The upstream and downstream pathways of osspl7 regulating rice plant architecture and its application
A kind of use and crop technology, applied in the field of plant genetic engineering, can solve the problems of rice quality improvement and so on
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Embodiment 1
[0180] Cloning of embodiment 1 OsSPL7 gene
[0181] The sequence number of the OsSPL7 gene is LOC_Os04g46580 (http: / / rice.plantbiology.msu.edu / ). In the present invention, the nucleotide sequence of the OsSPL7 gene is shown in SEQ ID NO.:1. In the present invention, the OsSPL7 gene was cloned and constructed into the overexpression plasmid p1301-35SNos, the tagged vector p1305-3*FLAGNOS, and the RNAi vector DS1301, respectively.
Embodiment 2
[0182] Example 2 OsSPL7 gene overexpression and RNAi phenotype
[0183]The inventors cloned the rice OsSPL7 gene, constructed the coding region of the gene into the overexpression plasmid p1301-35SNos to form the OsSPL7OE plasmid, and genetically transformed the plasmid into wild-type rice ZH11. The transgenic plants showed obvious phenotypes, which were mainly reflected in: ①The roots of the transgenic plants became thicker, and there were almost no lateral roots ( figure 1 B); ②The transgenic plants hardly had tillers ( figure 1 C); ③ transgenic plants are sterile ( figure 1 C).
[0184] Subsequently, an RNAi plasmid of the OsSPL7 gene was constructed and genetically transformed into wild-type rice ZH11. The transgenic plants showed obvious dwarf plant height and increased tiller number phenotype ( figure 2 ).
[0185] Combining the phenotypes of OsSPL7 gene overexpression and RNAi transgenic plants, the results showed that OsSPL7 gene is an important gene affecting ri...
Embodiment 3
[0186] Example 3 OsSPL7 gene is regulated by an upstream functional gene
[0187] Transgenic plants with down-regulated function of the upstream regulatory gene exhibited a phenotype very similar to OsSPL7OE, reflected in thicker roots and reduced tillering ( figure 1 A); and the transgenic plants with overexpression of the upstream regulatory gene show a very similar phenotype with OsSPL7RNAi, the plant height becomes shorter, and the number of tillers increases ( figure 2 ). Meanwhile, the expression of OsSPL7 was obviously up-regulated in the transgenic plants whose function of the upstream regulation gene was down-regulated, and the expression of OsSPL7 was obviously down-regulated in the transgenic plants whose function of the upstream regulation gene was up-regulated ( image 3 ). Therefore, from a functional point of view, it is proved that the OsSPL7 gene is negatively regulated by the upstream gene, and has the function of regulating rice plant height and tiller nu...
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