction of foreign genetic material using vectors, etc., which can solve the problem of uncertain function of P2 protein.
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Embodiment 1
[0037] Embodiment 1, the cultivation of turning P2 wheat
[0038] 1. Acquisition of recombinant vector p34 and plasmid pAM2001
[0039] 1. Preparation of recombinant vector p34
[0040] WY-8: 5′-CTCAACCATATGGTCGGCTCAGCTGAGG-3′, corresponding to 904~931nt of WYMV RNA2 (1~28nt of P2 protein gene fragment),
[0041] WY-9: 5′-ATGCGGATCCCTAATGGCCCGG-3′, complementary to 2881~2860nt of WYMV RNA2 (1978~1957nt of P2 protein gene fragment). The DNA molecule shown in the artificially synthesized sequence 1 was used as a template, and PCR amplification was performed with primers WY-8 and WY-9 to obtain a PCR product of 1978bp.
[0042] After sequencing, the PCR product has the nucleotides shown in sequence 1 in the sequence listing (the coding region is the 10th-1968 nucleotides from the 5' end of sequence 1 in the sequence listing), and is the full-length gene P2 of the P2 protein. The protein encoded by the gene is P2, and the amino acid sequence of the protein is sequence 2 in the ...
Embodiment 2
[0123] Embodiment 2, the anti-yellow mosaic virus identification of P2 protein wheat
[0124] According to the symptoms caused by wheat yellow mosaic virus damage to wheat in the field, the disease evaluation criteria were formulated:
[0125] Grade 0 (N0): no obvious symptoms;
[0126] Level 1 (N1): Mild mosaic, leaves do not produce shuttle pattern or yellow mosaic symptoms, plants do not dwarf;
[0127] Level 2 (N2): Obvious shuttles or yellow mosaic leaves, accounting for about 50% of the leaf area, and the plants appear dwarfed;
[0128] Grade 3 (N3): Severe shuttles or yellow mosaic leaves, accounting for 75% or more of the leaf area, twisted heart leaves, reduced tillers, and some leaves or the whole plant are dead.
[0129] Wheat yellow mosaic disease is caused by wheat yellow mosaic virus (WYMV).
[0130] The wheat yellow mosaic field in the Yushan area of the Lixiahe Agricultural Science Institute in Jiangsu Province was used as a disease nursery for the resista...
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