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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

Inactive Publication Date: 2008-10-08
XISHUANGBANNA TROPICAL BOTANICAL GARDEN CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far, the function of the small molecule of Arabidopsis miR396 has not been elucidated in the prior art, and the research on the improvement of the drought tolerance of plants by the small molecule of Arabidopsis miR396 has not been reported so far

Method used

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  • Use of Arabidopsis miR396 micromolecule
  • Use of Arabidopsis miR396 micromolecule
  • Use of Arabidopsis miR396 micromolecule

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 - Cloning of small molecules of miR396

[0026] There are two sites of miR396 in Arabidopsis, which exist on chromosome 2 and chromosome 5 respectively, and both sites can produce corresponding small molecules of miR396, namely miR396a and miR396b (they have only one base difference) (Jones-Rhoades and Bartel, 2004). Using PCR technology, the present invention amplifies and obtains 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 then cloned into a high expression vector containing a strong 35S promoter, which were recorded as 35S:MIR396a and 35S:MIR396b respectively.

Embodiment 2

[0027] Example 2——Screening of high-expression miR396 small molecule transgenic plants

[0028] The constructed high-expression fusion plasmid was transformed into Agrobacterium LBA4404, and the living plant Agrobacterium flower dipping method was used for transformation, and the obtained transgenic seeds were obtained by using kanamycin (50 μg ml -1 ) MS medium for screening to obtain T 1 Transgenic plants were further confirmed by northern blotanalysis. Test results such as figure 2 shown.

Embodiment 3

[0029] Embodiment 3——analysis of drought resistance

[0030] The high expression and wild-type Arabidopsis plants grown for 21 days were saturated with water for 1 day, and then rehydrated after 14 days of continuous drought. The experimental results are as follows image 3 shown. The average survival rate and standard error were obtained by calculating the results of 5 repeated experiments.

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Abstract

The invention discloses a usage of an Arabidopsis miR396 small molecular. The invention relates to a feasible and practical application method for improving the anti-arid ability of plants by utilizing Arabidopsis miR396 small molecular. The method comprises the following steps that the precursor of the Arabidopsis miR396 small molecular is prepared; the expressive fusion plasmid of the corresponding plants is constructed, corresponding agricultural bacillus are transformed; the transformation of the precursor of the Arabidopsis miR396 small molecular is mediated by adopting the recombination of the agricultural bacillus, the precursor of the miR396 small molecular cam be inducted into the Arabidopsis and the corresponding transgenic plant is obtained; and then the evaluation of anti-arid ability of the corresponding Arabidopsis transgenic plant is performed. The invention provides a method that the crops, the garden plants and the new pasture variety with anti-arid ability are cultivated by utilizing the Arabidopsis miR396 small molecular.

Description

technical field [0001] The invention relates to agricultural biotechnology mainly based on transgenic plant technology, and more specifically, the invention relates to improving the ability of plants to resist drought by using small molecules of Arabidopsis thaliana miR396. Background technique [0002] Since 2002, it was rated as the first of the top ten scientific and technological breakthroughs by Science, MicroRNAs (miRNAs) has become one of the hotspots of biological research today. miRNAs are small molecular RNAs with regulatory functions and a size of about 21 bases produced by processing single-stranded RNA precursors (about 70-90 bases in size) with a hairpin structure through Dicer enzymes (Bartel and Bartel, 2003; Bartel 2004). A common feature of miRNAs is that their sequences exist on the stem of a stem-loop structure (Jones-Rhoades and Bartel, 2004). This stem-loop structure is usually an incomplete hairpin structure composed of more than 70 nucleotides, with...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/29C12N15/82A01H1/00C12N15/113
Inventor 余迪求刘冬梅
Owner XISHUANGBANNA TROPICAL BOTANICAL GARDEN CHINESE ACAD OF SCI
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