Paddy cool injury gene and application

A technology for rice and cold damage, applied in the field of plant biology, can solve the problems of reduced cold sensitivity, increased cold sensitivity, and increased metastasis of yeast mutant drs2

Inactive Publication Date: 2007-09-19
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, expression of the ALA1 gene encoding an aminophospholipid translocase leads to increased translocation of aminophospholipids in recombinant yeast membrane vesicles, produces membrane lipid asymmetry, and reduces cold sensitivity in the yeast mutant drs2, while antisense expression in Arabidopsis ALA1 increases cold sensitivity

Method used

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  • Paddy cool injury gene and application
  • Paddy cool injury gene and application
  • Paddy cool injury gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Rice material

[0026] Rice (Oryza sativa) mutant osgpat1, the original wild-type material is "Zhonghua 11" japonica rice variety. The acquisition of this mutant is to use Agrobacterium-mediated genetic transformation to obtain a large number of Zhonghua No. 11 strains with T-DNA insertion tags, and the vector used for transformation is pFX-E24.2-R15 (Wu et al., Development of enhancer trap lines for functional analysis of the rice genome. Plant J. 2003, 35(3): 418-27.), the transformant was obtained by screening. Transformed plants are now T 0 Generation, T 0 T 1 Substitute plant, this plant is planted in paddy field by the interval of 5 cun * 8 cun, carries out field management by conventional paddy planting method. Through screening, a rice mutant Osgpat1 with stable phenotype was obtained, which was slightly shorter, had fewer tillers, and was very sensitive to low temperature.

[0027] 2. Isolation of T-DNA insertion flanking sequences.

[0028] The T-DNA ...

Embodiment 2

[0155] Embodiment 2: Mutant and control seedling low temperature stress test

[0156] Seeds of the Osgpat1 mutant and wild type were soaked at 25°C for 3 days, germinated at 37°C for 2 days, and then planted in paddy soil for about 20 days. There were 17 strains in the mutant mutants that had obvious changes, with curled leaves and severe withering, while 20 wild-type plants had no change (Fig. 4). This shows that the mutation of OsGPAT1 gene has a very important impact on plant frost resistance.

Embodiment 3

[0157] Example 3: Overexpression of the Osgpat1 gene

[0158] The vector used for overexpression is pU1301 constructed in our laboratory. pU1301 is the basis of pCAMBIA1301 (Sun et al., 2004, Xa26, a gene conferring resistance to Xanthomonas oryzae pv. oryzae in rice, encoding a LRR receptorkinase-like protein. Plant Journal. 37: 517-527), a commonly used plant genetic transformation vector in the world (Figure 6), an Agrobacterium-mediated genetic transformation vector carrying a maize ubiquitin promoter with constitutive and overexpression characteristics. The pCAMBIA1301 vector was kindly provided by the Australian CAMBIA Laboratory (Center for the Application of Molecular Biology to International Agriculture).

[0159] PCR primers gpatoef and gpatoer were designed to amplify the complete ORF of OsGPAT1 gene from the rice genome, and Kpn I and BamH I restriction sites were added to the PCR primers respectively.

[0160] The sequences of gpatoef and gpatoer are:

[0161] ...

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Abstract

The invention discloses a rice cold injury gene and application thereof, a separaterd protein of rice cold injury gene, provides a nucleic acid sequence of the gene and protein sequence thereof. The inactivating gene can cause decreasing low temperature-resistant of rice, and the gene also affects rice normal growth, mutant of the gene is a bit short, decreases tillering and declines fertility. The overexpression of the gene can improve cold resistance of rice. By low temperature stress test, the gene has good application foreground in rice cold resistance breeding.

Description

technical field [0001] The invention relates to the field of plant biotechnology. The present invention uses T-DNA tagging (T-DNA tagging) technology to identify and clone a new rice chilling damage-related gene OsGPAT1 from rice low-temperature-sensitivity mutants. The inactivation of this gene can reduce the low-temperature tolerance of rice. It also affects the normal growth and development of rice. The mutant is slightly shorter, with reduced tillers and reduced fertility. Analysis of the amino acid sequence encoded by the gene proves that the protein encoded by the gene is a glycerol-3-phosphateacyltransferase (GPAT), which plays an important role in the response of plants to resist low temperature stress. The invention provides the nucleic acid sequence of the gene and its protein sequence, and also relates to the application of the gene in rice cold-resistant breeding. Low temperature stress experiments showed that the OsGPAT1 gene plays an important role in the frost...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415
Inventor 张启发李向俊吴昌银
Owner HUAZHONG AGRI UNIV
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