Gene for controlling paddy tillering and usage
A gene and rice technology, applied in the direction of genetic engineering, plant gene improvement, application, etc., can solve the problems of unclear molecular mechanism of plant tillering, hovering rice production, etc., and achieve fast and high efficiency
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Embodiment 1
[0042] Example 1: Isolation and cloning of the Osa-MIR156e gene
[0043] 1. Create a mutant library (T0 generation)
[0044] The vector pSMR-J18R used was donated by the Australian CAMBIA Laboratory (Center for the Application of Molecular Biology to International Agriculture). As shown in Figure 1, the japonica rice variety Zhonghua 11 (Oryza sativa L. cv. Zhonghua 11) Randomly insert T-DNA (including Enhancer Trap) into the genome (Springer P S. Gene Traps: Tools for plant development and genomics. Plant Cell, 2000, 12: 1007-1020) to create a mutant library. The mutant library was constructed according to the method described by Wu et al. (Wu C et al., Development of enhancer trap lines for functional analysis of the rice genome. Plant J, 2003, 35: 418-427). About 100,000 independent transformants have been obtained so far.
[0045] 2. Field screening of T1 generation tillering traits
[0046] 3000 copies of T1 generation seeds were selected from the T0 generation mutant ...
Embodiment 2
[0071] Example 2: Functional Verification and Application of Osa-MIR156 Gene
[0072] 1. Construction of genetic transformation vector
[0073] The vector used is pU1301 constructed in our laboratory. pU1301 is a commonly used plant genetic transformation vector pCAMBIA1301 in the world (Sun et al., 2004, Xa26, a gene conferring resistance to Xanthomonas oryzaepv.oryzae in rice, encoding a LRR receptor kinase-like protein. Plant Journal.37: 517-527) An Agrobacterium-mediated genetic transformation vector carrying a maize ubiquitin promoter with constitutive and overexpression characteristics rebuilt on the basis of Agrobacterium (Fig. 7). The pCAMBIA1301 vector was kindly provided by the Australian CAMBIA Laboratory (Center for the Application of Molecular Biology to International Agriculture). Using the PCR method, directly amplify the target gene SEQ.ID.No.1 from the rice genome, the total volume of the PCR reaction system is 20μl, the rice genome DNA template 1ul (about 5...
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