Newborn tissue-specific promoter p-ema1 in rice and its application

A technology of recombinant vectors and recombinant plasmids, applied in the fields of application, recombinant DNA technology, angiosperms/flowering plants, etc., can solve problems such as genetic bottlenecks, and achieve broad application and market prospects

Active Publication Date: 2015-10-28
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 that in wild rice, which leads to the genetic bottleneck (genetic bottleneck) problem faced by the current rice variety breeding

Method used

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  • Newborn tissue-specific promoter p-ema1 in rice and its application
  • Newborn tissue-specific promoter p-ema1 in rice and its application
  • Newborn tissue-specific promoter p-ema1 in rice and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1, Discovery of rice neonatal tissue-specific promoter p-EMA1

[0030] 1. Discovery of the EMA1 gene

[0031] An early-maturing introgression line YIL23 was isolated from an introgression line population constructed with Orzya rufipogon Griff. as a donor parent and an indica rice variety Teqing (Oryza sativa ssp. About 28 days after heading, the early-maturing gene EMA1 was cloned by map-based cloning strategy. Analysis of the expression of the EMA1 gene in the parents showed that the expression of the EMA1 gene in the leaves gradually increased at the seedling stage and decreased rapidly after reaching the apex, but the expression of the EMA1 gene increased faster in the introgression line YIL23 than in Teqing ( See figure 1 , TQ stands for Teqing, YIL23 stands for the precocious introgression line YIL23). That is, at the seedling stage, the expression of EMA1 in the introgression line YIL23 was stronger than that in the green, and as the seedlings grew up, ...

Embodiment 2

[0035] Embodiment 2, the acquisition and identification of p-EMA1 transgenic rice

[0036] 1. Construction of plant expression vectors

[0037] 1. Synthesize the double-stranded DNA molecule shown in sequence 1 of the sequence listing.

[0038]2. Using the double-stranded DNA molecule synthesized in step 1 as a template, perform PCR amplification with a primer pair composed of P1 and P4 to obtain a PCR amplification product.

[0039] P1: 5'-AT GGATCC ATCACCGCAAATACTTCTCCC-3';

[0040] P4: 5'-GC AAGCTT ACGCAGAGATCCAGCTTATTCC-3'.

[0041] In P1, the underlined base is the recognition site of the restriction endonuclease BamHI;

[0042] In P4, the underlined base is the recognition site of the restriction endonuclease HindIII.

[0043] 3. Digest the PCR amplified product in step 2 with restriction endonucleases BamHI and HindIII, and recover the digested product.

[0044] 3. The plant expression vector pCambia1381 was double digested with restriction endonucleases BamH...

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Abstract

The present invention discloses a rice newly-growing tissue specific promoter p-EMA1 and an application thereof. According to the present invention, the promoter is derived from O.rufipogon Griff., and can be the following 1) or 2) or 3) or 4): 1) DNA molecule represented by the 5' terminal nucleotide from site 1 to site 1841 in the sequence 1, 2) DNA molecule represented by the sequence 1, 3) DNA molecule capable of being subjected to hybridization with the DNA sequence limited by the sequence 1 or the sequence 2 and having a promoter function, and 4) DNA molecule having more than 90% homology with the DNA sequence limited by the sequence 1 or the sequence 2 and having a promoter function; important theoretical and practical significance is provided for research on the rice pre-maturing molecular mechanism and breeding of the rice pre-maturing variety; important theoretical and practical significance is provided for research on the molecular mechanism of special rice newly-growing tissue expression and breeding of the special rice newly-growing tissue expression characteristic molecule; and broad application and market prospects are provided in the agriculture field.

Description

technical field [0001] The invention relates to a new rice tissue-specific promoter p-EMA1 and its application. Background technique [0002] Plant new tissues (organs) are all differentiated and developed from primordia, and plants have the ability to continuously develop new tissues (organs), so the growth and development of new tissues is crucial to the growth and development of the entire plant. [0003] Rice is one of the most important food crops in the world, and its seeds are edible by humans. The determinants of rice grain yield mainly include effective panicles, seed setting rate, number of grains per panicle and grain weight, and the number of effective panicles and the number of grains per panicle are closely related to the growth state of new tissue. Therefore, the design of new tissue-specific expression promoters in rice is crucial for molecular breeding of rice yield. [0004] Common wild rice is the ancestor of cultivated rice in Asia. During the evolution...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 孙传清谢道昕彭文谭禄宾叶盛张永生单晓昳刘凤霞
Owner CHINA AGRI UNIV
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