Rice blast gene related with virulence and its use
A rice blast fungus and gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems such as affecting growth and yield, and seedling death.
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[0035] 1. Obtaining of T-DNA insertion mutants of MgURO-D gene:
[0036] This mutant was obtained by the Agrobacterium tumefaciens-mediated transformation method (ATMT) (Caroline et al., 2005).
[0037] 1.1 Cultivation of Agrobacterium
[0038] Pick a single colony of Agrobacterium tumefaciens strain Agl-1 (Mullins, 2001) containing the plasmid pBHt2 (Mullins, 2001), minimal medium (MM; Hooykaas et al., 1979), 50ug / ml kanamycin, 10ug / ml rifampicin, 250rpm, shake culture at 28°C for 48h; centrifuge at 4000rpm for 5 minutes, discard the supernatant, resuspend in induction medium (IM; Bundock et al., 1995), centrifuge at 4000rpm for 5 minutes, discard the supernatant; IM (containing 300 uM acetosyringone) medium resuspended, 28 ° C, 250 rpm shaking shake bacteria 6h.
[0039] 1.2 Acquisition of Magnaporthe grisea spores
[0040] In this study, the pathogenic strain-Y34 (provided by China Agricultural University) isolated from the field of Yunnan, China was selected as the res...
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