Use of Arabidopsis miR396 micromolecule
A kind of Arabidopsis, small molecule technology, applied in the field of agricultural biology, can solve the problems that have not yet been reported, and achieve the effect of improving drought tolerance and broad application prospects
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[0025] Example 1-Cloning of miR396 small molecule
[0026] miR396 has two sites in Arabidopsis thaliana, which exist on chromosome 2 and chromosome 5, and both of these sites can produce corresponding miR396 small molecules, namely miR396a and miR396b (they have only one base Difference) (Jones-Rhoades and Bartel, 2004). Using PCR technology, the present invention amplifies miR396a and miR396b precursor sequences from Arabidopsis genomic DNA, and then clones them into pUCm-T Vector (Fermentas). Through related software ( http: / / www.bioinfo.rpi.edu / applications / mfold / rna / forml.cgi ) To obtain its secondary structure see figure 1 . It was confirmed by sequencing that it was the precursor sequence of miR396 and cloned into a 35S promoter high expression vector containing a strong promoter, which was denoted as 35S: MIR396a and 35S: MIR396b, respectively.
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[0027] Example 2-Screening of small-molecule transgenic plants with high expression of miR396
[0028] The constructed high-expression fusion plasmid was transformed into Agrobacterium LBA4404, and the live plant Agrobacterium soaking method was used for transformation. The obtained transgenic seeds were transformed with kanamycin (50μg ml -1 ) MS medium to obtain T 1 Transgenic plants were further confirmed by northern blot analysis. The test results are as figure 2 Shown.
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[0029] Example 3-Drought resistance analysis
[0030] High expression and wild-type Arabidopsis plants grown for 21 days, saturated with water for 1 day, and then rewatered after 14 days of continuous drought, the experimental results are as follows image 3 Shown. The average survival rate and standard error are calculated from the results of 5 repeated experiments.
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