A rice semi-dominant brittle stalk control gene, molecular marker and application
A semi-dominant, rice technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as affecting the mechanical strength of plants and affecting the synthesis of secondary cell walls.
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Embodiment 1
[0079] Phenotypic analysis of sdbc1 mutants
[0080] Using heavy ions 12 C 6+ Mutagenesis (energy 80MeV, dose 120Gy) japonica rice variety Wuyunjing 7 (WYJ7) was used to obtain the sdbc1 mutant, and the F1 generation plants were obtained by crossing the sdbc1 mutant with the wild type WYJ7, and the F1 generation was selfed to obtain the F2 generation population, as shown in the table As shown in 1, in the segregation of the F2 generation, the segregation ratio of wild type: slightly brittle: extra brittle is 1:2:1, so it can be determined that the mutant is a semi-dominant brittle culm mutant.
[0081] Table 1 is the genetic analysis table of sdbc1 mutants
[0082]
[0083] 2. Determination of agronomic traits
[0084] The agronomic traits of sdbc1 mutants, wild-type WT, and F1 generations were determined, such as Figure 1-Figure 11 As shown, compared with the wild type, the mutant has no significant difference in plant height, but the stems and leaves are very easy to...
Embodiment 2
[0098] Fine mapping of SDBC1 gene in map-based cloning
[0099] 1. Positioning group construction
[0100] The sdbc1 mutant was crossed with the indica rice variety 93-11, and the F1 generation was all of the slightly brittle phenotype, and the F1 generation was self-crossed, and the extra-fragile individual plants and normal individual plants in the segregation population of the F2 generation were selected as the positioning population. About 100 mg of leaves were taken from each plant for DNA extraction.
[0101] 2. Simple sequence repeat (SSR, Simple Sequence Repeat) polymorphism screening
[0102] The sdbc1 mutant and 93-11 were screened for polymorphisms using the reported SSR primers evenly distributed on 12 rice chromosomes, and polymorphic SSR primers were obtained for the next step.
[0103] 3. Preliminary mapping of SDBC1 gene
[0104] For the mapping population consisting of 50 ultra-fragile individuals in the F2 population, the SSR primers obtained by screening ...
Embodiment 3
[0123] Mutations in sdbc1 do not affect its expression pattern and subcellular localization
[0124] 1. sdbc1 mutation does not affect its expression pattern
[0125] The expression pattern of a gene determines the function of that gene. Using qRT-PCR to analyze the expression level of SDBC1 gene in different tissues of wild type, sdbc1 mutant and F1, the results are as follows Figure 26 As shown, the gene was expressed at the same level in the three materials, indicating that the sdbc1 mutation did not affect its expression pattern. The primers used in the above qRT-PCR analysis are as follows:
[0126] qSDBC1-F: 5'-GAGCGCTTTCTCTACATCAACACC-3'
[0127] qSDBC1-R: 5'-AACTCGCAAACGTGCTAATCGG-3'
[0128] 2. sdbc1 mutation does not change its subcellular localization
[0129] The subcellular localization of a protein determines its function. Using the cDNA of wild-type and sdbc1 mutants, and the following primers to amplify the relevant fragments and insert them into pCAMBIA...
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