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Small-molecule ribonucleic acid (RNA) Osa-miR393 for improving rice tillering and application

A rnaosa-mir393, small molecule technology, applied in the field of plant genetic engineering, can solve problems such as unseen miR393

Inactive Publication Date: 2012-07-04
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some studies have been done on the function of miR393, they all focus on the induction of plant response to stress. There is no report that miR393 regulates plant tillering, and there is no report that miR393 regulates plant tillering by regulating the expression of auxin receptor protein gene mRNA.

Method used

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  • Small-molecule ribonucleic acid (RNA) Osa-miR393 for improving rice tillering and application
  • Small-molecule ribonucleic acid (RNA) Osa-miR393 for improving rice tillering and application
  • Small-molecule ribonucleic acid (RNA) Osa-miR393 for improving rice tillering and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Cloning of small molecule RNA Osa-miR393

[0050] through the website http: / / www.mirbase.org Find the sequence of Osa-miR393, and enter this microRNA sequence into the microRNA design application website http: / / wmd3.weigelworld.org / cgi-bin / webapp.cgi? page=Home; project=stdwmd Four primers were automatically synthesized and amplified by multiplex PCR to amplify the PCR product fragment expressing the stem-loop structure of Osa-miR393.

Embodiment 2

[0051] Example 2: Verification and application of Osa-miR393 function

[0052] 1. Construction of genetic transformation vector

[0053] The vector used for overexpression is Ami393 constructed in our laboratory. Ami393 is a commonly used plant genetic transformation vector pCAMBIA1301 in the world (Sun et al.Xa26, a gene conferring resistance to Xanthomonas oryzae pv.oryzaein rice, encoding a LRR receptor kinase-like protein. Plant Journal. 2004, 37: 517-527 ) based on Agrobacterium-mediated genetic transformation vector carrying the maize 35S promoter with constitutive and overexpression characteristics ( figure 2 ). The cloned Osa-miR393 fragment was ligated into the pGEMT-easy vector, digested with AflII and BglII and ligated into the binary expression vector pCambia1301, and the Gus expression gene inside the vector was replaced. The constructed final vector was named Ami393.

[0054] 2. Genetic Transformation

[0055] The genetic transformation method mediated by A...

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Abstract

The invention belongs to the field of plant genetic engineering, particularly relates to small-molecule ribonucleic acid (RNA) Osa-miR393 for improving rice tillering, and simultaneously relates to application of the small-molecule RNA and target genes acted by the small-molecule RNA in transgenic rice. The transgenic rice with over-expression Osa-miR393 is obtained through a genetic engineering method. The over-expression rice stably shows that significance of tillering number is higher than a compared phenotype so as to confirm functions of Osa-miR393 genes. Expression analysis of combined genes of the small-molecule RNA Osa-miR393 shows that the Osa-miR393 has a plant tillering regulation and control function by influencing expression of the target genes LOC_Os05g05800.1 and LOC_Os04g32460. Expression of the Osa-miR393 in the rice is improved or lowered through the gene engineering technology, the rice tillering can be regulated and controlled, and plant types of plants can be controlled so as to achieve the purpose of high yield.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. It specifically relates to a small molecule microRNA Osa-miR393 that controls rice tillering and its target genes LOC_Os05g05800.1 and LOC_Os04g32460, and also involves the application of the microRNA and target genes in transgenic rice. In the present invention, a small molecule RNA Osa-miR393 belonging to bHLB transcription factor was cloned by using the microRNA database on the Internet, and after it was introduced into the pNW55 vector, it was connected into the binary expression vector pCambial301 by enzyme digestion to construct the overexpression vector Ami393, and transferred into Rice Zhonghua11. After the microRNA was overexpressed, the T0, T1 and T2 transgenic plants all stably showed a significant increase in the number of tillers. The gene and its mode of interaction with the target gene have important application value for elucidating the growth and development process of ri...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00
Inventor 夏快飞王忍区晓劲张明永
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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