Molecular marker for black rot of rape as well as acquisition method and application of molecular marker
A technology of molecular markers and molecular markers, applied in the field of molecular biology, can solve problems such as no fully immune varieties, no black rot, and no identified disease-resistant genes
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Embodiment 1
[0088] Example 1: Black rot pathogen inoculation and identification of resistant material ZS9mXccR-1
[0089] According to the normal growth conditions of rapeseed, 357 natural rapeseed varieties were collected in this laboratory through international and domestic cooperation in greenhouses (59 spring varieties, 255 semi-winter varieties, winter varieties 43; divided by geographical location, 257 Asian varieties, 60 European varieties, 16 Oceania, 13 South America, and 11 African and incomplete varieties), of which 41 were obtained by chemical mutagenesis and multi-generation self-breeding. . The 357 materials were thinned one week after germination, so that about 15 seedlings with strong growth were retained for each variety. Three biological replicates were set during inoculation, and each replicate was inoculated with 5 seedlings. When the seedlings grow to 3-4 unfolded leaves, start to prepare the bacterial solution for inoculation. First, the black rot Xcc bacteria were...
Embodiment 2
[0091] Example 2: Identification of other agronomic traits of resistant material ZS9mXccR-1
[0092] At the mature harvest stage of rapeseed, the resistant variety ZS9mXccR-1 and its source susceptible material ZS9 were copied for multiple traits. The result is as image 3 shown. All data in the figures are mean ± standard deviation of 3 biological replicates (10 plants per replicate). Asterisks indicate significant differences between the two lineages. It can be seen from the figure that the main inflorescence length and 1000-grain weight of the resistant variety ZS9mXccR-1 were significantly higher than those of the susceptible variety ZS9, indicating that the yield of the resistant variety ZS9mXccR-1 was better than that of the susceptible variety ZS9.
Embodiment 3
[0093] Example 3: Submicrostructural identification of the resistant material ZS9mXccR-1
[0094] Eight days after inoculation, leaf tissue at the junction of diseased and healthy was carefully cut into small squares of 3 × 3 mm and immediately fixed in a buffer solution (pH 7.2) containing 2.5% (v / v) glutaraldehyde and 0.1 M phosphate middle. The collected samples were then vacuumed until the blades sank to the bottom and were fully fixed. The fixed samples were sent to the electron microscope platform of the Institute of Virology, Chinese Academy of Sciences for subsequent processing and transmission electron microscope observation. The results are as follows: Figure 4 shown.
[0095] From the results of transmission electron microscopy, it can be seen that after 5 days of inoculation with black rot pathogenic bacteria, the susceptible material ZS9 has a slight plasmolysis of the mesophyll cells compared with the disease-resistant material ZS9mXccR-1, and a few chloroplas...
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