Method for preserving rice black-streaked dwarf virus indoor living bodies
A technology of rice black bar dwarfing and preservation method, which is applied in the field of agricultural science, can solve problems such as the difficulty of breeding disease-resistant varieties, and achieve the effects of broadening the identification time of variety resistance, overcoming the difficulty of long-term preservation of plants, and overcoming technical bottlenecks
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[0019] Embodiment 1, indoor living body preservation method of rice black-streaked dwarf virus
[0020] ① Obtained from fresh plants carrying rice black-streaked dwarf virus.
[0021] In September 2010, rice plants with typical "green dwarf" symptoms were collected in the severe disease area of Ganyu County, Jiangsu Province. The total RNA of rice was extracted according to the instructions of Trizol (Reagent). Primers were designed according to the published RBSDV-S5-ORF2 sequence (primers were synthesized by Shanghai Handsome Biotechnology Co.):
[0022] F: 5′-TAATCAGCGCGATGTCTAC-3′
[0023] R: 5'-CCCCCGCATCTAAGGAG-3'.
[0024] RT-PCR was used to determine whether the diseased plants carried rice black-streaked dwarf virus. Reverse transcription was performed according to the instruction of M-MLV reverse transcriptase (Promega). The PCR amplification system is: 3.0 μL of cDNA template, 0.5 μL of Taq plus enzyme, 0.5 μL of dNTP (10 mmol. -1) 1.0 μL, 1.0 μL of 3’ end p...
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