Cold induced promoter p-LTT1 for rice and application thereof

A technology of promoters and DNA molecules, applied in the field of rice cold-inducible promoter p-LTT1, can solve problems such as genetic bottlenecks, and achieve broad application and market prospects

Active Publication Date: 2014-06-04
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

According to statistics, the number of alleles in cultivated rice is about 60% of that in wild

Method used

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  • Cold induced promoter p-LTT1 for rice and application thereof
  • Cold induced promoter p-LTT1 for rice and application thereof
  • Cold induced promoter p-LTT1 for rice and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0029] Example 1. Discovery of rice cold-induced promoter p-LTT1

[0030] 1. Discovery of cold-induced genes

[0031] First, the cold-tolerant rice line IL112 and the control parent Guichao No. 2 rice were used as materials for the chip (the chip is the rice whole genome chip from Affimetrix ( Rice Genome Array), item number: 900599) hybridization, and on the basis of chip data analysis, combined with comparative genomics analysis, screening cold tolerance related genes, after genome comparison, cold tolerance correlation analysis, and finally the cold tolerance gene LTT1. Under cold treatment conditions, the expression of this gene in rice cold-tolerant line IL112 increased significantly (see figure 1 ) Is a cold-induced gene.

[0032] 2. The discovery of cold-induced promoter p-LTT1

[0033] Use primer3 primer design software to design primers T1GUSF and T1GUSR, and introduce restriction endonuclease SalI and HindIII recognition sites and protective bases at both ends of the primer...

Example Embodiment

[0040] Example 2. Functional verification of p-LTT1

[0041] 1. Construction of plant expression vector

[0042] 1. Using the genomic DNA of cold-tolerant line IL112 rice as a template, PCR amplification was performed with a primer pair composed of T1GUSF and T1GUSR to obtain PCR amplification products.

[0043] PCR reaction conditions: first 94℃5min; then 94℃30sec, 58℃45sec, 72℃2min, total 30 cycles; finally 72℃10min.

[0044] 2. Double digest the PCR amplified product obtained in step 1 with restriction enzymes SalI and HindIII, and recover the digested product.

[0045] 3. Double-cut the plant expression vector pCambia1381 with restriction enzymes SalI and HindIII, and recover the vector backbone (about 10650bp).

[0046] 4. Connect the digested product of step 2 and the vector backbone of step 3 to obtain the recombinant plasmid pCambia1381-pLTT1. According to the sequencing results, the structure of the recombinant plasmid pCambia1381-pLTT1 is described as follows: the DNA fragment...

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Abstract

The invention discloses a cold induced promoter p-LTT1 for rice and application thereof. The invention provides a DNA fragment having the function of the promoter, the DNA fragment is derived from oryzae rufipogon (O.rufipogon Griff.), and is a DNA molecule of the following 1), 2) or 3): 1) a DNA molecule expressed by sequence 1 of a sequence table; 2) a DNA molecule capable of hybridizing with the DNA sequence limited by the 1) under strict conditions and having the function of the promoter; and 3) a DNA molecule having over 90 percent of homology with the DNA sequence limited by the 1) and having the function of the promoter. The DNA fragment has the function of the promoter, comprises ABRE and MYB and other stress response elements, is specifically expressed in rice bud, stigma and anther, and has the characteristics of low temperature induced expression. The cold induced promoter p-LTT1 has important theoretical and practical significance for researches on a cold-resistant molecular mechanism of the rice and cold-resistant molecule breeding of the rice. The promoter has broad application and market prospect in the field of agriculture.

Description

technical field [0001] The invention relates to a rice cold-induced promoter p-LTT1 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 that in wild rice, which leads to the genetic bottleneck problem faced by the current rice va...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 刘凤霞付永彩谭禄宾朱作峰顾凭才宏伟孙传清彭文谢道昕
Owner CHINA AGRI UNIV
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