A kind of rice grain shape related protein gif1 and its coding gene and application
A technology of related proteins and coding genes, applied in rice grain shape-related protein GIF1 and its coding gene and application field, can solve the problems of unclear and incomplete regulation mechanism of grain shape in rice, complex QTL cloning of quantitative traits, etc.
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Embodiment 1GI
[0058] Embodiment 1 GIF1 gene rice transgenic test
[0059] 1. Construction of GIF1 gene overexpression vector
[0060] In this example, the expression vector PHB (Mao et al., 2005, PNAS 102: 12270-12275, commercially available) was used as the rice transgene carrier. The vector encodes a bacterial origin of replication (ori), a kanamycin resistance gene (Kan r ), hygromycin resistance gene (Hyg r ), herbicide resistance gene (Bar r ), 2×35S promoter, NOS gene termination signal sequence and multiple cloning site (MCS) for connecting target fragments. The GIF1 gene or its fragment can be inserted into the restriction endonuclease site to construct a transgenic plasmid.
[0061] Using the normal wild-type Nipponbare RNA as a template, the first-strand cDNA was synthesized, and the oligonucleotides at the 5' and 3' ends of the GIF1 gene ORF sequence were used as PCR primers (SEQ ID NO:3 and SEQ ID NO:4), Amplify with high-fidelity enzyme, obtain the full-length cDNA (SEQ ID...
Embodiment 2
[0070] Example 2 GIF1 gene function analysis
[0071] The method described in embodiment 1 obtains the transgenic plant of rice GIF1 overexpression, is divided into 5 groups, named respectively OE1-1, OE1-6, OE2-5, OE4-1, OE4-5, and normal control group control , Observing rice grain phenotypes with transgenic T1 generation plants. The result is as Figure 2-6 as shown, image 3 middle bar=1cm, Figure 3-6 Among them, ** is significant at the 0.01 level. It can be seen from the figure that compared with the control group, the grain length, grain width and thousand-grain weight of the plants OE1-1, OE1-6, OE2-5, OE4-1, and OE4-5 transfected with GIF1 overexpression genes were higher than those of the control group. There was a significant increase. It indicated that the overexpression of GIF1 would increase the grain length and grain width, and then lead to the increase of 1000-grain weight.
[0072] From the above analysis, it can be seen that GIF1 can control rice grain...
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