Rice ageing control gene OsCDC48E and coded protein thereof
A protein and gene technology, applied in the field of plant genetic engineering
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Embodiment 1
[0024] Example 1: Isolation and genetic analysis of mutants
[0025] Through the screening of the rice (IR64) mutant library induced by EMS, a rice premature senescence mutant W95 was isolated and its phenotype can be inherited stably, and the premature senescence phenotype began to appear at the tillering stage figure 1 As shown in 1, 2), the premature senescence phenotype is very obvious at the heading stage (such as figure 1 Shown in 3, 4). The mutant W95 was crossed with Moroberekan to obtain F1. The F1 plants had normal leaf phenotype and growth period. The segregation ratio of the wild-type and mutant phenotypes in the F2 generation was consistent with the segregation ratio of the Mendelian single recessive mutation, indicating that the mutant The progeria phenotype of W95 is controlled by a single recessive gene.
Embodiment 2
[0026] Example 2: Fine Mapping of Mutant Genes
[0027] According to the published molecular genetic maps of japonica and indica rice, select SSR primers that are approximately evenly distributed on each chromosome for polymorphism detection and preliminary location analysis, and the F2 population obtained by crossing mutant W95 with Moroberekan has a mutant phenotype A total of 3429 F2 individuals with mutant phenotype were identified, and 1192 of them were used as the initial mapping population. The preliminary mapping results showed that the mutant gene was located between RM14394 and RM14408 on the short arm of chromosome 3.
[0028] The primers RM6829, HQN195, HQN32, RM14395 and HQN141 that were polymorphic between the parents between RM14394 and RM14408 were screened, and these 7 pairs of primers were used to analyze the mutant individuals in 2237 F2 populations, and the obtained information was used to construct a physical map. Finally, the mutated gene was finely ma...
Embodiment 3
[0029] Example 3: Gene prediction and sequence comparison analysis
[0030] According to the above results, the sequencing analysis of the three predicted genes in this interval was carried out. It was found that the gene encoding cell division control protein 48 homolog E protein was compared with the wild type, the 2347th position of the CDS nucleotide series of the gene in the mutant W95 was changed from C to T, resulting in the premature termination of the protein encoded by this nucleotide , so that the function of the protein is affected, so the gene is designated as a candidate gene and named as OsCDC48E .
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