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Application of miR1156f in regulating rice root and tiller growth

A technology of mim156f and rice, applied in the field of crop genetics, can solve problems such as the role cannot be underestimated

Inactive Publication Date: 2013-11-13
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the proportion of microRNA in the total RNA of cells is very small, because it can efficiently regulate all mRNAs with target sites, the role of microRNA in crop quality improvement cannot be underestimated.

Method used

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  • Application of miR1156f in regulating rice root and tiller growth
  • Application of miR1156f in regulating rice root and tiller growth
  • Application of miR1156f in regulating rice root and tiller growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Rice mutant up-regulated by embodiment 1miR156f

[0066] Preparation of rice T-DNA insertion mutant population:

[0067] The rice used for transformation is: japonica rice variety Zhonghua 11 (Oryza Sativa L. subsp. Japonica cv. Zhonghua No. 11).

[0068] The plasmid used for transformation is the commercially available pCAMBIA1300 vector, and the maize Ds transposable element is inserted into the T-DNA region of the pCAMBIA1300 vector. At the same time, the Ds contains the bar gene of the herbicide-resistant PPT to obtain the pDsBar1300 plasmid.

[0069] The commercially available Agrobacterium EHA105 was transformed by freeze-thaw method, and the immature embryo tissue of Zhonghua 11 was infected. The resistant callus was obtained by screening on the medium containing 50mg / L hygromycin, and the seedlings were obtained on the differentiation medium. Thus a rice T-DNA insertion mutant population is obtained.

[0070] A dwarf multi-tiller mutant (abbreviated as a bush ...

Embodiment 2

[0074] Example 2 Target mimicry technology to construct transgenic plants with down-regulated miR156f competitive function

[0075] 1. By PCR method, use primers ipsF: GTGGATCCaagaaaaatggccatcccctagc (SEQ ID NO.: 3) and ipsR: CTGGAGCTCgaggaattcactataaagagaatcg (SEQ ID NO.: 4) to amplify to obtain the cDNA sequence of Arabidopsis IPS gene with a length of 522 bp,

[0076] 2. Through BamHI and SacI digestion, clone the fragment into PBSK (pBluescript II SK(+)) vector, Figure 8 The structure of the vector PBSK vector is shown, and the complete sequence of the vector is shown in SEQ ID NO.: 7; spare.

[0077] 3. Then pass the primer MIM156f-I:

[0078] cgaagctTGACAGAAGAtagaAGTGAGCATtttctagaggggagataa (SEQ ID NO.: 5) matches ipsF;

[0079] Primer MIM156f-II:

[0080] cctctagaaaATGCTCACTTCTATCTTCTGTCAagcttcggttcccctcg (SEQ ID NO.: 6) matches ipsR;

[0081] 4. Overlap extension (overlap extension) PCR method [method reference (An et al., 2005)], the miR156f competitive small RNA...

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Abstract

The invention relates to an application of miR1156f in regulating rice root and tiller growth. Specifically, miR1156f can not only regulate rice root growth and / or control rice tiller but also regulate characters such as rice panicle type, grain shape, output, etc. Not only are applications of miR1156f, or miR1156f active segments, or miR1156f antagonist, or miR1156f agonist provided by the invention, but also a method for improving the rice crop is provided.

Description

technical field [0001] The invention belongs to the field of crop genetics, in particular, the invention relates to the application of miR156f in regulating rice root and tiller development. Background technique [0002] Rice is the most important food crop, and more than one-third of the world's population uses rice as a staple food. Because rice has the advantages of small genome, fine genetic and physical maps, relatively easy transgenic technology, and collinearity with other grass crops, rice has always been an excellent model plant. With the completion of rice genome sequencing, human beings have entered the post-genome era, and the comprehensive research on rice functional genome and gene annotation has become the frontier field of life science. Therefore, the study of rice functional genes is of great significance to socioeconomic development and biological research. [0003] In order to solve the contradiction between population growth and the reduction of cultiva...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/11C12N15/63A01H5/00
Inventor 时振英王江戴争妍张景六李琳
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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