LTP protein related to plant cold resistance, and coding gene and applications thereof

A technology encoding genes and cold resistance, applied in the field of LTP protein and its encoding genes and applications, can solve problems such as genetic bottlenecks, and achieve the effects of simple operation, suitable for popularization and application, and short cycle

Active Publication Date: 2014-11-12
CHINA AGRI UNIV
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  • 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 Gr

Method used

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  • LTP protein related to plant cold resistance, and coding gene and applications thereof
  • LTP protein related to plant cold resistance, and coding gene and applications thereof
  • LTP protein related to plant cold resistance, and coding gene and applications thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1, Discovery of cold tolerance-related gene LTP

[0069] First, Jiangxi Dongxiang common wild rice was backcrossed and self-crossed with the ultra-high-yielding rice variety Guichao 2, and an introgression line population (one of which was IL112) with Guichao 2 as the genetic background was constructed. The identification method of cold tolerance at the seedling stage is as follows: after the seeds are sterilized with 20% sodium hypochlorite, they are placed in a glass test tube with a bottom diameter of 5 cm and a height of 12 cm, and the seedlings are germinated in a tissue culture room at 25°C. Cultivate with distilled water in the early stage, and start to use 1 / 3B after about 13 days 5 culture medium. Put the neat seedlings with 2 leaves and 1 heart in a glass test tube with a bottom diameter of 5cm and a height of 18cm, and add 1 / 3B about 3cm deep 5 The culture medium was placed in a 4°C incubator for 6 days of low temperature treatment, and then moved t...

Embodiment 2

[0071] Embodiment 2, the acquisition of LTP transgenic rice and its cold tolerance identification

[0072] 1. Construction of recombinant expression vector Super1300-LTP

[0073]Primers were designed according to the full-length cDNA sequence of the Nipponbare gene LOC_Os10g36160, and restriction endonuclease KpnI and SacI recognition sites and protective bases were respectively introduced at both ends of the primers. The primer sequences are as follows:

[0074] Lt10F: 5'- CGGGGTACC AGCTCGAGCAATGGTGAAGT-3' (the underlined part is the recognition site of restriction endonuclease Kpn I and the protected base, and the following sequence is the 1st-20th position of sequence 2);

[0075] Lt10R: 5'- CGAGCTC CCAAAACGAATGTACACACAGC-3' (the underlined part is the recognition site of the restriction endonuclease Sac I and the protective base, and the following sequence is the reverse complementary sequence of positions 498-519 of Sequence 2).

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Abstract

The invention discloses an LTP protein related to plant cold resistance, and a coding gene and applications thereof. The applications specifically relate to applications in following a1) or a2) of the protein formed by an amino acid sequence shown as a sequence 1 in a sequence table: a1) regulation and control of plant cold resistance; and a2) breeding of cold-resistant plant varieties. Experiments show that: the coding gene of the protein formed by the amino acid sequence shown as the sequence 1 in the sequence table is utilized for breeding of cold-resistant plant, and the breeding method has characteristics of simple operation and short period and is suitable for popularization and application. The survival rate of LTP-transgenic rice obtained by the protein and the coding gene after low-temperature processing can also reach 100%. The method has important theoretical and practical meaning for research of plant cold resistance molecular mechanisms, breeding of cold-resistant varieties, and plant cold-resistant molecular breeding, and provides an economical, rapid and effective route for improving the plant cold resistance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an LTP protein related to plant cold tolerance, its coding gene and application. Background technique [0002] Rice is an important food crop. Chilling injury is one of the important factors affecting rice production in the early rice planting areas in the middle and lower reaches of the Yangtze River, the northeast and northwest rice areas, and the one-crop rice area in the Yunnan-Guizhou Plateau. If the rice seedlings are exposed to chilling damage during the germination and seedling stages, the growth of rice seedlings will be slowed down and the number of tillers will be reduced. In severe cases, there will even be a large area of ​​stiff and dead seedlings, which will eventually lead to a significant reduction in rice yield. Therefore, it is urgent to cultivate Cold-tolerant rice varieties. Common wild rice is the ancestor of cultivated rice in Asia. During the evolution of wild ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
CPCC07K14/415C12N15/8273
Inventor 刘凤霞付永彩谭禄宾朱作峰顾凭才宏伟孙传清
Owner CHINA AGRI UNIV
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