Herbicide-resistant brassica plants and methods of use
A herbicide and herbicide-tolerant technology, applied in botany equipment and methods, biochemical equipment and methods, plant products, etc., can solve problems that depend on the availability of imidazolinone-resistant varieties
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Embodiment 1-A
[0224] Example 1 - AHAS in vitro enzyme test
[0225]The AHAS enzyme test is a rapid chromogenic method that was used to quantify the level of drug resistance in different samples by determining the level of AHAS enzyme activity in the presence of an AHAS inhibitor, as described by Singh et al. (Anal. Biochem. 171: 173-179, 1988). Two types of tests were used: a basal test using only one inhibitor and an exact test requiring the use of both inhibitors. Both tests indicate the level of imidazolinone resistance; the exact test is able to detect subtle differences in the level of resistance between some plant lines. The AHAS test stock solution contains: 0.2M monobasic sodium phosphate + 0.2M dibasic sodium phosphate + 50M 1,1 cyclopropane dicarboxylic acid (CPCA) + full strength Murashige & Skoogs alkaline salts + 1mM imazethapyr (AC299 , 263 industrial grade)+5%H 2 SO 4 + 2M NaOH + 2.5% alpha-naphthol + 0.25% sarcosine in 1M phosphate buffer at pH 6.0.
[0226] The final A...
Embodiment 2
[0240] Example 2 - Herbicide Resistance Test in Greenhouse
[0241] The first experiment was designed to determine imidazolinone herbicide resistance in Brassica napus lines containing one gene (bR or PM2) and two genes (bR / PM2) relative to rapeseed lines containing two genes (PM1 / PM2) gender difference.
[0242] Six individual plants from each line were subjected to each spray treatment. Apply 1x (17g ai / acre), 2x (34g ai / acre) and 3x (51g ai / acre) imidazolinone herbicides at the 2-3 leaf stage Spray at the 2-3 leaf stage about 14 days after planting. Set the spray gun to 40 psi and the speed to '80' (34.98 L / ac). The following calculation was performed to prepare 25ml of Odyssey stock solution: 17*0.025 / 34.98=0.1215g of Odyssey granules. This is based on the following numerical estimates: the amount of Odyssey required per acre is 17g, and one advance of the spray gun delivers 8.33ml of solution. Add at an application rate of 0.5L / 100L or 0.000125L or 125μL After spr...
Embodiment 3
[0258] Example 3 - Herbicide resistance test in the field
[0259] Four mustard entries and one canola entry were tested for herbicide resistance (see Table 4 for different treatments) and tested for yield in a randomized split-plot design experiment (4 replicates) at four locations in North Dakota. The plots are a minimum size of 1.5x5m, each plot is swathed and harvested at maturity. Of the 4 mustard entries, 1 is a PM1 / PM2 mustard line, which is the introgression of the PM1 and PM2 mutations from rapeseed into mustard by conventional backcrossing techniques, followed by two generations of selfing to generate homozygous mustard PM1 / PM2. The other three mustard entries were different mustard genotypes containing the B genome bR mutation and stacked PM2 mutations. All bR / PM2 mustard lines were homozygous for both mutations. Rapeseed entries are Homozygous for the PM1 / PM2 mutation in a commercially available control variety. Crop injury ratings (% phytotoxicity) were per...
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