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Rice mTERF transcription termination factor gene V14 and application thereof

A technology of transcription termination and V14, applied in the field of plant biology, to achieve the effect of improving cold stress tolerance

Active Publication Date: 2018-01-30
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Rice cold stress is a quantitative trait, and there are multiple regulatory sites, but there are few known genes that regulate cold tolerance in rice. This invention explores the methods that affect the regulation of cold tolerance genes and their applications in rice, and uses molecular breeding technology for directional transformation Improving cold tolerance in rice by controlling key genes for cold tolerance

Method used

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  • Rice mTERF transcription termination factor gene V14 and application thereof
  • Rice mTERF transcription termination factor gene V14 and application thereof
  • Rice mTERF transcription termination factor gene V14 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Rice V14 Gene

[0029] The inventor previously reported that the mTERF gene with the sequence 0s07g0583200 in rice may regulate the transcription function of some chloroplast genes in the three-leaf stage of chloroplast development, and identified it as a gene related to rice chloroplast development V14 , which in the indica type V14-I and japonica V14-J The nucleotide sequence of is shown in the sequences SEQ ID NO: 1 and SEQ ID NO: 2.

[0030] The present invention is constructed by following examples V14 Gene expression vectors for further validation V14 The function of the gene.

Embodiment 2

[0031] Construction of embodiment 2 expression vector pV14-J

[0032] Through the BLAST tool in the NCBI (http: / / www.ncbi.nlm.nih.gov / ) website V14 Nucleic acid sequence and amino acid sequence homology analysis, comparison V14 The structural characteristics of the gene, by comparing with the genome sequences of the rice japonica variety Nipponbare and the rice indica variety Huanghuazhan, a total of 6977bp sequences from the upstream 3227bp to the downstream 914bp of the coding region were selected, and the primer pair SEQ ID NO: 5: 5'-GTAGACTGATGGATATAC-3' and SEQ ID NO.6: 5'-AGTTACTTCCTCCGTTTCAC-3' The nucleotide sequence amplified from the genomic DNA of the rice japonica variety Taichung 65 is shown in SEQ ID NO: 2 ( V14-J ), clone the amplified sequence into the plant transformation vector, pick the clone containing the target fragment of V14-J that has been synthesized, and culture it in 5 mL LB liquid medium containing Kna antibiotic, and culture it in a shaker at 37...

Embodiment 3

[0033] The genetic transformation of embodiment 3 expression vector pV14-J

[0034] (1) Preparation of Agrobacterium Competent Cells

[0035]Agrobacterium tumefaciens EHA105 stored at -80°C was taken, streaked on a YEB plate containing rifampicin (50ug / mL), and cultured at 28°C for 2 days. A single colony of Agrobacterium was picked from the YEB plate, inoculated into 5 mL of YEB liquid medium, placed on a shaker at 200 rpm, and cultured overnight at 28°C with shaking. Take 2 mL of the overnight cultured bacterial solution, inoculate it into 50 ml of YEB liquid medium, and culture it with shaking at 200 rpm at 28°C until the OD600 reaches 0.5. Transfer the bacterial solution to a sterile centrifuge tube, place in an ice bath for 30 minutes, then centrifuge at 5000 rpm for 10 minutes, discard the supernatant, and collect the bacterial cells. The collected bacteria were then suspended in 10 mL of 0.15 mol / L pre-cooled NaCl, centrifuged at 5000 rpm for 10 min, and the supernata...

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Abstract

The invention discloses a rice mTERF transcription termination factor gene V14. A nucleotide sequence of indica type rice V14-I is shown as SEQ ID NO:1, and an encoded amino acid sequence is shown asSEQ ID NO:3; a nucleotide sequence of japonica rice type V14-J is shown as a SEQ ID NO:2, and an encoded amino acid sequence is shown as SEQ ID NO:4; rice chloroplast development is ensured by normalexpression of a V14 gene; both the V14-J and V14-I can revert a whitening phenotype of a v14 mutant; moreover, the V14 is a novel rice response environmental temperature gene, which can regulate and control the cold tolerance of rice; the gene has important significance to cultivation of novel cold-tolerant plant varieties, in particular to cultivation of novel cold-tolerant rice varieties; a novel gene resource is provided for researching of a rice breeding and developing mechanism, and the gene has an application value on the aspects of regulation and control of the cold tolerance of indicatype rice.

Description

technical field [0001] The invention belongs to the field of plant biotechnology. More specifically, rice mTERF transcription termination factor gene V14 and its application. Background technique [0002] Rice is a cold-sensitive crop originating in the tropics or subtropics. Rice varieties are mainly cultivated in Asia, and are divided into indica ( Oryza saliva L. subsp. indica ) and Japonica ( Oryza saliva L. subsp. japanica ). In terms of cold stress resistance, most of the cold-resistant genes come from the more cold-tolerant japonica rice, while indica rice belongs to the cold-sensitive type. [0003] Rice cold stress is a quantitative trait, and there are multiple regulatory sites, but there are few known genes that regulate cold tolerance in rice. This invention explores the methods that affect the regulation of cold tolerance genes and their applications in rice, and uses molecular breeding technology for directional transformation Improve cold tolerance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
Inventor 张群宇刘耀光周峰王曼裴蕾薛德星谢敏江燕翔
Owner SOUTH CHINA AGRI UNIV
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