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Rice leaf width adjustment and control related microRNA (ribonucleic acid) and encoded nucleic acid molecule thereof as well as application

A technology for nucleic acid molecules and encoding nucleic acids, applied in the direction of DNA/RNA fragments, applications, recombinant DNA technology, etc., can solve problems such as research lag

Inactive Publication Date: 2013-10-23
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although great progress has been made in theoretical research, miRNA also has the potential to provide abundant gene resources for plant genetic improvement, but the current research on its application in production is relatively lagging behind

Method used

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  • Rice leaf width adjustment and control related microRNA (ribonucleic acid) and encoded nucleic acid molecule thereof as well as application
  • Rice leaf width adjustment and control related microRNA (ribonucleic acid) and encoded nucleic acid molecule thereof as well as application
  • Rice leaf width adjustment and control related microRNA (ribonucleic acid) and encoded nucleic acid molecule thereof as well as application

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Example 1. Discovery and expression analysis of miRNAs and their coding genes related to regulation of rice leaf width

[0057] 1. Discovery of rice leaf width regulation-related miRNAs and their coding genes

[0058] A reverse genetics study was conducted on multiple rice miRNA genes pre-screened from the japonica rice variety Nipponbare (Oryza sativa L.subsp.japonica cv.Nipponbare). The main research strategy was to analyze the overexpression of the target gene. During phenotypic analysis of transgenic rice populations, a miRNA and its coding gene related to rice leaf width regulation were discovered and identified.

[0059] The nucleotide sequence of the miRNA is sequence 1 of the sequence table, and belongs to a miRNA family conserved in plants—miRNA319 family. There are two members of the miR319 family in rice, namely osa-miR319a and osa-miR319b. The sequences of the mature osa-miR319b and osa-miR319a are identical, both consisting of 20 nucleotides (sequence 1 o...

Embodiment 2

[0076] Example 2, Obtaining and phenotypic analysis of transgenic rice overexpressing Osa-MIR319a and Osa-MIR319b genes and gene fragments

[0077] Theoretically, the biological effects produced by overexpressing miRNA genes and gene fragments with complete stem-loop structure are roughly the same. In order to study the biological function of the rice MIR319 gene, Osa-MIR319aFL (referring to the full-length cDNA of Osa-MIR319a), Osa-MIR319aST (referring to the gene fragment containing the stem-loop structure of Osa-MIR319a), Osa-MIR319bFL (referring to Osa - the full-length cDNA of MIR319b) and Osa-MIR319bST (referring to the gene segment comprising the stem-loop structure of Osa-MIR319b) are respectively placed under the control of the Actin of rice or the Ubi promoter of corn to construct a binary vector ( Figure 5 ), and transformed into rice indica variety Kasalath (Oryza sativa L. subsp. indica cv. Kasalath).

[0078] 1. Preparation of recombinant plasmids

[0079] 1. ...

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Abstract

The invention discloses a rice leaf width adjustment and control related microRNA (ribonucleic acid) as well as an encoding gene and application thereof. The RNA provided by the invention is one of 1)-4), namely 1) RNA consisting of a nucleotide sequence shown in the sequence 1 of a sequence table, 2) RNA consisting of a nucleotide sequence shown in the sequence 4 of the sequence table, 3) RNA consisting of a nucleotide sequence shown in the sequence 5 of the sequence table and 4) RNA which is substituted and / or lost and / or added from the RNA sequence shown in 1) or 2) or 3) through one or more basic groups, is related to plant leaf width, and is derived by 1) or 2) or 3). Experiments prove that Osa-miR319 and encoding gene thereof are discovered, the width of rice leaves can be effectively adjusted and controlled by utilizing the gene through a genetic engineering method, and the leaf shapes of rice can be changed.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a microRNA related to the regulation of rice leaf width and its encoding nucleic acid molecule and application. Background technique [0002] MicroRNA (miRNA) is a class of endogenous non-coding RNA with regulatory functions found in eukaryotes and viruses, and its size is 19-24 nucleotides (nucleotides, nt). In plants, miRNAs are mostly encoded by independent genes. In the nucleus, miRNA genes are first transcribed into longer transcripts by RNA polymerase II, which are called primary miRNA transcripts (primary miRNA, pri-miRNAs). The structure of primary transcripts is similar to general mRNA, and most of them also have 5'-cap and 3'-poly(A) tail structures. Primary transcripts contain imperfectly matching stem-loop structures. Primary transcripts can be processed by the RNaseIII family enzyme Dicer like1 (DCL1) and some other related double-stranded RNA-binding proteins into mi...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/11C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 朱立煌李大勇杨春花刘雪
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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