Plant leaf width related protein KRDP1 and a coding gene thereof and application thereof
A plant leaf and plant technology, applied in the field of plant leaf width-related protein KRDP1 and its coding gene and application, can solve problems such as insect resistance and herbicide resistance
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Embodiment 1
[0059] Embodiment 1, discovery of gene
[0060] The inventor of the present invention identified a pair of near-isogenic lines with obvious differences in leaf width from the natural population of the Nongda 108 recombinant inbred line. Using the mapping population constructed by the near-isogenic lines, cloned the Maize KRDP1 gene encodes the protein shown in Sequence 1 of the Sequence Listing. The cDNA of the maize inbred line X178 has the DNA molecule shown in sequence 2 of the sequence table, wherein positions 1-2088 are the open reading frame of the KRDP1 gene.
[0061] details as follows:
[0062] Corn inbred line X178 was crossed with corn inbred line Huang C to obtain F1 generation (namely Nongda 108), and Nongda 108 was self-crossed to obtain F1 generation 2 Generation, F 2 F 6:7 , obtained multiple RILs, including near-isogenic lines W964 and W968. The leaf width of RIL population was normally distributed. The segregation ratio of leaf width in BC1 population w...
Embodiment 2
[0069] Example 2. Obtaining transgenic pure line KRDP1-1 by gene editing and selfing
[0070] 1. Construction of recombinant plasmids
[0071] The basic vector adopts pCAMBIA3301 vector. The selection marker of the pCAMBIA3301 vector in bacteria is the kanamycin resistance gene, which encodes aminoglycoside phosphotransferase. There is no report that its encoded product has toxic side effects on animals. Because it is located on the periphery of the T-DNA region, it will not transfer to plant cells, so it is not pathogenic, and the probability of becoming pathogenic is extremely low. The origin of replication of the pCAMBIA3301 vector, PBR ori, comes from Escherichia coli. Its main function is to help the vector replicate in E. coli. It is widely used in the construction of engineering vectors. It only exists as an origin of replication without encoding protein, and it is located in the T-DNA The periphery of the area cannot be transferred to plant cells, so it is not pathog...
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