Rice spikelet development regulation protein, encoding genes MS1 thereof and application
A technology for regulating proteins and rice, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as affecting yield
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Embodiment 1
[0046] 1. Rice material:
[0047] Rice (Oryzasativa L.) mutant ms1 (malformedspikelet1), the original wild-type material is the japonica rice variety "Zhonghua 11". The ms1 mutant comes from the mutation generated by the EMS (EthylMethylSulfonate) mutagenesis of Zhonghua 11 (such as figure 1 shown).
[0048] 2. Analyze and target groups:
[0049] The reciprocal cross experiment between the ms1 mutant and "Zhonghua 11" showed that the mutant was controlled by a single recessive gene. The homozygous ms1 mutant was crossed with the indica rice variety NJ06, F 1 Generation selfing, and from F 2 From the population, 962 individuals with abnormal glume and small grain phenotype were selected as the positioning population. At the heading stage, about 1 gram of young leaves were taken from each plant to extract the total DNA.
[0050] 3. DNA extraction
[0051]Genomic DNA for gene localization was extracted from rice leaves by a rapid extraction method of rice trace DNA. Abou...
Embodiment 2
[0062] Plant Transformation:
[0063] Utilize homologous recombination and PCR technology to introduce the SalI restriction site into the amplification primer, the amplification primer is: M34ORF-1F: ATGGGGCGAGGCAAGGTGGTG (SEQ ID NO: 22), M34ORF-1R: CTAGGCCATCCACTCAGGAGG (SEQ ID NO: 23), the annealing temperature is 60°C. The front and rear primers contain SalI restriction sites. Use this primer to PCR amplify the cDNA of Zhonghua 11, recover and purify a 720bp fragment after electrophoresis, and also use SalI to single-enzyme digest the 35S-GFP(S65T)-NOS(pCA1301) vector, and then recover and purify the product with the correct size of the fragment Connect to the digested vector for Escherichia coli transformation, and select positive single clones for sequencing. Obtain the correct transformation vector pCA1301-35S-MS1ORF (such as Figure 6 shown), the rice ms1 mutant was transformed into Agrobacterium tumefaciens strain LBA4404 by electric shock. We used the calli induce...
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