Plant cold tolerance-associated protein LTT9, coding genes thereof and applications

A coding and genetic technology, applied in protein LTT9 and its coding gene and application fields, can solve problems such as genetic bottlenecks, and achieve the effects of simple operation, high plant height, and improved cold resistance

Active Publication Date: 2014-12-03
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 problem faced by the current rice variety breeding

Method used

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  • Plant cold tolerance-associated protein LTT9, coding genes thereof and applications
  • Plant cold tolerance-associated protein LTT9, coding genes thereof and applications
  • Plant cold tolerance-associated protein LTT9, coding genes thereof and applications

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1. Discovery of cold tolerance-related protein LTT9 and genes

[0050] 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 SIL208) with Guichao 2 as the genetic background was constructed. Seedling cold tolerance identification, the specific identification method is: 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...

Embodiment 2

[0052] Embodiment 2, the acquisition and identification of transgenic LTT9 rice

[0053] 1. Construction of recombinant expression vector super1300-LTT9

[0054] 1. Design specific primer pairs

[0055] Primers were designed according to the full-length cDNA sequence of Nipponbare's LOC_Os09g32310 (genbank: AK074021), and restriction endonucleases Xba I and Kpn I recognition sites and protective bases were introduced at both ends of the primers respectively. Sequence and protection base) are as follows:

[0056] Ge9F: 5'- GCTCTAGA ATGCAGCTCAGTTTCTTCC-3' (carrying XbaI restriction recognition sequence);

[0057] Ge9R: 5'- GCGGTACC TCATCCTATGGTTATTTTCAGC-3' (carrying KpnI restriction recognition sequence).

[0058] 2. After the seedling stage plants of the rice cold-tolerant line SIL208 were treated at a low temperature of 4°C, the shoots were taken to extract total RNA.

[0059] 3. Using the total RNA extracted in step 2 as a template, perform reverse transcription usi...

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Abstract

The invention discloses a plant cold tolerance-associated protein, coding genes thereof and applications. The protein is characterized in that: (a) the protein is composed of an amino acid sequence shown as a sequence 1 in a sequence table; and (b) the protein is a at least one plant cold tolerance- associated protein, which is derived from the protein in (a), and of which the amino acid sequence of the sequence 1 in the sequence table is substituted and / or deleted and / or added by one or more amino acid residues. The protein provided by the invention is closely related to the plant cold tolerance, and a method for culturing cold resistant plants by the coding genes of the protein has characteristics of simple operation and short period, and is suitable for popularization and application. A survival rate of obtained LTT9 gene-transferring paddy rice after low temperature treatment can still reach 100%, and simultaneously, compared with the paddy rice without transgene, strain is higher, and fresh weight is heavier. The plant cold tolerance-associated protein, coding genes thereof and applications has important theoretical and practical significance on research of plant cold-tolerance molecular mechanism, seed selection of plant cold tolerance kinds and plant cold tolerance molecular breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a protein related to plant cold tolerance and its coding gene and application, in particular to a protein LTT9 derived from common wild rice (O.rufipogon Griff.) and related to plant cold tolerance and its coding genes and applications. Background technique [0002] Rice is an important food crop. Low temperature and chilling damage seriously affects the high and stable yield of rice. The optimum temperature for rice growth is 15-18°C to 30-33°C. Japonica rice and indica rice will suffer chilling damage below 15°C and 18°C ​​respectively. , which seriously restricts the sowing area and sowing range of rice. There are also various manifestations of low temperature chilling damage. Green deficiency is the initial performance of cold-resistant varieties when they encounter low temperature attack. Even if the temperature returns to normal, the green deficiency leaves cannot resume normal ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N15/82A01H5/00
Inventor 孙传清刘加勇刘凤霞谭禄宾付永彩朱作峰苏震
Owner CHINA AGRI UNIV
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