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Rice cold-inducible promoter p-ltt7 and its application

A technology of rice and fragments, applied in the fields of application, angiosperm/flowering plants, and the introduction of foreign genetic material using vectors, which can solve problems such as genetic bottlenecks

Inactive Publication Date: 2011-12-21
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to statistics, the number of alleles in cultivated rice is about 60% of wild rice (Sun C Q, Wang X K, Li Z C, Yoshimura A. Comparison of the genetic diversity of common wild rice (Oryza rufipogon Griff.) and cultivated rice (O.sativa L.) using RFLP markers.Theor Appl Genet, 2001, 102: 157-162), which leads to the genetic bottleneck (genetic bottleneck) problem faced by the current rice variety breeding

Method used

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  • Rice cold-inducible promoter p-ltt7 and its application
  • Rice cold-inducible promoter p-ltt7 and its application
  • Rice cold-inducible promoter p-ltt7 and its application

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Experimental program
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Effect test

Embodiment 1

[0030] Example 1, Discovery of rice cold-inducible promoter p-LTT7

[0031] 1. Discovery of cold-induced genes

[0032] First, the cold-tolerant line IL112 rice and the control parent Guichao 2 rice were used as materials for microarray (the chip was the rice whole genome chip (GeneChip) from Affimetrix Company. Rice Genome Array), Cat. No.: 900599) hybridization, and on the basis of chip data analysis, combined with comparative genomics analysis, screened cold tolerance related genes, after genome comparison and cold tolerance correlation analysis, finally obtained the cold tolerance gene LTT7, under cold treatment conditions, The expression of the gene was significantly increased ( figure 1 ), is a cold-inducible gene.

[0033] 2. Discovery of the cold-inducible promoter p-LTT7

[0034] Primers were designed using primer3 primer design software, and restriction endonuclease PstI and SmaI recognition sites and protective bases were introduced at both ends of the primers. ...

Embodiment 2

[0040] Embodiment 2, the acquisition and identification of p-LTT7 transgenic rice

[0041] 1. Construction of p-LTT7 plant expression vector

[0042] 1. Synthesize the DNA shown in sequence 1 of the sequence list, use the synthesized DNA as a template, and use specific primers to perform PCR amplification on T7GUS (T7GUSF / T7GUSR). After the reaction, perform 1% agarose on the PCR amplification product Gel electrophoresis detection, recovery and purification of the 1979bp DNA fragment (p-LTT7) (the recovery kit used is the DNA product purification kit of Tiangen Biochemical Technology Co., Ltd., catalog number: DP204-02).

[0043] 2. Digest the PCR product recovered in step 1 with restriction enzymes SmaI and PstI.

[0044] 3. Digest the plant expression vector pCambia1381 with restriction enzymes SmaI and PstI, and recover the vector skeleton.

[0045] 4. Ligate the digested product of step 2 with the vector backbone of step 3 to obtain a recombinant plasmid.

[0046] 5. Th...

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Abstract

The invention discloses a rice cold-inducible promoter p-LTT7 and its application. The promoter is the DNA molecule of the following 1) or 2) or 3) or 4): 1) the DNA molecule shown in the sequence 1 of the sequence listing from the 75th to the 2036th nucleotide at the 5' end; 2) in the sequence listing A DNA molecule shown in sequence 1; 3) a DNA molecule that hybridizes with the DNA sequence defined in 1) or 2) under stringent conditions and has a promoter function; 4) has a DNA molecule with the DNA sequence defined in 1) or 2) or 3) A DNA molecule with more than 90% homology and promoter function. The promoter contains stress-responsive elements such as ABRE, DRE and CBF, and is specifically expressed in rice shoots, stigmas and anthers. The promoter has important theoretical and practical significance for the research on the molecular mechanism of cold tolerance of rice and the molecular breeding of cold tolerance of rice. The invention will have broad application and market prospects in the agricultural field.

Description

technical field [0001] The invention relates to a rice cold-inducible promoter p-LTT7 and its application. Background technique [0002] Rice is an important food crop, and chilling injury is one of the important limiting factors affecting rice production. Chilling damage at the germination stage, seedling stage, and booting flowering stage will lead to stunted growth of rice seedlings, reduced tillering, and ultimately a significant reduction in rice yield. Therefore, it is urgent to discover cold-tolerant rice germplasm resources and breed cold-tolerant rice varieties. [0003] Common wild rice is the ancestor of cultivated rice in Asia. During the evolution of wild rice into cultivated rice, through natural selection and artificial selection, the genetic diversity and the number of alleles decreased. According to statistics, the number of alleles in cultivated rice is about 60% of wild rice (Sun C Q, Wang X K, Li Z C, Yoshimura A. Comparison of the genetic diversity of c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12N15/11A01H5/00
Inventor 孙传清刘凤霞谭禄宾付永彩朱作峰谢道昕苏震
Owner CHINA AGRI UNIV